Engineering Principles For Food Processing Technology and Product Realization - Sanet.st

ENGINEERING PRINCIPLES FOR
FOOD PROCESSING TECHNOLOGY

AND PRODUCT REALIZATION
ENGINEERING PRINCIPLES FOR
FOOD PROCESSING TECHNOLOGY
AND PRODUCT REALIZATION
Edited by
Leonardo Sepúlveda Torre, PhD
Porteen Kannan, PhD
A. K. Haghi, PhD
First edition published 2025
Apple Academic Press Inc. CRC Press
1265 Goldenrod Circle, NE, 2385 NW Executive Center Drive,
Palm Bay, FL 32905 USA Suite 320, Boca Raton FL 33431
760 Laurentian Drive, Unit 19, 4 Park Square, Milton Park,
Burlington, ON L7N 0A4, CANADA Abingdon, Oxon, OX14 4RN UK
© 2025 by Apple Academic Press, Inc.

Apple Academic Press exclusively co-publishes with CRC Press, an imprint of Taylor & Francis Group, LLC
Reasonable efforts have been made to publish reliable data and information, but the authors, editors, and publisher cannot assume
responsibility for the validity of all materials or the consequences of their use. The authors, editors, and publishers have attempted
to trace the copyright holders of all material reproduced in this publication and apologize to copyright holders if permission to
publish in this form has not been obtained. If any copyright material has not been acknowledged, please write and let us know so
we may rectify in any future reprint.
Except as permitted under U.S. Copyright Law, no part of this book may be reprinted, reproduced, transmitted, or utilized in any
form by any electronic, mechanical, or other means, now known or hereafter invented, including photocopying, microfilming, and
recording, or in any information storage or retrieval system, without written permission from the publishers.
For permission to photocopy or use material electronically from this work, access www.copyright.com or contact the Copyright
Clearance Center, Inc. (CCC), 222 Rosewood Drive, Danvers, MA 01923, 978-750-8400. For works that are not available on CCC
please contact [email protected]
Trademark notice: Product or corporate names may be trademarks or registered trademarks and are used only for identification
and explanation without intent to infringe.
Library and Archives Canada Cataloguing in Publication
CIP data on file with Canada Library and Archives
Library of Congress Cataloging-in-Publication Data
CIP data on file with US Library of Congress
ISBN: 978-1-77491-575-2 (hbk)

ISBN: 978-1-77491-576-9 (pbk)
ISBN: 978-1-03271-392-2 (ebk)
About the Editors
Leonardo Sepúlveda Torre, PhD

Professor, School of Chemistry at the Universidad Autónoma de Coahuila,
Saltillo, Coahuila, México
Leonardo Sepúlveda Torre, PhD, is a full Professor at the School of Chemistry

at the Universidad Autónoma de Coahuila, Saltillo, Coahuila, México, where
he is also a leader of the Bioprocesses and Microbial Biochemistry Group.
His postgraduate studies were related to topics on food biotechnology in the
Food Research Department of the UAdeC. In 2011, he made a research stay
at the Institute of Biotechnology and Bioengineering at the University of
Minho (Uminho), Braga, Portugal. He also worked as a collaborator at the
Center of Biological Engineering in Uminho, Braga, Portugal, with the link
project “Biotechnologies for Regional Food Biodiversity in Latin America.”
He has participated as author and co-author in 35 scientific articles in indexed
journals, several books, and 24 book chapters, and he holds two national
patents.
Porteen Kannan, PhD

Assistant Professor, Department of Veterinary Public Health,
Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences
University, India
Porteen Kannan, PhD, is an Assistant Professor in the Department of Veteri-

nary Public Health at Madras Veterinary College, Tamil Nadu Veterinary and
Animal Sciences University, India. The research activities of Dr. Kannan
include food safety and antimicrobial resistance. He did his postdoctoral
studies at the US Department of Agriculture, Maryland, USA, with a special-
ization in foodborne pathogens. He has published his work in both national
and international journals. He is actively involved in mentoring both MVSc
and PhD students.
vi About the Editors
A. K. Haghi, PhD
Research Associate, Department of Chemistry, University of Coimbra,
Portugal
A. K. Haghi, PhD, is a retired professor and has written, co-written, edited

or co-edited more than 1000 publications, including books, book chapters,
and papers in refereed journals with over 3800 citations and h-index of 32,
according to the Google Scholar database. He is currently a research asso-
ciate at the University of Coimbra, Portugal. Professor Haghi has received
several grants, consulted for several major corporations, and is a frequent
speaker to national and international audiences. He is the founder and former
Editor-in-Chief of the International Journal of Chemoinformatics and
Chemical Engineering and Polymers Research Journal. Professor Haghi has
worked as an editorial board member of many international journals. He has
served as a member of the Canadian Research and Development Center of
Sciences and Cultures. He has supervised several PhD and MSc theses at the
University of Guilan (UG) and cosupervised international doctoral projects.
Professor Haghi holds a BSc in urban and environmental engineering from
the University of North Carolina (USA) and holds two MSc degrees, one
in mechanical engineering from North Carolina State University (USA)
and another one in applied mechanics, acoustics, and materials from the
Université de Technologie de Compiègne (France). He was awarded a PhD
in engineering sciences at the Université de Franche-Comté (France). He is
a persistent reviewer of leading international journals.
Contents
Contributors.............................................................................................................ix
Abbreviations .........................................................................................................xiii
Preface .................................................................................................................. xvii
SECTION 1: Food Packaging and Storage Engineering ...................................1

1. Advances in the Development of Hydrogels as an Intelligent
Food Packaging for Loading and Release of Natural Compounds ............3
Teresa Darlen Carrillo-Castillo, José Eduardo Urquizo-Contreras,
Cristian Torres León, and Ayerim Y. Hernández-Almanza
2. Bionanocomposites Based on Thermoplastic Starch for the

Food Industry ................................................................................................37
Juan M. Tirado-Gallegos, Emilio Ochoa-Reyes, Juan C. Bustillos-Rodríguez,
Julio C. Tafolla-Arellano, Francisco Hernández-Centeno, and
Juan Buenrostro-Figueroa
3. Release Studies to Improve the Mechanical Properties of the

Biopolymer, Polylactic Acid (PLA) for Food Packaging Applications .....67
Y. K. Reyes Acosta, W. E. Cruz Martinez, A. V. Reyes Acosta,
L. Sepúlveda Torre, C. N. Aguilar Gonzales, and R. I. Narro Cespedes
4. Thermal Properties by Adding Natural Oils, Foods,

Organic Materials, Fibers, and Nanocomposites in
the PLA, and Applications in 3D Printing ..................................................87
Y. K. Reyes Acosta, W. E. Cruz Martinez, A. V. Reyes Acosta, V. A. Cepeda Tovar,
J. C. Contreras Esquivel, C. N. Aguilar Gonzales, R. I. Narro Cespedes, and
R. Reyna Martinez
5. Antioxidant and Antimicrobial Properties of Moringa

oleifera in Food Preservation ..................................................................... 111
Alaín Martínez-Pérez, Eliseo Sanchez-Loredo, Diana B. Muñiz Márquez, and
Jorge E. Wong-Paz
6. Storage of Fruits and Vegetables in Controlled Atmospheres and

Its Application in the Preservation of White Onion Quality...................135
Celia Chávez-Mendoza and Alexandro Guevara-Aguilar
viii Contents
SECTION 2: Food Biotechnology .....................................................................165

7. Larrea tridentata Extracts and the Capacity of Its Biocompounds
in Medicine and Agriculture ......................................................................167
Jolanta E. Marszalek, Daniela M. Sánchez-Pérez, Erika Flores Loyola,
Agustina Ramírez-Moreno, and Lucio Rodríguez-Sifuentes
8. Perspectives for Innovation of New Food Preservatives from

Euphorbiaceae.............................................................................................197
Rebeca Villanueva Chávez, Luis Enrique Cobos-Puc,
Elda Patricia Segura-Ceniceros, Anna Iliná, and Sonia Yesenia Silva Belmares
9. Biotechnology and Food Safety: Zoonotic Perspectives ..........................221

A. Arun Prince Milton, G. Bhuvana Priya, Samir Das, Sandeep Ghatak, and
K. Srinivas
SECTION 3: Case Studies..................................................................................247

10. Functional Properties of Agroindustrial Wastes from
Pomegranate Peel (Punica granatum) .......................................................249
Ana L. Izábal-Carvajal, Cristian Torres-León, Leonardo Sepúlveda,
Mónica L. Chávez González, Cristóbal N. Aguilar, and J. A. Ascacio-Valdés
11. Foodborne Zoonoses: Current Status and Control Strategies................265

M. Suman Kumar, Himani Dhanze, S. S. Reddy Vakamalla,
A. R. Chandni, and S. Wilfed Ruban
12. Omics Technologies: An Overview of Application of Proteomics

in Meat Quality and Safety Assurance......................................................303
Kiran Mohan, Naveena Basappa Maheswarappa, S. Wilfred Ruban, and
Rituparna Banerjee
13. Food Waste Revalorization as a Functional Food or Ingredient ............321

Estrada-Gil L. E., N. D. Cerda-Cejudo, Chávez-González M. L.,
Flores-Gallegos A. C., M. Govea-Salas, Contreras-Esquivel J. C.,
C. N. Aguilar-González, and J. A. Ascacio-Valdés
Index .....................................................................................................................337
Contributors
Y. K. Reyes Acosta
Facultad de Ciencias Químicas de la Universidad Autónoma de Coahuila, Saltillo, Coahuila, México
Cristóbal N. Aguilar
Bioprocesses and Bioproducts Group, Food Research Department, School of Chemistry,
Universidad Autónoma de Coahuila, Saltillo, Coahuila, México
C. N. Aguilar Gonzales
Bioprocesses & Bioproducts Group, Food Research Department, School of Chemistry,
J. A. Ascacio-Valdés
Rituparna Banerjee
ICAR-National Research Centre on Meat, Chengicherla, Hyderabad, Telangana, India
Sonia Yesenia Silva Belmares

Department of Food Research, School of Chemical Sciences, Autonomous University of Coahuila,
Saltillo, Coahuila, Mexico
Juan Buenrostro-Figueroa
Research Center in Food and Development, Cd. Delicias, Chihuahua, México
Juan C. Bustillos-Rodríguez
Tecnológico Nacional de México, Instituto Tecnológico de Cd. Cuauhtémoc, Cd. Cuauhtémoc,
Chihuahua, México
Teresa Darlen Carrillo-Castillo

School of Biological Science, Universidad Autonoma de Coahuila, Carretara Torreón-Matamoros,
Torreón, Coahuila, Mexico
N.D. Cerda-Cejudo
R. I. Narro Cespedes
A. R. Chandni
Division of Veterinary Public Health, ICAR-IVRI, Izatnagar, Uttar Pradesh, India
Rebeca Villanueva Chávez

Department of food research, School of Chemical Sciences, Autonomous University of Coahuila,
M. L. Chávez-González
x Contributors
Celia Chávez-Mendoza
Centro de Investigación en Alimentación y Desarrollo A C, Coordinación Tecnología de Productos
Hortofrutícolas y Lácteos, Delicias, Chihuahua, México
Luis Enrique Cobos-Puc

Department of Food Research, School of Chemical Sciences, Autonomous University of Coahuila,
J. C. Contreras-Esquivel
Samir Das
ICAR Research Complex for NEH Region, Umiam, Meghalaya, India
Himani Dhanze
L.E. Estrada-Gil
A. C. Flores-Gallegos
Sandeep Ghatak
M. Govea-Salas
Laboratory of Nanobiociences, School of Chemistry, Universidad Autónoma de Coahuila,
Ing. J. Cárdenas Valdéz S/N, República, Saltillo, México
Alexandro Guevara-Aguilar
Centro de Investigación en Alimentación y Desarrollo A C, Coordinación Tecnología de Productos
Hortofrutícolas y Lácteos, Delicias, Chihuahua, México
Ayerim Y. Hernández-Almanza
Francisco Hernández-Centeno
Universidad Autónoma Agraria Antonio Narro, Department of Food Science and Technology, Saltillo,
Coahuila, México
Anna Iliná
Ana L. Izábal-Carvajal
M. Suman Kumar
Contributors xi
Cristian Torres León

Erika Flores Loyola

Facultad de Ciencias Biológicas, Universidad Autónoma de Coahuila, Coahuila, México
Naveena Basappa Maheswarappa
ICAR-National Research Centre on Meat, Chengicherla, Hyderabad, Telangana, India
Diana B. Muñiz Márquez

Universidad Autónoma de San Luis Potosí, Facultad de Estudios Profesionales de la Zona Huasteca,
San Luis Potosí, México
Jolanta E. Marszalek
W. E. Cruz Martinez
R. Reyna Martinez
Colegio de Estudios Científicos y Tecnológicos del Estado de Coahuila Plantel San Antonio de las
Alazanas, Arteaga, Coahuila, México
Alaín Martínez-Pérez
Tecnológico Nacional de México/Instituto Tecnológico de Durango, Durango, México
A. Arun Prince Milton

Kiran Mohan
Department of Livestock Products Technology, Veterinary College, KVAFSU, Bidar, Karnataka, India
Emilio Ochoa-Reyes
Research Center in Food and Development, Cd. Delicias, Chihuahua, México
G. Bhuvana Priya
Agustina Ramírez-Moreno
Lucio Rodríguez-Sifuentes
S. Wilfred Ruban
Department of Livestock Products Technology, Veterinary College Bengaluru, KVAFSU, Bidar,
Karnataka, India
Veterinary College, KVA&FSU, Hebbal, Bengaluru, Karnataka, India
Eliseo Sanchez-Loredo
Bioprocesses and Bioproducts Research Group, Food Research Department, School of Chemistry,
Universidad Autónoma de Coahuila, Saltillo, Coahuila, Mexico
Daniela M. Sánchez-Pérez
Instituto Tecnológico de Torreón (ITT)-DEPI, Carretera Torreón-San Pedro, Torreón, Coahuila, México
xii Contributors
Elda Patricia Segura-Ceniceros

Leonardo Sepúlveda
K. Srinivas
Julio C. Tafolla-Arellano
Universidad Autónoma Agraria Antonio Narro, Department of Food Science and Technology, Saltillo,
Coahuila, México
Juan M. Tirado-Gallegos
Tecnología de Alimentos de Origen Animal, Facultad de Zootecnia y Ecología, Universidad Autónoma
de Chihuahua, Chihuahua, Chihuahua, México
L. Sepúlveda Torre
Cristian Torres-León
Research Center and Ethno biological Garden, Universidad Autónoma de Coahuila, Unidad Torreón,
Viesca, Coahuila, México
V. A. Cepeda Tovar
José Eduardo Urquizo-Contreras

S. S. Reddy Vakamalla
Jorge E. Wong-Paz
Universidad Autónoma de San Luis Potosí, Facultad de Estudios Profesionales de la Zona Huasteca,
San Luis Potosí, México
Abbreviations
ABF alkali-treated bamboo fibers

AC azodicarbonamide
AF agave fibers
AFB1 aflatoxin-sensitive aptamer
AFM atomic force microscopy
AFW agricultural food wastes
BF bamboo fibers
BHA butylated hydroxyanisole
BHT butylated hydroxytoluene
BW beeswax
CA controlled atmosphere
CCA conventional CA
CDC Centers for Disease Control and Prevention
ChNC chitin nanocrystals
ChNCs chitin nanocomposites
CM compression molded
CMC carboxy methyl cellulose
CNC cellulose nanocrystals
CNT carbon nanotubes
CPC centrifugal partition chromatography
CRV carvacrol
DAEC diffuse adhering Escherichia coli
DCS dynamic control system
DE diatomaceous earth
DMA dynamic thermogravimetric study
DMTA dynamic mechanical thermal analysis
DSC differential scanning calorimetry
ECSO epoxidized cottonseed oil
EIEC enteroinvasive
ELO epoxidized linseed oil
EPEC enteropathogenic
ETEC enterotoxigenic
EVA ethylene vinyl acetate
EVOH ethanol vinyl alcohol
FAO Food and Agriculture Organization
FCCA fast conventional CA
FCNs flax cellulose nanocrystals
xiv Abbreviations
FDM fused deposition modeling

FEN fennel
FE-SEM field-emission scanning electron microscopy
FG fish gelatin
GAE gallic acid equivalents
GMA glycidyl methacrylate
GRAS generally recognized as safe
GSLCs glioma stem-like cells
GT genotypes
HDPE high-density polyethylene
HEV hepatitis E virus
HHDP hexahydroxydiphenic acid
HLCNCs high lignin containing cellulose nanocrystals
HPC hydroxypropyl cellulose
HPLC high-performance liquid chromatography
HPMC hydroxypropyl methylcellulose
HSCC high-speed countercurrent chromatography
HSPs heat shock proteins
HUS hemolytic uremic syndrome
HUVEC human umbilical vein endothelial cells
IPG immobilized pH gradient
LA lactic acid
LC liquid chromatography
LCST lowest critical solution temperature
LECA low-ethylene CA
LEO lavender essential oil
LO linseed oil
LT Larrea tridentata
MAE microwave-assisted extraction
MC methylcellulose
MD machine direction
MFLC microfibrillated lignocellulose
MFW microfibrillated wood
MIC minimal inhibitory concentration
MLPC medium pressure liquid chromatography
MMT montmorillonite
MRSA methicillin-resistant Staphylococcus aureus
MS mass spectrometry
NCC nanocrystalline cellulose
NDGA nordihydroguaiaretic acid
NFS nanocomposite-forming solutions
NG nanographene
OEO orange essential oil
Abbreviations xv
OHF olive husk flour

OMMT organomontmorillonite
PAAc poly(acrylic acid)
PAAm polyacrylamide
PAMPS poly( 2-acrylamide-2-methylpropanesulfonic
PBH polyhydroxybutyrate
PET polyethylene terephthalate
PLA Polylactic acid
PO Posidonia oceanica
PP pomegranate peel
PPE pomegranate peel extract
PS polystyrene
PUF polyurethane foam
PUFA polyunsaturated fatty acids
R-BC rotomolded biocomposites
REO rosemary essential oil
RH relative humidity
RM rotomolded
RTKs receptor tyrosine kinase
RVA Rapid Visco Analyser
SAP superabsorbent polymer
SBO soybean oil
SCB sugar bagasse pulp
SE solvent extraction
SEM scanning electron microscopy
SFA saturated fatty acids
SFE supercritical fluid extraction
SLS static light scattering
SPH superporous hydrogels
SSF solid-state fermentation
STEC Shiga-toxin-producing Escherichia coli
STEC Shigatoxigenic Escherichia coli
SWC subcritical water extraction
TBHQ tertiary butylated hydroquinone
TC crystallization temperature
TD transverse direction
TEM transmission electron microscopy
Tg glass transition temperatures
TGA thermogravimetric analysis
Tm melting temperature
TP tea polyphenols
TPS thermoplastic starch
TVB-N total volatile compounds
xvi Abbreviations
UAE ultrasound-assisted extraction

UBF untreated bamboo fibers
ULO ultra-low oxygen
ULOCA ultra-low oxygen CA
VLF venlafaxine
WF wood flour
WHC water holding capacity
WPC wood-structured composites
WVP water vapor permeability
Preface
This title is a comprehensive research-oriented book designed to cover essen-

tial elements of food process engineering, and it discusses the most critical
achievements in food science along with innovations that are changing the
food industry. It links the key concepts of food engineering and science.
All chapters have been updated to provide information about the modern
methods for engineering processing along with food safety control and their
properties. It combines engineering and product quality/safety concepts with
emphasis on practical usefulness of preservation processes as well as process
control.
This title focuses on the chemical features of food products, food micro-
biology, packaging, processing, distribution of quality foods, and preserva-
tion. It also reviews the most practical approaches into food product design
with practical discussions.
This book is divided into three sections in different areas of food process
engineering:
Section 1 covers a wide range of food packaging and storage engineering
processes.
Section 2 describes food biotechnology with an emphasis on novel food
processes.
Section 3 contains case studies to illustrate the engineering application
of technologies that are discussed.
It is an indispensable reference book for food industry professionals and

is a valuable research-oriented volume for postgraduate students in food
science and technology.
SECTION 1
FOOD PACKAGING AND STORAGE
ENGINEERING
CHAPTER 1
Advances in the Development of

Hydrogels as an Intelligent Food
Packaging for Loading and Release of
Natural Compounds
TERESA DARLEN CARRILLO-CASTILLO,
JOSÉ EDUARDO URQUIZO-CONTRERAS, CRISTIAN TORRES LEÓN, and
AYERIM Y. HERNÁNDEZ-ALMANZA
School of Biological Science, Universidad Autonoma de Coahuila,
Carretara Torreón-Matamoros, Torreón, Coahuila, Mexico
ABSTRACT
Hydrogels are networks formed by polymeric chains, cross-linked chemically

or by physical interaction. This three-dimensional structure allows them to
absorb large amounts of water or other fluids giving them a stable structure
and a moist consistency. These functional properties of hydrogels make them
suitable for countless practical applications. From tissue engineering to food
science, the ability of hydrogels to respond to stimuli has been harnessed,
which can be designed and modified. Similarly, its interconnected networks
allow extracts, oils, and other molecules to be encapsulated and subsequently
released at specific sites that improve the shelf life of foods. When targeting
the food industry, the main concern is to use naturally derived polymer-
based hydrogels for food safety and to reduce the environmental impact of
single-use packaging. Therefore, this chapter focuses on materials of natural
origin that have proven to be more appropriate to form polymeric hydrogels
with application in the food industry. The application is oriented toward
the loading and release of active compounds that provide benefits to food
Engineering Principles for Food Processing Technology and Product Realization. Leonardo Sepúlveda Torre,
Porteen Kannan, A. K. Haghi, PhD (Eds.)
© 2025 Apple Academic Press, Inc. Co-published with CRC Press (Taylor & Francis)
4 Engineering Principles for Food Processing Technology and Product Realization
improving its organoleptic characteristics, shelf life, and slowing down the
growth of pathogenic microorganisms.
1.1 INTRODUCTION
Intelligent food packaging is a new type of material to packaging that
includes protective barrier for food, detection, and recording functions
(Yang et al., 2021). Currently, attention to this packaging is increasing due
to the advantages it represents for the food industry, as well as its friendly
characteristics with the environment. They can communicate the conditions
of the packaged product on real time, but they do not interact with the food.
However, the real-time monitoring requires the use of biosensors that allow
the information to be transmitted to the consumer (Yang et al., 2021; Müller
and Schmid, 2019). In food industry, hydrogels are a great alternative to be
used in intelligent packaging system or like carriers of natural compounds
(Batista et al., 2019). Although currently most hydrogels are produced from
synthetic polymers that guarantee greater physical-chemical stability than
natural ones, hydrogels based on natural compounds are making their way
into the industry because they are renewable, have biological properties,
present versatile structures, and in addition to that they can be produced on
a large scale due to the abundance of the raw material (Batista et al., 2019).
This chapter describes the physicochemical characteristics of hydrogels as
well as their potential use in food packaging systems. On the other hand, the
advantages that these intelligent systems have shown in various applications
for food preservation are discussed.
1.1.1 HYDROGELS CHARACTERISTICS AND THEIR APPLICATION

AREAS
Hydrogels are cross-linked, hydrophilic polymer networks with a wide range
of applications due to their high biocompatibility, hydrophilicity, adaptability,
and responsiveness (Heimbuck et al., 2018). They are classified depending
on their parent materials, their physical properties, the nature of swelling,
their method of polymerization, the ionic charges on the network, the rate
of biodegradation, and the nature of crosslinking (Ullah et al., 2015). The
three-dimensional (swollen) hydrogels structure of hydrogels comprises a
solid phase, based on a polymer or mixtures of polymers, and a liquid phase,
usually water or biological fluids (Mahinroosta et al., 2018). In the swollen
state exposed to certain pressures, its three-dimensional structure allows the
Advances in the Development of Hydrogels as an Intelligent Food Packaging 5
absorption of large quantities of the target fluids without changes in its struc-
ture (Hu et al., 2019). They are also known to be very versatile materials as
they can self-assemble into a variety of shapes including micelles, nanogels,
microgels, beads, films, membranes, fibers, and sponges (Crini et al., 2019).
1.1.1.1 CHEMICAL AND PHYSICAL CROSSLINKING

When hydrogels are formed by chemical crosslinking, the polymer chains
interact with each other to form permanent bonds through the action of
crosslinking agents (Hu et al., 2019). To initiate chemical crosslinking, it is
necessary to introduce a low molecular weight crosslinking agent together
with a polymer into the reaction mixture. In the absence of crossover points,
hydrophilic linear polymer chains dissolve in water, due to the thermody-
namic compatibility of the polymer chain and water (Ullah et al., 2015).
Thanks to these cross-linked structures, hydrogels do not disintegrate during
swelling. The covalent bonds formed in chemical crosslinking are irrevers-
ible and because they are interconnected, they are very strong; therefore,
they cannot be dissolved again and are thermally stable (Crini et al., 2019).
The main methods used to achieve chemical crosslinking are radical
polymerization, reaction of complementary groups, grafting, and enzymatic
reactions (Nezhad-Mokhtari et al., 2019). The hydrophilicity of the network
is due to the presence of hydrophilic groups such as –NH2, –COOH, –OH,
–CONH2, –CONH– and –SO3H, capillary effect, and osmotic pressure.
Crosslinking agents are generally small multifunctional molecules that react
with polymers, favoring interaction with their previously activated functional
groups. Chemical cross-linking results in permanent three-dimensional struc-
tures, giving the hydrogel thermal stability and mechanical strength (Peppas
and Hoffman, 2020). These hydrogels are not degradable and cannot be easily
recycled, and since after polymerization they need to be purified to remove
residues of crosslinking agents and other chemical compounds; they have not
been considered for application in the food industry, due to a possible toxicity.
Hydrogels obtained by physical methods are called hydrogels complexed
with polyelectrolytes since they result from ionic interactions between poly-
mers (Barroso et al., 2019). That is, crosslinks are formed by physical domain
bonds: hydrogen bonds, hydrophobic interaction, and/or ionic complexation.
For example, polymer chains containing amino groups ensure the cationic
nature required to interact with the anionic group of other polyelectrolytes
and form mixtures with a wide range of macromolecules, especially poly-
saccharides and proteins (Mujtaba et al., 2019). These types of physical
interactions are very suitable because they allow the melting of solvents,
the modification of the volume after the process, they are easy to manufac-
ture, remodel, biodegrade and, since they do not use chemical cross-linking
agents, they do not present toxicity, which makes them biocompatible (Liu
et al., May 2019). The reversibility of physical bonds, that is, their ability to
intertwine and then flow again, is caused by some specific stimulus, to which
the polymer chains respond. This behavior is what makes them ideal for use
in dynamic systems such as food packaging. However, achieving this phys-
ical crosslinking crucially depends on two conditions: (1) the interactions
between the polymer chains must be strong enough to favor the existence
of binding sites during the formation of the three-dimensional network; and
(2) the assembled three-dimensional network must allow the absorption of a
considerable amount of water in the polymeric network (Mahinroosta et al.,
2018). Without these two conditions, the hydrogel will not form and would
not be useful for use, despite being nontoxic and environmentally friendly.
The choice of the type of crosslinking to be used (chemical or physical)
is based on the intended application of the hydrogel obtained, since, in
addition to its reversibility, modifying the degree of crosslinking modulates
the mechanical resistance, its absorption capacity, and its electrical charge.
Hydrogels can also acquire a neutral, ionic, or amphiphilic charge, which
will influence their capacity to retain absorbed water (Batista et al., 2019).
That is why in hydrogel science, the general aspects to take into account
are: the base polymers, the fabrication of the structure, the response of the
hydrogel, and the applications of the hydrogel (Li et al., 2021).
1.1.1.2 SUPERPOROUS AND SUPERABSORBENT HYDROGELS

Superporous and superabsorbent hydrogels represent an innovative next-
generation category developed for hydrogels, but they still represent disad-
vantages to be applied in the food area, since their conformation requires high
polymer density and chemical agents such as crosslinkers (Batista et al., 2019).
Mention is made of them, since their properties can be very useful, once a
methodology that does not include toxic agents is developed in the future.
Superporous hydrogels (SPH) were initially developed as a drug delivery
system to be applied in the gastric environment and for the drug to remain
inside the pores without being completely released. Its mechanism is to swell
instantly by rapidly absorbing water and maintaining its integrity by the
osmotic pressure generated by its porous structure, while slowly releasing
the active ingredient with which it was loaded (Kumari et al., 2020).
Superabsorbent hydrogel, also named superabsorbent polymer (SAP),
is a type of hydrogel that can absorb water up to 500–1500 g/g, while the
absorption capacity of ordinary hydrogels does not exceed 1000 g/g, and
retains it even under mechanical pressure (Zhang et al., 2011). This occurs
because when the hydrophilic groups of the polymer chains come into
contact with water, they become ionized, which promotes electrostatic
repulsion leading to a higher degree of swelling (Ma and Wen, 2020). The
crosslinked network prevents water loss and thus maintains the expansion
equilibrium, but the swelling process is limited by the mechanical strength
of the matrix because the interactions between the polymer chains become
weaker. In some cases, the increase in temperature improves the degree of
swelling because it allows a separation of the chains.
1.1.1.3 APPLICATIONS AREAS

Hydrogels are currently the subject of considerable scientific research due to
their potential in high-tech applications in the biomedical, pharmaceutical,
biotechnological, bioseparation, biosensor, agricultural, oil recovery, and
cosmetic fields (Ullah et al., 2015). During the evolution of their applications,
the traditional use of hydrogels in the food area has been active packaging
for moisture control generated by fresh fruits, vegetables, and meat products
(Bodbodak and Rafiee, 2016). But the development of new materials and
their ability to encapsulate compounds in their network has opened up more
applications in packaging systems such as the release of bioactive agents and
sensors, among others (Figure 1.1).
FIGURE 1.1 Application of hydrogels in the food field.

The method of making films for the application of hydrogels in food

packaging involves melting the polymer, generally without the addition of
chemical crosslinking agents, but during the process, temperature is the key
physical agent that must be controlled (Kalia, 2015).
The inclusion of active biocompounds and nanoparticles in the films has
been for the purpose of enhancing food preservation by taking advantage
of the antimicrobial and antioxidant capacity that these compounds and
nanoparticles possess. Studies have shown that its inclusion in the polymeric
structure of the hydrogel has also had an effect on improving the mechanical
strength of the film and its permeability to water of the hydrogel (Batista et
al., 2019).
The use of hydrogels as a carrier for flavorings or as part of an intel-
ligent packaging system with direct application in food matrices has been
developed in the work of some researchers:
Food freshness indicator (Baek et al., 2017).
• Information generated about the freshness of food.
• Based on the production of metabolites in food.
• Detection of changes in pH, chemical degradation, or microbial
growth.
• Stability and retention of volatile substances (Kwan and Davidov-
Pardo, 2017).
• Encapsulation of flavors (nanoemulsions in hydrogels).
• Controlled release occurs by pH modification (e.g., in contact with
saliva).
Improvement of the bioavailability of lipophilic compounds (Park et al.,
2017).
• Incorporation of lipophilic bioactive compounds (e.g., β-carotene)
in the food matrix, improving its bioavailability.
1.2 HYDROGELS BASED ON BIOPOLYMERS TO REDUCE

ENVIRONMENTAL IMPACT
Due to their versatility and low cost, synthetic polymers have been widely
used in all areas of industry; nevertheless, this and the poor waste manage-
ment of these materials have resulted in a wide variety of environmental
problems (Geyer, 2020). Consequently, many alternatives have emerged to
mitigate these negative effects. One of these alternatives consists of using
polymers of biological origin instead of those produced synthetically, since

they are widely distributed in nature and, due to their characteristics, are
environmentally friendly (Shen et al., 2020). These types of polymers, also
called biopolymers, are macromolecules made from monomers synthesized
in biological systems such as plants, animals, and microorganisms. Therefore,
biopolymers represent a biodegradable, biocompatible, and easily renewable
material (Varghese et al., 2019). The use of natural polymers as a basis for
the creation of hydrogels is a field with enormous potential in areas such as
medicine, the food industry, and cosmetics. Mainly because in these areas the
use of synthetic polymers has been limited due to their poor biodegradability
and potential toxicity (Bao et al., 2019). Natural polymers can be classified
into six groups: polysaccharides, proteins, polynucleotides, polyisoprenes,
polyesters, and lignins (Varghese et al., 2019). Some of these natural polymers
used in the elaboration of hydrogels are discussed in the following section.
1.2.1 PROTEINS
Proteins are biomolecules composed of a chain of amino acids linked by
peptide bonds; these are essential macromolecules found in all living organ-
isms and can represent up to fifty percent of the dry weight of their cells.
In the food industry, proteins have been widely used due to their functional
properties, biodegradability, nontoxicity, and nutritional value. One of these
functional properties is the ability to form gels at different scales ranging
from micro to macro and has applications in preserving or increasing
the bioavailability of functional and bioactive compounds in food (Bourbon
et al., 2019). The main proteins used in the elaboration of hydrogels are
described below.
1.2.1.1 ANIMAL-DERIVED PROTEINS

Animals and their products represent a well-known source of protein. Table
1.1 describes the main types of animal proteins to produce hydrogels.
1.2.1.2 PLANT-DERIVED PROTEINS

In the food industry, plant-based proteins represent a cheap, abundant, biode-
gradable, and nontoxic source of natural polymers with fewer environmental
implications than animal-based proteins. This is due to the large amounts of
water and feed needed to produce the proteins in animals (Lopes et al., 2022).
TABLE 1.1 Biopolymers Derived from Animal Proteins to Form Hydrogels.

Biopolymer Characteristics Application
Collagen Protein present in the extracellular matrix Colorimetric sensor. Deliver
and the most abundant in the animal bioactive compounds (Shi et al.,
kingdom. It is obtained from tissues 2018).
from marine, bovine, and porcine species
(Sarrigiannidis et al., 2021).
Gelatin Protein derived through the denaturation of Encapsulation of essential oils
collagen either by chemical or by physical with antimicrobial activity
ways (Irastorza et al., 2021). (Goudoulas et al., 2022).
Caseins Protein present in milk. They are negatively Encapsulation of active
charged phosphorylated proteins, rich in compounds (Sadiq et al., 1965).
the amino acid proline and resistant to heat
denaturation (Panahi and Baghban-Salehi,
2014).
Whey Protein derived from milk. This fraction of Have been reported to have the
protein milk proteins is composed of α-lactalbumin ability to form hydrogels that
(α-Lg), β-lactoglobulin (β-Lg), and bovine can be used in the food industry
serum albumin (Sadiq et al., 1965). for encapsulation and delivery
of bioactive compounds (Abaee
et al., 2017).
1.2.1.2.1 Soybean Proteins
The main component of soybeans is protein (35–40%). This fraction is

composed of glycinin (11s globulins) and β-conglycinin (7s globulins). Soy
protein is used in industry and research as soy protein isolate, which is a
purified and concentrated version of the protein (Sadiq et al., 1965). This
biomolecule is widely used due to its low cost, availability, and nutritional
properties. However, these gels exhibit weak mechanical characteristics and
rapid degradation. In response to this effect, the use of chemical and enzy-
matic crosslinkers represents a viable solution to improve the characteristics
of these materials (Maltais et al., 2009).
1.2.1.2.2 Pea Proteins
The seeds of this legume contain about 30% protein consisting of legumin
11s and vicilin 7s globulins. Like soy protein, pea protein isolate is the
derived product used in the food industry (Djoullah and Saurel, 2021). This
protein has been used in the development of hydrogels with food applications
mainly because of its sustainable production, non-allergenic status, and

tunable properties (Meng and Cloutier, 2014).
1.2.2 POLYSACCHARIDES
These polymers are carbohydrates and composed by the union of monosac-

charides through glycosidic bonds (Liu et al., 2021). Polysaccharides as base
biomaterials in the elaboration of hydrogels represent a nontoxic, biodegrad-
able, biocompatible, and stable alternative. Furthermore, they have good
mechanical properties (Varghese et al., 2019). Consequently, animals, plants,
algae, and microorganisms have attracted attention in recent years because
it is possible to isolate a wide variety of polysaccharides from them (Liu et
al., 2021).
1.2.2.1 PLANT-DERIVED POLYSACCHARIDES
The use of plants as a source of polysaccharides is probably one of the most

characteristic approaches in biotechnology. Plant-derived polysaccharides
are considered a viable alternative for obtaining raw materials that can be
used in a wide variety of biotechnological processes such as the production
of gels. This is due to their potential to be produced sustainably, their high
availability, and their low cost (Bahú et al., 2022).
1.2.2.1.1 Cellulose
Cellulose is the most abundant polysaccharide in nature. In the food industry,

this polymer is characterized by its versatility, which is a consequence of the
wide variety of cellulose derivatives used in this field, such as methylcellulose
(MC), carboxy methyl cellulose (CMC), hydroxypropyl cellulose (HPC), or
hydroxypropyl methylcellulose (HPMC) (Ghosh and Katiyar, 2019). Hydro-
gels made from cellulose and its derivatives are mainly used for packaging,
although their ability as delivery carriers of bioactive compounds and their
capacity to respond to stimuli have also been studied (Yan et al., 2020).
However, these materials still represent a series of significant challenges
to be solved. The main one is finding a suitable solvent for this material
that is insoluble in water (Ghosh and Katiyar, 2019). Despite this drawback,
hydrogels have been developed for the production of smart packaging, such
as the work carried out by Shaghaleh et al., who obtained a hydrogel with
pH/thermo-responsive behavior and antimicrobial properties, made from
TEMPO oxidized nano fibrillated cellulose and food-grade cationic-modi-
fied poly(N-isopropyl acrylamide-co-acrylamide). The hydrogel was able to
release preservative agents in response to stimuli generated by a climacteric
fruit (Shaghaleh et al., 2021).
1.2.2.1.2 Starch
This biopolymer is found in granules form consisting of amylose and amylo-

pectin (Zhu, 2017). Starch is characterized by its ability to form gels in the
presence of water and heat. For this reason, this polysaccharide is widely
used as a food additive or as a packaging material. However, processes such
as starch retrogradation and the loss of mechanical properties in hydrogels
caused by low temperatures represent a series of drawbacks that need to be
resolved to be used (Yang et al., 2021). Despite this, the use of hydrogels
based on starch obtained from different sources is studied in the elaboration
of systems to encapsulate bioactive compounds which are also present in
hydrogels as copolymers together with other polysaccharides or proteins
(Ghosh and Katiyar, 2019).
1.2.2.1.3 Algae Polysaccharides
In recent years, algae have been the source for obtaining a great variety
of important biopolymers such as agarose, alginate, or carrageenans, to
mention a few. The main advantage of these polysaccharides over those
obtained from other sources is their abundance, since even due to this fact
these biomolecules are considered a base resource in the next generation of
energy and bioproduct generation (Lin and Jiao, 2012).
1.2.2.1.4 Alginate
Alginate is a polysaccharide synthesized mainly by brown seaweeds,

although it is known that some microorganisms are also capable of producing
it. This heteropolysaccharide is formed by α-D-mannuronic acid and
β-L-guluronic acid. Furthermore, one of the most significant properties of
this biopolymer is its ability to produce hydrogels in the presence of divalent
cations (generally Ca2+), a process known as ionotropic gelation (Dalla-

bona et al., 2020). Hydrogels obtained from alginate are probably the best
known and most widely implemented due to the facilities that their creation
process involves. In this context, alginate hydrogels have been widely used
at nano-, micro-, and macro- scales in the encapsulation of different bioac-
tive compounds and the creation of food packaging (films and coatings)
(Osojnik Črnivec and Poklar Ulrih, 2019). An example of its application
in the development of smart packaging was the work of Sutthasupa et al.;
they elaborated a hydrogel with a mixture of alginate and methylcellulose
cross-linked with Ca2+. This packaging functioned as a spoilage indicator
for ground pork. This effect was achieved by incorporating bromothymol
blue dye, which is sensitive to pH, into the hydrogel matrix. The result was
a hydrogel with the ability to react with a color change in the presence of
total volatile compounds (TVB-N) when the pH becomes basic, working
in this way as a viable alternative in the field of active packaging and with
application in meat products (Sutthasupa et al., 2021).
1.2.2.1.5 Carrageenans
Carrageenans are another representative polysaccharide of this group and
are produced by red seaweeds. This polysaccharide is characterized by being
sulfated and having six different forms depending on its composition and
origin. These forms are kappa, iota, lambda, mu, un, and theta (Lin and Jiao,
2012). However, only the kappa, lambda, and iota variants are studied for
the elaboration of hydrogels due to their gelling properties (Berton et al.,
2020). The gelation of these polymers generally occurs through cationic
crosslinkers such as metal ions or other polymers. This process, as with
alginate hydrogels, is a relatively simple process (Zhang et al., 2016).
1.2.2.2 ANIMAL-DERIVED POLYSACCHARIDES

Chitosan is the main example of biopolymers of animal origin which have
properties to form hydrogels. Chitin is an amino polysaccharide that can
be obtained from the exoskeleton of insects, crustaceans, and mollusks
and in the cell wall of fungi. Chemically, chitin is made up of N-acetyl
glucosamine units; however, chitosan is formed by the deacetylation of this
polymer, which is a linear polymer made up of N-acetyl-D-glucosamine
and D-glucosamine units (Maddaloni and Vassalini, 2020). This polysac-
charide is the most important derivative of chitin and is widely used in
the food industry as a material to produce films, coatings, and hydrogels.

These hydrogels have great potential for applications due to their safe and
innocuous nature, their response to pH, and their ability to encapsulate and
release natural compounds in their matrix (Qu and Luo, 2020).
1.2.2.3 MICROORGANISM-DERIVED POLYSACCHARIDES
Microbial polysaccharides, due to their diversity and properties, have

positioned themselves commercially as a resource of great importance
in different processes; xanthan gum is probably the most representative
polymer of the group. It is an extracellular polysaccharide produced by
the Gram-negative bacterium Xanthomonas campestris by fermentation of
sugars (Osojnik Črnivec and Poklar Ulrih, 2019). It has the ability to form
hydrogels by physical and chemical crosslinking. However, due to their
stability, chemically cross-linked hydrogels are the most studied, mainly in
the encapsulation and controlled release of compounds (Tao et al., 2016).
1.2.3 SYNERGIES BETWEEN BIOPOLYMERS TO ENHANCE

HYDROGEL PROPERTIES
Throughout this section, the importance of natural polymers in the devel-

opment of hydrogels was reviewed, highlighting their great variety, safety,
biodegradability, and high availability. However, the main drawbacks in the
use of these materials were also raised, where the mechanical properties,
instability to external factors, and water retention capacity are the main limi-
tations in the use and manufacture of these gels. Consequently, in response
to these problems, the combination of two or more natural polymers has
emerged as a viable alternative for the improvement of these materials (Liu
et al., 2021). Therefore, on this basis, it is possible to create hydrogels using
protein–protein, protein–polysaccharide, polysaccharide–polysaccharide
mixtures, and even systems containing three of these biopolymers in addition
to hydrogels that embed fibers in their structure, which can be food grade
for further application in this field (Khalesi et al., 2021). These associations
between polymers are possible thanks to the bonds that can form between
them and their ability to form complexes through noncovalent and covalent
interactions. The examples of these systems include soy protein–whey
protein, gelatin–whey protein, cellulose–starch, and casein–carrageenans,
among many other hydrogels (Liu et al., 2021).
1.2.4 RELEVANCE OF THE RESIDUAL ORIGIN OF BIOPOLYMERS
It is a reality that the use of biopolymers for the elaboration of hydrogels

is a promising and environmentally friendly approach. Furthermore, due to
the characteristics shown, these materials have significant advantages over
synthetic polymers. In this context, the use of natural polymers in the produc-
tion of hydrogels can also have another positive effect on environmental
awareness if we consider the fact that these biopolymers can be obtained
from waste produced by the food industry itself. In fact, according to FAO
data, around 1.3 billion tons of food are discarded each year, representing a
potential wasted source of biomolecules such as proteins and carbohydrates
(Ranganathan et al., 2020). Therefore, the use of biopolymers in the produc-
tion of hydrogels not only represents an alternative to environmental care,
providing materials that in the long term do not present the same problems
as synthetic polymers but also making greater use of raw materials obtained
from the food industry waste that compromise the environment. Thus,
biopolymers represent a diverse, safe, nonpolluting source of raw materials
to produce food-grade hydrogels with the potential to be produced in a
sustainable manner.
1.3 SMART HYDROGELS, THEIR DESIGN TO RESPOND TO

STIMULI
Smart hydrogels are those that show significant physicochemical change

in response to small changes in the environment. These stimuli include
physical environmental conditions, such as temperature, electric and
magnetic fields, light, pressure, and sounds, and the chemical conditions of
the system, such as pH, ionic strength, solvent composition, and molecular
species (Dehshahri et al., 2021). These are stimuli that change the degree
of swelling or deswelling of the hydrogel in the form of volume collapse or
phase transition (Figure 1.2). Such changes are reversible; therefore, hydro-
gels are capable of returning to their initial state as soon as the stimulus is
removed. That is why they can be "reusable." These responses of hydrogels
to the environment provide a variety of applications that can be exploited in
the field of food packaging.
FIGURE 1.2 Response of smart hydrogels to stimuli.
Physical stimuli change the molecular interactions at the critical starting

points of the hydrogel, thus undergoing changes in its growth actions,
network structure, mechanical strength, and permeability. For their part,
chemical stimuli change the interactions between polymer chains and
between polymer chains and solvents at the molecular level.
1.3.1 DESIGN OF SMART HYDROGELS
Smart hydrogels have been designed to swell or shrink in response to physical,

chemical, or biological cues from the environment or even a combination of
these. The development of these smart hydrogels is a meticulous process
that takes into account polymer type, polymer molecular weight, crosslinker
molecule, and crosslink density to suit a specific application and provide
a combination of the properties listed above. The stimuli disturb the inter-
molecular interactions of the hydrogels formed in an aqueous environment,
resulting in changes in the conformation of the polymeric chains, which
causes macroscopic swelling or contractions. To make these chains respond
to stimuli, they are designed using functional groups, adapting the blocks of
different types of polymers, type and quantity of the crosslinking molecule,
and the integration of nanoparticles to broaden their functionalities and the
scope of their application (Ding et al., 2020).
1.3.1.1 THERMOSENSITIVE HYDROGELS
Its conformational change is based on the phase transition behavior of thermo-

sensitive polymers. Hydrogels with inverse thermosensitivity are insoluble
in water at room temperature, that is, above their lowest critical solution
temperature (LCST). This causes the aggregation of unsolvated polymeric
chains (Taylor et al., 2017), as happens with poly (N-isopropylacrylamide)
(PNIPAAm), when the temperature is above its LCST (32°C), its chains
they shrink due to their decreased solubility in water (Heskins and Guillet,
1968). The example of a polymer with a higher critical solution temperature
(UCST) response is polyacrylamide (PAAm), when the ambient temperature
is above its UCST; the strength of the water–polymer interactions caused
by the bonds of hydrogen becomes stronger than the polymer–polymer
interactions, causing an increase in the solubility of the hydrogel (Seuring
and Agarwal, 2012). If it is chosen to combine polymers of the two types,
a hydrogel is achieved that has a dual response to temperatures, above the
UCST (15°C) and below the LCST (41°C) (Chang et al., 2009).
1.3.1.2 PH-SENSITIVE HYDROGELS
These types of hydrogels are developed with polyelectrolytes containing

weak acidic or basic functional groups. These functional groups only allow
the passage of ions with opposite charges, creating an osmotic pressure
difference in the hydrogel network, inducing its deformation (Firestone
and Siegel, 1991). pH-sensitive hydrogels are classified as acidic or basic
depending on the pendant group they have on their chains. Acids possess
carboxylic groups, phosphonic acid groups, or boronic acid groups (Wu et
al., 2010). For their part, basic hydrogels have basic groups such as amines
or amide groups (Mukhopadhyay et al., 2014).
When the pH value surrounding the acid hydrogel is greater than the
dissociation constant (Ka), the acid pendant groups release protons, obtaining
a negative charge, causing an increase in the hydrophilic nature of the hydro-
gels, and allowing the network to swell. When the pH decreases, the effect
is reversible. In the case of the basic hydrogel, its pendant groups accept
protons at a pH below the dissociation constant (Kb), remaining positively
charged. This increases the hydrophilic nature causing an electrostatic repul-
sion between the polymer chains, causing the hydrogel to expand (Buenger
et al., 2012). Amphoteric hydrogels have also been created, with groups of
both acidic and basic monomers, which allows the hydrogel to only remain
swollen in a specific pH range, and contract at any other value (Abou Taleb,
2013).
1.3.1.3 SALT-SENSITIVE HYDROGELS
These hydrogels change when the salinity in the solution varies; the poly-
electrolyte networks expand in pure water and contract in saline solutions. Its
response mechanism is similar to that of pH-sensitive hydrogels. When the
hydrogel is in low salinity solutions, its ionizable groups dissociate causing
a high charge density. This leads to an intermolecular repulsion between
the chains of the hydrogel, causing a volumetric expansion. When the salt
concentration increases, counterions are being added, which attenuate the
intermolecular repulsion, causing the collapse of the polymeric network (Xu
et al., 2008).
1.3.1.4 ELECTROSENSITIVE HYDROGELS
They are a type of electroactive polymers that, when receiving a voltage,

result in a nonuniform distribution of ions on the hydrogel layer, which
causes an osmotic pressure difference, resulting in changes in curvature of the
hydrogel layer. This electrical responsiveness of hydrogels can be engineered
using ionizable functional groups. Some known electroactive polymers are
poly(acrylic acid) (PAAc) (Moschou et al., 2004), poly(4-hydroxybutyl
acrylate) (poly(4-HBA)) (Kwon et al., 2010), and poly(2-acrylamide-
2-methylpropanesulfonic) (PAMPS) (Yang et al., 2017). Ionizable functional
groups are located both in the backbone of the chain and in the side chains,
which provide abundant fixed charges that will react rapidly to electric
fields, resulting in a short response time (approx. 0.1 s) (Mirfakhrai et al.,
2007). If, in addition, ionic compounds are added to the hydrogel matrix, the
concentration of mobile ions increases, inducing an even faster response of
the hydrogel to electric fields (Ding et al., 2020).
1.3.1.5 PHOTOSENSITIVE HYDROGELS
This type is designed by incorporating photoactive fractions within the

hydrogel matrix (Li et al., 2019), which leads the hydrogel to have a photo-
thermal excitation or a reversible crosslinking reaction. For the reversible
crosslinking mechanism, photoisomerization (ring opening/closing reac-
tions) and photocleavage (induced by photolabile protecting groups) are
two predominant approaches that generate sequential steps of crosslinking,
resulting in reversible contraction–expansion of polymeric chains (Kloxin et

al., 1979).
Dehydration–hydration processes are also a physical response in the
hydrogel that can be achieved by selective lighting to cause localized
heating. Similarly, if photosensitive nanomaterials are incorporated into the
hydrogel matrix, dehydration processes can be accelerated and intensified.
These photothermal nanomaterials convert light irradiation into heat dissipa-
tion, inducing hydrogel deformation. Some known nanomaterials, which
have already been used for their photothermal capacity, are gold (Aida et al.,
2018), carbon-based materials (Yang et al., 2019) and organic compounds
such as cyanines (Zhang et al., 2016).
1.4 RESPONSE OF HYDROGELS TO CHANGES IN FOOD
When a smart hydrogel is used as part of a smart packaging system, its main
purpose is to provide information about the freshness of the contained food
products or as part of a simple detection method to determine the presence
of contaminants. Aflatoxin, for example, is a mycotoxin that can develop
in many agricultural products. Zhao et al designed a smart hydrogel by
cross-linking the polymer chains with an aflatoxin-sensitive aptamer (AFB1)
and by loading the hydrogel with urease. When the system is exposed to
concentrations of AFB1, the hydrogel collapses releasing urease into solu-
tion, causing changes in pH. This pH change can be easily measured, and
indirectly determines the concentration of AFB1 present (Zhao et al., 2018).
1.4.1 FOOD CHARACTERISTICS THAT ARE IMPROVED WITH THE

APPLICATION OF SMART HYDROGELS
Hydrogels are ideal for improving the characteristics of foods because of

their softness, elasticity, absorbent nature, flexibility, and hygroscopic
nature (Manzoor et al., 2022). Hydrogels can be used in food preservation,
pharmaceuticals, agriculture, and food packaging. Intelligent hydrogels
alter their structure and change phase to outside stimuli and can be used for
innovative food applications. Food characteristics that hydrogels primarily
influence are antimicrobial, gas barrier, nutritional, functional, flavor, and
color properties.
1.4.1.1 ANTIMICROBIAL PROPERTIES
Food preservation's primary objective has been controlling microbial popu-

lations, specifically pathogenic microorganisms (Torres-León et al., 2022).
Food safety is a big problem for humans, especially bacteria that cause
foodborne diseases (Torres-León et al., 2022). One way to inhibit the growth
of microorganisms in food is by implementing antimicrobial food packaging
systems. Hydrogels formulated with biopolymers and nanomaterials have
been used in food packaging to improve microbial stability and prolong the
shelf life (Beigmohammadi et al., 2016). The carrageenan-based hydrogel
films with metallic nanoparticles exhibited antibacterial activity against
Escherichia coli, and Listeria monocytogenes (Oun and Rhim, 2017).
1.4.1.2 GAS BARRIER PROPERTIES
Nowadays, the short shelf life of fruit and vegetables is one of the biggest
trading problems (Torres-León et al., 2018). The edible coatings have been
broadly used to extend the shelf life of fruits, vegetables, cheese, mush-
rooms, rainbow trout fillets, chicken fillets, and processed potatoes (Chacon
et al., 2022). Hydrogels can control moisture loss and transpiration produced
by the food (Leyva-Jiménez et al., 2023). Multicomponent hydrogel films
(composed of agar, κ-carrageenan, konjac glucomannan powder, and nano-
clay) applied to fresh spinach packaging reveal a high potential for the use
as an antifogging film for packaging highly respiring agricultural produce
(Rhim and Wang, 2013).
1.4.1.3 NUTRITIONAL PROPERTIES
Hydrogels may also be used to encapsulate and control the release of

nutrients (Yang et al., 2022). Oil in water-hydrogelled emulsions and
ethylcellulose organogels replaced beef fat in emulsion-type meat batters.
Reformulated products improved the fatty acid profile (decrease in saturated
fatty acids (SFA) from 11.8% to ≈2% and an increase in polyunsaturated
fatty acids (PUFA) from 0.3% to ≈5%) (Alejandre et al., 2019). Dos Santos
et al. (2020) demonstrated that emulsion gels based on pork skin, inulin,
α-cyclodextrin, polydextrose, and bamboo fiber are alternative ingredients
to replace pork back fat in manufacturing emulsified meat products. The
fat content in sugar-snap cookies was reduced by replacing a commercial
shortening with a hydrogel formed by mixing a canola oil/candelilla oleogel

(5 g/100 g) and a gelatinized corn starch hydrogel (5 g/100 g) in a 1:1 ratio
(Barragán-Martínez et al., 2022).
1.4.1.4 FUNCTIONAL PROPERTIES
The hydrogels in food are a vehicle system for the antimicrobials, antioxi-
dants, flavorings, colorants, and bioactive components. The hydrogel loaded
with 5% pomegranate peel extract showed good antioxidant (81.13%) and
antimicrobial activities (Guo et al., 2019). In recent years, polyphenols (as
naturally derived antioxidants) have gained growing interest in the food
industry. Phenolic compounds are secondary metabolites produced by
plants and exhibit a wide range of functional properties (Torres-león et al.,
2017). Trifković et al. (2014) designed thyme polyphenols loaded chitosan
hydrogels to prolong the release of polyphenols in simulated gastrointestinal
conditions. The release of encapsulated polyphenols in simulated gastroin-
testinal fluids was extended to 3 h.
1.4.1.5 FLAVOR PROPERTIES
The hydrogels can also be used to encapsulate and control the release of
flavor compounds in food. Soluble-filled hydrogels can encapsulate a
flavor and release the flavor in the presence of artificial saliva. pH-sensitive
hydrogels can dissociate in the presence of saliva; this leads to the release
and perception of encapsulated flavors (Amiryousefi et al., 2016). Filled
hydrogels were determined to encapsulate and release flavor (Kwan and
Davidov-Pardo, December 2017).
1.4.1.6 COLOR PROPERTIES
Food additives are used for color enrichment in the food industry. The use of
a hydrogel formed by mixing a canola oil/candelilla oleogel (5 g/100 g) and a
gelatinized corn starch hydrogel (5 g/100 g) in the cookie preparation reduced
the effective content of lipids, reducing in this way the extent of redness and
yellowness reactions, which was reflected in the color parameters (Barragán-
Martínez et al., 2022). Hydrogels are also used as a biosensor in innova-
tive packaging. Intelligent hydrogel packaging may contain a colorimetric
indicator that changes color in response to a change in pH in the food, which

is related to quality attributes. Lu et al. (2020) formulated sugarcane bagasse
nanocellulose hydrogel as a colourimetric freshness indicator for intelligent
food packaging.
1.5 THE HYDROGELS’ ABILITY TO BE LOADED WITH NATURAL

COMPOUNDS
As has been described so far, hydrogels are a versatile material, which is

why they also have the ability to be loaded with natural compounds, to later
release them in a controlled manner, thus expanding their applications. Due
to the amphiphilic characteristic of their chains and the polarity of their
networks, hydrogels can be loaded with both hydrophilic and hydrophobic
compounds. Essential oils can be incorporated by means of emulsifying
agents, which stabilize them in the hydrogel matrix. Natural aqueous-based
extracts have a natural affinity with the hydrophilic part of the hydrogel, so
their incorporation can be done by simply incorporating it into the aqueous
medium that will serve to swell the hydrogel. This is in order to develop
intelligent systems for the delivery of bioactive compounds in food when
a change occurs in the microenvironment of the packaging or outside of it.
1.5.1 LOADED WITH ESSENTIAL OILS
In addition to their utility as flavorings and food additives, essential oils can
exhibit excellent antibacterial and antimicrobial activity. Its incorporation in
hydrogels has been for the purpose of creating systems that can help food
packaging to prolong the storage life of food by inhibiting bacterial growth
and avoiding oxidative processes.
From the research carried out, using hydrogels loaded with oils, which
had antibacterial properties, Goudoulas et al. developed gelatin hydrogels
containing microdroplets of rosemary essential oil (REO) and orange essen-
tial oil (OEO). They stabilized the droplets using the surfactant Tween®80.
The gels/REO inhibited the growth of Bacillus cereus and Escherichia coli
(contaminating meat products bacteria); meanwhile, the gels/OEO inhib-
ited the growth of B. cereus, although to a minor degree than REO. These
hydrogels loaded with microdispersed essential oils are a product that has the
potential to be used as a coating for meat products and prolong their storage
life (Goudoulas et al., 2022).
Lavender essential oil (LEO) also has antibacterial properties, but

its application is limited due to its high volatility. Therefore, Deng et al.
investigated the possibility of using hydrogel particles to encapsulate and
control the release of LEO. The oil was encapsulated in oil-in-water emul-
sions, which were then loaded onto hydrogel particles to finally cross-link
them with Ca2+. The results were that the hydrogel improved the stability of
LEO during storage and controlled its release by changes in pH. The authors
suggested that this system can be incorporated into food packaging as a new
aroma delivery system with controlled release of essential oils (Deng et al.,
2020).
On the other hand, the essential oils of garlic (GEO) and holy basil
(HBEO) are commonly used in food products for their antimicrobial activity,
but there are studies in which the synergistic actions of two oils improve
potency and spectrum of activity antimicrobial. A common problem with
these oils is their chemical deterioration due to light, temperature, and expo-
sure to oxygen. To overcome these drawbacks, Torpol et al. encapsulated
a mixture of GEO and HBEO in pectin beads, which were coated with
chitosan and finally cross-linked with CaCl2 using the ionic gelation method.
The beads achieved a mean encapsulation efficiency of 70% of the oil mix
and cumulative release after storage at 5°C for 15 days. The inhibitory effect
was tested by depositing the pearls (4.4 g) in cellulose bags and applying it
to Bacillus cereus, Clostridium perfringens, Escherichia coli, Pseudomonas
florescens, Listeria monocytogenes, and Staphylococcus aureus, demon-
strating an inhibitory effect on all microorganisms. In view of these results,
they suggest that pearls can be included in storage bags or food packaging to
provide microbiological safety (Torpol et al., 2019).
1.5.2 LOADED WITH NATURAL EXTRACTS
Due to their biocompatible properties, hydrogels are ideal for the delivery
of phenolic compounds extracted from plant material since they can
maintain the stability of the compounds that are incorporated into them,
improving the nutritional value, color, and aroma of the food product. For
example, curcumin is a polyphenol with antimicrobial, anticancer, and anti-
inflammatory activity, but with low solubility; therefore, encapsulation in
vehicles such as hydrogels can increase its solubility, keep it stable, and
allow controlled and targeted delivery to the food of interest (Corkovi et
al., 2021). Thus, many of the phenolic and volatile compounds of interest
that have an antioxidant and antimicrobial effect can be used to prolong

the shelf life of foods, as long as their properties are maintained. This is
how the strategy of preserving them within hydrogel matrices seems to be
a good strategy.
Malaysian herbal extract is high in phenols and exhibits antioxidant
activities. Wan Yajana et al. extracted these biocomposites and added them
to an edible film developed with semirefined carrageenan and glycerol. Once
the film was obtained, meat burgers were wrapped with it and stored in the
refrigerator (4 ± 2°C) for 14 days. Lipid deterioration was significantly less
than the product that was not film-wrapped with the herbal extract. Due to
this evidence and that of other tests they carried out, they concluded that this
film can be used as a packaging material to improve the quality characteris-
tics of meat (Wan Yahaya et al., 2019).
Pomegranate extract is another biocompound that has been studied and
successfully incorporated into hydrogel matrices. As in the work of Maroufi
et al., who created a polymeric hydrogel pad based on chitosan, guar gum, and
modified dialdehyde, the pad had an absorption capacity of up to 12,000% of
its dry weight and loading with 5% pomegranate peel extract (PPE) showed
good antioxidant activity (81.13%) and antimicrobial efficacy against gram-
negative bacteria of Escherichia coli and gram-positive of Staphylococcus
aureus. Taking these results into consideration, they concluded that the
pomegranate extract-loaded hydrogel was a potentially suitable material for
absorbing liquid at the bottom of fresh meat, poultry, and fish package trays
to increase shelf life (Maroufi et al., 2021).
Tea polyphenols (TP) are the main active ingredient in tea, recognized
for their antioxidant, antiviral, and antibacterial biological activities.
These extracts have been widely used as additives in the pharmaceutical
and food industries because they have stable chemical properties and
good reactivity with biological macromolecules (Matsui, 2015). That is
why it has been studied as an active ingredient in polysaccharide-based
films. Gao et al. developed a film for the use in the food industry using
pectin, chitosan, and tea polyphenols, cross-linked with calcium ions.
They determined its antioxidant activity in vitro and its antiseptic and
color-protective capacity was evaluated in freshly cut beef cubes. The
film exhibited ability to scavenge free radicals, inhibitory ability against
bacteria and to prevent deterioration in meat color. These results led to the
conclusion that the film may have the value to be used as a packaging film
and slow release material, with the application in the food and pharmaceu-
tical industries (Gao et al., 2019).
1.5.3 EXTRACTS THAT FUNCTION AS INDICATORS
Some extracts have the property of responding colorimetrically to changes in

the chemical environment of the food. That is why, in addition to their anti-
oxidant and antimicrobial effects, they can be used as indicators in real time
by incorporating them into the hydrogels that are part of a food container.
Taking advantage of this peculiarity, Pereira et al. used a mixture of chitosan
polymers and polyvinyl alcohol to create a film for food packaging. This
film was added with anthocyanins extracted from red cabbage (Brassica
oleracea) to develop a time-temperature indicator. They carried out color
variation tests on the film in relation to the change in pH. They applied the
indicator to a sample of pasteurized milk and the film showed evident changes
in its color when the storage temperature increased, indicating changes in the
chemical composition of the milk as evidence of its deterioration (Pereira et
al., 2015).
Another novel colorimetric indicator used the ability of the Arnebia
euchroma root extract to change color in the presence of volatile basic
nitrogen (TVB-N), which manifests itself as the fish meat deteriorates
(increase in pH), being an indicator of its freshness in real time. The dye
was incorporated into a red algae agar matrix to form a film that was applied
directly to Wuchang goldfish (Megalobrama amblycephala). The test was
carried out at refrigeration temperatures (4°C) and at room temperature
(25°C). The results showed that the indicator film exhibited a noticeable
color change consistent with the deterioration threshold of TVB-N content.
This system appears to be a simple, nondestructive, and visual method to
estimate the freshness of fish during storage and transportation (Huang et
al., 2019).
Finally, some articles mention that in the food industry most of the hydro-
gels used are based on proteins and carbohydrates, since the use of synthetic
polymers has been related to possible cytotoxicity. In a following section, the
types of biopolymers that have been used to create hydrogels with applica-
tion in smart packaging for food packaging will be detailed.
1.6 FUTURE PERSPECTIVES FOR HYDROGELS IN THE FOOD

INDUSTRY
Despite the multiple works carried out to develop hydrogels suitable for
application in the food industry and all the natural materials that have been
used, it is still an area rich in opportunities for the creation of new types
of packaging, innovative for the entire range of food and at the same time,
friendly to the environment.
1.6.1 OTHER WAYS TO CREATE HYDROGEL PLATFORMS
The synthesis of hydrogels-type nanogels and microgels has been exten-

sively studied for controlled release of drugs, but could be adapted to food
preservation to release active compounds.
Packaging labels can be printed using injectable hydrogels, which can
be created from natural materials such as pectin. The color can be added to
the hydrogel and these colorants can be completely of natural origin, so that
their contact with the food does not have toxic effects. These natural dyes
could naturally degrade over time along with the shelf life of the food so that
the consumer is informed in real time that they should consume it before the
packaging acquires a certain intensity of color.
1.6.2 OTHER PARTICLES THAT CAN BE INCLUDED IN HYDROGELS
Although this chapter focused on materials of natural origin and biocompos-

ites, graphene and graphene nanomaterials are another very good alternative
because they are used in biomedicine for being a non-cytotoxic and biocom-
patible material. Therefore, it could be included in smart food packaging
since it is classified as an active and smart material.
In the same way, silver nanoparticles have wide applications in water
treatment, agriculture, medicine, drug administration, among others. The
activity that has attracted the most attention of these nanoparticles is their
antimicrobial function, which is why polysaccharide hydrogels can be
developed together with silver nanoparticles, for the production of new
antibacterial films with biocompatibility and stability of silver nanoparticles.
1.6.3 OTHER STIMULI TO WHICH HYDROGELS CAN RESPOND
Hydrogels that respond to the presence of bacterial metabolites (sensitive to

enzymes) are beginning to be explored. Its sensitivity is specific, so it can
be a qualitative and quantitative method. The current disadvantage is that it
takes a relatively long time, so the area of interest is to develop systems that
do it in a short time, since bacterial contamination in food represents a great

threat to human health. The sensitivity of hydrogels can also focus on the
separation of proteins and lipids. Its hydrophobic character can selectively
adsorb a hydrophobic target under certain conditions and thereby make the
separation. This answer can be applied to the benefit of the food industry.
If the field outside of food packaging is opened up a bit, thermally stable
hydrogels may find applications in food processing. These hydrogels can be
used for the separation of oil and water and other waste produced during food
processing. They also help masking of unpleasant tastes and the prevention
of scaling and boilers, pipes and heat exchange surfaces. The combination of
thermoresistant hydrogels has very unusual rheological and thermal proper-
ties and therefore resists much higher temperatures without melting.
Magnetic response is another attribute of smart hydrogels, but much
research remains to be done for practical application in the food area.
1.7 CONCLUSIONS
The large amount of work that has been carried out, using smart hydrogels,
has obtained promising results to be a safe and sustainable alternative to
plastic materials that are currently still used in excess, such as food pack-
aging. The long-term goal must be to achieve 100 percent biodegradable
packaging. Their elimination does not cause burden to the environment, they
can be created with materials obtained from agro-industrial waste and they
can be reincorporated into the environment in a natural way.
Something that is important to consider is that, despite all the advan-
tages that the use of biomaterials for food packaging can represent and the
additional benefits provided by the inclusion of bioactive components, it
cannot be ignored how it can affect the basic functionality of the packaging.
A package must have mechanical resistance, permeability to water and
oxygen, protection against light, and preferably transparency to be attractive
to the consumer. The alteration of the packaging can affect the brightness,
appearance, flavor, and even the crispness of the food, something that can be
negative in consumer preference.
Another disadvantage that must be overcome is that, although stimuli-
responsive hydrogels are already widely known and elaborated, as well as
the way to load them with bioactive compounds, it is necessary to develop
suitable manufacturing methods to produce these hydrogels commercially, a
scale that is economically profitable for the industry.
KEYWORDS
• hydrogel
• food packaging
• polymers
• active compounds
• microorganisms
• food safety
REFERENCES
Abaee, A.; Mohammadian, M.; Jafari, S. M. Whey and Soy Protein-Based Hydrogels and
Nano-Hydrogels as Bioactive Delivery Systems. Trends Food Sci. Technol. 2017, 70,
69–81. https://doi.org/10.1016/j.tifs.2017.10.011
Abou Taleb, M. F. Radiation Synthesis of Multifunctional Polymeric Hydrogels for Oral
Delivery of Insulin. Int. J. Biol. Macromol. 2013, 62, 341–347. https://doi.org/10.1016/j.
ijbiomac.2013.09.004
Aida, T.; Sun, Z.; Yamauchi, Y.; Araoka, F.; Kim, Y. S.; Bergueiro, J.; Ishida, Y.; Ebina, Y.;
Sasaki, T.; Hikima, T. An Anisotropic Hydrogel Actuator Enabling Earthworm-like Directed
Peristaltic Crawling. Angew. Chem. 2018, 130 (48), 15772–15776. https://doi.org/10.1002/
ange.201810052
Alejandre, M.; Astiasarán, I.; Ansorena, D.; Barbut, S. Using Canola Oil Hydrogels and
Organogels to Reduce Saturated Animal Fat in Meat Batters. Food Res. Int. 2019, 122,
129–136. https://doi.org/10.1016/J.FOODRES.2019.03.056
Amiryousefi, M. R.; Mohebbi, M.; Golmohammadzadeh, S.; Koocheki, A. Encapsulation
of Caffeine in Hydrogel Colloidosome: Optimization of Fabrication, Characterization
and Release Kinetics Evaluation. Flavour Fragr. J. 2016, 31 (2), 163–172. https://doi.
org/10.1002/FFJ.3297
Baek, S.; Kim, D.; Jeon, S. L.; Seo, J. Preparation and Characterization of PH-Responsive
Poly(N,N-Dimethyl Acrylamide-Co-Methacryloyl Sulfadimethoxine) Hydrogels for
Application as Food Freshness Indicators. React. Funct. Polym. 2017, 120 (September),
57–65. https://doi.org/10.1016/j.reactfunctpolym.2017.09.003
Bahú, J. O.; de Andrade, L. R. M.; de Melo Barbosa, R.; Crivellin, S.; da Silva, A. P.; Souza,
S. D. A.; Cárdenas Concha, V. O.; Severino, P.; Souto, E. B. Plant Polysaccharides in
Engineered Pharmaceutical Gels. Bioengineering 2022, 9 (8), 376. https://doi.org/10.3390/
bioengineering9080376
Bao, Z.; Xian, C.; Yuan, Q.; Liu, G.; Wu, J. Natural Polymer-Based Hydrogels with Enhanced
Mechanical Performances: Preparation, Structure, and Property. Adv. Healthc. Mater. 2019,
8 (17), 1–11. https://doi.org/10.1002/adhm.201900670
Barragán-Martínez, L. P.; Román-Guerrero, A.; Vernon-Carter, E. J.; Alvarez-Ramirez,
J. Impact of Fat Replacement by a Hybrid Gel (Canola Oil/Candelilla Wax Oleogel and
Gelatinized Corn Starch Hydrogel) on Dough Viscoelasticity, Color, Texture, Structure, and
Starch Digestibility of Sugar-Snap Cookies. Int. J. Gastron. Food Sci. 2022, 29, 100563.
https://doi.org/10.1016/J.IJGFS.2022.100563
Barroso, N.; Guaresti, O.; Pérez-álvarez, L.; Ruiz-rubio, L. Self-Healable Hyaluronic Acid/
Chitosan Polyelectrolyte Complex Hydrogels and Multilayers. Eur. Polym. J. 2019, 120
(September), 109268. https://doi.org/10.1016/j.eurpolymj.2019.109268
Batista, R. A.; Espitia, P. J. P.; de Quintans, J. S. S.; Freitas, M. M.; Cerqueira, M. Â.;
Teixeira, J. A.; Cardoso, J. C. Hydrogel as an Alternative Structure for Food Packaging
Systems. Carbohydr. Polym. 2019, 205, 106–116. https://doi.org/https://doi.org/10.1016/j.
carbpol.2018.10.006
Batista, R. A.; Perez Espitia, P. J.; Quintans, J. D. S. S.; Machado, M.; Ângelo, M.; António, J.;
Cordeiro, J. Hydrogel as an Alternative Structure for Food Packaging Systems. Carbohydr.
Polym. 2019, 205, 106–116.
Beigmohammadi, F.; Peighambardoust, S. H.; Hesari, J.; Azadmard-Damirchi, S.;
Peighambardoust, S. J.; Khosrowshahi, N. K. Antibacterial Properties of LDPE
Nanocomposite Films in Packaging of UF Cheese. LWT - Food Sci. Technol. 2016, 65,
106–111. https://doi.org/10.1016/J.LWT.2015.07.059
Berton, S. B. R.; de Jesus, G. A. M.; Sabino, R. M.; Monteiro, J. P.; Venter, S. A. S.; Bruschi, M.
L.; Popat, K. C.; Matsushita, M.; Martins, A. F.; Bonafé, E. G. Properties of a Commercial
κ-Carrageenan Food Ingredient and Its Durable Superabsorbent Hydrogels. Carbohydr Res
2020, 487, 107883. https://doi.org/10.1016/j.carres.2019.107883.
Bodbodak, S.; Rafiee, Z. Recent Trends in Active Packaging in Fruits and Vegetables; Elsevier
Inc., 2016. https://doi.org/10.1016/B978-0-12-804313-4.00003-7
Bourbon, A. I.; Pereira, R. N.; Pastrana, L. M.; Vicente, A. A.; Cerqueira, M. A. Protein-based
Nanostructures for Food Applications. Gels 2019, 5 (1), 1–18. https://doi.org/10.3390/
gels5010009
Buenger, D.; Topuz, F.; Groll, J. Hydrogels in Sensing Applications. Prog. Polym. Sci. 2012,
37, 1678–1719. https://doi.org/10.1016/j.progpolymsci.2012.09.001
Chacon, W.; Paz-arteaga, S.; Torres-león, C.; Ayala Valencia, G. Gums-Based Coatings
Applied to Extend the Shelf Life of Foods: A Review. J. Polym. Environ. 2022, 2020.
Chang, Y.; Chen, W.; Yandi, W.; Shih, Y.; Chu, W. Dual-Thermoresponsive Phase Behavior
of Blood Compatible Zwitterionic Copolymers Containing Nonionic Poly (N-Isopropyl
Acrylamide). Biomacromolecules 2009, 10 (8), 2092–2100.
Corkovi, I.; Pichler, A.; Šimunovi, J.; Kopjar, M. Hydrogels: Characteristics and Application
as Delivery Systems of Phenolic and Aroma Compounds. Foods 2021, 10 (6), 1252.
Crini, G.; Torri, G.; Lichtfouse, E.; Kyzas, G. Z.; Wilson, L. D.; Morin-Crini, N. Dye Removal
by Biosorption Using Cross-Linked Chitosan-Based Hydrogels. Environ. Chem. Lett. 2019,
17 (4), 1645–1666. https://doi.org/10.1007/s10311-019-00903-y
Dalponte Dallabona, I.; de Lima, G. G.; Cestaro, B. I.; Tasso, I. D. S.; Paiva, T. S.; Laureanti,
E. J. G.; Jorge, L. M. D. M.; da Silva, B. J. G.; Helm, C. V.; Mathias, A. L.; Jorge, R. M. M.
Development of Alginate Beads with Encapsulated Jabuticaba Peel and Propolis Extracts to
Achieve a New Natural Colorant Antioxidant Additive. Int. J. Biol. Macromol. 2020, 163,
1421–1432. https://doi.org/10.1016/j.ijbiomac.2020.07.256
Dehshahri, A.; Kumar, A.; Madamsetty, V. S.; Uzieliene, I.; Tavakol, S.; Azedi, F.; Fekri,
H. S.; Zarrabi, A.; Mohammadinejad, R.; Thakur, V. K. New Horizons in Hydrogels for
Methotrexate Delivery. Gels 2021, 7 (1), 1–20. https://doi.org/10.3390/gels7010002
Deng, X.; Chen, J.; Chen, W. Hydrogel Particles as a Controlled Release Delivery System for
Lavender Essential Oil Using PH Triggers. Colloids Surf. A 2020, 603, 125134. https://doi.
org/10.1016/j.colsurfa.2020.125134
Ding, M.; Jing, L.; Yang, H.; Machnicki, C. E.; Fu, X.; Li, K.; Wong, I. Y.; Chen, P.
Multifunctional Soft Machines Based on Stimuli-Responsive Hydrogels: From Freestanding
Hydrogels to Smart Integrated Systems. Mater. Today Adv. 2020, 8, 1–19. https://doi.
org/10.1016/j.mtadv.2020.100088
Djoullah, A.; Saurel, R. Controlled Release of Riboflavin Encapsulated in Pea Protein
Microparticles Prepared by Emulsion-Enzymatic Gelation Process. J. Food Eng. 2021,
292. https://doi.org/10.1016/j.jfoodeng.2020.110276
Dos Santos, M.; Ozaki, M. M.; Ribeiro, W. O.; Paglarini, C. D. S.; Vidal, V. A. S.; Campagnol,
P. C. B.; Pollonio, M. A. R. Emulsion Gels Based on Pork Skin and Dietary Fibers as
Animal Fat Replacers in Meat Emulsions: An Adding Value Strategy to Byproducts. LWT
2020, 120, 108895. https://doi.org/10.1016/J.LWT.2019.108895
Firestone, B. A.; Siegel, R. A. Kinetics and Mechanisms of Water Sorption in Hydrophobic,
Ionizable Copolymer Gels. J. Appl. Polym. Sci. 1991, 43, 901–914.
Gao, H. X.; He, Z.; Sun, Q.; He, Q.; Zeng, W. C. A Functional Polysaccharide Film Forming
by Pectin, Chitosan, and Tea Polyphenols. Carbohydr. Polym. 2019, 215, 1–7. https://doi.
org/10.1016/j.carbpol.2019.03.029
Geyer, R. Production, Use, and Fate of Synthetic Polymers; Elsevier Inc., 2020. https://doi.
org/10.1016/b978-0-12-817880-5.00002-5
Ghosh, T.; Katiyar, V. Cellulose-Based Hydrogel Films for Food Packaging; 2019. https://doi.
org/10.1007/978-3-319-77830-3_35
Goudoulas, T. B.; Vanderhaeghen, S.; Germann, N. Micro-Dispersed Essential Oils Loaded
Gelatin Hydrogels with Antibacterial Activity. LWT - Food Sci. Technol. 2022, 154, 1–10.
https://doi.org/10.1016/j.lwt.2021.112797
Guo, J. S.; Fang, F.; Yan, P.; Chen, Y. P. Sludge Reduction Based on Microbial Metabolism
for Sustainable Wastewater Treatment. Bioresour. Technol. 2020, 297 (September 2019),
122506. https://doi.org/10.1016/j.biortech.2019.122506
Heimbuck, A. M.; Priddy-Arrington, T. R.; Sawyer, B. J.; Caldorera-Moore, M. E. Effects
of Post-Processing Methods on Chitosan-Genipin Hydrogel Properties. Mater. Sci. Eng. C
2019, 98 (April 2018), 612–618. https://doi.org/10.1016/j.msec.2018.12.119
Heskins, M.; Guillet, J. E. Solution Properties of Poly (N-Isopropylacrylamide). J. Macromol.
Sci.: Part A- Chem. 1968, 2, 1441–1455.
Hu, W.; Wang, Z.; Xiao, Y.; Zhang, S.; Wang, J. Advances in Crosslinking Strategies of
Biomedical Hydrogels. Biomater. Sci. 2019, 7 (3), 843–855. https://doi.org/10.1039/
c8bm01246f
Huang, S.; Xiong, Y.; Zou, Y.; Dong, Q.; Ding, F.; Liu, X. A Novel Colorimetric Indicator Based
on Agar Incorporated with Arnebia Euchroma Root Extracts for Monitoring Fish Freshness.
Food Hydrocoll. 2019, 90, 198–205. https://doi.org/10.1016/j.foodhyd.2018.12.009
Irastorza, A.; Zarandona, I.; Andonegi, M.; Guerrero, P.; de la Caba, K. The Versatility of
Collagen and Chitosan: From Food to Biomedical Applications. Food Hydrocoll. 2021,
116, 1–11. https://doi.org/10.1016/j.foodhyd.2021.106633
Kalia, S. Polymeric Hydrogels as Smart Biomaterials (Springer Series on Polymer and
Composite Materials); Springer International Publishing, 2015.
Khalesi, H.; Lu, W.; Nishinari, K.; Fang, Y. Fundamentals of Composites Containing Fibrous
Materials and Hydrogels: A Review on Design and Development for Food Applications.
Food Chem. 2021, 364 (June), 130329. https://doi.org/10.1016/j.foodchem.2021.130329
Kloxin, A. M.; Kasko, A. M.; Salinas, C. N.; Anseth, K. S. Photodegradable Hydrogels for
Dynamic Tuning of Physical and Chemical Properties. Science (1979) 2013, 324 (2009),
59–63. https://doi.org/10.1126/science.1169494
Kumari, P. V. K.; Sharmila, M.; Rao, Y. S. Super Porous Hydrogels: A Review. J. Pharm. Res.
Int. 2020, 153–165. https://doi.org/10.9734/jpri/2020/v32i1330595
Kwan, A.; Davidov-Pardo, G. Controlled Release of Flavor Oil Nanoemulsions Encapsulated
in Filled Soluble Hydrogels. Food Chem. 2018, 250 (December 2017), 46–53. https://doi.
org/10.1016/j.foodchem.2017.12.089
Kwon, G. H.; Choi, Y.; Park, Y.; Woo, H.; Lee, B.; Kim, H.; Lee, S. Electrically-Driven
Hydrogel Actuators in Microfluidic Channels: Fabrication, Characterization, and Biological
Application. Lab-on-a-Chip 2010, 10, 1604–1610. https://doi.org/10.1039/b926443d
Leyva-Jiménez, F. J.; Oliver-Simancas, R.; Castangia, I.; Rodríguez-García, A. M.;
Alañón, M. E. Comprehensive Review of Natural Based Hydrogels as an Upcoming
Trend for Food Packing. Food Hydrocoll. 2023, 135, 108124. https://doi.org/10.1016/J.
FOODHYD.2022.108124
Li, J.; Jia, X.; Yin, L. Hydrogel: Diversity of Structures and Applications in Food Science
Hydrogel: Diversity of Structures and Applications in Food Science. Food Rev. Int. 2021,
37 (3), 313–372. https://doi.org/10.1080/87559129.2020.1858313.
Li, L.; Scheiger, J. M.; Levkin, P. A. Design and Applications of Photoresponsive Hydrogels.
Adv. Mat. Res. 2019, 31, 1–17. https://doi.org/10.1002/adma.201807333
Lin, J.; Jiao, G.; Kermanshahi-pour, A. Algal Polysaccharides-Based Hydrogels: Extraction,
Synthesis, Characterization, and Applications. Mar. Drugs 2012, 253–264. https://doi.
org/10.1201/b13868-22
Liu, K.; Chen, Y. Y.; Zha, X. Q.; Li, Q. M.; Pan, L. H.; Luo, J. P. Research Progress on
Polysaccharide/Protein Hydrogels: Preparation Method, Functional Property and
Application as Delivery Systems for Bioactive Ingredients. Food Res. Int. 2021, 147, 1–16.
https://doi.org/10.1016/j.foodres.2021.110542
Liu, Z.; Liu, C.; Sun, X.; Zhang, S.; Yuan, Y.; Wang, D.; Xu, Y. Fabrication and Characterization
of Cold-Gelation Whey Protein-Chitosan Complex Hydrogels for the Controlled Release
of Curcumin. Food Hydrocoll. 2020, 103 (May 2019), 105619. https://doi.org/10.1016/j.
foodhyd.2019.105619
Lopes, P. M. P.; Moldovan, D.; Moldovan, M.; Carpa, R.; Saroşi, C.; Păşcuţă, P.; Moldovan,
A. M.; Fechete, R.; Popescu, V. New Composite Hydrogel Based on Whey and Gelatin
Crosslinked with Copper Sulphate. Materials 2022, 15 (7), 1–21. https://doi.org/10.3390/
ma15072611
Lu, P.; Yang, Y.; Liu, R.; Liu, X.; Ma, J.; Wu, M.; Wang, S. Preparation of Sugarcane Bagasse
Nanocellulose Hydrogel as a Colourimetric Freshness Indicator for Intelligent Food
Packaging. Carbohydr. Polym. 2020, 249 (August), 116831. https://doi.org/10.1016/j.
carbpol.2020.116831
Maddaloni, M.; Vassalini, I.; Alessandri, I. Green Routes for the Development of Chitin/
Chitosan Sustainable Hydrogels. Sustain. Chem. 2020, 1 (3), 325–344. https://doi.
org/10.3390/suschem1030022
Mahinroosta, M.; Jomeh, Z.; Allahverdi, A.; Shakoori, Z. Hydrogels as Intelligent Materials:
A Brief Review of Synthesis, Properties and Applications. Mater. Today Chem. 2018, 8,
42–55. https://doi.org/10.1016/j.mtchem.2018.02.004
Maltais, A.; Remondetto, G. E.; Subirade, M. Soy Protein Cold-Set Hydrogels as Controlled
Delivery Devices for Nutraceutical Compounds. Food Hydrocoll. 2009, 23 (7), 1647–1653.
https://doi.org/10.1016/j.foodhyd.2008.12.006
Manzoor, A.; Dar, A. H.; Pandey, V. K.; Shams, R.; Khan, S.; Panesar, P. S.; Kennedy, J.
F.; Fayaz, U.; Khan, S. A. Recent Insights into Polysaccharide-Based Hydrogels and
Their Potential Applications in Food Sector: A Review. Int. J. Biol. Macromol. 2022, 213,
987–1006. https://doi.org/10.1016/J.IJBIOMAC.2022.06.044
Maroufi, L. Y.; Tabibiazar, M.; Ghorbani, M.; Jahanban-Esfahlan, A. Fabrication and
Characterization of Novel Antibacterial Chitosan/Dialdehyde Guar Gum Hydrogels
Containing Pomegranate Peel Extract for Active Food Packaging Application. Int. J. Biol.
Macromol. 2021, 187 (July), 179–188. https://doi.org/10.1016/j.ijbiomac.2021.07.126
Matsui, T. Condensed Catechins and Their Potential Health-Bene Fi Ts. Eur. J. Pharmacol.
2015, 765, 495–502. https://doi.org/10.1016/j.ejphar.2015.09.017
Meng, Y.; Cloutier, S. Gelatin and Other Proteins for Microencapsulation; Elsevier Inc.,
2014. https://doi.org/10.1016/b978-0-12-404568-2.00020-0
Mirfakhrai, T.; Madden, J. D. W.; Baughman, R. H. Polymer Artificial. Materials Today,
2007, 10 (4), 30–38
Moschou, E. A.; Peteu, S. F.; Bachas, L. G.; Madou, M. J.; Daunert, S. Artificial Muscle
Material with Fast Electroactuation under Neutral PH Conditions. Chem. Mater. 2004, 16,
2499–2502.
Mujtaba, M.; Morsi, R. E.; Kerch, G.; Elsabee, M. Z.; Kaya, M.; Labidi, J.; Khawar, K. M.
Current Advancements in Chitosan-Based Film Production for Food Technology; A Review.
Int. J. Biol. Macromol. 2019, 121, 889–904. https://doi.org/10.1016/j.ijbiomac.2018.10.109
Mukhopadhyay, P.; Sarkar, K.; Bhattacharya, S.; Bhattacharyya, A.; Mishra, R.; Kundu, P. P. PH
Sensitive N-Succinyl Chitosan Grafted Polyacrylamide Hydrogel for Oral Insulin Delivery.
Carbohydr. Polym. 2014, 112, 627–637. https://doi.org/10.1016/j.carbpol.2014.06.045
Müller, P.; Schmid, M. Intelligent. Packaging in the Food Sector: A Brief Overview. Foods
2019, 8 (1), 16.
Nezhad-Mokhtari, P.; Ghorbani, M.; Roshangar, L.; Soleimani Rad, J. Chemical Gelling
of Hydrogels-Based Biological Macromolecules for Tissue Engineering: Photo- and
Enzymatic-Crosslinking Methods. Int. J. Biol. Macromol. 2019, 139, 760–772. https://doi.
org/10.1016/j.ijbiomac.2019.08.047
Osojnik Črnivec, I. G.; Poklar Ulrih, N. Nano-Hydrogels of Alginate for Encapsulation of
Food Ingredients; 2019. https://doi.org/10.1016/B978-0-12-815663-6.00013-6
Oun, A. A.; Rhim, J. W. Carrageenan-Based Hydrogels and Films: Effect of ZnO and CuO
Nanoparticles on the Physical, Mechanical, and Antimicrobial Properties. Food Hydrocoll.
2017, 67, 45–53. https://doi.org/10.1016/J.FOODHYD.2016.12.040
Panahi, R.; Baghban-Salehi, M. Protein-Based Hydrogels; 2014. https://doi.
org/10.4032/9789814463218
Park, S.; Mun, S.; Kim, Y. R. Effect of Xanthan Gum on Lipid Digestion and Bioaccessibility
of β-Carotene-Loaded Rice Starch-Based Filled Hydrogels. Food Res. Int. 2018, 105
(August 2017), 440–445. https://doi.org/10.1016/j.foodres.2017.11.039
Peppas, N. A.; Hoffman, A. S. Hydrogels. In Biomaterials Science, 4th ed.; 2020, No. 1; pp
153–166. https://doi.org/10.1016/B978-0-12-816137-1.00014-3.
Pereira, V. A.; de Arruda, I. N. Q.; Stefani, R. Active Chitosan/PVA Films with Anthocyanins
from Brassica Oleraceae (Red Cabbage) as Time–Temperature Indicators for Application in
Intelligent Food Packaging. Food Hydrocoll. 2015, 43, 180–188. https://doi.org/10.1016/j.
foodhyd.2014.05.014
Qu, B.; Luo, Y. Chitosan-Based Hydrogel Beads: Preparations, Modifications and Applications
in Food and Agriculture Sectors—A Review. Int. J. Biol. Macromol. 2020, 152, 437–448.
https://doi.org/10.1016/j.ijbiomac.2020.02.240
Ranganathan, S.; Dutta, S.; Moses, J. A.; Anandharamakrishnan, C. Utilization of Food
Waste Streams for the Production of Biopolymers. Heliyon 2020, 6 (9), 1–13. https://doi.
org/10.1016/j.heliyon.2020.e04891
Rhim, J. W.; Wang, L. F. Mechanical and Water Barrier Properties of Agar/κ-Carrageenan/
Konjac Glucomannan Ternary Blend Biohydrogel Films. Carbohydr. Polym. 2013, 96 (1),
71–81. https://doi.org/10.1016/J.CARBPOL.2013.03.083
Sadiq, U.; Gill, H.; Chandrapala, J. Casein Micelles as an Emerging Delivery System
for Bioactive Food Components. Foods 2021, 10 (8), 1965. https://doi.org/10.3390/
foods10081965
Sarrigiannidis, S. O.; Rey, J. M.; Dobre, O.; González-García, C.; Dalby, M. J.; Salmeron-
Sanchez, M. A Tough Act to Follow: Collagen Hydrogel Modifications to Improve
Mechanical and Growth Factor Loading Capabilities. Mater. Today Bio. 2021, 10, 1–22.
https://doi.org/10.1016/j.mtbio.2021.100098
Seuring, J.; Agarwal, S. First Example of a Universal and Cost-E Ff Ective Approach:
Polymers with Tunable Upper Critical Solution Temperature in Water and Electrolyte
Solution. 2012.
Shaghaleh, H.; Hamoud, Y. A.; Xu, X.; Liu, H.; Wang, S.; Sheteiwy, M.; Dong, F.; Guo,
L.; Qian, Y.; Li, P.; Zhang, S. Thermo-/PH-Responsive Preservative Delivery Based on
TEMPO Cellulose Nanofiber/Cationic Copolymer Hydrogel Film in Fruit Packaging. Int.
J. Biol. Macromol. 2021, 183, 1911–1924. https://doi.org/10.1016/j.ijbiomac.2021.05.208
Shen, M.; Song, B.; Zeng, G.; Zhang, Y.; Huang, W.; Wen, X.; Tang, W. Are Biodegradable
Plastics a Promising Solution to Solve the Global Plastic Pollution? Environ. Pollut. 2020,
263, 114469. https://doi.org/10.1016/j.envpol.2020.114469.
Shi, M.; Shi, Y. L.; Li, X. M.; Yang, R.; Cai, Z. Y.; Li, Q. S.; Ma, S. C.; Ye, J. H.; Lu, J. L.;
Liang, Y. R.; Zheng, X. Q. Food-Grade Encapsulation Systems for (-)-Epigallocatechin
Gallate. Molecules 2018, 23 (2), 1–17. https://doi.org/10.3390/molecules23020445
Sutthasupa, S.; Padungkit, C.; Suriyong, S. Colorimetric Ammonia (NH3) Sensor Based
on an Alginate-Methylcellulose Blend Hydrogel and the Potential Opportunity for the
Development of a Minced Pork Spoilage Indicator. Food Chem. 2021, 362, 130151. https://
doi.org/10.1016/j.foodchem.2021.130151
Tao, Y.; Zhang, R.; Xu, W.; Bai, Z.; Zhou, Y.; Zhao, S.; Xu, Y.; Yu, D. Rheological Behavior and
Microstructure of Release-Controlled Hydrogels Based on Xanthan Gum Crosslinked with
Sodium Trimetaphosphate. Food Hydrocoll. 2016, 52, 923–933. https://doi.org/10.1016/j.
foodhyd.2015.09.006
Taylor, M. J.; Tomlins, P.; Sahota, T. S. Thermoresponsive Gels. Gels 2017, 3 (4), 1–31.
https://doi.org/10.3390/gels3010004
Torpol, K.; Sriwattana, S.; Sangsuwan, J.; Wiriyacharee, P.; Prinyawiwatkul, W. Optimising
Chitosan—Pectin Hydrogel Beads Containing Combined Garlic and Holy Basil Essential
Oils and Their Application as Antimicrobial Inhibitor. Food Sci. Technol. 2019, 54 (6),
2064–2074. https://doi.org/10.1111/ijfs.14107
Torres-León, C.; Aguilar, C. Food Preservation. In Quantitative Methods and Analytical

Techniques in Food Microbiology; Sepulveda, L., N. Aguilar, C., Porteen, K., Haghi, A.,
Eds.; Apple Academic Press Inc.: Abingdon, Oxon, OX14 4RN UK, 2022; pp 39–57.
Torres-León, C.; Sepulveda, L.; Aguilar, C. N. Food and Diseases: What to Know in the
Fight to Ensure Food Safety. In Quantitative Methods and Analytical Techniques in Food
Microbiology; Leonardo, S. T., Cristóbal, N. A., Porteen Kannan, A. K. H., Ed.; Apple
Academic Press Inc.: Abingdon, Oxon, OX14 4RN UK, 2022; pp 57–75.
Torres-león, C.; Ventura-Sobrevilla, J.; Serna-Cock, L.; Ascacio-Valdés, J. A. J. A.; Contreras-
Esquivel, J.; Aguilar, C. N. Pentagalloylglucose (PGG): A Valuable Phenolic Compound
with Functional Properties. J. Funct. Foods 2017, 37, 176–189. https://doi.org/10.1016/j.
jff.2017.07.045
Torres-León, C.; Vicente, A. A.; Flores-López, M. L.; Rojas, R.; Serna-Cock, L.; Alvarez-
Pérez, O. B.; Aguilar, C. N. Edible Films and Coatings Based on Mango (Var. Ataulfo)
by-Products to Improve Gas Transfer Rate of Peach. LWT - Food Sci. Technol. 2018, 97,
624–631. https://doi.org/10.1016/j.lwt.2018.07.057
Trifković, K. T.; Milašinović, N. Z.; Djordjević, V. B.; Krušić, M. T. K.; Knežević-Jugović,
Z. D.; Nedović, V. A.; Bugarski, B. M. Chitosan Microbeads for Encapsulation of Thyme
(Thymus Serpyllum L.) Polyphenols. Carbohydr. Polym. 2014, 111, 901–907. https://doi.
org/10.1016/J.CARBPOL.2014.05.053
Ullah, F.; Othman, M. B. H.; Javed, F.; Ahmad, Z.; Akil, H. M. Classification, Processing and
Application of Hydrogels: A Review. Mater. Sci. Eng. C 2015, 57, 414–433. https://doi.
org/10.1016/j.msec.2015.07.053
Varghese, S. A.; Rangappa, S. M.; Siengchin, S.; Parameswaranpillai, J. Natural Polymers
and the Hydrogels Prepared from Them; Elsevier Inc., 2019. https://doi.org/10.1016/
B978-0-12-816421-1.00002-1
Wan Yahaya, W. A.; Yazid, N. A.; Mohod Azman, N. A.; Almajano, M. P. Antioxidant Activities
and Total Phenolic Content of Malaysian Herbs as Components of Active Packaging Film
in Beef Patties. Antioxidants 2019, 8 (7), 1–15.
Wu, W.; Mitra, N.; Yan, E. C. Y.; Zhou, S. Multifunctional Hybrid Nanogel for and Self-
Regulated Insulin Release at Physiological PH. ACS Nano 2010, 4 (8), 4831–4839.
Xu, Y.; Bolisetty, S.; Drechsler, M.; Fang, B.; Yuan, J.; Ballauff, M.; Mu, A. H. E. PH and Salt
Responsive Poly (N, N-Dimethylaminoethyl Methacrylate ) Cylindrical Brushes and Their
Quaternized Derivatives. Polymer (Guildf) 2008, 49, 3957–3964. https://doi.org/10.1016/j.
polymer.2008.06.051
Yan, L.; Wang, L.; Gao, S.; Liu, C.; Zhang, Z.; Ma, A.; Zheng, L. Celery Cellulose Hydrogel
as Carriers for Controlled Release of Short-Chain Fatty Acid by Ultrasound. Food Chem.
2020, 309, 125717. https://doi.org/10.1016/j.foodchem.2019.125717
Yang, C.; Liu, Z.; Chen, C.; Shi, K.; Zhang, L.; Ju, X.-J.; Wang, W.; Xie, R.; Chu, L.-Y.
Reduced Graphene Oxide-Containing Smart Hydrogels with Excellent Electro-Response
and Mechanical Property for Soft Actuators. ACS Appl. Mater. Interfaces, 2017, 9,
15758–15767.
Yang, J.; Shen, M.; Luo, Y.; Wu, T.; Chen, X.; Wang, Y.; Xie, J. Advanced Applications
of Chitosan-Based Hydrogels: From Biosensors to Intelligent Food Packaging System.
Trends Food Sci. Technol. 2021, 110, 822–832. https://doi.org/https://doi.org/10.1016/j.
tifs.2021.02.032
Yang, M.; Yuan, Z.; Liu, J.; Fang, Z.; Fang, L.; Yu, D.; Li, Q. Photoresponsive Actuators
Built from Carbon-Based Soft Materials. Adv. Opt. Mater. 2019, 7 (16), 1–30. https://doi.
org/10.1002/adom.201900069
Yang, Z.; Chen, L.; McClements, D. J.; Qiu, C.; Li, C.; Zhang, Z.; Miao, M.; Tian, Y.; Zhu, K.;
Jin, Z. Stimulus-Responsive Hydrogels in Food Science: A Review. Food Hydrocoll. 2022,
124, 107218. https://doi.org/10.1016/j.foodhyd.2021.107218
Yang, Z.; Chen, X.; Xu, Z.; Ji, N.; Xiong, L.; Sun, Q. Anti-Freezing Starch Hydrogels with
Superior Mechanical Properties and Water Retention Ability for 3D Printing. Int. J. Biol.
Macromol. 2021, 190, 382–389. https://doi.org/10.1016/j.ijbiomac.2021.08.235
Zhang, J.; Liu, S.; Hui, X.; Xie, Z.; Jing, X. Cyanine-Curcumin Assembling Nanoparticles for
Near-Infrared Imaging and Photothermal Therapy. ACS Biomater. Sci. Eng. 2016, 2 (11),
1942–1950. https://doi.org/10.1021/acsbiomaterials.6b00315
Zhang, Z.; Zhang, R.; Chen, L.; McClements, D. J. Encapsulation of Lactase (β-Galactosidase)
into κ-Carrageenan-Based Hydrogel Beads: Impact of Environmental Conditions on Enzyme
Activity. Food Chem. 2016, 200, 69–75. https://doi.org/10.1016/j.foodchem.2016.01.014
Zhao, M.; Wang, P.; Guo, Y.; Wang, L.; Luo, F.; Qiu, B.; Guo, L.; Su, X.; Lin, Z.; Chen,
G. Detection of a Fl Atoxin B 1 in Food Samples Based on Target-Responsive Aptamer-
Cross-Linked Hydrogel Using a Handheld PH Meter as Readout. Talanta 2018, 176, 34–39.
https://doi.org/10.1016/j.talanta.2017.08.006
Zhu, F. Encapsulation and Delivery of Food Ingredients Using Starch Based Systems. Food
Chem 2017, 229, 542–552. https://doi.org/10.1016/j.foodchem.2017.02.101
CHAPTER 2
Bionanocomposites Based on
Thermoplastic Starch for the Food
Industry
JUAN M. TIRADO-GALLEGOS1, EMILIO OCHOA-REYES2,
JUAN C. BUSTILLOS-RODRÍGUEZ3, JULIO C. TAFOLLA-ARELLANO4,
FRANCISCO HERNÁNDEZ-CENTENO4, and
JUAN BUENROSTRO-FIGUEROA2
1
Tecnología de Alimentos de Origen Animal, Facultad de Zootecnia y
Ecología, Universidad Autónoma de Chihuahua, Chihuahua, Chihuahua,
México
Research Center in Food and Development, Cd. Delicias, Chihuahua,
2
México
3
Tecnológico Nacional de México, Instituto Tecnológico de Cd.
Cuauhtémoc, Cd. Cuauhtémoc, Chihuahua, México
4
Universidad Autónoma Agraria Antonio Narro, Department of Food
Science and Technology, Saltillo, Coahuila, México
ABSTRACT
Food packaging plays an important role in the determination of shelf life

of food. The principal functions are to contain and protect the food during
their storage and distribution. Also, the packaging informs the consumer
about the food properties and conditions of use. For this purpose, most of
the materials used as food packaging are the synthetic packaging materials.
However, these materials are nonbiodegradable, and their excessive use
promotes ecological problems. For this reason, there is an increasing interest
in the development of biodegradable packaging from starch alone or as part

of a composite, which offers great potential to replace the conventional food
packaging due to its versatility, availability, and low cost. Nevertheless,
starch thermoplastic materials present poor mechanical and barrier properties
compared with synthetic polymers. Research groups have tried to overcome
these limitations adding additives. In this context, the nanotechnology repre-
sents a great potential to design new packaging materials with improved
physicochemical properties. In this chapter, we analyze and summarize the
recent advances in the use of nanomaterials like nanocrystals, nanoparticles,
nanoclays, among others on the design of bionanocomposites obtained from
thermoplastic starch and their application as food packaging.
2.1 INTRODUCTION
Nanotechnology is an important tool to develop and improve advanced mate-

rials. Nanotechnology implies to work on a scale of about 1–100 nm to the
characterization, preparation, and influence of material structures, devices,
and systems with new properties according to their nanostructures (Azeredo,
2009; Ashfaq et al., 2022; Sharma et al., 2020). As a result, nanocomposited
materials have a large surface-to-volume ratio and surface activity, which
allows adding to compatible polymers matrix with different loadings, and this
linkage will improve the mechanical, thermal stability, and electrical conduc-
tivity. For that, nanomaterials are favorable to use in food packaging materials
to produce smart and active packaging (Youssef and El-Sayed, 2018).
The environmental problems derived from petroleum-based polymers,
biopolymers (naturally occurring polymers obtained from renewable
resources) have emerged as renewable packaging materials such as a
diverse set of lipids (wax and free fatty acids), proteins (casein, gluten, and
whey), polysaccharides (alginates, chitosan, cellulose, and starch derivates),
polyphenols, and polymers produced by bacteria, fungi, plants, and animals
(Kaplan, 1998). Within the biopolymers produced from polysaccharides, the
use of starch and its derivates has been especially interesting.
Starch is a biopolymer used to obtain biocomposite materials with several
advantages: is renewable source, completely biodegradable, nontoxicity,
easily available, and low cost (Mendes et al., 2016; Fahmy et al., 2020; Ahari
et al., 2021; Gamage et al., 2022). Thermoplastic starch (TPS) is obtained
after disruption and plasticization of native starch using plasticizers such as
glycerol, water, and sorbitol. TPS present poor mechanical properties and a
Bionanocomposites Based on Thermoplastic Starch for the Food Industry 39
high-water sensibility; however, these disadvantages can be overcome using

reinforcing agents in the starch matrix, such as cellulosic fibers, whiskers,
and nanofibers (Babaee et al., 2015; Nasseri and Mohammadi, 2014). In this
chapter, recent advances in research and development of bionanocomposites
obtained from thermoplastic starch with nanomaterials are analyzed and
summarized, as well as their application in food packaging.
2.2 PREPARATION AND CHARACTERIZATION OF NANOCRYSTALS

AND NANOPARTICLES
2.2.1 SYNTHESIS OF NANOCRYSTALS

Usually, the term “nanocrystals” involved particles in a crystalline state,
in most of the reported cases, in the strict sense, an amorphous nanopar-
ticle should not be referred to as nanocrystal (Malamatari et al., 2018).
The changing physical is the most important properties of materials in a
nano-size regime, being this a major challenge in the nanomaterials science
directly related with processing method to modify the most important
characteristics of nanocrystal systems such as size and shape (Byrappa et
al., 2008; Mohanraj and Chen, 2006). Nanoparticles derived from natural
resources, specifical polysaccharides, such as starch, cellulose, and chitin,
are sources of nanocrystals for food and nonfood applications. These poly-
saccharides are obtained at a low cost from biomass available in large quanti-
ties, renewable, biodegradable, some properties like their thermal behavior,
processability, and ability to assemble into ordered structures provide the
potential for compatibilizing nanocrystals and directing their assembly,
being these advantages of natural polysaccharide nanocrystals over inorganic
nanoparticles (Skaff and Emrick, 2004; Labet et al., 2007; Lin et al., 2012;
Boufi et al., 2018). In the case of nanocrystals produced from starches, the
remnants nanocrystals differ in size and shape in function of the amylose/
amylopectin ratio, the chain-length distribution of amylopectin, and the crys-
talline arrangement (X-ray diffraction pattern) (de la Concha et al., 2018).
The polymer nanocrystals can be ranging in crystal size from about tens to
hundred nanometers, to obtain crystals uniform is crucial the standardization
of the nanocrystallization processes to further control the size, distribution
and the shape (Oikawa et al., 2004). Besides, the synthesis, development,
and material characterization of novel material to conform nanocrystals
have attracted the research interest of diverse engineering areas as physic
and chemistry (Tjong, 2013). The nanocrystals from polysaccharides are
obtained mainly through chemical modifications, for example, the acid

hydrolysis by hydrochloric or sulfuric acid as reagents. First, the biomass
is generally submitted to a bleaching treatment with NaOH to purify the
cellulose, starch, or chitin by removing other constituents as the amorphous
region, the bleached material is then disintegrated in water, and the resulting
suspension is submitted to a hydrolysis treatment which produces nanocrys-
tals which have high crystallinity and nanoscale platelet morphology. By
another side, the conditions of hydrolysis during the extraction process (such
as the type of acid, acid concentration, temperature, and time) affect the
size and yield of nanocrystals, including chemical modification, triggered
changes in solubility or polarity, as well as surface and thermal properties.
Despite this, acid hydrolysis is mostly utilized in the preparation of polysac-
charide nanocrystals (Lin et al., 2012; Lin et al., 2011; Dufresne, 2008). On
the other hand, during acid hydrolysis, abundant hydroxyl groups are gener-
ated on the surface of nanocrystals which contribute to their positive surface
chemical properties and are therefore an essential route to altering the surface
structure, regulating surface properties, and developing functional materials
(Huang et al., 2015). In this sense, the hierarchical organization of polysac-
charides such as cellulose, starch, and chitin in natural materials along with
their semi-crystalline structure has been a key factor in the preparation of
nanocrystals through controlled acid hydrolysis (Haaj et al., 2016). Finally,
nanocrystals originated from biopolymers have drawn considerable attention
because they may be utilized as novel and biofunctional materials in diverse
industries (Kim et al., 2013).
2.2.2 SYNTHESIS OF NANOPARTICLES

The nanotechnology has gained high interest for its versatile applications
in various fields, because many polysaccharide nanoparticles have recently
been intensively studied (Huh et al., 2017; Korbekandi and Iravani, 2013).
Polysaccharides are natural biopolymers which are widely used for applica-
tions in food, medical, agricultural, and environmental industries, which
have been recognized because of their outstanding biocompatible, biode-
gradable, and renewable properties; on the other hand, these present partly
crystalline structures conferring some attributes to conform nanoparticles
(Le Corre et al., 2010; Wang et al., 2017). Likewise, the use of natural
polymers is promising for solving resource shortages and minimizing the
environmental problems compared with conventional plastics (Jiang et
al., 2016). Diverse polymeric materials for preparing nanoparticles have
been used such as chitosan, starch, alginate, and pectin, among others,
through mechanical, physical, and chemical methods. Among them are
acid hydrolysis, ultrasonic treatment, supercritical fluid extraction, solvent
evaporation, precipitation, and emulsion crosslinking. Basically, these
processes involve the conversion of solid to solid or liquid to solid or gas
to solid with the aim of making polymeric particles in specific size ranges.
Nowadays, identification of new sources of biopolymers and new types of
preparation method has become relevant to fabricate biopolymer particles
with novel or improved functional attributes. For example, the new methods
are combining physical treatments with chemical reaction (Byrappa et al.,
2008; Boufi et al., 2018; de la Concha et al., 2018; El-Sheikh, 2017; Liu et
al., 2008; Joye and McClements, 2014). Acid hydrolysis has been widely
used for the preparation of nanoparticles from polysaccharides, mainly due
to simplicity and easy to control, typically using hydrochloric or sulfuric
acid. However, this technique is time-consuming with nanoparticles yield
relatively low (Kim et al., 2015). Ultrasonic treatment has been employed
to physical polymers modification of microparticles into nanoparticles in a
range from 30 to 100 nm by vibration fracturing; this method is effective
and environmentally friendly avoiding any chemical additive. Furthermore,
this treatment can be used in combination with acid hydrolysis to prevent
the formation of aggregates during the hydrolysis; on the other hand, the
use of high-intensity ultrasound for the generation and application of nano-
sized materials has become increasingly widespread and widely adopted for
particle size reduction in emulsification, dispersion, and deagglomeration
as well as milling (Boufi et al., 2018; Chang et al., 2017; Haaj et al., 2013).
Some methods such as solvent extraction-evaporation, solvent diffusion,
and organic phase separation require the use of organic solvents which are
hazardous to the environment and physiological systems. Another important
tool of materials processing in the last two decades has been the supercritical
fluid technology, usually defined as a solvent at a temperature above its
critical temperature, where the fluid remains a single phase regardless of
pressure (Byrappa et al., 2008; Mohanraj and Chen, 2006). The precipitation
method is simplicity and high reproducibility, involves a successive addition
of a dilute polysaccharide solution into a nonsolvent, or inversely, however,
there are some disadvantages such as the generation of wastes highly diluted
polysaccharide solutions and larger volume of non-solvents affecting the
efficiency negatively and increasing the cost (Chang et al., 2017; Chin et al.,
2011).
2.2.3 CHARACTERIZATION OF NANOCRYSTALS AND

NANOPARTICLES
Recently, the design of particles with specific properties has recently

been driven by the application of nanotechnology principles to foods;
the dimensions of a biopolymer particle alter its functional performance
in foods, for example, particle size influences the bulk physicochemical
properties of foods (optics, rheology, and stability), and encapsulation
characteristics (e.g., loading, retention, and release); however, some
characteristics remain unknown or not well understood (Joye and McCle-
ments, 2014). Many techniques have been explored to elucidate the
structure and spectroscopic characters of nanocrystals and nanoparticles.
However, as the basic parameter, size and morphology are first considered
in many studies and are measured by either light scattering or microscopy
methods (Malamatari et al., 2018; Joye and McClements, 2014). The char-
acterization of shape and size distribution of polysaccharide nanocrystals
and nanoparticles can be investigated using static light scattering (SLS)
that relies on the measurement of the intensity of scattered light waves
as a function of scattering angle, and then, utilization of an appropriate
mathematical model to convert the scattering pattern into a particle size
distribution. The models used assume that the scattering particles are
spherical, homogeneous, and noninteracting with a well-defined refrac-
tive index. However, in practice, many types of biopolymer particles do
not conform to these assumptions. Besides, sample preparation often
requires dilution and stirring, which may alter the integrity or aggregation
state of biopolymer particles. The microscopy methods have been conve-
nient to characterize nanocrystal and nanoparticles due to the variety of
morphology and geometrical dimensions as length, diameter, thickness,
and aspect ratio of polysaccharide due to the different sources and
extracting methods. Among them are transmission electron microscopy
(TEM), atomic force microscopy (AFM), and field-emission scanning
electron microscopy (FE-SEM). Besides, the use of scanning electron
microscopy (SEM) provides a more extensive morphological inspection
of polysaccharide nanocrystal reinforced which leads to conclusions about
the homogeneity of the composite, the presence of voids, the dispersion
level of the nanoparticles within the continuous matrix, the presence of
aggregates, sedimentation, and the possible orientation of particles (Lin et
al., 2012; Dufresne, 2008; Joye and McClements, 2014).
TABLE 2.1 Obtention and Application of Nanocrystals and Nanoparticles of

Polysaccharides.
Source of extraction Application Author
of nanocrystals and
nanoparticles
Waxy corn starch As reinforces in the waterborne Wang et al. (2010)
Cotton linter pulp polyurethanes matrix.
Sago starch The synthesis method of precipitation is Chin et al. (2011)
fast and easy to perform, the mean sizes of
starch nanoparticles could be modulated by
the synthesis parameters such as the use of
surfactants during the synthesis process.
Cellulose Pea starch Drug release. Lin et al. (2011)
Standard corn starch The use of a physical method as Haaj et al. (2013)
Waxy corn starch ultrasonication in nanoparticles formation
leads to a high yield in nanoparticles and
does not require any chemical treatment.
Standard corn starch The combination of acid hydrolysis at low Kim et al. (2013)
Waxy corn starch temperature and ultrasonic treatment is an
effective process for the mass production of
crystalline starch nanoparticles
Cellulose Ingredient to gels formation Hu et al. (2014)
Standard Corn starch As improvers in the mechanical properties Dai et al. (2015)
Taro starch and thermal stability of films.
Potato starch As a filler in composite films. Water vapor Gujral et al.,
transition rate was reduced. (2021)
Amadumbe starch As improver in thermal stability, tensile Mukurumbira et
strength and decreased water vapor al., (2017)
permeability of films.
Waxy corn starch Acid hydrolysis was used for nanocrystal de la Concha et al.
Amaranth starch production, the arrangement of starch (2018)
components in the crystalline lamella is
related to starch granule size, producing
nanocrystals with different structures.
2.3 TECHNIQUES FOR THE DEVELOPMENT OF NANOCOMPOSITES
2.3.1 CASTING
Several techniques have been developed to nanocomposites elaboration of
natural polymers such as starch and cellulose. Starch processing by casting
and extrusion are the most used at the laboratory and industrial scales.
The casting is the main technique at the laboratory level of production of

starch-based films in an aqueous medium due to simplicity, ensuring the
proper mixing of the nanofillers with a starch matrix. Basically, it consists in
pouring on small plates or molds an aqueous suspension of gelatinized starch
by heating, plasticizer, and other additives, followed by a drying process
to eliminate the water to obtain a solid nanocomposite film. The resulting
average size of films is associated with both masses of suspension poured it
and mold or plate (García et al., 2015).
Several studies have reported particular parameters both gelatinization
and drying varying temperatures and times, which influenced and showed
detrimental effects on film formation. Thus, the mixing absorption and time
are significant to ensure homogeneous dispersion of the fillers in the starch
matrix (Madhumitha et al., 2018). For example, biodegradable starch/clay
nanocomposite films for food packaging applications were made by casting
since film blowing presented various issues such as low melt toughness and
films became brittle and sticky straight after production due to the high-water
content (Avella et al., 2005). In this sense, thermoplastic starch-waxy maize
starch nanocrystals nanocomposites were processed by casting, mixing
native waxy maize starch granules, glycerol, and distilled water. Then, the
mixture was heated at 150°C for 10 min in a reactor, under pressure and
mechanical stirring. The suspension of waxy maize starch nanocrystals was
added at 40°C. Finally, the mixture was evaporated in a nonventilated oven
at 40°C for 24 h (Angellier et al., 2006).
Besides, starch-based nanocomposites reinforced with flax cellulose nano-
crystals (FCNs) were produced by casting solution process which consisted
in gelatinization at 100°C for 30 min, after cooling down to about 70°C;
the resulting paste was degassed under vacuum to remove the remaining air
and cast in a polystyrene square Petri dish, followed by drying in a vented
oven at 50°C. The results showed that the FCNs played an important role
in improving the mechanical properties and water resistance of the starch-
based materials (Cao et al., 2008). Also, the casting process was employed
to sago starch biocomposite filled with nanocrystalline cellulose (NCC)
from rattan biomass, where starch, glycerol, and acetic acid were mixed and
heated at 70°C for 30 min to gelatinize. The water was evaporated at room
temperature overnight after that solution poured into an acrylic mold. The
film was formed uniformly and removed smoothly (Nasution et al., 2018).
Bio-nanocomposite films based on cassava starch or chitosan with a
constant thickness, reinforced with montmorillonite or bamboo nanofibers,
were prepared by the casting technique. The solution was drying at 32 ± 2°C
for approximately 24 h, in an oven with forced air circulation, after all films
were stored at a controlled relative humidity of 75% for not less than 48 h at
25°C. The nanofibers increased the tensile strength in 50% of starch films.
Also, cassava starch films showed a better response to nanostructure process
through the mechanical properties as well as good interaction between the
polymer matrix and bamboo nanofibers (Llanos and Tadini, 2018).
Although there is the extensive utilization of this technique, the main
limiting factor is the scalability from laboratory to industrial levels due to
control on the sample film thickness and uniformity. The polymer needs to
be soluble in the same solvent used for dispersion and swelling of the filler,
also it is slow and expensive. For large-scale manufacture, an extrusion
process is usually used.
2.3.2 EXTRUSION
Extrusion compounding is the most interesting method for industrial

processing due to easy scale-up and the possibility of further molding of the
materials; it is also an energy-efficient system achieving breakdown of the
starch granule structure through a combination of high shear, temperature,
and pressure to successfully melt starch. Typical single- or doublé-screw
extruders in general form consist of a hopper, barrel, feed screw, thermo-
couples, and dies (García et al., 2015).
The extrusion can be classified into two types: reactive extrusion and
extrusion-cooking. Reactive extrusion is a concurrent reaction in extru-
sion processing of polymers (Kim et al., 2015). This method is an efficient,
versatile, cost-effective, and environmentally-friendly method to produce
new materials with enhanced performance properties such as the interface
interactions between used components (Formela et al., 2018). For example, a
novel extrusion method for the preparation of TPS and TPS/CNF considered
as green nanocomposites was reported by Ghanbari et al. (2018), consisting of
a two-step process. First, the main components of the matrix were manually
premixed and extruded by a co-rotating twin-screw extruder, with a screw
speed of 80 rpm. The barrel temperatures of the extruder were controlled at
different temperatures while the temperature of the die was set at 125°C. The
extruded strand was passed through a water bath, granulated, and dried at
105°C for 24 h. In the second step, the resulting granules were subsequently
compression-molded under following conditions: press temperature 190°C,
pressure during heating 3.5 MPa, heating time 10 min, and cooling time 5 min.
The results strongly showed the advantages of reinforcing TPS with CNFs, a
natural, cheap, and abundant green nanocomposite using extrusion process.
Moreover, biodegradable polylactide and thermoplastic starch blends
were prepared with drug release purposes in which the granulates form
was blown using a single-screw extruder with screw diameter D = 20 mm
(L:D = 30:1) connected to blown film unit with a ring-shaped die to prepare
biodegradable films. The blown film extrusion process was carried out at
a temperature range from 155°C to 175°C and a screw speed of 44 rpm.
Films in the range 50–90 mm of thickness, which could be used for drug
delivery systems (Karolina et al., 2018) were obtained. On another side, the
extrusion-cooking commonly is related to food industry and used in direct
extrusion snacks, cereal flakes, and diverse breakfast foods produced from
cereal, among others (García et al., 2015). However, it has been employed
in foamed starch-based materials under various conditions: temperature,
humidity, type of the die, screw rotational speed, various raw materials, and
additives blends. For example, the mixtures based on potato starch and with
the addition the foaming agent Plastron foam PDE and poly(vinyl) alcohol
PVA presented the best properties and processing costs under this method
(Combrzyński et al., 2018). Thus, with these methods we can improve nano-
composites elaboration of natural polymers and its properties.
2.4 INFLUENCE OF INCORPORATION OF NANOMATERIALS IN THE

PROPERTIES OF STARCH NANOCOMPOSITES
The thermoplastic starch packaging designed at the laboratory presents
poor properties compared with polymers derived from petroleum. For this
reason, most of the time some additives must be added to the formulation to
enhancing their functional properties (Nasseri and Mohammadi, 2014). The
additives mots used in the formulation of bioplastics are lipids, surfactants,
antioxidants, antimicrobials, and recently, nanomaterials. There are many
materials with nanomaterials incorporated (nanocomposites) food packages
commercialized in the world, while others are in development. The addi-
tion of nanomaterials to polymers can improve the mechanical, barrier,
and thermal properties, among others (Babaee et al., 2015; Ahmed et al.,
2012). However, according to Anglès and Dufresne (2000) it is not easy
to model the properties of starch nanocomposites because they are formed
by different components: starch, nanomaterial, plasticizer, and water. These
four components can be present in crystalline, amorphous, or liquid phase,
which promotes different competitive interactions between them. These
factors should be considered in the study of rheological, mechanical, barrier,

and thermal properties
2.4.1 RHEOLOGICAL AND PASTING PROPERTIES
Rheological properties offer information to understand the relation between

the nanomaterials and thermoplastic starch (TPS) matrix in solution. More-
over, for the potential scaling of films production, these properties play a
key factor in the fluid transport and therefore in the pumps and impellers
design. On the other hand, the pasting properties of starch dispersions are
commonly evaluated by rapid visco analyzer (RVA); this technique involves
subjecting the starch dispersion to heating–cooling controlled cycles under
constant agitation.
Most of the studies of bionanocomposites have been conducted to final
product properties. Therefore, there are limited studies that measured the
evolving dynamic rheological properties of nanocomposite forming solu-
tions (NFS) during and after gelatinization. Chiou et al. (2005) studied the
dynamic rheological properties of high solids starch–nanoclay formulations
containing different types of starches (wheat, potato, corn, and waxy corn)
and hydrophilic (Cloisite Na+) and hydrophobic (Cloisite 30B, 10A, and 15A)
nanoclays. The nanocomposite with Cloisite Na+ presented the more gel-like
behavior at higher clay concentrations than the other hydrophobic samples.
The authors suggested that during the gelatinization process, the lixiviated
amylose established strong interactions with the Cloisite Na+. Montmoril-
lonite (MMT) is the most used nanoclay for the design of films with food
applications. In a recent study, Romero-Bastida et al. (2018) evaluated the
effect of MMT on the rheological properties of nanocomposite-forming
solutions (NFS) based on corn starch with different amylose contents (30 and
70%). The flow curves evidenced a non-Newtonian behavior with flow index
lower than one (n < 1, pseudoplastic fluid) with a shear thinning behavior,
characterized by a reduction of apparent viscosity when increasing the shear
rate. When the amylose content increases, the flow index (n) decreased
as a consequence of interactions between starch components. Regarding
dynamic properties, the storage module (G′) was higher than loss modulus
(G″) suggesting a gel-like behavior. The addition of MMT increased the
storage modulus, which indicated reinforcing of starch matrix.
On the other hand, there are reports about how the inclusion of nanomate-
rials in NFS affect their pasting properties. In this sense, Chang et al. (2010)
reported that the pasting peak viscosity of nanocomposite-forming solu-

tions (NFS) decreased with the addition of chitin-NP. The pasting viscosity
increased as the chitin-NP content was higher. The authors suggest that it
was the result of higher interactions between chitin-NP and starch. Similar
results were observed in plasticized starch bionanocomposites reinforced
with chitosan nanoparticles (Chang et al., 2010).
2.4.2 MECHANICAL PROPERTIES
The nanocomposites consist of two fractions, one in mayor percent designed

as “matrix,” and the other minor percent known as “charge.” The presence
of nanomaterials promotes nanocomposites with a great interfacial area,
which affects their mechanical properties. The main mechanical properties
of materials are tensile strength (TE, the maximum force required to rupture
the material), Young’s modulus (MY, proportionality coefficient between
applied stress and strain), and elongation at the break (EB, is the ratio
between changed length and initial length after breakage).
There are many reports about the influence of incorporation of nanoma-
terials of different sources on the mechanical properties of thermoplastic
starch films. For instance, the addition of cellulose–starch nanocomposites
promoted increments up to 1052 (MPa) and 2909 (GPa) in TS and YM,
respectively. In contrast, the EB decreased around 79% (Grande et al., 2009).
Lu et al. (2006) observed increments in YM and TS from 56 to 480 MPa and
from 2.8 to 6.9 MPa when the content of ramie crystallites increased from
0 to 20% wt in the wheat TPS matrix. This behavior was associated with
the great number of three-dimensional networks of intermolecular hydrogen
bonding interactions between the same cellulose crystals and cellulose crys-
tals with the TPS matrix. Similar results were reported by Ma et al. (2009)
in the TPS pea matrix reinforced with ZnO nanoparticles, as the nano-ZnO
content varied from 0 to 4 wt%, the TS and YM increased from 3.94 to 10.80
MPa and from 49.80 to 137.00 MPa, respectively.
Other authors reported starch nanoparticles and nanocrystals for obtaining
nanocomposites, as the same form that the above studies mentioned; the
inclusion of these nanomaterials promotes increments in the TS and YM
(Angellier et al., 2006; Viguié et al., 2007). According to the authors, the
chemical similarity between starch nanocrystals and TPS matrix allows
strong interactions between starch nanocrystals/matrix and nanocrystals/
nanocrystals. In studies with chitin nanoparticles (Chitin-NP) used for
obtaining starch nanocomposites, there was an increment in the TS from

2.84 to 7.79 MPa when the concentration of Chitin-NP varied from 0 to
5%. However, the EB decreased from 59.3% to 19.3% (Chang et al., 2010).
As the same form that when starch nanoparticles were used as filler, the
Chitin-NP establishes strong interfacial interactions with the TPS matrix as
a consequence of the similar structure of chitin and starch. The decrement in
the EB is a consequence of the decreased flexibility of the starch molecules.
Another material used as filler in the TPS matrix is the nanoclays (Layered
silicates), which are economical, environmentally friendly, and naturally
abundant (Li et al., 2006). This material can form competitive interactions
in the starch matrix. The plasticizer can be absorbed on the clay surface,
affecting the mechanical properties of the nanocomposite (Chivrac et al.,
2008). Chung et al. (2010) observed that the addition of montmorillonite
or laponite to the TPS matrix enhances the YM and TS until 65 and 30%,
respectively, compared with the unfilled starch material. This was attributed
to the good dispersion of clay in the starch matrix.
2.4.3 BARRIER PROPERTIES
The barrier properties of starch films depend on their microstructure and

chemical nature of components. Compared with synthetic polymers, thermo-
plastic starch matrix presents high water vapor permeability (WVP) values.
This disadvantage is the result of the great number of hydroxyl groups
present in the matrix components (starch and glycerol), which interact with
the migrating water molecules. For food applications, the moisture exchange
with the environment must be minimal.
García et al. (2009) observed that the addition of waxy starch nanocrystals
(2.5%wt) in nanocomposites of cassava starch reduced the WVP by 40%.
This behavior was associated with the fact that the nanocrystals were distrib-
uted into the TPS matrix, generating a tortuous path for the water vapor
diffusion. Then, the same research group (García et al., 2011) prepared bion-
anocomposites made from thermoplastic waxy maize starch reinforced with
waxy maize starch nanocrystals at the same quantity previously evaluated
(2.5%wt). The results were opposite to that observed using a cassava starch
matrix, the addition of 2.5%wt waxy maize starch nanocrystals increased the
WVP by 79% (from 3.8 to 6.8 × 10−10 g s−1 m−1 Pa−1). This phenomenon
was a consequence of the formation of preferential paths for water diffusion
with high concentrations of OH where the nanocrystals are glue–glycerol
bonded, which increased the WVP. The addition and concentration of mont-
morillonite in corn starch films plasticized with glycerol reduced the WVP
(Slavutsky et al., 2012; Wang et al., 2009). The author mentioned that it
was the result of the strong interactions of montmorillonite with the starch
and the formation of a tortuous path. The addition of chitosan nanoparticles
(chitosan-NP) from 0 to 6%wt into TPS films from potato starch reduced the
WVP from 5.8 to 3.5 × 10–10 g m–1 s–1 Pa–1. However, when the chitosan-NP
concentration increased up to 8%wt, the WVP slightly increased (Chang et
al., 2010). The addition of chitosan-NP at low levels allows good disper-
sion of nanoparticles in the TPS matrix, which promotes a tortuous path
for the water molecules. These observations were similar to those reported
by the same group research (Chang et al., 2010) in TPS nanocomposites
with chitin nanocrystals (chitin-NC) from 0 to 5%wt. The WVP decreased
with the chitin-NC increment from 1 to 4%wt, but when the concentration
of nanocrystals increased until 5%wt, the WVP increased. Low levels of
chitin-NC dispersed well in the TPS matrix; however, at high concentra-
tions, the chitin-NC form aggregates that facilitie water vapor permeation.
On the other hand, the addition of cellulose nanofibers isolated from kenaf
bast fibers (Hibiscus cannabinus) improved the WVP by the production of
denser and less porous materials. Moreover, compared with the neat TPS,
the interactions between cellulose nanofibers and TPS matrix restrict the
moisture diffusion rate (Babaee et al., 2015).
The permeability to gases such as oxygen plays a key role in food
packaging since this gas causes deterioration of food by oxidation. With
the incorporation of the nanofiller, starch-based materials generally are
improved. For instance, the addition of starch nanocrystals (2.5 and 5 wt%)
reduced the oxygen permeance in the TPS matrix (González et al., 2015).
This behavior has also been reported in bionanocomposites reinforced with
natural and an organically modified montmorillonite (Zeppa et al., 2009). In
another hand, compared with the neat matrix, the incorporation of chitin-NC
(up to 20 wt%) decreased by about 30% the oxygen permeability (Salaberria
et al., 2015). According to the authors, the increase in the intermolecular
interactions in the matrix due to the addition of nanofillers leads to a high
oxygen barrier.
2.4.4 THERMAL PROPERTIES

The study of thermal properties allows understanding the processing
temperature range and use of materials. The glass transition temperature (Tg)
is the most important thermal property of polymers. Materials under their

Tg behave as brittle. Thermal glass transition can be measured by dynamic
mechanical thermal analysis (DMTA) or differential scanning calorimetry
(DSC). Thermoplastic starch plasticized by polyols like glycerol results in a
complex and heterogeneous system with a rich phase of glycerol dispersed
in another phase rich in starch (continuous phase), where each of them has
its own Tg value (Lu et al., 2006; Dufresne and Castaño, 2017). These values
are affected by the type and concentration of nanomaterial used as filler. Cao
et al. (2008) studied nanocomposites from pea starch reinforced with cellu-
lose nanocrystals of flax fiber (5, 10, 15, 20, 25, and 30 wt%). The authors
reported that increments in the loading of cellulose nanocrystal promoted
variations from 43.3 to 48.8 in Tg2 (starch-rich phase), whereas the Tg1
(glycerol-rich phase) remained without variations. This behavior was similar
to that previously reported by Lu et al. (2006); they observed increments
in Tg2 from 26.8 to 55.7°C as a consequence of the incorporation of ramie
nanocrystallites (0–40%wt) into the thermoplastic wheat starch matrix. In
general, the addition of cellulose nanocrystals increases the glass transition
temperature. This phenomenon is a consequence of the strong molecular
interactions by hydrogen bonding between starch and nanocrystals, which
results in low mobility of starch chains (Chang et al., 2010; Lu et al., 2006;
Dufresne and Castaño, 2017; Cao et al., 2008).
Nanoclays are nanomaterials widely used for food packaging design;
among them, the most used is the montmorillonite (Chung et al., 2010;
Sinha Ray and Okamoto, 2003). González et al. (2015) prepared thermo-
plastic starch bionanocomposites reinforced with starch nanocrystals. The
authors reported that the thermal stability of bionanocomposites decreased
with starch nanocrystals added. It was explained by the high affinity between
glycerol and waxy starch nanocrystals. Moreover, after acid hydrolysis for
nanocrystal production, some acidic sulfate ester groups should be remained
attached to waxy starch nanocrystals surface, which seems to act as cata-
lyzers of the thermal decomposition, decreasing the degradation temperature
for nanocomposites with the filled matrix.
Another thermal property of materials is their thermal stability, which
is evaluated by thermogravimetric analysis (TGA) (Chang et al., 2010).
Chang et al. (2010) added nanoparticles of chitosan (chitosan-NP) to the
TPS matrix at concentrations from 0 to 4 wt%. They reported that the addi-
tion of chitosan-NP reduced the mass loss at the onset temperature. The Tmax
increased with chitosan-NP increments, obtaining 315, 321, and 328°C, for
0, 2, and 4 wt.% CN, respectively. The authors attributed the higher thermal
stability in the TPS matrix with chitosan-NP to the strong interfacial interac-
tions established between them. González et al. (2015) prepared thermo-
plastic starch bionanocomposites reinforced with starch nanocrystals. The
thermal stability of bionanocomposites decreased with starch nanocrystals
added. This was explained by the high affinity between glycerol and waxy
starch nanocrystals. Moreover, after acid hydrolysis for nanocrystal produc-
tion, some acidic sulfate ester groups should be remained attached to waxy
starch nanocrystals surface, which seems to act as catalyzers of the thermal
decomposition, decreasing the degradation temperature for nanocomposites
with filled matrix (García et al., 2011; González et al., 2015).
2.5 APPLICATION OF STARCH NANOCOMPOSITES AS FOOD

PACKAGING
In recent decades, the demand for “ready-to-eat,” “ready-to-cook,” or
“ready-to-use” products has grown considerably without ever stopping the
aspect of food security implicit in its production, commercialization, and
consumption (He and Hwang, 2016; Carbone et al., 2016). In the specific
case of food packaging and preservation, applications have been envisioned
ranging from a simple cover, as a mechanical barrier to moisture or damping
to prevent or reduce physical damage, to active and intelligent packaging,
which interact with the food to extend its shelf life. For example, combining
materials with antioxidant or antimicrobial compounds. On another hand,
it is used to communicate information to the consumer through sensory
messages, such as color change, which may indicate changes in the integrity
of the matrix food or chemical composition, whether favorable or unfavor-
able for the consumption of the product. Thus, nanotechnology promises to
revolutionize the food packaging (Chellaram et al., 2014).
2.5.1 COATINGS
The formulation and use of starch-based covers combined with other nanom-
eter-scale materials that could be used to preserve the sensory and nutritional
quality of foods, especially those marketed fresh, or minimally processed.
Falguera et al. (2011) reported that these covers have to meet certain char-
acteristics that make them practical for their use, such as a good moisture
barrier with certain permeability to gases and above all to microorganisms
that corrupt foods (Valdés et al., 2015). They should also comply with
the damping function to reduce physical damage due to friction between
the product units and the packaging, and even during their handling and
commercialization in the logistics chain (Han, 2014). Also, additional char-
acteristics are required such as be edible or easily removed for subsequent
safe consumption and not causing or with reducing the negative impact on
the environment (Cazón et al., 2017; De Ancos et al., 2015).
The covers of starch composites have been used for the conservation
of fresh or minimally processed fruits and vegetables (Valdés et al., 2015;
Quintero et al., 2010); they can act as an alternative to modified atmospheres,
regulating the respiration of vegetables depending on the properties of the
cover (Park et al., 2014). Also, the adhesive properties have been explored
in some food packaging, such as that used in hamburger wrappers and other
products (Mbey et al., 2012; Hamad et al., 2018). The success of this type
of materials is markedly conditioned by the handling of the products; an
inadequate handling can seriously compromise the covers protection to the
food by damaging or removing them prematurely from the surface of the
product, even, due to the nature of the materials, for example the starch does
not favor the functionality of the coatings in fresh foods, resulting in an
opposite effect on the expected (Quintero et al., 2010).
2.5.2 RIGID PACKAGING
The current consumer is very familiar with rigid packaging, and packaging,
have been extensively used, for example, the transparent plastic boxes, to
preserved and reduce physical damage due to rough handling. However,
most containers and packaging of this type are commonly made from petro-
leum-based plastics and are generally used once, promoting environmental
problems (Cavero and Arroyo, 2017). To face these issues, thermoplastic
starch (TPS) nanocomposites have been developed from TPS in combination
with nanometric structures, such as silica nanoparticles (Arayaphan et al.,
2020; de Azêvedo et al., 2021), nanoclays (Aouada et al., 2013; Müller et al.,
2012; de Jara et al., 2020), organic nanofibers (Babaee et al., 2015), among
others, in order to achieve rigid packaging (Salaberria et al., 2014). The
result is promising, according to important properties such as the optical,
mechanical, and biodegradability (Mohammadi Nafchi et al., 2013; Pelissari
et al., 2017). An interesting food industry area is the packaging of fruits and
vegetables for fresh consumption, as has been reported by Campos-Requena
et al. (2017). However, it cannot be ruled out all other uses of packaging of
rigid material made with conventional plastics especially about dry products
(for example, cookies, nuts, dried meat, cheeses, etc.) (Ghoshal, 2018) and
ready-to-eat products (De Ancos et al., 2015).
Another type of rigid materials based on starch nanocomposites are
plastic foams (Ahmadzadeh et al., 2017). These materials can be applied in
the design of individual containers and buffers for foods such as fresh whole
fruits, to prevent and reduce the mechanical damages caused by the handling
of the same from the harvest to the consumption, reducing some negative
effects in their maturation process to allow proper transpiration.
2.5.3 FLEXIBLE PACKAGING

At present day, there are no containers that are used more than those that
are flexible, which, together with the rigid containers, make up most of the
universe of packaging used in the food production industry (González and
Alvarez Igarzabal, 2015; Podshivalov et al., 2017). Plastic films are the main
example of this, and because of the problems posed by its wide use and
origin (petroleum derivatives) (Mittal et al., 2016), they have been devel-
oping alternatives based on starch nanocomposites with other materials, such
as organic nanofibers (González et al., 2015), other polysaccharides such
as chitosan (Chang et al., 2010), talc nanoparticles (Castillo et al., 2019).
All these materials with encouraging results in terms of their chemical and
mechanical properties and with high advantages in terms of their biodegrad-
ability (Kim et al., 2015) make these types of materials an economical and
promising option for the substitution of the use of flexible plastics derived
from conventional nonrenewable sources (Ayana et al., 2014).
The applications of films starch‐based nanocomposites go from individual
packages (Nathalie et al., 2017) to packaging with greater capacity for the
transport of food, such as traditional plastic bags (López et al., 2015). They
could be used in the vacuum packaging of fresh meat and dairy products,
such as cheeses, once the problem of hydrophilicity and low mechanical
resistance has been solved, characteristics of starch films (Mendes et al.,
2016). Due to their gases and water vapor barrier properties achieved due
to the combination of materials, they can also be used in packaging of mini-
mally processed vegetables (De Ancos et al., 2015).
2.5.4 ACTIVE AND INTELLIGENT PACKAGING

Active and intelligent packagings have been developed and evaluated in
recent decades as one of the complete options aimed at extending the shelf
life of products and all their intrinsic properties (flavor, color, nutrients,
texture, etc.) (Barzegar et al., 2014; Goudarzi et al., 2017), and at the same
time interact with the consumer, providing information in real time about the
state of conservation of the packaged food product (Mihindukulasuriya and
Lim, 2014).
For the formulation of materials for active starch-based packaging,
including nanocomposites, antimicrobial agents, antioxidants, or double
action have been included, mainly compounds of plant origin, such as essen-
tial oils of different species (Atarés and Chiralt, 2016; Ribeiro-Santos et al.,
2017). In this sense, there are reposts with rosemary (Estevez-Areco et al.,
2019), oregano (Aguilar-Sánchez et al., 2019), peptides, such as nisin (Meira
et al., 2017), as well as inorganic compounds, such as ZnO nanoparticles
(Arifin et al., 2022), TiO2 (Rong et al., 2021), silver (Srikhao et al., 2021),
and gold (Rovina et al., 2021), although traditional conservatives, such as
potassium sorbate, have not been neglected (Barzegar et al., 2014).
Its applications are aimed especially at packaging food susceptible to
contamination with pathogenic microorganisms, such as fresh or minimally
processed meat and fish (Wu et al., 2010), soft cheeses (Meira et al., 2016),
foods of lower aqueous activity, susceptible to attack by molds, such as fresh
bread (Barzegar et al., 2014). The applications have also been made in the
trapping of harmful compounds in foods, such as cholesterol, for which
tests have been carried out on the packaging of milk (González and Alvarez
Igarzabal, 2015). Another application that has been explored is that of starch
films with antioxidants used in butter preservation (Hernández et al., 2017).
Some intelligent packaging has been designed to reduce certain
compounds, such as blue methylene or methyl orange (Mallakpour and
Rashidimoghadam, 2017) and some natural pigments, such as anthocyanins
(Choi et al., 2017), which usually indicate an alteration in the pH of the food,
due to exposure to an inadequate temperature or microbial development
(Goudarzi et al., 2017), indicating the freshness of the packaged food (Imran
et al., 2010), thus warning the consumer about the convenience or not of
acquiring or ingesting it. The applications of this type of material could be in
a very broad sense; the materials described above can be functionalized to be
used as active and/or intelligent packaging (Ranjan et al., 2014; Usman et al.,
2016). They have been successfully tested, for example, in the packaging of
pork (Choi et al., 2017), fresh pasteurized milk (Liu et al., 2017), and fresh
fish (Zhai et al., 2017).
The advantages that this would imply, from the prevention of food waste,
the increase of its shelf-life, the prevention of foodborne diseases, and
the reduction of environmental contamination (Othman, 2014) by having

materials of accelerated biodegradability and that it is possible to produce
at low cost by being based on materials of an organic nature (Carbone et al.,
2016). However, even with all the benefits reported in the various studies,
there are others that warn about the inappropriate use of this type of material
in food packaging, since the active ingredients used could also affect the
health of the consumers if they are used in inappropriate concentrations,
especially those that include heavy metals in their composition (Ranjan
et al., 2014; Dimitrijevic et al., 2015), although the base materials are not
toxic for humans or animals (Woehl et al., 2010), so it is necessary to study
exhaustively the impact of these materials on the health of consumers and
their regulation before adopting them in a massive way.
2.6 CONCLUSIONS
Throughout this chapter, it has become evident that the nanotechnology has
gained high interest for its versatile applications for preparing starch food
packaging. Also, it has been indicated that the starch can be considered a
suitable natural polymer for preparing biodegradable matrices of thermo-
plastic starch reinforced with nanoparticles (nanocomposites). Several
techniques have been developed for nanocomposites elaboration of natural
polymers, such as casting and extrusion are the most used at the laboratory
and industrial scales with the purpose of improving the production of natural
polymer nanocomposites. There are few studies regarding the rheological
characterization of nanocomposite forming solutions because it is not easy
to model the properties of nanocomposites, as it consists of different
components that come in different forms. However, there are starch-based
nanocomposite films combined with various materials that do not present
problems of hydrophobicity and little mechanical resistance and can be used
in the food industry.
KEYWORDS
• thermal properties
• starch nanoparticles
• mechanical properties
• cellulose nanocrystals
REFERENCES
Aguilar-Sánchez, R.; Munguía-Pérez, R.; Reyes-Jurado, F.; Navarro-Cruz, A. R.; Cid-Pérez,

T. S.; Hernández-Carranza, P.; Beristain-Bauza, S. D.; Ochoa-Velasco, C. E.; Avila-Sosa,
R. Structural, Physical, and Antifungal Characterization of Starch Edible Films Added
with Nanocomposites and Mexican Oregano (Lippia berlandieri Schauer) Essential Oil.
Molecules [Online], 2019.
Ahari, H.; Anvar, A. A.; Ataee, M.; Naeimabadi, M. Employing Nanosilver, Nanocopper, and
Nanoclays in Food Packaging Production: A Systematic Review. Coatings [Online], 2021.
Ahmadzadeh, S.; Nasirpour, A.; Keramat, J.; Desobry, S. Powerful Solution to Mitigate the
Temperature Variation Effect: Development of Novel Superinsulating Materials. In Food
Packaging and Preservation, Handbook of Food Bioengineering, 1st ed.; Grumezescu, A.
M.; Holban, A. M., Eds.; Elsevier. International.: 2017; Vol. 9, p 137.
Ahmed, J.; Tiwari, B. K.; Imam, S. H.; Rao, M. A. Starch as Biopolymer and Nanocomposite.
In Starch-based Polymeric Materials and Nanocomposites: Chemistry, Processing, and
Applications; Ahmed, J., Tiwari, B. K., Imam, S. H., Rao, M. A., Eds.; CRC Press: Boca
Raton, FL, 2012.
Angellier, H.; Molina-Boisseau, S.; Dole, P.; Dufresne, A. Thermoplastic Starch−Waxy Maize
Starch Nanocrystals Nanocomposites. Biomacromolecules 2006, 7 (2), 531–539.
Anglès, M. N.; Dufresne, A. Plasticized Starch/Tunicin Whiskers Nanocomposites. 1.
Structural Analysis. Macromolecules 2000, 33 (22), 8344–8353.
Aouada, F. A.; Mattoso, L. H.; Longo, E. A Simple Procedure for the Preparation of
Laponite and Thermoplastic Starch Nanocomposites: Structural, Mechanical, and Thermal
Characterizations. J. Thermoplast. Compos. Mater. 2013, 26 (1), 109–124.
Arayaphan, J.; Boonsuk, P.; Chantarak, S. Enhancement of Water Barrier Properties of
Cassava Starch-based Biodegradable Films Using Silica Particles. Iran. Polym. J. 2020,
29 (9), 749–757.
Arifin, H. R.; Djali, M.; Nurhadi, B.; Azlin-Hasim, S.; Masruchin, N.; Vania, P. A.; Hilmi,
A. Corn Starch-Based Bionanocomposite Film Reinforced With ZnO Nanoparticles and
Different Types of Plasticizers. Front. Sustain. Food Syst. 2022, 6.
Ashfaq, A.; Khursheed, N.; Fatima, S.; Anjum, Z.; Younis, K. Application of Nanotechnology
in Food Packaging: Pros and Cons. J. Agric. Food Res. 2022, 7, 100270.
Atarés, L.; Chiralt, A. Essential Oils as Additives in Biodegradable Films and Coatings for
Active Food Packaging. Trends Food Sci. Technol. 2016, 48, 51–62.
Avella, M.; De Vlieger, J. J.; Errico, M. E.; Fischer, S.; Vacca, P.; Volpe, M. G. Biodegradable
Starch/Clay Nanocomposite Films for Food Packaging Applications. Food Chem. 2005, 93
(3), 467–474.
Ayana, B.; Suin, S.; Khatua, B. B. Highly Exfoliated Eco-Friendly Thermoplastic Starch
(TPS)/poly (lactic acid)(PLA)/Clay Nanocomposites Using Unmodified Nanoclay.
Carbohydr. Polym. 2014, 110, 430–439.
Azeredo, H. M. C. D., Nanocomposites for Food Packaging Applications. Food Res. Int.
2009, 42 (9), 1240–1253.
Babaee, M.; Jonoobi, M.; Hamzeh, Y.; Ashori, A. Biodegradability and Mechanical Properties
of Reinforced Starch Nanocomposites Using Cellulose Nanofibers. Carbohydr. Polym.
2015, 132, 1–8.
Barzegar, H.; Azizi, M. H.; Barzegar, M.; Hamidi-Esfahani, Z. Effect of Potassium Sorbate on
Antimicrobial and Physical Properties of Starch–Clay Nanocomposite Films. Carbohydr.
Polym. 2014, 110, 26–31.
Boufi, S.; Haaj, S. B.; Magnin, A.; Pignon, F.; Impéror-Clerc, M.; Mortha, G. Ultrasonic
Assisted Production of Starch Nanoparticles: Structural Characterization and Mechanism
of Disintegration. Ultrason. Sonochem. 2018, 41, 327–336.
Byrappa, K.; Ohara, S.; Adschiri, T. Nanoparticles Synthesis Using Supercritical Fluid
Technology–Towards Biomedical Applications. Adv. Drug Deliv. Rev. 2008, 60 (3),
299–327.
Campos-Requena, V. H.; Rivas, B. L.; Pérez, M. A.; Figueroa, C. R.; Figueroa, N. E.;
Sanfuentes, E. A. Thermoplastic Starch/Clay Nanocomposites Loaded with Essential Oil
Constituents as Packaging for Strawberries—In Vivo Antimicrobial Synergy Over Botrytis
Cinerea. Postharvest Biol. Technol. 2017, 129, 29–36.
Cao, X.; Chen, Y.; Chang, P. R.; Muir, A. D.; Falk, G. Starch-based Nanocomposites
Reinforced with Flax Cellulose Nanocrystals. Express Polym. Lett. 2008, 2 (7), 502–510.
Cao, X.; Chen, Y.; Chang, P.; Muir, A.; Falk, G. Starch-based Nanocomposites Reinforced
with Flax Cellulose Nanocrystals. Express Polym. Lett. 2008, 2 (7), 502–510.
Carbone, M.; Donia, D. T.; Sabbatella, G.; Antiochia, R. Silver Nanoparticles in Polymeric
Matrices for Fresh Food Packaging. J. King Saud Univ. Sci. 2016, 28 (4), 273–279.
Castillo, L. A.; López, O. V.; García, M. A.; Barbosa, S. E.; Villar, M. A. Crystalline Morphology
of Thermoplastic Starch/Talc Nanocomposites Induced by Thermal Processing. Heliyon
2019, 5 (6), e01877.
Cavero, H. A.; Arroyo, E. Obtencion de Biopolimeros de Papa Como Una Alternativa al
Desarrollo de Materiales Inocuos al Medio Ambiente. Revista Cientifica TECNIA 2017,
24 (1), 13–19.
Cazón, P.; Velazquez, G.; Ramírez, J. A.; Vázquez, M. Polysaccharide-Based Films and
Coatings for Food Packaging: A REVIEW. Food Hydrocoll. 2017, 68, 136–148.
Chang, P. R.; Jian, R.; Yu, J.; Ma, X. Fabrication and Characterisation of Chitosan
Nanoparticles/Plasticised-Starch Composites. Food Chem. 2010, 120 (3), 736–740.
Chang, P. R.; Jian, R.; Yu, J.; Ma, X. Starch-based Composites Reinforced with novel chitin
Nanoparticles. Carbohydr. Polym. 2010, 80 (2), 420–425.
Chang, Y.; Yan, X.; Wang, Q.; Ren, L.; Tong, J.; Zhou, J. High Efficiency and Low Cost
Preparation of Size Controlled Starch Nanoparticles Through Ultrasonic Treatment and
Precipitation. Food Chem. 2017, 227, 369–375.
Chellaram, C.; Murugaboopathi, G.; John, A. A.; Sivakumar, R.; Ganesan, S.; Krithika, S.;
Priya, G. Significance of Nanotechnology in Food Industry. APCBEE Procedia 2014, 8,
109–113.
Chin, S. F.; Pang, S. C.; Tay, S. H. Size Controlled Synthesis of Starch Nanoparticles by a
Simple Nanoprecipitation Method. Carbohydr. Polym. 2011, 86 (4), 1817–1819.
Chiou, B.-S.; Yee, E.; Glenn, G. M.; Orts, W. J. Rheology of Starch–Clay Nanocomposites.
Carbohydr. Polym. 2005, 59 (4), 467–475.
Chivrac, F.; Gueguen, O.; Pollet, E.; Ahzi, S.; Makradi, A.; Averous, L. Micromechanical
Modeling and Characterization of the Effective Properties in Starch-based Nano-
Biocomposites. Acta Biomater. 2008, 4 (6), 1707–1714.
Choi, I.; Lee, J. Y.; Lacroix, M.; Han, J. Intelligent pH Indicator Film Composed of Agar/
Potato Starch and Anthocyanin Extracts from Purple Sweet Potato. Food Chem. 2017, 218,
122–128.
Chung, Y.-L.; Ansari, S.; Estevez, L.; Hayrapetyan, S.; Giannelis, E. P.; Lai, H.-M. Preparation
and Properties of Biodegradable Starch–Clay Nanocomposites. Carbohydr. Polym. 2010,
79 (2), 391–396.
Combrzyński, M.; Mościcki, L.; Mitrus, M.; Kupryaniuk, K.; Oniszczuk, A. Application of
Extrusion-cooking Technique for Foamed Starch-based Materials. BIO Web Conf. 2018,
10, 1–6.
Dai, L.; Qiu, C.; Xiong, L.; Sun, Q. Characterisation of Corn Starch-based Films Reinforced
with Taro Starch Nanoparticles. Food Chem. 2015, 174, 82–88.
De Ancos, B.; González-Peña, D.; Colina-Coca, C.; Sánchez-Moreno, C. Uso de Películas/
Recubrimientos Comestibles en los Productos de IV y V Gama. Revista Iberoamericana de
Tecnología Postcosecha 2015, 16 (1).
de Azêvedo, L. C.; Rovani, S.; Santos, J. J.; Dias, D. B.; Nascimento, S. S.; Oliveira, F.
F.; Silva, L. G. A.; Fungaro, D. A. Study of Renewable Silica Powder Influence in the
Preparation of Bioplastics from Corn and Potato Starch. J. Polym. Environ. 2021, 29 (3),
707–720.
de Jara, E. M.; García-Hernández, E.; Quequezana-Bedregal, M.; Arrieta-González, C.;
Salgado-Delgado, R.; Lastarria-Tapia, H.; Castañón-Vilca, J. Potato Starch-based Films:
Effects of Glycerol and Montmorillonite Nanoclay Concentration. Rev. Mex. Ing. Quím.
2020, 19 (2), 627–637.
de la Concha, B. B. S.; Agama-Acevedo, E.; Nuñez-Santiago, M. C.; Bello-Perez, L. A.;
Garcia, H. S.; Alvarez-Ramirez, J. Acid Hydrolysis of Waxy Starches with Different
Granule Size for Nanocrystal Production. J. Cereal Sci. 2018, 79, 193–200.
Dimitrijevic, M.; Karabasil, N.; Boskovic, M.; Teodorovic, V.; Vasilev, D.; Djordjevic, V.;
Kilibarda, N.; Cobanovic, N. Safety Aspects of Nanotechnology Applications in Food
Packaging. Proc. Food Sci. 2015, 5, 57–60.
Dufresne, A. Polysaccharide Nano Crystal Reinforced Nanocomposites. Can. J. Chem. 2008,
86 (6), 484–494.
Dufresne, A.; Castaño, J. Polysaccharide Nanomaterial Reinforced Starch Nanocomposites: A
Review. Starch-Stärke 2017, 69 (1–2), 1500307.
El-Sheikh, M. A. New Technique in Starch Nanoparticles Synthesis. Carbohydr. Polym. 2017,
176, 214–219.
Estevez-Areco, S.; Guz, L.; Famá, L.; Candal, R.; Goyanes, S. Bioactive Starch Nanocomposite
Films with Antioxidant Activity and Enhanced Mechanical Properties Obtained by
Extrusion Followed by Thermo-Compression. Food Hydrocoll. 2019, 96, 518–528.
Fahmy, H. M.; Salah Eldin, R. E.; Abu Serea, E. S.; Gomaa, N. M.; AboElmagd, G. M.;
Salem, S. A.; Elsayed, Z. A.; Edrees, A.; Shams-Eldin, E.; Shalan, A. E. Advances in
Nanotechnology and Antibacterial Properties of Biodegradable Food Packaging Materials.
RSC Adv. 2020, 10 (35), 20467–20484.
Falguera, V.; Quintero, J. P.; Jiménez, A.; Muñoz, J. A.; Ibarz, A. Edible Films and Coatings:
Structures, Active Functions and Trends in their Use. Trends Food Sci. Technol. 2011, 22
(6), 292–303.
Fernández Valdés, D.; Bautista Baños, S.; Fernández Valdés, D.; Ocampo Ramírez, A.;
García Pereira, A.; Falcón Rodríguez, A. Películas y Recubrimientos Comestibles: Una
Alternativa Favorable en la Conservación Poscosecha de Frutas y Hortalizas. Rev. Cienc.
Téc. Agropecu. 2015, 24 (3), 52–57.
Formela, K.; Zedler, Ł.; Hejna, A.; Tercjak, A. Reactive Extrusion of Bio-based Polymer
Blends and Composites-Current Trends and Future Developments. Express Polym. Lett.
2018, 12 (1).
Gamage, A.; Thiviya, P.; Mani, S.; Ponnusamy, P. G.; Manamperi, A.; Evon, P.; Merah, O.;
Madhujith, T. Environmental Properties and Applications of Biodegradable Starch-based
Nanocomposites. Polymers [Online], 2022.
García, N. L.; Famá, L.; D’Accorso, N. B.; Goyanes, S. Biodegradable Starch Nanocomposites
BT—Eco-friendly Polymer Nanocomposites: Processing and Properties. Thakur, V. K.,
Thakur, M. K., Eds.; Springer India: New Delhi, 2015; pp 17–77.
García, N. L.; Ribba, L.; Dufresne, A.; Aranguren, M. I.; Goyanes, S. Physico-Mechanical
Properties of Biodegradable Starch Nanocomposites. Macromol. Mater. Eng. 2009, 294
(3), 169–177.
García, N. L.; Ribba, L.; Dufresne, A.; Aranguren, M.; Goyanes, S. Effect of Glycerol on the
Morphology of Nanocomposites Made from Thermoplastic Starch and Starch Nanocrystals.
Carbohydr. Polym. 2011, 84 (1), 203–210.
Ghanbari, A.; Tabarsa, T.; Ashori, A.; Shakeri, A.; Mashkour, M. Preparation and
Characterization of Thermoplastic Starch and Cellulose Nanofibers as Green
Nanocomposites: Extrusion Processing. Int. J. Biol. Macromol. 2018, 112, 442–447.
Ghoshal, G. Recent Trends in Active, Smart, and Intelligent Packaging for Food Products
A2—Grumezescu, Alexandru Mihai. In Food Packaging and Preservation; Holban, A. M.
G. a. A. M. Ed.; Academic Press, 2018; pp 343–374.
González, A.; Alvarez Igarzabal, C. I. Nanocrystal-reinforced Soy Protein Films and their
Application as Active Packaging. Food Hydrocoll. 2015, 43, 777–784.
González, K.; Retegi, A.; González, A.; Eceiza, A.; Gabilondo, N. Starch and cellulose
nanocrystals together into thermoplastic starch bionanocomposites. Carbohydr. Polym.
2015, 117, 83–90.
Goudarzi, V.; Shahabi-Ghahfarrokhi, I.; Babaei-Ghazvini, A. Preparation of Ecofriendly
UV-Protective Food Packaging Material by Starch/TiO2 Bio-nanocomposite:
Characterization. Int. J. Biol. Macromol. 2017, 95, 306–313.
Grande, C. J.; Torres, F. G.; Gomez, C. M.; Troncoso, O. P.; Canet-Ferrer, J.; Martínez-Pastor,
J. Development of Self-assembled Bacterial Cellulose–Starch Nanocomposites. Mater. Sci.
Eng. C 2009, 29 (4), 1098–1104.
Gujral, H.; Sinhmar, A.; Nehra, M.; Nain, V.; Thory, R.; Pathera, A. K.; Chavan, P. Synthesis,
Characterization, and Utilization of Potato Starch Nanoparticles as a Filler in Nanocomposite
Films. Int. J. Biol. Macromol. 2021, 186, 155–162.
Haaj, S. B.; Magnin, A.; Pétrier, C.; Boufi, S. Starch Nanoparticles Formation via High Power
Ultrasonication. Carbohydr. Polym. 2013, 92 (2), 1625–1632.
Haaj, S. B.; Thielemans, W.; Magnin, A.; Boufi, S. Starch Nanocrystals and Starch
Nanoparticles from Waxy Maize as Nanoreinforcement: A Comparative Study. Carbohydr.
Polym. 2016, 143, 310–317.
Hamad, A. F.; Han, J.-H.; Kim, B.-C.; Rather, I. A. The Intertwine of Nanotechnology with
the Food Industry. Saudi J. Biol. Sci. 2018, 25 (1), 27–30.
Han, J. H. Chapter 9—Edible Films and Coatings: A Review. In Innovations in Food
Packaging, 2nd ed.; Han, J. H. Ed.; Academic Press: San Diego, 2014; pp 213–255.
He, X.; Hwang, H.-M. Nanotechnology in Food Science: Functionality, Applicability, and
Safety Assessment. J. Food Drug Anal. 2016, 24 (4), 671–681.
Hernández, A. I. M.; González, R. A. O.; Padrón, P.; Araya, Q.; Wexler, G.; Cubero, C.
Biodegradable Films with Bioactive Properties. Rev. Venez. Cienc. Tecnol. Aliment 2017,
8 (1), 57–89.
Hu, Z.; Cranston, E. D.; Ng, R.; Pelton, R. Tuning Cellulose Nanocrystal Gelation with
Polysaccharides and Surfactants. Langmuir 2014, 30 (10), 2684–2692.
Huang, J.; Chang, P. R.; Dufresne, A. Polysaccharide Nanocrystals: Current Status and
Prospects in Material Science. In Polysaccharide-Based Nanocrystals; Huang, J., Chang, P.
R., Dufresne, A., Eds.; Wiley-VCH: Weinheim, Germany, 2015; Vol. 1, pp 1–14.
Huh, M. S.; Lee, E. J.; Koo, H.; Yhee, J. Y.; Oh, K. S.; Son, S.; Lee, S.; Kim, S. H.; Kwon,
I. C.; Kim, K. Polysaccharide-based Nanoparticles for Gene Delivery. Topics Curr. Chem.
2017, 375 (2), 31.
Imran, M.; Revol-Junelles, A.-M.; Martyn, A.; Tehrany, E. A.; Jacquot, M.; Linder, M.; Desobry,
S. Active Food Packaging Evolution: Transformation from Micro-to Nanotechnology. Crit.
Rev. Food Sci. Nutr. 2010, 50 (9), 799–821.
Jiang, S.; Liu, C.; Wang, X.; Xiong, L.; Sun, Q. Physicochemical Properties of Starch
Nanocomposite Films Enhanced by Self-assembled Potato Starch Nanoparticles. LWT -
Food Sci. Technol. 2016, 69, 251–257.
Joye, I. J.; McClements, D. J. Biopolymer-based Nanoparticles and Microparticles:
Fabrication, Characterization, and Application. Curr. Opin. Colloid Interf. Sci. 2014, 19
(5), 417–427.
Kaplan, D. L. Introduction to Biopolymers from Renewable Resources. In Biopolymers from
Renewable Resources; Kaplan, D. L., Ed.; Springer Berlin Heidelberg: Berlin, Heidelberg,
1998; pp 1–29.
Karolina, L.; Anna, T.-H.; Damian, S.; Justyna, O.; Piotr, T.; Jakub, B. Biodegradable
Polylactide and Thermoplastic Starch Blends as Drug Release Device—Mass Transfer
Study. 2018, 20, 75–75.
Kim, H.-Y.; Park, D. J.; Kim, J.-Y.; Lim, S.-T. Preparation of Crystalline Starch Nanoparticles
Using Cold Acid Hydrolysis and Ultrasonication. Carbohydr. Polym. 2013, 98 (1), 295–301.
Kim, H.-Y.; Park, S. S.; Lim, S.-T. Preparation, Characterization and Utilization of Starch
Nanoparticles PRUEBA Y. Colloids Surf. B: 2015, 126, 607–620.
Korbekandi, H.; Iravani, S. Biological Synthesis of Nanoparticles Using Algae. In Green
Biosynthesis of Nanoparticles: Mechanisms and Applications; Rai, M., Posten, C., Eds.;
2013; pp 53–60.
Labet, M.; Thielemans, W.; Dufresne, A. Polymer Grafting Onto Starch Nanocrystals.
Biomacromolecules 2007, 8 (9), 2916–2927.
Le Corre, D.; Bras, J.; Dufresne, A. Starch Nanoparticles: A Review. Biomacromolecules
2010, 11 (5), 1139–1153.
Li, T.; Turng, L. S.; Gong, S.; Erlacher, K. Polylactide, Nanoclay, and Core–Shell Rubber
Composites. Polym. Eng. Sci. 2006, 46 (10), 1419–1427.
Lin, N.; Huang, J.; Chang, P. R.; Anderson, D. P.; Yu, J. Preparation, Modification, and
Application of Starch Nanocrystals in Nanomaterials: A Review. J. Nanomater. 2011, 2011,
1–13.
Lin, N.; Huang, J.; Chang, P. R.; Feng, L.; Yu, J. Effect of Polysaccharide Nanocrystals on
Structure, Properties, and Drug Release Kinetics of Alginate-based Microspheres. Colloids
Surf. B: Biointerfaces 2011, 85 (2), 270–279.
Lin, N.; Huang, J.; Dufresne, A. Preparation, Properties and Applications of Polysaccharide
Nanocrystals in Advanced Functional Nanomaterials: A Review. Nanoscale 2012, 4 (11),
3274–3294.
Liu, B.; Xu, H.; Zhao, H.; Liu, W.; Zhao, L.; Li, Y. Preparation and Characterization of
Intelligent Starch/PVA Films for Simultaneous Colorimetric Indication and Antimicrobial
Activity for Food Packaging Applications. Carbohydr. Polym. 2017, 157, 842–849.
Liu, Z.; Jiao, Y.; Wang, Y.; Zhou, C.; Zhang, Z. Polysaccharides-based Nanoparticles as Drug
Delivery Systems. Adv. Drug Deliv. Rev. 2008, 60 (15), 1650–1662.
Llanos, J. H. R.; Tadini, C. C. Preparation and Characterization of Bio-nanocomposite
Films Based on Cassava Starch or Chitosan, Reinforced with Montmorillonite or Bamboo
Nanofibers. Int. J. Biol. Macromol. 2018, 107, 371–382.
López, O.; Castillo, L. A.; García, M. A.; Villar, M. A.; Barbosa, S. E. Food Packaging Bags
based on Thermoplastic Corn Starch Reinforced with Talc Nanoparticles. Food Hydrocoll.
2015, 43, 18–24.
Lu, Y.; Weng, L.; Cao, X. Morphological, Thermal and Mechanical Properties of Ramie
Crystallites—Reinforced Plasticized Starch Biocomposites. Carbohydr. Polym. 2006, 63
(2), 198–204.
Ma, X.; Chang, P. R.; Yang, J.; Yu, J. Preparation and Properties of Glycerol Plasticized-Pea
Starch/Zinc Oxide-Starch Bionanocomposites. Carbohydr. Polym. 2009, 75 (3), 472–478.
Madhumitha, G.; Fowsiya, J.; Mohana Roopan, S.; Thakur, V. K. Recent Advances in Starch–
Clay Nanocomposites. Int. J. Polym. Anal. Charact. 2018, 1–15.
Malamatari, M.; Taylor, K. M. G.; Malamataris, S.; Douroumis, D.; Kachrimanis, K.
Pharmaceutical Nanocrystals: Production by Wet Milling and Applications. Drug Discov.
Today 2018, 23 (3), 534–547.
Mallakpour, S.; Rashidimoghadam, S. Starch/MWCNT-vitamin C Nanocomposites:
Electrical, Thermal Properties and Their Utilization for Removal of Methyl Orange.
Carbohydr. Polym. 2017, 169, 23–32.
Mbey, J. A.; Hoppe, S.; Thomas, F. Cassava Starch–Kaolinite Composite Film. Effect of
Clay Content and Clay Modification on Film Properties. Carbohydr. Polym. 2012, 88 (1),
213–222.
Meira, S. M. M.; Zehetmeyer, G.; Scheibel, J. M.; Werner, J. O.; Brandelli, A. Starch-
halloysite Nanocomposites Containing Nisin: Characterization and Inhibition of Listeria
Monocytogenes in Soft Cheese. LWT - Food Sci. Technol. 2016, 68, 226–234.
Meira, S. M. M.; Zehetmeyer, G.; Werner, J. O.; Brandelli, A. A Novel Active Packaging
Material based on Starch-halloysite Nanocomposites Incorporating Antimicrobial Peptides.
Food Hydrocoll. 2017, 63, 561–570.
Mendes, J. F.; Paschoalin, R. T.; Carmona, V. B.; Sena Neto, A. R.; Marques, A. C. P.;
Marconcini, J. M.; Mattoso, L. H. C.; Medeiros, E. S.; Oliveira, J. E. Biodegradable
Polymer Blends based on Corn Starch and Thermoplastic Chitosan Processed by Extrusion.
Carbohydr. Polym. 2016, 137, 452–458.
Mihindukulasuriya, S. D. F.; Lim, L. T. Nanotechnology Development in Food Packaging: A
Review. Trends Food Sci. Technol. 2014, 40 (2), 149–167.
Mittal, A.; Garg, S.; Kohli, D.; Maiti, M.; Jana, A. K.; Bajpai, S. Effect of Cross Linking of
PVA/Starch and Reinforcement of Modified Barley Husk on the Properties of Composite
Films. Carbohydr. Polym. 2016, 151, 926–938.
Mohammadi Nafchi, A.; Moradpour, M.; Saeidi, M.; Alias, A. K. Thermoplastic Starches:
Properties, Challenges, and Prospects. Starch-Stärke 2013, 65 (1-2), 61–72.
Mohanraj, V.; Chen, Y. Nanoparticles-A Review. Tropi. J. Pharmaceut. Res. 2006, 5 (1),
561–573.
Mukurumbira, A. R.; Mellem, J. J.; Amonsou, E. O. Effects of Amadumbe Starch Nanocrystals
on the Physicochemical Properties of Starch Biocomposite Films. Carbohydr. Polym. 2017,
165, 142–148.
Müller, C. M. O.; Laurindo, J. B.; Yamashita, F. Composites of Thermoplastic Starch and
Nanoclays Produced by Extrusion and Thermopressing. Carbohydr. Polym. 2012, 89 (2),
504–510.
Nasseri, R.; Mohammadi, N. Starch-based Nanocomposites: A Comparative Performance
Study of Cellulose Whiskers and Starch Nanoparticles. Carbohydr. Polym. 2014, 106,
432–439.
Nasution, A. H.; Harahap, H.; Al Fath, M. T. Physical Properties of Sago Starch Biocomposite
Filled with Nanocrystalline Cellulose (NCC) from Rattan Biomass: The Effect of Filler
Loading and Co-plasticizer Addition. IOP Conf. Ser.: Mater. Sci. Eng. 2018, 309 (1),
12033–12033.
Nathalie, G.; Stéphane, P.; Jose, M. L.; Yolanda, E.; Carole, G. Nanotechnologies for Active
and Intelligent Food Packaging: Opportunities and Risks. In Nanotechnology in Agriculture
and Food Science; Axelos, M. A.. Voorde, M. H. V. D., Eds.; Wiley-VCH Verlang GmbH
& Co. KGaA. 2017; pp 177–196.
Oikawa, H.; Masuhara, A.; Kasai, H.; Mitsui, T.; Sekiguchi, T.; Nakanishi, H. Organic and
Polymer Nanocrystals: Their Optical Properties and Function. In Handai Nanophotonics;
Masuhara, H.; Kawata, S., Eds.; Elsevier: New York, USA, 2004; Vol. 1, pp 205–224.
Othman, S. H. Bio-nanocomposite Materials for Food Packaging Applications: Types of
Biopolymer and Nano-sized Filler. Agric. Agric. Sci. Procedia 2014, 2, 296–303.
Park, H. J.; Byun, Y. J.; Kim, Y. T.; Whiteside, W. S.; Bae, H. J. Processes and Applications for
Edible Coating and Film Materials from Agropolymers A2—Han, Jung H. In Innovations in
Food Packaging, 2nd ed.; Han, J. H., Ed.; Academic Press: San Diego, 2014; pp 257–275.
Pelissari, F. M.; Andrade-Mahecha, M. M.; Sobral, P. J. D. A.; Menegalli, F. C. Nanocomposites
based on Banana Starch Reinforced with Cellulose Nanofibers Isolated from Banana Peels.
J. Colloid Interface Sci. 2017, 505, 154–167.
Podshivalov, A.; Zakharova, M.; Glazacheva, E.; Uspenskaya, M. Gelatin/Potato Starch
Edible Biocomposite Films: Correlation between Morphology and Physical Properties.
Carbohydr. Polym. 2017, 157, 1162–1172.
Quintero, C.; Falguera, V.; Muñoz, H. Films and Edible Coatings: Importance, and Recent
Trends in Fruit and Vegetable Value Chain. Rev. Tumbaga 2010, 5, 93–118.
Ranjan, S.; Dasgupta, N.; Chakraborty, A. R.; Samuel, S. M.; Ramalingam, C.; Shanker,
R.; Kumar, A. Nanoscience and Nanotechnologies in Food Industries: Opportunities and
Research Trends. J. Nanopart. Res. 2014, 16 (6), 2464.
Ribeiro-Santos, R.; Andrade, M.; Melo, N. R. D.; Sanches-Silva, A. Use of Essential Oils in
Active Food Packaging: Recent Advances and Future Trends. Trends Food Sci. Technol.
2017, 61, 132–140.
Romero-Bastida, C. A.; Chávez Gutiérrez, M.; Bello-Pérez, L. A.; Abarca-Ramírez, E.;
Velazquez, G.; Mendez-Montealvo, G. Rheological Properties of Nanocomposite-Forming
Solutions and Film Based on Montmorillonite and Corn Starch with Different Amylose
Content. Carbohydr. Polym. 2018, 188, 121–127.
Rong, L.; Shen, M.; Wen, H.; Ren, Y.; Xiao, W.; Xie, J. Preparation and Characterization of
Hyacinth Bean Starch Film Incorporated with TiO2 Nanoparticles and Mesona Chinensis
Benth Polysaccharide. Int. J. Biol. Macromol. 2021, 190, 151–158.
Rovina, K.; Vonnie, J. M.; Mantihal, S.; Joseph, J.; Halid, N. F. A. Development of Films
based on Tapioca Starch/Gold Nanoparticles for the Detection of Organophosphorus
Pesticides. J. Consum. Prot. Food Safety 2021, 16 (2), 143–152.
Salaberria, A. M.; Diaz, R. H.; Labidi, J.; Fernandes, S. C. M. Role of Chitin Nanocrystals
and nanofibers on Physical, Mechanical and Functional Properties in Thermoplastic Starch
Films. Food Hydrocoll. 2015, 46, 93–102.
Salaberria, A. M.; Labidi, J.; Fernandes, S. C. M. Chitin Nanocrystals and Nanofibers as
Nano-sized Fillers into Thermoplastic Starch-based Biocomposites Processed by Melt-
mixing. Chem. Eng. J. 2014, 256, 356–364.
Sharma, R.; Jafari, S. M.; Sharma, S. Antimicrobial Bio-nanocomposites and their Potential
Applications in Food Packaging. Food Cont. 2020, 112, 107086.
Sinha Ray, S.; Okamoto, M. Polymer/Layered Silicate Nanocomposites: A Review from
Preparation to Processing. Prog. Polym. Sci. 2003, 28 (11), 1539–1641.
Skaff, H.; Emrick, T. Semiconductor Nanoparticles. In Nanoparticles: Building Blocks for
Nanotechnology; Rotello, V., Ed.; Kluwer Academic/Plenum Publishers: New York, USA,
2004; pp 27–46.
Slavutsky, A. M.; Bertuzzi, M. A.; Armada, M. Water Barrier Properties of Starch-Clay
Nanocomposite Films. Braz. J. Food Technol. 2012, 15 (3), 208–218.
Srikhao, N.; Kasemsiri, P.; Ounkaew, A.; Lorwanishpaisarn, N.; Okhawilai, M.; Pongsa, U.;
Hiziroglu, S.; Chindaprasirt, P. Bioactive Nanocomposite Film based on Cassava Starch/
Polyvinyl Alcohol Containing Green Synthesized Silver Nanoparticles. J. Polym. Environ.
2021, 29 (2), 672–684.
Tjong, S. C. Nanocrystalline Materials: Their Synthesis-Structure-Property Relationships
and Applications, 2nd ed.; Newnes, 2013; p 418.
Usman, A.; Hussain, Z.; Riaz, A.; Khan, A. N. Enhanced Mechanical, Thermal and
Antimicrobial Properties of poly(vinyl alcohol)/Graphene Oxide/Starch/Silver
Nanocomposites Films. Carbohydr. Polym. 2016, 153, 592–599.
Viguié, J.; Molina-Boisseau, S.; Dufresne, A. Processing and Characterization of Waxy Maize
Starch Films Plasticized by Sorbitol and Reinforced with Starch Nanocrystals. Macromol.
Biosci. 2007, 7 (11), 1206–1216.
Wang, C.; Gao, X.; Chen, Z.; Chen, Y.; Chen, H. Preparation, Characterization and Application
of Polysaccharide-Based Metallic Nanoparticles: A Review. Polymers 2017, 9 (12), 689.
Wang, N.; Zhang, X.; Han, N.; Bai, S. Effect of Citric Acid and Processing on the Performance
of Thermoplastic Starch/Montmorillonite Nanocomposites. Carbohydr. Polym. 2009, 76
(1), 68–73.
Wang, Y.; Tian, H.; Zhang, L. Role of Starch Nanocrystals and Cellulose Whiskers in
Synergistic Reinforcement of Waterborne Polyurethane. Carbohydr. Polym. 2010, 80 (3),
665–671.
Woehl, M. A.; Canestraro, C. D.; Mikowski, A.; Sierakowski, M. R.; Ramos, L. P.; Wypych,
F. Bionanocomposites of Thermoplastic Starch Reinforced with Bacterial Cellulose
Nanofibres: Effect of Enzymatic Treatment on Mechanical Properties. Carbohydr. Polym.
2010, 80 (3), 866–873.
Wu, J. G.; Wang, P. J.; Chen, S. C. Antioxidant and Antimicrobial Effectiveness of Catechin‐
Impregnated PVA—Starch Film on Red Meat. J. Food Qual. 2010, 33 (6), 780–801.
Youssef, A. M.; El-Sayed, S. M. Bionanocomposites Materials for Food Packaging

Applications: Concepts and Future Outlook. Carbohydr. Polym. 2018, 193, 19–27.
Zeppa, C.; Gouanvé, F.; Espuche, E. Effect of a Plasticizer on the Structure of Biodegradable
Starch/Clay Nanocomposites: Thermal, Water, Sorption, and Oxygen-Barrier Properties. J.
Appl. Polym. Sci. 2009, 112 (4), 2044–2056.
Zhai, X.; Shi, J.; Zou, X.; Wang, S.; Jiang, C.; Zhang, J.; Huang, X.; Zhang, W.; Holmes, M.
Novel Colorimetric Films based on Starch/Polyvinyl Alcohol Incorporated with Roselle
Anthocyanins for Fish Freshness Monitoring. Food Hydrocoll. 2017, 69, 308–317.
CHAPTER 3
Release Studies to Improve the

Mechanical Properties of the Biopolymer,
Polylactic Acid (PLA) for Food Packaging
Applications
Y. K. REYES ACOSTA1, W. E. CRUZ MARTINEZ1, A. V. REYES ACOSTA2,
L. SEPÚLVEDA TORRE1, C. N. AGUILAR GONZALES1, and
R. I. NARRO CESPEDES1
1
Facultad de Ciencias Químicas de la Universidad Autónoma de Coahuila,
Saltillo, Coahuila, México
2
Facultad de Sistemas de la Universidad Autónoma de Coahuila, Zona
Centro, Arteaga, Coahuila, México
ABSTRACT
Polylactic acid (PLA) is a biodegradable thermoplastic and aliphatic poly-
ester obtained from renewable and sustainable starch-rich materials. Due to
its biodegradability, biocompatibility, and high mechanical properties, such
as tensile strength and Young’s modulus, comparable to those of polystyrene
(PS) and polyethylene terephthalate (PET), PLA has been considered one
of the most important biopolymers. However, due to its lower impact resis-
tance, many materials have been used to improve its properties. Materials
such as food and its derivatives, natural oils, organic material, different types
of fibers, nanoparticles, and others have been incorporated into the PLA
matrix with the aim of showing improved mechanical properties. This paper
presents a review of the research on PLA and the use of enhancers, as well
as the results for the improvement of mechanical properties and applications
of food containers.
3.1 INTRODUCTION
In recent years, there has been a growing scientific and industrial interest in
the use of compounds obtained from renewable sources and with biodegrad-
ability properties that reduce the environmental effect (Scaffaro et al., 2018).
One of the methods recently introduced to resolve this issue is the use of
biopolymers that are naturally occurring in living organisms (Suryanegara et
al., 2017). The use of biopolymers as raw materials would be of great benefit
to environmental protection efforts compared to the use of fossil-based mate-
rials due to less adverse effects (Suryanegara et al., 2017).
Biopolymers have important physicochemical and biological properties.
These properties are targeted for applications in a broad range of fields, espe-
cially in the packaging, textile, agricultural, pharmaceutical, electronics, and
biomedical sectors (Hammiche et al., 2019). Among biodegradable polymers,
poly(lactic acid) (PLA) derived from sugar feedstock, corn, etc. has gained
growing attention as a sustainable and readily biodegradable biomaterial.
This aliphatic polyester, known as GRAS (Generally Recognized As Safe),
was used for food packaging applications (Nilsuwan et al., 2018).
PLA has an immense potential to replace petroleum-based plastics due to
its high mechanical properties, which are comparable to polystyrene (PS) and
polyethylene terephthalate (PET) polymers currently used in the packaging
industry (Herrera et al., 2016; Miao et al., 2019; Suryanegara et al., 2017).
PLA has gained a lot of attention due to its mechanical properties such as
high tensile strength and high Young’s modulus (Carbonell-Verdu et al.,
2017) and its relatively strong biocompatibility (Ge et al., 2018). However,
due to the inherent fragility and lower impact resistance of PLA, a number of
efforts have been made to develop its properties in order to achieve compat-
ibility with thermoplastic processing and manufacturing, thereby enabling it
to compete with commodity polymers (Suryanegara et al., 2017).
Many materials have been used to increase PLA properties. Materials that
can be considered food or derivates: fish gelatin (FG), (Nilsuwan et al., 2018),
carvacrol (CRV) (Scaffaro et al., 2018), beeswax (BW) (Reis et al., 2018),
sugarcane bagasse pulp (SCB) (Suryanegara et al., 2018), olive husk flour
(OHF) (Hammiche et al., 2019). Natural oils like epoxidized cottonseed oil
(ECSO) (Carbonell-Verdu et al., 2017), epoxidized (ELO, ESBO) and non-
epoxidized linseed oil (LO) and soybean oil (SBO) (Orue et al., 2018), and
fennel (FEN) oil (Miao et al., 2019). Organic material as leaves of Posidonia
oceanica (PO) (Scaffaro, Lopresti, and Botta, 2018), wood flour (WF) (Ge et
al., 2018; Holcapkova et al., 2017), wood fiber (F) (Asadi et al., 2018) and
diatomaceous earth (DE) (Aggarwal et al., 2019) can also be considered.
Release Studies to Improve the Mechanical Properties 69
Different kinds of fiber like bamboo fibers (BF) (Li et al., 2019; Long
et al., 2019), Agave fibers (AF) (Cisneros-López et al., 2018), ramie fibers
(Debeli et al., 2019), and cellulose fibers (Laadila et al., 2017) are used.
Nanocomposites as carbon nanotubes (CNT) (Liu et al., 2019) and cellulose
nanocrystals (CNC) (Mariano et al., 2017; Wei et al., 2018). Finally, mate-
rials such as carbon fiber, microfibrillated wood (MFW), microfibrillated
lignocellulose (MFLC) (Winter et al., 2018), and diatomaceous earth (DE)
(Aggarwal et al., 2019) have been used in 3D printing.
3.2 FOOD DERIVATIVES USED TO IMPROVE PLA MECHANICAL

PROPERTIES
Polymeric materials for packaging applications have been cautiously

considered due to the limitations and regulations relating to primary and
post-consumer plastic waste management (Nilsuwan et al., 2018). Ecological
concern and the challenge of recycling packaging materials have stimulated
the production of biodegradable materials (Reis et al., 2018). Mechanical
properties of biobased materials used in food packaging structures are
critical criteria for their performance and functionality. The mechanical
properties of PLA biopolymers can vary from soft plastics to rigid elastics
and high strength materials, depending on a wide variety of factors, such
as molecular weight, stereochemical spine makeup. As a result, different
materials considered as food or food derivatives were used to improve the
mechanical properties of PLA: FG and PLA bilayer films were developed by
casting, exhibiting lower tensile strength but higher elongation at break than
PLA film (Nilsuwan et al., 2018), PLA/CRV nanofiber membranes were also
prepared by electrospinning, adding 28 wt% of CRV caused an increment of
E by ~70%, σm by ~450% and εb by ~1350% (Scaffaro, Lopresti, and Botta,
2018). In addition, attempts have been made to achieve sufficient mechanical
and barrier properties. Thermoplastic starch (TPS) with PLA trays coated
with beeswax (BW) emulsion was prepared by Reis and coworkers. The
trays coated with 1% BW had higher tensile strength than the trays with a
higher BW emulsion content (Reis et al., 2018).
Another reinforcement agent, sugar bagasse pulp (SCB), was used.
It was found that the tensile strength decreased due to an increase in the
quality of the filler. ‘Young’s modulus, however, increased by up to 13–20
wt % (Suryanegara et al., 2018). Polylactic acid and olive husk flour (PLA/
OHF) composites were prepared by extrusion, finding that Young’s modulus
increased by 18% , the tensile strength and the elongation at break decreased
as compared to neat PLA (Hammiche et al., 2019).
TABLE 3.1 Reinforcement Used in Polylactic Acid Food Packing Material.

Reinforcement
Concentration Methodology Results
agent
σm = 31.79 MPa (PLA)
σm = 24.23 MPa (FG)
Layer thickness σm = 27.23 MPa (FG/PLA5:5)
Fish Gelatin (FG) ratios of 9:1, ɛb = 8.89% (PLA)
(Nilsuwan et al., 8:2, 7:3, 6:4 Casting
2018) and 5:5 (FG/ ɛb = 36.27% (FG)
LPA) ɛb = 11.6% (FG/PLA 5:5)
σm = 6.81 MPa (PLA/CRV 28%)
ɛb = 7.8% (PLA)
Carvacrol (CRV) Stirring ɛb = 115.2% (PLA/CRV 28%)

(Scaffaro et al., 28 wt % solution/ Eb = 57.1 MPa (PLA)
2018) electrospinning E = 98 MPa (PLA/CRV 28%)
σm = 10.3 MPa (TPS/PLA)

σm = 11.5 MPa (TPS/PLA−BW1)
Thermoplastic ɛb = 27% (TPS/PLA)
starch (TPS) using ɛb = 22% (TPS/PLA − BW1)
1, 2 and 3 wt % Mixing/
beeswax (BW)
of BW extrusion E = 294 MPa (TPS/PLA)
emulsion to coat
(Reis et al., 2018) E = 292 MPa (TPS/PLA − BW1)
σm = 38.3 MPa (PLA/pulp5)
εb = 15.6% (PLA)
εb = 5.5% (PLA/pulp 5)
Sugarcane bagasse E = 1.18 GPa (PLA)
pulp (SCB) 5, 10 and 20 Stirring
E = 1.32 GPa (PLA/pulp 5)
(Suryanegara et al., wt % solution
2018) σm = 75 MPa (PLA)
σm = 64 MPa (PLA/untreated OHF)
σm = 71 MPa (PLA/treated OHF)
εb = 6.8% (PLA)
TABLE 3.1 (Continued)

Reinforcement
agent
εb= 3.2% (PLA/untreated OHF)
Untreated and
treated olive εb = 3.5% (PLA/treated OHF)
husk flour (OHF) 20 wt % Extrusion E = 3101 GPa (PLA)
(Hammiche et al., E = 3635 GPa (PLA/ untreated OHF)
2019)
E = 3757 GPa (PLA/ treated OHF)
σm = 24.23 MPa (FG)
Layer thickness σm = 27.23 MPA (FG/PLA 5:5)
Fish Gelatin (FG) ratios of 9:1, εb = 8.89% (PLA)
(Nilsuwan et al., 8:2, 7:3, 6:4 Casting
2018) and 5:5 (FG/ εb = 36.27% (FG)
LPA) εb = 11.6% (FG/PLA 5:5)
σm = 6.81 MPa (PLA/CRV 28%)
εb = 7.8% (PLA)
Carvacrol (CRV) Stirring εb = 115.2% (PLA/CRV 28%)

(Scaffaro et al., 28 wt % solution/ E = 57.1 MPa (PLA)
2018) electrospinning E = 98 MPa (PLA/CRV 28%)
σm=10.3 MPa (TPS/PLA)

σm = 11.5 MPa (TPS/PLA‒BW1)
Thermoplastic εb = 27% (TPS/PLA)
starch (TPS) using εb = 22% (TPS/PLA‒BW1)
1, 2 and 3 wt % Mixing/
beeswax (BW)
of BW extrusion E = 294 MPa (TPS/PLA)
emulsion to coat
(Reis et al., 2018) E = 292 MPa (TPS/PLA‒BW1)
σm = 38.3 MPa (PLA/pulp 5)
εb = 15.6% (PLA)
εb = 5.5% (PLA/pulp 5)
Sugarcane bagasse E = 1.18 GPa (PLA)
pulp (SCB) 5, 10 and 20 Stirring
E = 1.32 GPa (PLA /pulp 5)
(Suryanegara et al., wt % solution
2018) σm = 75 MPa (PLA)
σm = 64 MPa (PLA/untreated OHF)
σm = 71 MPa (PLA/treated OHF)
εb = 6.8% (PLA)

Reinforcement
agent
εb = 3.2% (PLA/untreated OHF)
Untreated and
treated olive εb = 3.5% (PLA/treated OHF)
husk flour (OHF) 20 wt % Extrusion E = 3101 GPa (PLA)
(Hammiche et al., E = 3635 GPa (PLA/ untreated OHF)
2019)
E = 3737 GPa (PLA/ treated OHF)
3.3 NATURAL OILS USED AS PLA REINFORCEMENT AGENT
The use of compatible plasticizers for PLA may be an effective solution

to minimize the fragility of PLA, since plasticizers lead to a drop in the
glass transfer temperature of PLA resulting in more durable materials.
Increased interest in low migration and low toxicity plasticizers focus
on bio-based plasticizers manufactured from vegetable oils, citrus, and
sugar derivatives as an alternative to common plasticizers. Epoxidized
oils are known as reactive plasticizers due to the reactivity between the
epoxy functional groups and the –OH and –COOH PLA polymer groups.
Epoxidized vegetable oils can be produced by epoxidizing C=C bonds of
oils and plasticizers with zero volatility and exceptionally low migration
patterns (Orue et al., 2018).
In order to boost its mechanical properties, certain oils have been
applied to PLA: the melt extrusion was used to plasticize PLA formula-
tions with various ECSO contents in the range of 0–10 wt. %. The 10 wt.%
composition of PLA demonstrates a surprising improvement in mechanical
ductile properties (Carbonell et al., 2017). PLA was also plasticized with
epoxidized and non-epoxidized linseed and soybean oils, and the effect
of different plasticizers was assessed. Epoxidized oils have been shown
to have slightly higher tensile modulus values than non-epoxidized oils,
particularly for those blends plasticized with ELO (Orue et al., 2018). In
addition, a biodegradable film based on PLA and PHB was produced with
the addition of FEN oil, finding that the complete mechanical properties of
PLA-PHB film were comparable to those of EVOH film (Miao et al., 2019),
which are summarized in Table 3.2.
TABLE 3.2 Organic Materials Addition in PLA: Effect in the Stability Thermal.
Reinforcement agent Concentration Methodology Results
Exporized cottonseed oil 2.5, 5, 7.5 and Mixing/ σm = 63.7 MPa (PLA)
(ECSO) (Carbonell et 10 wt % extrusion σm = 49.5 MPa (PLA-10wt. %ECSO)
al., 2017) E = 3.592 GPa (PLA)
E = 3.99 GPa (PLA-10wt. %ECSO)
εb = 9.06% (PLA)
εb = 110.5% (PLA-10 wt. % ECSO)
FM = 3.296 GPa (PLA)
FM = 3.361 GPa (PLA-10 wt. % ECSO)
FS = 116.3 MPa (PLA)
FS = 53.3 MPa (PLA-10 wt. % ECSO)
Exporized (ELO, ESBO) 5, 10 y 20 wt % Melt mixing εb = increased 119% (PLA-20 wt. % oils)
and non-exporized a increased 100% (PLA-20 wt. % oils)
Linseed Oil (LO) and
soybean oil (SBO) (Orue
et al., 2018)
Fennel (FEN) Oil and 24.46 wt % PHB Extrusion σm = 30.38 MPa (EVOH)
polyhydroxybutyrate and 6 wt %FEN σm = 41.20 MPa (PLA-PBH)
(PBH). Using ethanol oil
vinyl alcohol (EVOH) σm = 41.49 MPa (PLA-PBH-FEN)
(Miao et al., 2019) εb = 212.7% (EVOH)
εb = 4.93% (PLA-PBH)
εb = 6.90% (PLA-PBH-FEN)
3.4 ORGANIC MATERIAL IN PLA-BASED COMPOSITES

Recently, various functional groups or fillers have been introduced into poly-
meric networks to add new functionalities and boost efficiency and preserve
reusability. In this sense, increasing attention has been paid to natural fibers
and particles as a reinforcement or filler agent in PLA-based composites. These
materials allowed the recovery of agricultural or marine waste, providing goods
with final properties suitable for a range of applications (Scaffaro et al., 2018).
Many materials, which can be called organic, have been used as rein-
forcement or filler in PLA composites. Leaves of Posidonia oceanica (PO)
have been used for the preparation of PLA-based biocomposites through
melting. The findings revealed that the increase in PO material improved both
the tensile and the flexural modulus (Scaffaro et al., 2018). Also, the PLA
composites produce 20 wt% of the wood flour (WF) was prepared by melting.
The inclusion of ZIKA did not influence the final mechanical properties of
the composite PLA/WF-ZIKA (Holcapkova et al., 2017). Wood flour, PLA,
and other ingredients have also been combined. The effect of four factors:
heat temperature, molding pressure, holding pressure time, and azodicarbon-
amide (AC) content was investigated. The optimal conditions were a heating
temperature of 178°C, a molding pressure of 7 MPa, and a holding pressure
of 25 s. In addition, the integration of nanographene (NG) into PLA/wood
fiber composites strengthened the tensile and bending properties (Asadi et
al., 2018). The findings showed that the composites made of PLA68.5/F30/
NG1.5 have the best tensile and bending properties. And a hybrid electrospun
membrane of electroactive PLLA polymer with glass-reinforced hydroxyapa-
tite (Bonelike®) microparticles has been processed. It has been found that the
addition of Bonelike® particles contributes to a decline in the Young modulus
and to tension at breakage (Santos et al., 2017). As well as applying Diato-
maceous Earth (DE) to the PLA, there has been a decrease in the mechanical
properties of the filaments (Aggarwal et al., 2019).
TABLE 3.3 Organic Material in PLA-Based Composites.

Leaves of Posidonia 10 and 20 wt % Melt mixing E = 1.891 GPa (PLA)
oceanica (PO) with PO particles E = 2.071 GPa (PLA/PO150 ‒ 10)
(Scaffaro, Lopresti, size <150 μm and
and Botta, 2018) 150–300 μm ϵb = 3.66% (PLA)
ϵb = 2.46% (PLA/PO150 ‒ 10)
σm = 39.8 MPa (PLA/PO150 ‒ 10)
FM = 3.39 GPa (PLA)
FM = 3.97 GPa (PLA/PO150 ‒ 10)
FS = 82 MPa (PLA)
FS = 65.6 MPa (PLA/ PO150 ‒ 10)
Maple Wood Flour The effects of Mixing Optimum conditions: heating
(WF) (Ge et al., 2018) heating tempera- temperature of 178°C, molding
ture, molding pressure of 7 MPa, and
pressure, holding holding pressure time of 25 s.
pressure time, and
AC foaming agent
Wood Fiber (F) and 15 and 30 wt.% of Mixing Highest σm for PLA68.5/F30/NG1.5
Nanographene (NG) F with 0.75, 1.5, Highest FM for PLA68.5/F30/NG1.5
(Asadi et al., 2018) and 3 wt.% of NG
Highest for PLA83.5//F15/NG1.5

Wood flour (WF) 20 wt.% of WF Melt mixing σm = 62.9 MPa (PLA)
and ZIKA additive σm = 55.4 MPa (PLA/WF‒ZIKA)
(Holcapkova et al.,
2017) ϵb = 1.02% (PLA)
ϵb = 3.5% (PLA/WF‒ZIKA)
E = 2.48 GPa (PLA)
E = 2.73 GPa (PLA/WF‒ZIKA)
Diatomaceous Earth 5, 10 and 15 wt.% Extrusion σm = 56.14 MPa (PLA)
(DE) (Aggarwal et al., σm = 44.46 MPa (PLA/DE ‒ 5%)
2019)
E = 2.25 GPa (PLA)
E = 2.16 GPa (PLA /DE ‒ 5%)
Bonelike® and PLLA 80 wt.% Stirring E = 18 MPa (PLLA)
(Santos et al., 2017) solution/ E = 5.4 MPa (Bonelike® PLLA)
electrospin-
ning σm = 8 MPa (PLLA)
σm = 0.57 MPa (Bonelike® PLLA)
ϵb = 38% (PLLA)
ϵb = 41% (Bonelike® PLLA)
Leaves of Posidonia 10 and 20 wt.% Melt mixing E = 1.891 GPa (PLA)
oceanica (PO) with PO particles E = 2.071 GPa (PLA/PO150 ‒ 10)
(Scaffaro, Lopresti, size <150 μm and
and Botta, 2018) 150–300 μm ϵb = 3.66% (PLA)
ϵb = 2.46% (PLA/PO150 ‒ 10)
σm = 39.8 MPa (PLA/ PO150 ‒ 10)
FM = 3.39 GPa (PLA)
FM = 3.97 GPa (PLA/PO150 ‒ 10)
FS = 82 GPa (PLA)
FS = 65.6 MPA (PLA/PO150 ‒ 10)
3.5 NATURAL FIBERS IN PLA-BASED COMPOSITES

In recent decades, the use of plant fibers in composite materials has drawn
significant attention. Natural fibers can greatly enhance fracture strength,
crack resistance, stress performance, flexural properties, impact strength, and
fatigue behavior of composites (Long et al., 2019). However, the mechanical
properties of any fiber-reinforced composite depend on the direction of the
fibers, the quality of the fibers, and the form of matrix used to determine the
adhesion strength of the fiber–matrix interface (Debeli et al., 2019).
Various types of fibers have been integrated into PLA, e.g. bamboo fiber
(BF), which has been incorporated as reinforced by a twin-screw extruder
and injection molding. Compared to UBF/PLA composites, the mechanical
properties of ABF/PLA composites have improved (Long et al., 2019). In
addition, PLA/agave fiber biocomposites were prepared by dry-molding and
rotational molding to determine the suitability of this process for the manu-
facture of these materials. Tensile and flexural strengths decreased with fiber
content for both manufacturing processes, while tensile and flexural moduli,
as well as impact strength and stiffness, were above the matrix (PLA) value
of 10 wt % agave for R-BCs (Cisneros-López et al., 2018). In addition, the
properties of molded ramie fibers reinforced with PLA biocomposites have
been investigated. DAP was added to the surface of ramie fibers with and
without pretreatment prior to the preparation of composites. DAP-treated
ramie fiber reinforced composites improved mechanical strength (Debeli et
al., 2019).
In addition, the properties of PLA-g-BF/PLA composites developed by
blending with a three-component plasticizer, glycerol/formamide/tributyl
citrate, were investigated. The bending power and elongation at breakage
of PLA-g-BF/PLA composites improved by 19.3% and 30.1% compared to
ungreased composites (Li et al., 2019). And biocomposites from recycled
PLA and pulp and paper solid waste processed cellulosic fibers. Mechanical
properties of biocomposites with 2% (w/w) of treated cellulosic fiber have
been improved relative to recycled PLA (Laadila et al., 2017).
Many nanoparticles have been used to strengthen PLA, for example
by using electrospinning to produce PLA/CNTs/CS composite fibers, and
it has been observed that, as the CS concentration rose from 1 to 5 wt.%,
the tensile strength of the fiber steadily increased, while the elongation at
breakage decreased (Liu et al., 2019). Nanocomposite films primed with
melt compounding and film blowing have also been tested for packaging
applications. Mechanical properties of nanocomposite and reference films
are found to be higher in the machine direction (MD) compared to the
transverse direction (TD) (Herrera et al., 2016). In addition, high lignin-
containing cellulose nanocrystals (HLCNCs) and lignin-coated CNCs
(BLCNCs)/PLA composites have been made, showing substantial improve-
ments in both Young’s modulus and brittle elongation but minor decreases
in tensile strength (Wei et al., 2018). Two separate PLLA-based surfactants
have also been prepared. It consists of either the PEG block PEG-b-PLLA
or the Im-PLLA, which can communicate with the surface of the CNCs in
various ways. The findings reveal that the addition of both surfactants to
the PLA matrix induces a decrease in Young’s modulus, intensity, and even
elongation at split. The addition of CNCs has shown no significant improve-
ment in the mechanical properties (Mariano et al., 2017). In addition to the
tensile strength and elasticity modules of the PLA/CNC composite films,
the addition of 0.1 wt.% and 0.3 wt.% CNCs was improved (Wang et al.,
2019). And PLA/CNC bio-nanocomposite films have been developed using
a twin-screw extruder. The tensile strength and Young’s modulus increased
at both 1 and 3% CNCs (Sung et al., 2017).
TABLE 3.4 Natural Fibers in PLA.

Untreated bamboo 20 wt % Mixing and σm = 50.83 MPa (PLA/UBF)
fibers (UBF) and extrusion σm = 56.46 MPa (PLA/ABF)
Alkali-treated
Bamboo Fibers εB= 2.21% (PLA/UBF)
(ABF) (Long et εB= 2.491% (PLA/ABF)
al.,2019)
FM = 4.75 GPa (PLA/UBF)
FS = 4.85 GPa (PLA/ABF)
FS = 83.38 MPa (PLA/UBF)
FS = 82 MPa (PLA/ABF)
α = 2.59 kJ/m2 (PLA/UBF)
α = 3.49 kJ/m2 (PLA/ABF)
σM = 59 MPa (PLA)
σm = 6 MPa (PLA/AF40)
E = 1,908 MPa (PLA)
E = 352 MPa (PLA/AF40)
εb = 3.5% (PLA)
εb = 2.8% (PLA/AF10)
FM = 3.51 GPa (PLA)
FM = 3.59 GPa (PLA/AF40)

Agave Fibers (AF) 10,20,30 and Dry-blending FS = 93 MPa (PLA)
(Cisneros-López et 40 wt % and FS = 4 MPa (PLA/AF40)
al., 2018) compression-
molded (CM) σm = 60 MPa (PLA)
σm = 29 MPa (PLA/AF40)
E = 2.03 GPa (PLA)
E = 1.77 GPa (PLA/AF40)
εb = 3.5% (PLA)
εb = 2.1% (PLA/AF10)
FM = 3.73 GPa (PLA)
FM = 3.93 GPa (PLA/AF10)
FS = 92 MPa (PLA)
FS = 36 MPa (PLA/AF40)
σm = 58.0 MPa (Ram/PLA)
σm = 72.7 MPa (Ram-Alk/PLA)
σm = 57.2 MPa (Ram-Sil/PLA)
σm = 63.7 MPa (Ram-Alk-Sil/PLA)
FS = 46.6 MPa (PLA)
FS = 35.9 MPa (Ram/PLA)
Alklai treated, silane 30 wt % Mixing FS = 64.1 MPa (Ram-Alk/PLA)
treated, alkali-silane FS = 49.2 MPa (Ram-Sil/PLA)
treated, and no treated
ramie fiber (Debeli et FS = 57.8 MPa (Ram-Alk-Sil/PLA)
al., 2019) α = 4.9 kJ/m2 (PLA)
α = 10.4 kJ/m2 (Ram/PLA)
α = 12.1 kJ/m2 (Ram-Alk/PLA)
α = 9.4 kJ/m2 (Ram-Sil/PLA)
α = 7.9 kJ/m2 (Ram-Alk-Sil/PLA)

Alkali treated, silane 30 wt % Mixing σm = 58.3 MPa (Ram-Alk-DAP/PLA)
treated, alkali-silane σm = 53.4 MPa (Ram-DAP/PLA)
treated, and no treated
(Debeli et al., 2019) σm = 17.0 MPa (Ram-Sil-DAP/PLA)
σm = 38.7 MPa (Ram-Alk-Sil-DAP/PLA)
FS = 46.6 MPa (PLA)
FS = 38.2 MPa (Ram-Alk-DAP/PLA)
FS = 37.3 MPa (Ram-DAP/PLA)
FS = 25.8 MPa (Ram-Sil-DAP/PLA)
FS = 34.7 MPa (Ram-Alk-Sil-DAP/PLA)
α = 8.1 kJ/m2(Ram-Alk-DAP/PLA)
α = 8.7 kJ/m2 (Ram-DAP/PLA)
α = 5.2 kJ/m2 (Ram-Sil-DAP/PLA)
α = 6.5 kJ/m2 (Ram-Alk-Sil-DAP/PLA)
σm = 16.1 MPa (BF/PLA)
σm = 19.43 MPa (PLA-g-BF/PLA)
Bamboo Fiber (BF) - Mixing/ εb = 4.27 % (BF/PLA)
to obtain PLA-g-BF/ extrusion εb = 5.59 % (PLA-g-BF/PLA)
PLA and BF/PLA (Li
et al., 2019) E = 4.5 GPa (BF/PLA)
E = 5.1 GPa (PLA-g-BF/PLA)
E = 644.4 MPa (PLAr)
Treated cellulose 2, 5 and 10 Dry blending E = 887.8 MPa (PLAr+2% fibers)
fibers, using recycled wt % E = 649.1 MPa (PLAr+5% fibers)
PLA (PLAr) as matrix
(Laadila et al., 2017) E = 714.1 MPa (PLAr+10% fibers)
TABLE 3.5 Nanoparticles in PLA.

Carbon nanotubes, 1, 3, 5, 7 and 9 Stirring solution/ σm = 28.01 MPa (PLA/CNTs)
(CNTs), Chitosan (CS) wt. % of CS electrospinning σ = 38.45 MPa (PLA/CNTs/CS-5)
m
(Liu et al., 2019)
εb = 83.41% (PLA/CNTs)
εb = 73.57% (PLA/CNTs/CS-1)

Chitin nanocomposite Reference Extrusion/blown σm = 33 MPa (PLA)
(ChNCs) (Herrera et compound: PLA/ film machine σm = 35 MPa (PLA/ChNCs)
al., 2016) PBAT/GTA/ direction (MD)
E = 0.8 GPa (PLA)
Talc (60/24/10/6
wt %) and E = 0.7 GPa (PLA/ChNCs)
nanocomposite εb = 282% (PLA)
compound: PLA/ εb = 281% (PLA/ChNCs)
PBAT/GTA/
Extrusion/blown σm = 17 MPa (PLA)
Talc/ChNCs
film, transverse σm = 28 MPa (PLA/ChNCs)
(59/25/9/6/1
direction, (TD)
wt %) E = 0.4 GPa (PLA)
E = 0.7 GPa (PLA/ChNCs)
εb = 174% (PLA)
High lignin containing 0.5, 1, 2 and 5 Mixing/extrusion εb = 220% (PLA/ChNCs)
cellulose nanocrystals wt % σm = 64.4 MPa (PLA)
(HLCNCs) and
σm = 61.2 MPa (PLA/0.5%HLCNC)
commercial lignin
coated CNCs σm = 60.3 MPa (PLA/0.5%BLCNC)
(BLCNCs) (Wei et al., εb = 7.1% (PLA)
2018) εb = 11.4% (PLA/0.5%HLCNC)
εb = 8.2% (PLA/0.5%BLCNC)
E = 3.7 GPa (PLA)
E = 4.3 GPa (PLA/0.5%HLCNC)
E = 4.1 GPa (PLA/0.5%BLCNC)
σm = 40 MPa (PLA)
σm = 33.2 MPa (PLAPEG-b-PLLA)
σm = 31.7 MPa (PLAIm-PLLA)
Polyethylene glycol 20 wt % of CNC Extrusion σm = 33 MPa (PLAPEG-b- PLLA+20CNC)
(PEG) block (PEGb- σm = 14.8 MPa (PLAIm-PLLA+20CNC)
PLLA), Imidazolium
εb = 3.5% (PLA)
group (Im-PLLA) and
CNC (Mariano et al., εb = 2.6% (PLAPEG-b-PLLA)
2017) εb = 2.4% (PLAIm-PLLA)
εb = 1.9% (PLAPEG-b-PLLA+20CNC)
εb = 1.6% (PLAIm-PLLA+20CNC)
E = 2 GPa (PLA)
E = 1.7 GPa (PLAPEG-b-PLLA)
E = 1.9 GPa (PLAIm-PLLA)
E = 2.2 GPa (PLAPEG-b-PLLA+20CNC)

CNCs from waste 0.1, 0.3, 0.5 and Mixing E = 2.2 GPa (PLAIm-PLLA+20CNC)
cotton cloth cellulose 0.7 wt % σm = 30 MPa (PLA)
(Wang et al., 2019)
σm = 294 MPa (PLA-0.1% CNCs)
εb = 20.1% (PLA)
εb = 5.2% (PLA-0.1% CNCs)
E = 2.8 GPa (PLA)
E = 12.8 GPa (PLA-0.1% CNCs)
CNC (extracted form 1, 3 and 5 wt % Extrusion σm = 54.6 MPa (PLA)
coffee silverskin) σm = 56.2 MPa (PLA/CNC 1%)
(Sung et al., 2017)
εb = 6.3% (PLA)
εb = 6.4% (PLA/CNC 1%)
E = 1166 MPa (PLA)
E = 1343 MPa (PLA/CNC 5%)
TABLE 3.6 Materials en PLA.

Carbon fiber and - Extrusion σm = 28 MPa (PLA)
modified carbon fiber σm = 80 MPa (carbon fiber/PLA)
(Li et al., 2016)
σm = 91 MPa (modified carbon fiber/PLA)
FS= 53 MPa (PLA)
FS= 59 MPa (carbon fiber/ PLA)
FS= 156 MPa (modified carbon fiber/PLA)
Microfibrillated 1 wt % Extrusion E = 3.5 GPa (PLA)
wood (MFW) E = 3.6 GPa (PLA-MFW)
Microfibrillated
lignocellulose (MFLC) E = 3.3 GPa (PLA-MFLC)
(Winter et al., 2018) σm = 107 MPa (PLA-MFW)
σm = 91 MPa (PLA-MFLC)
α = 17.3 kJ/m2 (PLA)
α = 18.0 kJ/m2 (PLA-MFW)
α = 19.1 kJ/m2 (PLA-MFLC)
Diatomaceous Earth 5, 10 and 15 Extrusion σm = 41.69 MPa (PLA)
(DE) (Aggarwal et al., wt % σm = 38.65 MPa (PLA/DE-5%)
2019)
E = 941.61 MPa (PLA)
E = 990.40 MPa (PLA/DE-5%)
3.6 3D PRINTING AS A METHOD OF INCORPORATING

REINFORCEMENT AGENTS IN PLA MATRICES
The 3D printing method is extensively studied in the manufacturing of PLA

thermoplastic resins due to the good operating characteristics, low cost, and
no need for tooling or molding. But the quick procurement of fiber-reinforced
PLA composites with outstanding mechanical properties by 3D printing is
still a major challenge (Li et al., 2016).
Some materials were integrated into the PLA (matrix) and used in 3D
printing, e.g. the carbon fiber reinforced PLA composite was developed
through the rapid prototyping method to 3D printing (Li et al., 2016). The
findings showed that the tensile strength and flexicurity of the improved
carbon fiber reinforced composites are 13.8 and 164% higher than the
initial carbon fiber reinforced samples (Li et al., 2016). In addition, MFW
and MFLC filaments with PLA have been prepared. The bars printed with
PLA-MFW behaved similar to pure PLA (no significant difference), although
PLA-MFW demonstrated marginally increased impact strength (Winter et
al., 2018). Diatomaceous earth has also been used as a possible part of PLA.
Adding DE to the PLA created an improvement in the elasticity module
(Aggarwal et al., 2019).
3.7 CONCLUSIONS
Polylactic acid is at the forefront of biopolymers and introduces itself as an

alternative to traditional polymers. Despite its high mechanical properties,
many materials have been integrated into PLA to improve properties such
as impact resistance. This work provides a description of the use of various
types of reinforcement materials to improve the mechanical properties of
PLA and extrusion as a form of integration. It should be considered that
the integration of such reinforcement materials into the PLA can positively
increase the tensile strength and the Young’s composite modulus. This study
of the role of reinforcement agents in the mechanical properties of PLA
opens up a vision of the vast field of operation for the application of this
biopolymer by optimizing its properties in order to focus on a particular
application.
KEYWORDS
• mechanical properties
• food packaging
• biocompatibility
• biopolymers
• polylactic acid (PLA)
• organic materials
REFERENCES
Aggarwal, S.; Johnson, S.; Saloni, D.; Hakovirta, M. Novel 3D Printing Filament Composite
using Diatomaceous Earth and PolylacticAcid for Materials Properties and Cost Improvement.
Compos. B: Eng., 2019, 177. https://doi.org/10.1016/j.compositesb.2019.107310
Asadi, M.; Bazyar, B.; Hemmasi, A. H.; Ghsemi, I.; Talaeipoor, M. Assessment of Mechanical
and Morphological Properties of New Poly Lactic Acid (PLA)/Wood Fibers/Nanographene
Composite. Drvna Industrija, 2018, 69 (2).
Carbonell-Verdu, A.; Samper, M. D.; Garcia-Garcia, D.; Sanchez-Nacher, L.; Balart, R.
Plasticization Effect of Epoxidized Cottonseed Oil (ECSO) on Poly (Lactic Acid). Indus.
Crops Prod. 2017, 104. https://doi.org/10.1016/j.indcrop.2017.04.050
Cisneros-López, E. O.; Pérez-Fonseca, A. A.; González-García, Y.; Ramírez-Arreola, D.
E.; González-Núñez, R.; Rodrigue, D.; Robledo-Ortíz, J. R. Polylactic Acid–Agave Fiber
Biocomposites Produced by Rotational Molding: A Comparative Study with Compression
Molding. Adv. Polym. Technol. 2018, 37 (7). https://doi.org/10.1002/adv.21928
Debeli, D. K.; Zhang, Z.; Jiao, F.; Guo, J. Diammonium Phosphate-Modified Ramie Fiber
Reinforced Polylactic Acid Composite and Its Performances on Interfacial, Thermal, and
Mechanical Properties. J. Nat. Fibers 2019, 16 (3). https://doi.org/10.1080/15440478.201
7.1423255
Ge, Z.; Qi, Z.; Si, D.; Yu, M. The Effects of Processing Parameters and ac Foaming Agent
on the Mechanical Properties and Morphology of Foamed Wood-polylactic Acid (PLA)
Composites. BioRes. 2018, 13 (1). https://doi.org/10.15376/biores.13.1.1605-1618
Hammiche, D.; Boukerrou, A.; Azzeddine, B.; Guermazi, N.; Budtova, T. Characterization of
Polylactic Acid Green Composites and Its Biodegradation in a Bacterial Environment. Int.
J. Polym. Anal. Charact. 2019, 24 (3). https://doi.org/10.1080/1023666X.2019.1567083
Herrera, N.; Roch, H.; Salaberria, A. M.; Pino-Orellana, M. A.; Labidi, J.; Fernandes, S. C.
M.; Radic, D.; Leiva, A.; Oksman, K. Functionalized Blown Films of Plasticized Polylactic
Acid/Chitin Nanocomposite: Preparation and Characterization. Mater. Des. 2016, 92.
https://doi.org/10.1016/j.matdes.2015.12.083
Holcapkova, P.; Stloukal, P.; Kucharczyk, P.; Omastova, M.; Kovalcik, A. Anti-Hydrolysis
Effect of Aromatic Carbodiimide in Poly(Lactic Acid)/Wood Flour Composites. Compos.
Part A: Appl. Sci. Manuf. 2017, 103. https://doi.org/10.1016/j.compositesa.2017.10.003
Laadila, M. A.; Hegde, K.; Rouissi, T.; Brar, S. K.; Galvez, R.; Sorelli, L.; Cheikh, R. B.,
Paiva, M.; Abokitse, K. Green Synthesis of Novel Biocomposites from Treated Cellulosic
Fibers and Recycled Bio-Plastic Polylactic Acid. J. Clean. Prod. 2017, 164. https://doi.
org/10.1016/j.jclepro.2017.06.235
Li, W.; He, X.; Zuo, Y.; Wang, S.; Wu, Y. Study on the Compatible Interface of Bamboo Fiber/
Polylactic Acid Composites by In-Situ Solid Phase Grafting. Int. J. Biol. Macromol. 2019,
141. https://doi.org/10.1016/j.ijbiomac.2019.09.005
Liu, Y.; Wang, S.; Lan, W.; Qin, W. Fabrication of Polylactic Acid/Carbon Nanotubes/
Chitosan Composite Fibers by Electrospinning for Strawberry Preservation. Int. J. Biol.
Macromol. 2019, 121. https://doi.org/10.1016/j.ijbiomac.2018.09.042
Long, H.; Wu, Z.; Dong, Q.; Shen, Y.; Zhou, W.; Luo, Y.; Zhang, C.; Dong, X. Effect of
Polyethylene Glycol on Mechanical Properties of Bamboo Fiber-Reinforced Polylactic
Acid Composites. J. Appl. Polym. Sci. 2019, 136 (26). https://doi.org/10.1002/app.47709
Mariano, M.; Pilate, F.; de Oliveira, F. B.; Khelifa, F.; Dubois, P.; Raquez, J. M.; Dufresne,
A. Preparation of Cellulose Nanocrystal-Reinforced Poly(Lactic Acid) Nanocomposites
through Noncovalent Modification with PLLA-Based Surfactants. ACS Omega 2017, 2 (6).
https://doi.org/10.1021/acsomega.7b00387
Miao, L.; Walton, W. C.; Wang, L.; Li, L.; Wang, Y. Characterization of Polylactic Acids-
Polyhydroxybutyrate Based Packaging Film With Fennel Oil, and Its Application on
Oysters. Food Pack. Shelf Life 2019, 22. https://doi.org/10.1016/j.fpsl.2019.100388
Nilsuwan, K.; Benjakul, S.; Prodpran, T. Physical/Thermal Properties and Heat Seal Ability
of Bilayer Films Based on Fish Gelatin and Poly(Lactic Acid). Food Hydrocol. 2018, 77.
Orue, A.; Eceiza, A.; Arbelaiz, A. Preparation and Characterization of Poly(Lactic Acid)
Plasticized with Vegetable Oils and Reinforced with Sisal Fibers. Indus. Crops Prod. 2018,
112. https://doi.org/10.1016/j.indcrop.2017.11.011
Reis, M. O.; Olivato, J. B.; Bilck, A. P.; Zanela, J.; Grossmann, M. V. E.; Yamashita, F.
Biodegradable Trays of Thermoplastic Starch/Poly (Lactic Acid) Coated with Beeswax.
Indus. Crops Prod. 2018, 112. https://doi.org/10.1016/j.indcrop.2017.12.045
Santos, D.; Correia, C. O.; Silva, D. M.; Gomes, P. S.; Fernandes, M. H.; Santos, J. D.;
Sencadas, V. Incorporation of Glass-Reinforced Hydroxyapatite Microparticles into
Poly(Lactic Acid) Electrospun Fibre Mats for Biomedical Applications. Mater. Sci. Eng. C
2017, 75. https://doi.org/10.1016/j.msec.2017.03.007
Scaffaro, R.; Lopresti, F.; Botta, L. PLA Based Biocomposites Reinforced with
Posidonia Oceanica Leaves. Comp. B: Eng. 2018, 139. https://doi.org/10.1016/j.
compositesb.2017.11.048
Scaffaro, R.; Lopresti, F.; D’Arrigo, M.; Marino, A.; Nostro, A. Efficacy of Poly(Lactic Acid)/
Carvacrol Electrospun Membranes Against Staphylococcus Aureus and Candida Albicans
in Single and Mixed Cultures. Appl. Microbiol. Biotechnol. 2018, 102 (9). https://doi.
org/10.1007/s00253-018-8879-7
Sung, S. H.; Chang, Y.; Han, J. Development of Polylactic Acid Nanocomposite Films
Reinforced with Cellulose Nanocrystals Derived from Coffee Silverskin. Carbohydr.
Polym. 2017, 169. https://doi.org/10.1016/j.carbpol.2017.04.037
Suryanegara, L.; Kurniawan, Y. D.; Syamani, F. A.; Nurhamiyah, Y. Mechanical Properties
of Composites based on Poly(Lactic Acid) and Soda-Treated Sugarcane Bagasse Pulp. In
Sustainable Future for Human Security: Society, Cities and Governance; 2017. https://doi.
org/10.1007/978-981-10-5433-4_19
Wang, Z.; Yao, Z.; Zhou, J.; He, M.; Jiang, Q.; Li, A.; Li, S.; Liu, M.; Luo, S.; Zhang, D.
Improvement of Polylactic Acid Film Properties Through the Addition of Cellulose
Nanocrystals Isolated from Waste Cotton Cloth. Int. J. Biol. Macromol. 2019, 129. https://
doi.org/10.1016/j.ijbiomac.2019.02.021
Wei, L.; Agarwal, U. P.; Matuana, L.; Sabo, R. C.; Stark, N. M. Performance of High Lignin
Content Cellulose Nanocrystals in Poly(Lactic Acid). Polymer 2018, 135. https://doi.
org/10.1016/j.polymer.2017.12.039
Winter, A.; Mundigler, N.; Holzweber, J.; Veigel, S.; Müller, U.; Kovalcik, A.; Gindl-Altmutter,
W. Residual Wood Polymers Facilitate Compounding of Microfibrillated Cellulose with
Poly(Lactic Acid) for 3D Printer Filaments. Philos. Transac. R. Soc. A: Math. Phys. Eng.
Sci. 2018, 376 (2112). https://doi.org/10.1098/rsta.2017.0046
CHAPTER 4
Thermal Properties by Adding Natural

Oils, Foods, Organic Materials, Fibers,
and Nanocomposites in the PLA, and
Applications in 3D Printing
Y. K. REYES ACOSTA1, W. E. CRUZ MARTINEZ1, A. V. REYES ACOSTA2,
V. A. CEPEDA TOVAR1, J. C. CONTRERAS ESQUIVEL1,
C. N. AGUILAR GONZALES1, R. I. NARRO CESPEDES1, and
R. REYNA MARTINEZ3
1
Facultad de Ciencias Químicas de la Universidad Autónoma de Coahuila,
Blvd. V. Carranza s/n, Saltillo, Coahuila, México
2
Facultad de Sistemas de la Universidad Autónoma de Coahuila, Zona
Centro, Arteaga, Coahuila, México
3
Colegio de Estudios Científicos y Tecnológicos del Estado de Coahuila
Plantel San Antonio de las Alazanas, Arteaga, Coahuila, México
ABSTRACT
Polylactic acid (PLA) is a biodegradable thermoplastic and aliphatic polyester

derived from sustainable and sustainable starch rich materials with important
physicochemical and biological properties. These properties are targeted for
applications in a broad range of fields, especially in the packaging, textile,
agricultural, pharmaceutical, electronics, and biomedical sectors. PLA is one
of the most significant biopolymers due to its biodegradability, biocompat-
ibility, and thermal properties with glass transition temperature degradation
temperature, comparable to polystyrene (PS) and polyethylene terephthalate
(PET). However, due to their lower thermal properties, many materials have
been used to improve. By adding, such as food and its derivatives, natural
oils, organic material, various types of fibers, nanoparticles, and others have
been integrated into the PLA matrix with the objective of improved thermal
properties. This study presents a review of the research on PLA and the use
of enhancers, as well as the findings of the thermal property's improvement.
4.1 INTRODUCTION
Thermal properties are those properties of materials that are related to their
heat conductivity, in other words, the properties that a material exhibits when
it is heated.
The increment in molecular mobility by increasing temperature induces
changes in properties such as density, heat capacity, electrical properties,
thermal conductivity, optical or mechanical properties, and rheological
behavior. In addition, phase transitions can occur associated with dramatic
modifications within the structure of materials, as in the case of composite
fabrication.
Consequently, thermal behavior is essential to understand fundamental
aspects of the structure, kinetics of crystallization, and properties of a
polymer in its final application.
Below are presented some studies in which the thermal properties of
materials have been of great importance for the development of different
technologies. In addition to this, many government organizations in all
countries have focused their efforts in seeking environmental protection for
decades, seeking in this way the friendliest processing of resources with
ecosystems, which will achieve a better use of natural resources—fossil
source or its elimination in the future; with this, countries are forced to obtain
a more conscious environmental legislation that promotes the quality of the
environment in the near future. As a result, the sustainable and recyclable use
of resources plays an important role in solving the growing demand for wood
resources and the scarcity of forest resources, thereby achieving a reduction
in the pressure on the preservation of the environment. Plastic and wood
source composites have become the most optimal and sustainable alterna-
tives to replace green forest source wood, due to their very good capabilities
and lower cost, since the materials emerged on the market (Nilsuwan et al.,
2018). Some applications have used this improvement in obtaining materials
from sustainable raw materials, such as boards, flooring, public and private
infrastructure, components such as interior decorations, furniture material,
automotive trim, and of course, engineering applications (Carbonell-Verdu
Thermal Properties by Adding Natural Oils, Foods, Organic Materials, Fibers 89
et al., 2017; Reis et al., 2018)—due to properties such as high resistance to

putrefaction, decomposition, contact with insects and corrosion, followed
by lower moisture absorption, higher rigidity strength and thermal stability
(Carbonell-Verdu et al., 2017; Orue et al., 2018; Reis et al., 2018). In the
manufacturing process, it consists of mixing wood fibers with polymers
considered thermoplastic (Aggarwal et al., 2019; Santos et al., 2017); this
is how thermoplastic materials exert important influences on the capacity
of the resulting compounds. This is where the use of polylactic acid (PLA)
influences, as a precursor in the manufacture of new tangible goods consid-
ered important in the field of 3D printing engineering (Scaffaro et al., 2018).
In this sense, the different technologies that have been developed seek
a significant improvement in the properties of materials in general, with
thermal properties being those that directly influence the conformational
structure of the final products. In this way, it is important to consider how
their effect influences the finished products, specifically in the food, phar-
maceutical, and, of course, 3D printing areas. 3D printing is becoming very
popular due to its wide spectrum in the creation of new prototypes that seek
to meet the needs at low manufacturing cost and really short lead times (Li
et al., 2016; Sun et al., 2021; Wang et al., 2021). Likewise, the biopolymer
considered PLA and its respective PLA-wood compounds have been used
in the formulation of filaments for 3D printing with properties that have
been considered green technologies. PLA is considered to be one of the
biocomposites obtained from agricultural sources; it has a great potential to
be an alternative as a self-sustaining material coupled or in substitution for
petroleum-based polymers. Likewise, inorganic loads from wood fibers are
considered renewable and biodegradable; due to this, the implementation of
this compound on polymeric matrices has resulted in the production of new
materials that have significantly improved the development of new commer-
cial products (Huang et al., 2021; Jin et al., 2017; le Duigou et al., 2016;
Sun et al., 2021; Z. Wang et al., 2021; Zhou et al., 2015). Therefore, the
objective of this present review was to understand the behavior depending
on the thermal decomposition of biopolymers formed by the natural addition
of oils, organic materials, fibers, and PLA nanocomposites as fillers destined
for the process and future commercial applications in the field of 3D printing.
4.2 FOODS AND NATURAL OILS AS ADDITAMENTS TO PLA

A study carried out by Nilsuwan et al. (2018) showed that when formulating
bilayer films of fish gelatin (FG) and PLA in different concentrations, they
presented a glass transition temperature (Tg) higher than those of PLA

independently, demonstrating that the bilayer FG/PLA films could be used
as a promising food packaging as they have a higher Tg producing a high
resistance to an increment of temperature, and it is biodegradable.
Another study carried out by Altan et al. (2018) demonstrates an important
change in the thermal properties of materials in which they designed fibrous
films composed of zein and PLA by incorporating carvacrol in three different
concentrations or weight percentage (5, 10, and 20% wt) using the electros-
pinning technique. The results show an effect on carvacrol incorporation and
thermal stability of electrospun zein and PLA fibers. The thermogravimetric
analysis (TGA) curve for pure zein fiber shows two stages of weight loss as a
function of temperature. The first stage occurs in the range of 130–220 °C with a
weight loss of around 3.4%, while the second stage is observed between 240 and
400 °C. Both weight losses are attributed to the main degradation of zein, which
was observed to be relatively slow at first. Pure carvacrol completely evaporates
at around 170°C. Zein fibers show a further stage of weight loss between 75
and 150 °C after carvacrol incorporation. Carvacrol weight loss was calculated
as ~3.3, 6.1, and 9.2% for 5, 10, and 20% carvacrol, respectively, indicating
indirect confirmation of the presence of carvacrol in zein fibers. The maximum
temperatures at the highest rate of weight loss are 329, 330, and 326 °C.
These results show that the incorporation of carvacrol in zein does not
significantly affect the thermal degradation profile of zein fibers. The addi-
tion of carvacrol could act as a natural plasticizer and disrupt the interac-
tions between the zein molecules. Therefore, no significant improvement in
thermal stability was observed for the carvacrol-loaded zein fibers.
Another case is the study carried out by Reis et al. (2018). Biodegradable
trays of thermoplastic starch (TPS)/PLA blends were produced by flat extru-
sion, calendering, and thermopressing. The trays were coated by immersion
in beeswax (BW) emulsion (1, 2, and 3 g BW/100 g ethyl alcohol + Tween
80 solution) to improve their barrier properties. The thermal stability of the
coated and non-coated TPS/PLA trays was measured by TGA, and all the
samples had two weight-loss steps. The first step was related to the starch
and the second one corresponded to the thermal degradation of PLA. The
BW content (1, 2, and 3% wt) affected the thermal stability of glycerol,
although the thermal stability of starch and PLA remains unchanged in the
coated samples, showing no interactions between the materials and different
mechanisms of thermal degradation.
Some studies demonstrate the importance of thermal properties in poly-
mers when making composite products or mixtures with natural oils; for
example, Carbonell-Verdu et al. (2017) studied new formulations of envi-

ronmentally friendly plasticized PLA incorporating epoxidized cottonseed oil
(ECSO). The Tg dropped by about 5–6 °C for formulations with 7.5–10.0%
wt ECSO. This decrease in Tg evidenced an increase in free volume with a
positive effect on the mobility of the chain, so that both the Tg and the cold
crystallization temperature (Tc) move to lower values. Therefore, the results
obtained in this work suggest that highly environmentally friendly toughened
PLA formulations can be obtained using ECSO in the range of 5–10% wt.
Another case is the study carried out by Orue et al. (2018) in which are
prepared and characterized PLAplasticized with vegetable oils and reinforced
with sisal fibers; their DSC study show that PLA plasticized with epoxidized
oils have lower Tg values than PLA plasticized with non-epoxidized oils.
Although the addition of plasticizer accelerates the crystallization process
of PLA, the values of the degree of crystallinity increase slightly. After the
addition of sisal fibers in PLA systems plasticized with 10% wt of vegetable
oils, in addition to increase the degree of crystallinity of the samples at
least twice due to the nucleating effect of the sisal fibers, the values of Tg
and onset Tc decrease slightly. Regarding the cold Tc values, it is observed
that after the addition of plasticizers, the Tc decrease considerably, and the
crystallization process is accelerated due to the increase in the segmental
mobility of the PLA chains by plasticization.
There are several cases, such as those presented in Table 4.1, we show
some studies carried out and how the addition of various materials influences
the thermal properties of polymers.
4.3 ORGANIC MATERIALS INCORPORATING IN PLA

In the organic materials field, there exists applications in which the thermal
properties of the polymers vary depending on the complement in which
they work; this undoubtedly represents a great area of opportunity for the
development of new compounds. Below we mention some examples.
Aggarwal et al. (2019) used diatomaceous earth as a potential component
for PLA-based 3D printing composites. DSC analyzes show that the Tc in the
filament composites shows a clear shift toward a Tc lower for PLA-DE fila-
ments compared to 100% PLA; the authors show that diatomaceous earth can
be used as a filler for PLA to create a composite material for 3D printing fila-
ments, showing no major degradation of material properties after 3D printing.
Santos et al. (2017) incorporated glass-reinforced hydroxyapa-
tite macroparticles into PLA electrospun fiber meshes for biomedical
TABLE 4.1 Foods and Natural Oils Addition in PLA: Effect in the Stability Thermal.
Complement Concentration Method Results
Fish gelatin (FG) Layer thickness ratios Casting Tg = 33.25 °C (PLA)
(Nilsuwan et al., 2018) of 9:1, 8:2, 7:3, 6:4, Tg = 43.09 °C (FG/PLA 6:4)
and 5:5 (FG/PLA)
Tc = 88.7 °C (PLA)
Tc = 89.07 °C (FG/PLA 6:4)
Tmax = 156.28 °C (PLA)
Tmax = 145.11–156.28 °C (FG/PLA 6:4)
Carvacrol (CRV) (Altan 5%, 10%, and 20% wt Mixing/ TTGA = 346 °C (PLA)
et al., 2018) electros- TTGA = 377 °C (PLA-5 CRV)
pinning
Thermoplastic starch 1, 2, and 3 g BW/100 Flat TTGA = 360 °C (PLA)
(TPS) (Reis et al., 2018) g ethyl alcohol + extrusion TTGA = 360 °C (PLA-5 CRV)
Tween 80 solution
Epoxidized cottonseed 2.5%, 5%, 7.5%, and Mixing Tg = 66.7 °C (PLA)
oil (ECSO) (Carbonell- 10% wt Tg = 63.76 °C (PLA-2 ECSO)
Verdu et al., 2017)
Tc = 97.16 °C (PLA)
Tc = 91.17 °C (PLA-2 ECSO)
Tm = 169.94 °C (PLA)
Tm = 172.94 °C (PLA-2 ECSO)
TTGA = 335.5 °C (PLA)
TTGA = 340 °C (PLA-10 ECSO)
Epoxidized (ELO, 5%, 10%, and 20% wt Mixing Tg = 66.2 °C (PLA)
ESBO) and non- Tg = 53.46 °C (PLA-5 LO)
epoxidized linseed oil
(LO) and soybean oil Tc = 123.8 °C (PLA)
(SBO) (Orue et al., Tc = 114.4 °C (PLA-5 SBO)
2018) Tm = 154–157.9 °C (PLA)
Tm = 150.8–157.8 °C (PLA-5 SBO)
TTGA = 358.3 °C (PLA)
TTGA = 357.2 °C (PLA-10 SBO)
10% wt of oils with Tg = 56 °C (PLA/sF)
30% wt of fiber Tg = 52.4 °C (PLA/sBO/sF)
Tc = 102.2 °C (PLA/sF)
Tc = 115 °C (PLA/sBO/sF)
Tm = 149.2–158.7 °C (PLA/sF)
Tm = 148.4–154.9 °C (PLA/sBO/sF)
applications. In their study, differential scanning calorimetry was performed

on the electrospun fibrous mats to study the thermal properties of the mate-
rials. Electrospun pure PLLA fibers show a large overshoot in the Tg, around
60 °C, which is related to the enthalpy recovery of the sample stored at room
temperature, around 30 °C below Tg, and therefore subject to physical aging.
As the temperature increases, a cold crystallization process is observed
immediately after the glass transition, which shows up as a broad exothermic
peak between 70 and 110 °C with a maximum (Tc) of around 81 °C. Melting
begins at 130–160 °C. Microcomposite membranes present a glass transition
of the polymer shifted toward low temperatures, with a maximum around
51 °C, and the cold crystallization process that occurs immediately after Tg
shows a peak of ~73 °C. Finally, the authors concluded that the presence of
the ceramic microparticles plays an important role in the thermal degrada-
tion of the polymeric matrix, and a decrease in the activation energy of the
thermal degradation of 134 kJ.mol-1.
There are many studies like these, in which the importance of thermal
properties in materials is highlighted before generating a final product, which
are summarized in Table 4.2.
TABLE 4.2 Organic Materials Addition in PLA: Effect in the Stability Thermal.
Diatomaceous earth 5%, 10%, and Extrusion Tc = 115.3 °C (PLA)
(DE) (Aggarwal et 15% wt Tc = 109.2 °C (PLA/DE-5)
al., 2019)
Tm = 148.5 °C (PLA)
Tm = 148.3–157.9 °C (PLA/DE-5)
Bonelike® and PLLA 80% wt Add to Tg = 60 °C (PLLA)
(Santos et al., 2017) solution and Tg = 51 °C (Bonelike® PLLA)
electrospinning
Tc = 81 °C (PLLA)
Tc = 73 °C (Bonelike® PLLA)
Leaves of Posidonia 10% and 20% Mixing Tg = 56 °C (PLA)
oceanica (PO) wt with PO Tg = 54.8 °C (PLA/PO300-10)
(Scaffaro et al., 2018) particles size
<150 μm and Tc = 105.8 °C (PLA)
150–300 μm Tc = 101.1 °C (PLA/PO150-10)
Tm = 161.7–167.8 °C (PLA)
Tm = 161.6–169.8 °C (PLA/PO300-10)

Wood flour (WF) 20% wt of WF Mixing Tg = 59.1 °C (PLA)
and ZIKA additive Tg = 58.6 °C (PLA/WF)
(Holcapkova et al.,
2017) Tc = 99.6–155.8 °C (PLA)
Tc = 98.6–154.8 °C (PLA/WF)
Tm = 168.4 °C (PLA)
Tm = 167.4 °C (PLA-ZIKA)
TTGA (50%) = 358 °C (PLA)
TTGA (50%) = 359 °C (PLA-ZIKA)
Corn starch and wood 30% wt Extrusion Tg = 59.4 °C (PLA)
flour (WF) with Tg = 54.8 °C (PLA-1200)
artificial weathering
(300 h, 600 h, 900 h, Tg = 58.2 °C (PLA/starch)
and 1200 h) (Lv et Tg = 54.4 °C (PLA/starch-600)
al., 2018)
Tg = 58.1 °C (PLA/WF)
Tg = 56.9 °C (PLA/WF-900)
Tc = 99.6 °C (PLA)
Tc = 93.4 °C (PLA-900)
Tc = 90.1 °C (PLA/starch)
Tc = 89.5 °C (PLA/starch-1200)
Tc = 93.3 °C (PLA/WF)
Tc = 90.1 °C (PLA/WF-600)
Tm = 156.2 °C (PLA)
Tm = 154.7 °C (PLA-1200)
Tm = 151 °C (PLA/starch)
Tm = 154.3 °C (PLA/starch-900)
Tm = 154.4 °C (PLA/WF)
Tm = 153.4 °C (PLA/WF-1200)
4.4 FIBERS INCORPORATING IN PLA

In this field, there are recent studies, such as Long et al. (2019), who studied
the effect of polyethylene glycol on the mechanical properties of PLA
compounds reinforced with bamboo fiber (BF). The results regarding the
DSC analysis show that the crystallinity of PLA in composites with low
BF content is increased. The crystallinity of PLA was 44.19% when the
BF content was 15%. However, PLA crystallinity decreased when the BF
content continued to increase. It can be concluded that a small amount of

BF acted as a nucleating agent and accelerated crystallization rates; on the
other hand, the excess of fiber hindered the crystallization of the matrix and
reduced the thermal stability of the composites.
Another study carried out by Cisneros-López et al. (2018) looked into
the development of biocomposites of agave fiber and PLA produced by
rotational molding; in the thermal analysis, a Tg of 60–61 °C was obtained
and a Tm of 168–169 °C, which could, again, indicate that both rotomolding
and compression molding do not produce significant degradation as optimal
processing conditions were used. The crystallinity of the rotomolded
biocomposites (R-BC) increases 22% (from 18% for pure PLA to 22% with
10% wt of agave fiber). However, higher fiber content decreased crystallinity
by up to 11% (16–40% wt). Compression molded biocomposites (CM-BC)
present higher crystallinity than R-BC for all fiber contents. The level of
crystallinity increased from 20% for PLA to 26–30% wt.
W. Li et al. (2019) conducted a study on the compatible interface of BF/
PLA composites by in situ solid phase grafting. The DSC thermograms of
the BF/PLA and PLA-g-BF/PLA systems show that the relative changes in
the crystal structure of the BF/PLA and PLA-g-BF/PLA compounds were
detected by the DSC test. The PLA-g-BF/PLA compounds show 170.63 °C
values, which were higher than those of the unmodified compound (163.25
°C). The Tc increased considerably, and the crystallization process slowed
down due to the higher mobility of the PLA chains. On the contrary, the BF/
PLA compounds show a double crystallization peak, low and high tempera-
ture endotherm. It is attributed to the melting mechanism, recrystallization,
and melting behaviors of the samples. It was concluded that the nucleation
effect of the PLA matrix is enhanced and that crosslinking of the interface is
enhanced. The thermal degradation of the compounds was divided into three
stages with temperature ranges of 50–90, 240–375, and 375–550 °C. The
first stage was the evaporation of water and other BF extracts. In the second
stage, hemicellulose, cellulose, a part of the lignin in the BF, and most of the
PLA were thermally decomposed. In the third stage (370 °C), the remaining
lignin and PLA were decomposed. In addition, the carbon residue content of
the PLA-g-BF/PLA composites at 550 °C is found to be marginally lower
than that of the unmodified composites. The terminal temperature of thermal
decomposition of the second stage for the composites prepared by treatment
with graft polymers is 278.4 °C, higher than the 252.3 °C of the unmodified
composites. The second thermal decomposition phase explained that the
BF grafting treatment strengthened the interfacial interactions between the
filler BF and the PLA matrix and reduced the mobility of the PLA molecular
chain. However, in contrast to the present results, it has also been inferred in
the above text that the BF modification improved the thermal stability of BF/
PLA. Table 4.3 shows some progress in this area.
TABLE 4.3 Fibers Incorporating in PLA: Effect in the Stability Thermal.

Alkali-treated 20% wt Mixing and Tc = 110.76 °C (PLA)
Bamboo Fibers extrusion Tc = 110.93 °C (ABF/PLA (10/90))
(ABF) (Long et al.,
2019) Tc = 110.98 °C (ABF/PLA/PEG (19/76/5))
Tm = 173.08 °C (PLA)
Tm = 173.19 °C (ABF/PLA (10/90))
Tm = 171.56 °C (ABF/PLA/PEG (19/76/5))
Agave fibers (AF) 10%, 20%, 30%, Dry-blending Tg = 60 °C (PLA)
(Cisneros-López et and 40% wt and roto- Tg = 61 °C (PLA/AF10)
al., 2018) molded (RM)
Tc = 102 °C (PLA)
Tc = 104 °C (PLA/AF30)
Tm = 169 °C (PLA)
Tm = 168 °C (PLA/AF20)
Dry-blending Tg = 60 °C (PLA)
and Tg = 61 °C (PLA/AF20)
compression-
molded (CM) Tc = 104 °C (PLA)
Tc = 103 °C (PLA/AF10)
T m = 169 °C (PLA)
T m = 168 °C (PLA/AF20)
Bamboo fiber (BF) - Graft reaction/ Tc = 167.36 °C (PLA)
to obtain PLA-g-BF/ extrusion Tc = 163.25 °C (BF/PLA)
PLA and BF/PLA (W.
Li et al., 2019) Tc = 170.63 °C (PLA-g-BF/PLA)
Alkali treated, silane 30% wt Mixing Tg = 54.9 °C (PLA)
treated, alkali-silane Tg = 62.4 °C (Ram-Sil/PLA)
treated, and no
treated ramie fiber, Tg = 60.3 °C (Ram-Alk/PLA)
no soaked and soaked Tc = -- °C (PLA)
in DAP solution Tc = 99.9 °C (Ram-Sil/PLA)
(Debeli et al., 2019)
Tc = 93.1 °C (Ram-Alk/PLA)
Tm = 164.76 °C (PLA)
Tm = 163.54 °C (Ram/PLA)
Tm = 163.48 °C (Ram-Alk-Sil-DAP/PLA)
4.5 INCORPORATING THE NANOCOMPOSITES IN PLA
Herrera et al. (2016) evaluated bionanocomposite films prepared with

compounds in melt and blown film for packaging applications. Their thermal
analyzes show that the Tg and melting temperature (Tm) of the nanocom-
posite were slightly higher (33 and 172 °C) compared to the values of the
reference films, which were 29 and 171 °C, respectively. The results indicate
that the addition of 1% wt of Chitin nanocrystals (ChNC) slightly improved
the thermal stability and the interaction between the components of the
nanocomposite, which hinders the mobility of the polymer chain. The degree
of crystallinity (Xc) of the nanocomposite films was slightly higher (56%)
compared to the reference (52%), indicating higher nucleation due to chitin
nanocrystals, which is consistent with our previous results. The higher crys-
tallinity results in a higher enthalpy of fusion (ΔHm) of the nanocomposite.
Thus, they concluded that the addition of ChNC to the compound increased
its viscosity, modified the melt strength, and improved the thermal stability
and crystallinity of the compound. The tear and puncture resistance of the
reference films was increased by the addition of the nanocrystals and thus
the polymer was reinforced.
Wei et al. (2018) evaluated the performance of cellulose nanocrystals with
high lignin content as a filler in PLA; DSC analysis showed that the Tg was
not affected by the incorporation of cellulose nanocrystals with high lignin
content. lignin content (HLCNC), or commercial cellulose nanocrystals
(BLCNC) in PLA. This indicated that the relatively small number and large
sizes of CNCs are not enough to change the polymer chain mobility within
the glass transition region. In the current study, the degree of crystallinity
(Xc) was found to increase by adding HLCNC or BLCNC, because CNCs
act as a nucleating agent for PLA compounds and facilitate PLA crystal-
lization. When concentrations of CNC were increased from 2% to 5%, the
Xc was slightly reduced. It is likely that this was due to a relatively poorer
dispersion of the CNCs in the polymeric matrix and to the higher fraction
of large agglomerates, which slightly inhibited the nucleation effect of the
CNCs. It is interesting the representation of the double melting peaks (repre-
senting low to high temperature peaks) of the PLA/HLCNC compounds. It is
speculated that there could be two types of crystal structures present in PLA/
HLCNC nanocomposites. The lower temperature endotherm could be attrib-
uted to the melting of the original PLA crystals, while the higher temperature
could be due to the melting of secondary crystallization resulting from the
rearrangement of the crystallizable amorphous polymer segments around the
nanolignin particles. This further confirmed the higher cold crystallization

enthalpies, ∆Hcc (J/g), of PLA/HLCNCs compared to pure PLA and the
corresponding PLA/BLCNC system.
Incorporation of BLCNC or HLCNC resulted in significant improve-
ments in both Young’s modulus and elongation at break, but small decreases
in tensile strength. When BLCNC (or HLCNC) increased from 2% to 5%, no
additional increase in Young's modulus was observed. Similarly, the degree
of PLA crystallinity (χc%) increased for both BLCNC and HLCNC. Dynamic
mechanical analysis showed that the storage modulus of the nanocomposites
increased by adding CNC, especially HLCNC. A broader tanδ peak indicated
a better interaction between the HLCNCs and the polymeric matrix. It can
be concluded that HLCNC derived from hydrothermally treated wood can
be used as excellent fillers and/or reinforcing agents to improve the physical
and thermal properties of polymeric nanocomposites. However, approaches
to improve the dispersion of cellulose nanomaterials in PLA are still needed.
Table 4.4 shows a summary of the area.
TABLE 4.4 Addition the Nanocomposites in PLA and the Stability Thermal.
Chitin nanocrystals Reference compound: Extrusion Tg = 29 °C (reference)
(ChNCs) (Herrera PLA/PBAT/GTA/Talc Tg = 33 °C (nanocomposite)
et al., 2016) (60%/24%/10%/6%
wt) and nanocomposite Tm = 171 °C (reference)
compound: PLA/PBAT/ Tm = 172 °C (nanocomposite)
GTA/Talc/ChNCs TTGA = 327 °C (reference)
(59%/25%/9%/6%/1%
wt) TTGA = 326 °C (nanocomposite)
High lignin 0.5%, 1%, 2%, and Extrusion Tg = 60.8 °C (PLA)
containing 5% wt Tg = 60.9 °C (PLA/2 HLCNC)
cellulose
nanocrystals Tg = 60.8 °C (PLA/0.5 BLCNC)
(HLCNCs) and Tc = 117.4 °C (PLA)
commercial lignin Tc = 115.8 °C (PLA/0.5 HLCNC)
coated CNCs
(BLCNCs) (Wei et Tc = 115.8 °C (PLA/0.5 BLCNC)
al., 2018) Tm = 149 °C (PLA)
Tm = 148–154 °C (PLA/2 HLCNC)
Tm = 148.4 °C (PLA/0.5 BLCNC)

Polyethylene 20% wt of CNC Extrusion Tg = 61.5 °C (PLA)
glycol (PEG) block Tg = 58.6 °C (PLA/PEG-b-PLLA)
(PEG-b-PLLA),
Imidazolium group Tg =56.4 °C (PLA/Im-PLLA)
(Im-PLLA), and Tg = 59.9 °C (PLA/PEG-b-PLLA+10CNC)
CNC (Mariano et
Tg = 56.3 °C (PLA/Im-PLLA+10CNC)
al., 2017)
Tm = 111.9 °C (PLA)
Tm = 97.4 °C (PLA/PEG - b -PLLA)
Tm = 93.1 °C (PLA/Im-PLLA)
Tm = 99.2 °C (PLA/PEG - b
-PLLA+10CNC)
Tm = 94.9 °C (PLA/Im-PLLA+10CNC)
CNCs from waste 0.1%, 0.3%, 0.5%, and Mixing Tg = 60.1 °C (PLA)
cotton cloth 0.7% wt Tg = 63.6 °C (PLA-0.5 CNCs)
cellulose (Z. Wang
et al., 2019) Tc = 114 °C (PLA)
Tc = 134 °C (PLA-0.7 CNCs)
Tm = 167.8 °C (PLA)
Tm = 166.5 °C (PLA-0.7 CNCs)
CNC (extracted 1%, 3%, and 5% wt Extrusion Tg = 61.4 °C (PLA)
form coffee Tg = 61.1 °C (PLA/CNC 1)
silverskin) (Sung et
al., 2017) Tc = 127.8 °C (PLA)
Tc = 126.2 °C (PLA/CNC 1)
Tm = 148.7 °C (PLA)
Tm = 149.6 °C (PLA/CNC 1)
TiO2 nanoparticles 1%, 5%, 10%, and Mixing TTGA (5%) = 331.9 °C (PLA)
(González et al., 20% wt TTGA (5%) = 341.9 °C (PLA/TiO2 -21-10)
2018)
4.6 3D PRINTING AS A METHOD OF INCORPORATING

REINFORCEMENT AGENTS IN PLA MATRICES
Today, in the scientific and technological field, many wood-based materials

have been used, which are commonly valid for their various applications,
to mention construction. However, some related to the properties of wood
such as dimensional instability, biodegradability by microbial agents
(fungi), and/or insects, restrict the use of materials in certain applications.
These undesirable properties could be improved by chemical treatments, as

reported in the literature (Temiz et al., 2010, 2016). The general applications
of composites structured by wood source fillers are decks, docks, design of
garden items, and fences increase the wettability of composites, as cited in
the literature (Morrell et al., 2006; Sommerhuber et al., 2016).
Bio-based polymers are susceptible to biodegradation under long periods
of exposure in outdoor conditions, citing wood as a sustainable organic
source (Khavkine et al., 2000). Kamdem et al. (2004) studied the feasibility
of wood-structured composites (WPC) filled with CCA-treated wood parti-
cles mixed with raw high-density polyethylene (HDPE) and another from a
recycled source, of which they reported an increase in physical properties,
specifically resistance, biological durability, and photo protection. It is in
this sense that scientific studies have been carried out that focus on the use
of waste from wood sources as organic loads within polymer matrices from
a sustainable source, such is the case of PLA.
As mentioned above, the use and exploitation of biodegradable polymers
and biomaterials in the synthesis of plastics as an option for the protection
and preservation of the environment, as well as the reduction of the use of
petrochemical polymers, has attracted special attention in the scientific and
industrial field. To exemplify, poly(lactic acid) or commonly known as PLA has
become an important biodegradable and bio-based thermoplastic that is fully
biodegradable, biocompatible, and produced from renewable source resources,
mainly corn kernel starch or cob husks and of course sugar cane through highly
sustainable bioconversion processes and polymerization routes (Carmona et al.,
2015; Chaiwutthinan et al., 2015; Chuayjuljit et al., 2017; Ibrahim et al., 2017;
Moura et al., 2014; van Cong et al., 2012; Zhang et al., 2019).
Currently, this biopolymer has been marketed at an increasing level in
many countries to solve environmental problems caused by waste from
synthetic sources (plastics). PLA disintegrates into practically harmless
components for the environment, such as carbon dioxide (CO2), water
(H2O), and other non-toxic small molecules that are safely released in land-
fills (Carmona et al., 2015; Petersson et al., 2007). In addition to this, it
can be used in numerous applications, which provides advantages such as
high tensile strength, elastic modulus and transparency, good processing and
reaction performance when it is synthesized, recyclability, and of course its
good availability in the market sustainable environment (Jiang et al., 2011;
Mihai and Ton-That, 2019; Mohapatra et al., 2014; Syazeven Effatin Azma
Mohd Asri et al., 2021). In addition, PLA is manufactured by many stan-
dard plastic processes, including injection, compression and blow molding,
extrusion, thermoforming, and film formation (Makhetha, 2016). Next, the

most common processes for the synthesis of this biopolymer are described,
as well as an explanation of this compound charged by species that cause
modifications in some of its physical, chemical, mechanical, and thermal
properties.
In this concept, certain researchers modified PLA by incorporating it in
a copolymerization process other polymers of synthetic and semi-synthetic
origin, as a second polymeric phase, for example: poly-caprolactone
(Chaiwutthinan et al., 2019), poly(butylenesuccinate) (Chaiwutthinan et
al., 2015), thermoplastic polyurethane (Ibrahim et al., 2017), ethylene vinyl
acetate (EVA) copolymer (Makhetha, 2016), poly(adipate-co-butylene tere-
phthalate) (Chaiwutthinan et al., 2019), epoxidized natural rubber (Syazeven
Effatin Azma Mohd Asri et al., 2021), and linear low density polyethylene,
LDPE (Jiang et al., 2011), which manage to improve both the physical
and mechanical properties and therefore its high processability. Among
these polymers, EVA is a good candidate to blend with PLA due to its high
flexibility and excellent impact resistance (Salehi et al., 2017). The wide
range of applications that its use involves has interested in the scientific and
technological field, one of them is 3D printing, mainly due to its easy avail-
ability and great use as an economical, viable, and sustainable product. It is
commonly known as a mainly additive or sequential manufacturing tech-
nique or for the formation of products or prototypes (X. Wang et al., 2017).
The three-dimensional entities that can be rapidly elaborated under the
control of the computer and software capable of producing a formed or
preformed product (Xie et al., 2017). Fused deposition modeling (FDM)
is the fastest, most widely used and easily accessible diffusion prototyping
technology for the manufacture of next-generation polymer products (Tian
et al., 2016), widely used for the manufacture of prototypes with certain
complex structures and shapes using thermoplastic polymers, among these
polymers, research has been carried out on acrylonitrile butadiene styrene
(ABS) and, of course, PLA (Jin et al., 2017; le Duigou et al., 2016). In this
sense, adding certain components that provide an improvement in properties
benefits the polymer that is intended to be printed in 3D, thus giving it the best
conditions before and after obtaining it as a finished product. With increasing
environmental change and low availability of natural source resources,
biodegradable materials have become an important topic in current research.
PLA is considered a highly biodegradable and environmentally friendly
biopolymer, which is why its use and disposal as a material for 3D printing
has been considered (Bhattacharjee and Bajwa, 2017; Zhou et al., 2015). The
PLA monomer is lactic acid (LA), produced by fermentation of corn and

other crops (Dong et al., 2014; Lai and Lan, 2013).
Increasing the use of PLA reduces the use of resources or fossil source
species in an impressive proportion, according to the latest reports cited in
the literature (Guo et al., 2018). The combination of PLA allows to improve
the relatively low performance in question at its cost (Lee and Wang, 2006;
X. Liu et al., 2017). Likewise, the by-products obtained from conventional
wood are considered as they are quite cheap and easily available, useful for
the formulation of compounds of sustainable origin, added as a by-product
of the conventional tree processing industry (Pilla et al., 2009). These resi-
dues have advantages such as: low density, high rigidity, and, of course,
degradability, as well as low cost (Xia et al., 2015). As reported, they have
been designed to improve the toughness of PLA composites, including
polymer blending and grafting (Rasal et al., 2010). Polyurethane-derived
polymers (TPU) are the most widely used thermoplastic elastomers, ideal as
a recyclable material and respectful of the ecosystem, allowing the forma-
tion of materials with a certain degree of crosslinking, thus providing good
mechanical properties, biocompatibility, and biodegradability, respectively
(Dogan et al., 2017; Ferri et al., 2016).
Just as the mixing of PLA with other polymers is very efficient, fillers as
reinforcement provide an improvement in these systems of organic origin.
However, the hydrophilicity that wood powders present, the compatibility
when they are mixed mechanically, is poor, the polymeric matrices of hydro-
phobic character, which results in a somewhat deficient interfacial adhesion
and therefore mechanical properties with little efficiency (D. Li et al., 2011;
N. Wang et al., 2007). One of the most outstanding experimental develop-
ments in research to improve the compatibility between the polymer matrix
and the reinforcements from wood residue is the use of compatibilizers, espe-
cially PLA grafted with glycidyl methacrylate (GMA), in the composites is
relatively efficient and environmentally friendly, compared to conventional
chemical treatments where only and exclusively these reinforcements are
added (Hill et al., 1998; Mishra et al., 2003; Ray et al., 2001).
Among the compatibilizers, the compounds that are used found in the
literature handle GMA, since it provides a relatively better compatibility
between PLA and reinforcements such as wood dust or sawdust (Juntuek
et al., 2012; J. Liu et al., 2012; Y. N. Wang et al., 2014). Guo et al. (2018)
concluded that this addition of both functional groups in GMA is due to
the acrylic group and the epoxy group, present in the polymers and the
reinforcing fillers. The acrylic group can couple with the polymer chain and
form a structure with good chemical stability. Meanwhile, the epoxy group
can react with the hydroxyl group that is in fact present in the wood residue,
since in some cases it presents a certain humidity typical of conventional
wood.
Complementing the work carried out by Z. Wang et al. (2021) they designed
four varieties of wood flour compounds (powder considered sawdust or WF)
in the presence of the PLA biopolymer, modified together with organomont-
morillonite (OMMT) as reinforcement and moisture precursor, through
different modification process. These distributions were characterized by
testing the mechanical capabilities of these resulting compounds. When
analyzing the different processes modifying the OMMT, the mechanical
capacities of the compounds were clearly different, as reported by the authors
in the manuscript. However, an exception was presented in the toughness,
almost in its entirety the mechanical capacities showed improvements after
the modification by OMMT; the compound made of pure PLA (homopol-
ymer) together with the wood filler modified with OMMT behaved with the
best mechanical properties, while the compound made of pure WF and PLA
modified with OMMT showed a low performance in terms of mechanical
properties; Whether for OMMT-modified PLA or OMMT-modified WF-co-
PLA compounds, the existence of OMMT aggregates could cause damage
to mechanical properties. However, the addition of OMMT intercalated in
the content of the WF or the WF-co-PLA compound was beneficial for the
resulting physical-mechanical properties in all the tests.
Sun et al. (2021) carried out a dynamic thermogravimetric study (DMA)
to understand the thermal decomposition process of 3D printing filaments,
made from a mixture of PLA and starch, as a polymeric matrix adding wood
filler as reinforcement. With increasing starch content in the filament, the
onset of thermal decomposition for the filaments presented by the authors
of the manuscript gradually decreased from 272.4 to 155.1 °C, respectively.
The degree of thermal degradation became smaller, and the transition
temperature range became larger as the proportion of polysaccharide (starch)
content increased. For a better understanding of the thermal decomposition
behavior of starch-PLA wood composites, thermal stability calculations were
performed, employing more detailed studies that allowed the development
of starch/PLA-based filaments to achieve material suitable for 3D printing.
The boom that 3D printing shows today, reaches high levels leading to its
development and exponential growth, seeking a gradual improvement in
the design of new materials that replace conventional polymers, making use
of self-sustaining polymeric materials or obtained from waste of vegetable
sources, etc. Additive manufacturing is considered one of the new technolo-

gies that give a key point in the field of research for future high precision
manufacturing, benefiting in the first instance the industry, in the foundation
of buildings, development of finished products, innovation, and invention in
medicine and biomedicine, etc. Its growing applications mainly depend on
its significant merits of reduced weight, minimal materials used, high preci-
sion, and shorter production time (Sun et al., 2021; Z. Wang et al., 2021).
Furthermore, it is very crucial that such 3D printed components can
maintain the same or even better material performance and product quality
as those obtained by conventional manufacturing methods. In the present
work (Dong et al., 2018), successfully fabricated 3D-printed mechanical
components in structured PLA trials of wood-fiber-based composites. The
parameters that stood out in the work were the density of the applied fill, the
height of the layer and the number of treatments investigated, this study was
carried out through a design of experiments, among which the number of
tests was determined as the most significant factor. to maximize the physical
properties, especially the tensile strength of PLA samples in conjunction with
wood fibers. In addition, the experimental design evaluated the effect of the
type of material (i.e., the content of pure PLA and the content of PLA with
wood fibers), resistance to bending and impact of the samples. Based on the
results supported in this work, it was suggested that the type of material is
the only predominant factor to maximize all the mechanical forces that were
present in all the tests; however, they are consistently lower for the wood
fiber/PLA compared to pure PLA—this is due to the mechanical resistance
provided by the PLA homopolymer.
4.7 CONCLUSIONS
PLA is at the forefront of biopolymers and is presented as an alternative
to traditional polymers. Despite its high thermal properties, many materials
have been integrated into PLA to improve thermal properties such as TGA
(thermal stability), Tg, Tm (melting point) and TC. This paper describes the
use of various types of reinforcing materials to improve the thermal proper-
ties of PLA and the application in 3D printer. It should be noted that the
integration of these reinforcing materials into PLA can positively increase
the thermal stability, Tg, Melting Point, Tc. This study about the role of
reinforcing agents in the thermal properties of PLA opens an insight into the
vast field of operation for the application of this biopolymer by optimizing
its properties to focus on a particular application with 3D printing.
KEYWORDS
• food, organic materials

• thermal properties
• nanoparticles
• nanofibers
• natural oils
REFERENCES
Aggarwal, S.; Johnson, S.; Saloni, D.; Hakovirta, M. Novel 3D Printing Filament
Composite Using Diatomaceous Earth and Polylactic Acid for Materials Properties
and Cost Improvement. Compos. B: Eng. 2019, 177. https://doi.org/10.1016/j.
compositesb.2019.107310
Altan, A.; Aytac, Z.; Uyar, T. Carvacrol Loaded Electrospun Fibrous Films from Zein and
Poly(Lactic Acid) for Active Food Packaging. Food Hydrocol. 2018, 81. https://doi.
org/10.1016/j.foodhyd.2018.02.028
Bhattacharjee, S.; Bajwa, D. S. Feasibility of Reprocessing Natural Fiber Filled Poly(Lactic
Acid) Composites: An In-depth Investigation. Adv. Mater. Sci. Eng. 2017, 2017. https://doi.
org/10.1155/2017/1430892
Carbonell-Verdu, A.; Samper, M. D.; Garcia-Garcia, D.; Sanchez-Nacher, L.; Balart, R.
Plasticization Effect of Epoxidized Cottonseed Oil (ECSO) on Poly(Lactic Acid). Indus.
CropsProd. 2017, 104. https://doi.org/10.1016/j.indcrop.2017.04.050
Carmona, V. B.; Corrêa, A. C.; Marconcini, J. M.; Mattoso, L. H. C. Properties of A
Biodegradable Ternary Blend of Thermoplastic Starch (TPS), Poly(ε-Caprolactone)
(PCL) and Poly(Lactic Acid) (PLA). J. Polym. Env. 2015, 23(1). https://doi.org/10.1007/
s10924-014-0666-7
Chaiwutthinan, P.; Chauyjuljit, S.; Thipkham, N.; Kowalski, C. P.; Boonmahitthisud, A.
Poly(Lactic Acid)/Ethylene Vinyl Acetate Copolymer Blend Composites With Wood Flour
and Wollastonite: Physical Properties, Morphology, and Biodegradability. J. Vinyl Additive
Technol. 2019, 25 (4). https://doi.org/10.1002/vnl.21697
Chaiwutthinan, P.; Pimpan, V.; Chuayjuljit, S.; Leejarkpai, T. Biodegradable Plastics Prepared
from Poly(Lactic Acid), Poly(Butylene Succinate) and Microcrystalline Cellulose Extracted
from Waste-Cotton Fabric With A Chain Extender. J. Polym. Environ. 2015, 23 (1). https://
doi.org/10.1007/s10924-014-0689-0
Chuayjuljit, S.; Wongwaiwattanakul, C.; Chaiwutthinan, P.; Prasassarakich, P. Biodegradable
Poly(Lactic Acid)/Poly(Butylene Succinate)/Wood Flour Composites: Physical and
Morphological Properties. Polym. Compos. 2017, 38 (12). https://doi.org/10.1002/pc.23886
Cisneros-López, E. O.; Pérez-Fonseca, A. A.; González-García, Y.; Ramírez-Arreola, D.
E.; González-Núñez, R.; Rodrigue, D.; Robledo-Ortíz, J. R. Polylactic Acid–Agave Fiber
Biocomposites Produced by Rotational Molding: A Comparative Study with Compression
Molding. Adv. Polym. Technol. 2018, 37 (7). https://doi.org/10.1002/adv.21928
Debeli, D. K.; Zhang, Z.; Jiao, F.; Guo, J. Diammonium Phosphate-Modified Ramie Fiber
Reinforced Polylactic Acid Composite and Its Performances on Interfacial, Thermal, and
Mechanical Properties. J. Nat. Fibers 2019, 16 (3). https://doi.org/10.1080/15440478.201
7.1423255
Dogan, S. K.; Boyacioglu, S.; Kodal, M.; Gokce, O.; Ozkoc, G. Thermally Induced Shape
Memory Behavior, Enzymatic Degradation and Biocompatibility of PLA/TPU Blends:
“Effects of Compatibilization.” J. Mech. Behav. Biomed. Mater. 2017, 71. https://doi.
org/10.1016/j.jmbbm.2017.04.001
Dong, Y.; Ghataura, A.; Takagi, H.; Haroosh, H. J.; Nakagaito, A. N.; Lau, K. T. Polylactic
Acid (PLA) Biocomposites Reinforced with Coir Fibres: Evaluation of Mechanical
Performance and Multifunctional Properties. Compos. Part A: Appl. Sci. Manufac. 2014,
63. https://doi.org/10.1016/j.compositesa.2014.04.003
Dong, Y.; Milentis, J.; Pramanik, A. Additive Manufacturing of Mechanical Testing Samples
Based on Virgin Poly (Lactic Acid) (PLA) and PLA/Wood Fibre Composites. Adv. Manufac.
2018, 6 (1). https://doi.org/10.1007/s40436-018-0211-3
Ferri, J. M.; Fenollar, O.; Jorda-Vilaplana, A.; García-Sanoguera, D.; Balart, R. Effect of
Miscibility on Mechanical and Thermal Properties of Poly(Lactic Acid)/Polycaprolactone
Blends. Polym. Int. 2016, 65 (4). https://doi.org/10.1002/Pi.5079
González, E. A. S.; Olmos, D.; Lorente, M. Ángel, Vélaz, I.; González-Benito, J. Preparation
and Characterization of Polymer Composite Materials Based on PLA/TiO2 for Antibacterial
Packaging. Polymers 2018, 10 (12). https://doi.org/10.3390/polym10121365
Guo, R.; Ren, Z.; Bi, H.; Song, Y.; Xu, M. Effect of Toughening Agents on the Properties
of Poplar Wood Flour/Poly (Lactic Acid) Composites Fabricated with Fused Deposition
Modeling. Eur. Polym. J. 2018, 107. https://doi.org/10.1016/j.eurpolymj.2018.07.035
Herrera, N.; Roch, H.; Salaberria, A. M.; Pino-Orellana, M. A.; Labidi, J.; Fernandes, S. C.
M.; Radic, D.; Leiva, A.; Oksman, K. Functionalized Blown Films of Plasticized Polylactic
Acid/Chitin Nanocomposite: Preparation and Characterization. Mater. Design 2016, 92.
https://doi.org/10.1016/j.matdes.2015.12.083
Hill, C. A. S.; Khalil, H. P. S. A.; Hale, M. D. A Study of the Potential of Acetylation to
Improve the Properties of Plant Fibres. Indus. Crops Prod. 1998, 8 (1). https://doi.
org/10.1016/s0926-6690(97)10012-7
Holcapkova, P.; Stloukal, P.; Kucharczyk, P.; Omastova, M.; Kovalcik, A. Anti-Hydrolysis
Effect of AromaticCarbodiimide in Poly(Lactic Acid)/Wood Flour Composites. Compos. Part
A: Appl. Sci. and Manufac. 2017, 103. https://doi.org/10.1016/j.compositesa.2017.10.003
Huang, Y.; Löschke, S.; Proust, G. In the Mix: The Effect of Wood Composition on The
3D Printability and Mechanical Performance of Wood-Plastic Composites. Comp. Part C:
Open Access 2021, 5. https://doi.org/10.1016/j.jcomc.2021.100140
Ibrahim, N.; Wahab, M. K. A.; Uylan, D. N.; Ismail, H. Physical and Degradation Properties of
Polylactic Acid and Thermoplastic Starch Blends—Effect of Citric Acid Treatment on Starch
Structures. Bioresources 2017, 12 (2). https://doi.org/10.15376/Biores.12.2.3076-3087
Jiang, G.; Huang, H. X.; Chen, Z. K. Rheological Responses and Morphology of Polylactide/
Linear Low Density Polyethylene Blends Produced by Different Mixing Type. Polym.
Plast. Technol. Eng. 2011, 50 (10). https://doi.org/10.1080/03602559.2011.557822
Jin, Y.; Wan, Y.; Zhang, B.; Liu, Z. Modeling of the Chemical Finishing Process for Polylactic
acid Parts in Fused Deposition Modeling and Investigation of Its Tensile Properties. J.
Mater. Proc. Technol. 2017, 240. https://doi.org/10.1016/j.jmatprotec.2016.10.003
Juntuek, P.; Ruksakulpiwat, C.; Chumsamrong, P.; Ruksakulpiwat, Y. Effect of Glycidyl

Methacrylate-Grafted Natural Rubber on Physical Properties of Polylactic Acid and Natural
Rubber Blends. J. Appl. Polym. Sci. 2012, 125 (1). https://doi.org/10.1002/app.36263
Kamdem, D. P.; Jiang, H.; Cui, W.; Freed, J.; Matuana, L. M. Properties of Wood Plastic
Composites Made of Recycled HDPE and Wood Flour from CCA-Treated Wood Removed
from Service. Compos. Part A: Appl. Sci. Manufac. 2004, 35 (3). https://doi.org/10.1016/j.
compositesa.2003.09.013
Khavkine, M.; Kazayawoko, M.; Law, S.; Balatinecz, J. J. Durability of Wood Flour-
Thermoplastic Composites Under Extreme Environmental Conditions and Fungal
Exposure. Int. J. Polym. Mater. Polym. Biomater. 2000, 46 (1–2), 255–269. https://doi.
org/10.1080/00914030008054859
Lai, S. M.; Lan, Y. C. Shape Memory Properties of Melt-Blended Polylactic Acid (PLA)/
Thermoplastic Polyurethane (TPU) Bio-Based Blends. J. Polym. Res. 2013, 20 (5). https://
doi.org/10.1007/s10965-013-0140-6
Le Duigou, A.; Castro, M.; Bevan, R.; Martin, N. 3D Printing of Wood Fibre Biocomposites:
From Mechanical to Actuation Functionality. Mater. Des. 2016, 96. https://doi.org/10.1016/j.
matdes.2016.02.018
Lee, S. H.; Wang, S. Biodegradable Polymers/Bamboo Fiber Biocomposite with Bio-Based
Coupling Agent. Compos. A: Appl. Sci. Manufac. 2006, 37 (1). https://doi.org/10.1016/j.
compositesa.2005.04.015
Li, D.; Shentu, B.; Weng, Z. Morphology, Rheology, and Mechanical Properties of Polylactide/
Poly(Ethylene-Co-Octene) Blends. J. Macromol. Sci. Part B: Phys. 2011, 50 (10). https://
doi.org/10.1080/00222348.2011.557617
Li, W.; He, X.; Zuo, Y.; Wang, S.; Wu, Y. Study on the Compatible Interface of Bamboo Fiber/
Polylactic Acid Composites by In-Situ Solid Phase Grafting. Int. J. Biol. Macromol. 2019,
141. https://doi.org/10.1016/J.Ijbiomac.2019.09.005
Liu, J.; Jiang, H.; Chen, L. Grafting of Glycidyl Methacrylate onto Poly(Lactide) and
Properties of PLA/starch Blends Compatibilized by the Grafted Copolymer. J. Polym.
Environ. 2012, 20 (3). https://doi.org/10.1007/s10924-012-0438-1
Liu, X.; Lv, S.; Jiang, Y.; Shi, J.; Tan, H.; Gu, J.; Zhang, Y. Effects of Alkali Treatment on the
Properties of WF/PLA Composites. J. Adhes. Sci. Technol. 2017, 31 (10). https://doi.org/1
0.1080/01694243.2016.1248703
Long, H.; Wu, Z.; Dong, Q.; Shen, Y.; Zhou, W.; Luo, Y.; Zhang, C.; Dong, X. Effect of
Polyethylene Glycol on Mechanical Properties of Bamboo Fiber-Reinforced Polylactic
Acid Composites. J. Appl. Polym. Sci. 2019, 136 (26). https://doi.org/10.1002/app.47709
Lv, S.; Gu, J.; Tan, H.; Zhang, Y. Enhanced Durability of Sustainable Poly (Lactic Acid)-
Based Composites with Renewable Starch and Wood Flour. J. Clean. Prod. 2018, 203.
https://doi.org/10.1016/j.jclepro.2018.08.266
Makhetha, T. A. Preparation and Characterization of Ethylene Vinyl Acetate Copolymer/
Poly(Lactic Acid)/sugarcane Bagasse Composites for Water Purification; University of
The Free State, 2016. http://scholar.ufs.ac.za/xmlui/handle/11660/7811
Mariano, M.; Pilate, F.; De Oliveira, F. B.; Khelifa, F.; Dubois, P.; Raquez, J. M.; Dufresne,
A. Preparation of Cellulose Nanocrystal-Reinforced Poly(Lactic Acid) Nanocomposites
Through Noncovalent Modification with PLLA-Based Surfactants. ACS Omega 2017, 2
(6). https://doi.org/10.1021/acsomega.7b00387
Mihai, M.; Ton-That, M. T. Novel Bio-Nanocomposite Hybrids Made from Polylactide/

Nanoclay Nanocomposites and Short Flax Fibers. J. Thermoplast. Compos. Mater. 2019,
32 (1). https://doi.org/10.1177/0892705717743293
Mishra, S.; Mohanty, A. K.; Drzal, L. T.; Misra, M.; Parija, S.; Nayak, S. K.; Tripathy,
S. S. Studies on Mechanical Performance of Biofibre/Glass Reinforced Polyester
Hybrid Composites. Compos. Sci. Technol. 2003, 63 (10). https://doi.org/10.1016/
s0266-3538(03)00084-8
Mohapatra, A. K.; Mohanty, S.; Nayak, S. K. Study of Thermo-Mechanical and Morphological
Behaviour of Biodegradable PLA/PBAT/Layered Silicate Blend Nanocomposites. J.
Polym. Environ. 2014, 22 (3). https://doi.org/10.1007/s10924-014-0639-X
Morrell, J. J.; Stark, N. M.; Pendleton, D. E.; Mcdonald, A. G. Durability of Wood-Plastic
Composites. Wood Design Focus 2006, 16 (3).
Moura, I.; Botelho, G.; Machado, A. V. Characterization of EVA/PLA Blends When Exposed
to Different Environments. J. Polym. Environ. 2014, 22 (1). https://doi.org/10.1007/
s10924-013-0614-y
Nilsuwan, K.; Benjakul, S.; Prodpran, T. Physical/Thermal Properties and Heat Seal Ability
of Bilayer Films Based on Fish Gelatin and Poly(Lactic Acid). Food Hydrocol. 2018, 77.
Petersson, L.; Kvien, I.; Oksman, K. Structure and Thermal Properties of Poly(Lactic Acid)/
Cellulose Whiskers Nanocomposite Materials. Compos. Sci. Technol. 2007, 67 (11–12).
https://doi.org/10.1016/j.compscitech.2006.12.012
Pilla, S.; Gong, S.; O’Neill, E.; Yang, L.; Rowell, R. M. Polylactide-Recycled Wood Fiber
Composites. J. Appl. Polym. Sci. 2009, 111 (1). https://doi.org/10.1002/app.28860
Rasal, R. M.; Janorkar, A. V.; Hirt, D. E. Poly(Lactic Acid) Modifications. In Progress In Polymer
Science; Oxford, Vol. 35 (3), 2010. https://doi.org/10.1016/j.progpolymsci.2009.12.003
Ray, D.; Sarkar, B. K.; Rana, A. K.; Bose, N. R. Effect of Alkali Treated Jute Fibres on
Composite Properties. Bull. Mater. Sci. 2001, 24 (2). https://doi.org/10.1007/BF02710089
Reis, M. O.; Olivato, J. B.; Bilck, A. P.; Zanela, J.; Grossmann, M. V. E.; Yamashita, F.
Biodegradable Trays of Thermoplastic Starch/Poly (Lactic Acid) Coated with Beeswax.
Indus. Crops Prod. 2018, 112. https://doi.org/10.1016/j.indcrop.2017.12.045
Salehi, S.; Ehsani, M.; Khonakdar, H. A. Assessment of Thermal, Morphological, and
Mechanical Properties of Poly(Methyl Methacrylate)/Glass Flake Composites. J. Vinyl
Addit. Technol. 2017, 23 (1). https://doi.org/10.1002/vnl.21476
Santos, D.; Correia, C. O.; Silva, D. M.; Gomes, P. S.; Fernandes, M. H.; Santos, J. D.;
Sencadas, V. Incorporation of Glass-Reinforced Hydroxyapatite Microparticles into
Poly(Lactic Acid) Electrospun Fibre Mats for Biomedical Applications. Mater. Sci. Eng. C
2017, 75. https://doi.org/10.1016/j.msec.2017.03.007
Scaffaro, R.; Lopresti, F.; Botta, L. PLA Based Biocomposites Reinforced with
Posidonia Oceanica Leaves. Compos. B: Eng. 2018, 139. https://doi.org/10.1016/j.
compositesb.2017.11.048
Sommerhuber, P. F.; Wang, T.; Krause, A. Wood-Plastic Composites as Potential Applications
of Recycled Plastics of Electronic Waste and Recycled Particleboard. J. Clean. Prod. 2016,
121. https://doi.org/10.1016/j.jclepro.2016.02.036
Sun, Y.; Lee, D.; Wang, Y.; Li, S.; Ying, J.; Liu, X.; Xu, G.; Gwon, J.; Wu, Q. Thermal
decomposition behavior of 3D printing filaments made of wood-filled polylactic acid/
starch blend. J. Appl. Polym. Sci. 2021, 138 (9). https://doi.org/10.1002/app.49944
Sung, S. H.; Chang, Y.; Han, J. Development of Polylactic Acid Nanocomposite Films
Reinforced with Cellulose Nanocrystals Derived from Coffee Silverskin. Carbohydr.
Polym. 2017, 169. https://doi.org/10.1016/j.carbpol.2017.04.037
Syazeven Effatin Azma Mohd Asri, Zakaria, Z.; Hassan, A.; Kassim, M. H. M.; Arjmandi,
R. Exploring the Effects of Fermented Chitin Nanowhiskers on Tensile and Thermal
Properties of Poly(Ethylene Glycol) Modified Polylactic Acid Nanocomposites. Mal. J.
Fundam. Appl. Sci. 2021, 17 (2). https://doi.org/10.11113/mjfas.v17n2.2002
Temiz, A.; Akbas, S.; Aydin, I.; Demirkir, C. The Effect of Plasma Treatment on Mechanical
Properties, Surface Roughness and Durability of Plywood Treated with Copper-Based Wood
Preservatives. Wood Sci. Technol. 2016, 50 (1). https://doi.org/10.1007/s00226-015-0773-1
Temiz, A.; Alma, M. H.; Terziev, N.; Palanti, S.; Feci, E. Efficiency of Bio-Oil Against Wood
Destroying Organisms. J. Biobased Mater. Bioenergy 2010, 4 (4). https://doi.org/10.1166/
jbmb.2010.1092
Tian, X.; Liu, T.; Yang, C.; Wang, Q.; Li, D. Interface and Performance of 3D Printed
Continuous Carbon Fiber Reinforced PLA Composites. Compos. A: Appl. Sci. Manufac.
2016, 88. https://doi.org/10.1016/j.compositesa.2016.05.032
van Cong, D.; Hoang, T.; Giang, N. V.; Ha, N. T.; Lam, T. D.; Sumita, M. A Novel Enzymatic
Biodegradable Route for PLA/EVA Blends Under Agricultural Soil of Vietnam. Mater. Sci.
Eng. C 2012, 32 (3). https://doi.org/10.1016/j.msec.2011.12.012
Wang, N.; Yu, J.; Ma, X. Preparation and Characterization of Thermoplastic Starch/PLA
Blends by One-Step Reactive Extrusion. Polym. Int. 2007, 56 (11). https://doi.org/10.1002/
pi.2302
Wang, X.; Jiang, M.; Zhou, Z.; Gou, J.; Hui, D. (2017). 3D Printing of Polymer Matrix
Composites: A Review and Prospective. In Composites Part B: Engineering (Vol. 110).
https://doi.org/10.1016/J.Compositesb.2016.11.034
Wang, Y. N.; Weng, Y. X.; Wang, L. Characterization of Interfacial Compatibility of Polylactic
Acid and Bamboo Flour (PLA/BF) in Biocomposites. Polym. Test. 2014, 36. https://doi.
org/10.1016/j.polymertesting.2014.04.001
Wang, Z.; Wang, C.; Qin, Y.; Huang, A.; Liu, R. Organo-Montmorillonite Modified Wood
Flour/Poly (Lactic Acid) Composites via Different Modification Process. Polym. Compos.
2021, 42 (2). https://doi.org/10.1002/pc.25880
Wang, Z.; Yao, Z.; Zhou, J.; He, M.; Jiang, Q.; Li, A.; Li, S.; Liu, M.; Luo, S.; Zhang, D.
Improvement of Polylactic Acid Film Properties Through the Addition of Cellulose
Nanocrystals Isolated from Waste Cotton Cloth. Int. J. Biol. Macromol. 2019, 129. https://
doi.org/10.1016/J.Ijbiomac.2019.02.021
Wei, L.; Agarwal, U. P.; Matuana, L.; Sabo, R. C.; Stark, N. M. Performance of High Lignin
Content Cellulose Nanocrystals in Poly(Lactic Acid). Polymer 2018, 135. https://doi.
org/10.1016/j.polymer.2017.12.039
Xia, X.; Liu, W.; Zhou, L.; Liu, H.; He, S.; Zhu, C. Study on Flax Fiber Toughened Poly
(Lactic Acid) Composites. J. Appl. Polym. Sci. 2015, 132 (38). https://doi.org/10.1002/
app.42573
Xie, G.; Zhang, Y.; Lin, W. Plasticizer Combinations and Performance of Wood Flour-
Poly(Lactic Acid) 3D Printing Filaments. Bioresources 2017, 12 (3). https://doi.
org/10.15376/biores.12.3.6736-6748
Zhang, M.; Jia, H.; Weng, Y.; Li, C. Biodegradable PLA/PBAT Mulch on Microbial
Community Structure In Different Soils. Int. Biodeteriorat. Biodegrad. 2019, 145. https://
doi.org/10.1016/J.Ibiod.2019.104817
Zhou, Y.; Luo, L.; Liu, W.; Zeng, G.; Chen, Y. Preparation and Characteristic of PC/
PLA/TPU Blends by Reactive Extrusion. Adv. Mater. Sci. Eng. 2015, 2015. https://doi.
org/10.1155/2015/393582
CHAPTER 5
Antioxidant and Antimicrobial

Properties of Moringa oleifera in Food
Preservation
ALAÍN MARTÍNEZ-PÉREZ1, ELISEO SANCHEZ-LOREDO2,
DIANA B. MUÑIZ MÁRQUEZ3, and JORGE E. WONG-PAZ3
Tecnológico Nacional de México/Instituto Tecnológico de Durango,
1
Durango, México
2
Bioprocesses and Bioproducts Research Group, Food Research
Department, School of Chemistry, Universidad Autónoma de Coahuila,
3
Universidad Autónoma de San Luis Potosí, Facultad de Estudios
Profesionales de la Zona Huasteca, San Luis Potosí, México
ABSTRACT
The use of preservatives has become indispensable to extend the shelf life
of foods. However, uncertainty about the toxicity and safety of the use of
synthetic preservatives has led the population to demand more natural ingre-
dients for food preservation. There are naturally occurring substances in
plants with high antioxidant and antimicrobial capacities that could be used
as natural preservatives. Thus, extending the shelf life of perishable foods
and in turn providing phytochemical compounds, phenolics, and nutrients.
Moringa oleifera is a plant rich in phenolic compounds with antioxidant
and antimicrobial capacities that have been shown to inhibit the growth
of microorganisms such as Staphylococcus aureus, Escherichia coli, and
Listeria monocytogenes. The purpose of this chapter is to provide detailed
information on the use of M. oleifera extracts in the preservation of different

types of foods due to their antioxidant and antimicrobial capacity.
5.1 INTRODUCTION
The current trend in the population is the consumption of healthy and mini-
mally processed foods, coupled with the negative perception of food additives
such as preservatives, the demand for natural extracts as food preservatives
is increasing. Natural plant extracts contain phytochemicals and essential
oils consisting of a wide variety of bioactive compounds with antioxidant
and antimicrobial properties. Studies focused on the development of new
preservatives based on plant extracts are based on polyphenols and essential
oils with antimicrobial and antioxidant activity to be applied to perishable
foods such as meat, dairy, seafood, fruits, and vegetables (Debonne et al.,
2018; Efenberger-Szmechtyk et al., 2021; Gutiérrez-del-Río et al., 2018;
El-Saber Batiha et al., 2021). However, it is not known with certainty what
the mechanisms of action are, since, in most research, crude extracts are used
without purifying and/or identifying the compounds present in the extracts.
In general, extracts obtained from plants are a complex mixture of substances
such as carbohydrates, proteins, polyphenols, terpenoids, and others.
Among the plants most studied for their antioxidant and antimicrobial
properties is Moringa oleifera. This is a plant with a high nutritional value due
to its vitamin and mineral content. It also contains beneficial phytochemicals
with antioxidant, anticarcinogenic, antimicrobial, and anti-inflammatory
properties (Abd El-Hack et al., 2022; Sultana, 2020). Moreover, it has a
high protein content, especially the seeds (~52%), which in addition to their
nutritional value have multiple biological activities such as antioxidant,
antimicrobial, and antidiabetic (Kumar et al., 2022). M. oleifera polysac-
charides also possess biological activity such as antihyperlipidemic, antidia-
betic, immunomodulatory, antihypertensive, and gastrointestinal protection
(Sharma et al., 2022). The high nutritional value and content of bioactive
compounds make the moringa plant an important source of ingredients for
the development of functional foods, and its antimicrobial capacity makes it
important for food preservation (Saucedo-Pompa et al., 2018).
The uses of moringa are varied, which range from its consumption as
food to wastewater treatment due to the variety of bioactive compounds
present in the leaves, seeds, flowers, and root of moringa. Extraction plays an
important role in antimicrobial and antioxidant capacity since; depending on
the method and solvents used, different compounds with different activities
Antioxidant and Antimicrobial Properties of Moringa oleifera 113
will be extracted. Therefore, traditional methods to the use of emerging tech-

nologies will be addressed. Identifying the compounds present in moringa
extracts is important to know the mechanisms of action, as well as possible
synergistic interactions between the extracted compounds. Finally, the use
of moringa extracts (seeds, leaves, flowers, and roots.) applied in perishable
foods such as dairy products, meat, seafood, edible oils, and others, as a
preservative against microbial and oxidative deterioration will be addressed.
5.2 MORINGA USES

M. oleifera is a native plant mainly in India. It is a very large plant having
nutritional, medicinal, and many other outstanding uses both in the prepara-
tion of other food and industrial products. It is one of the most nutrient-rich
plants discovered today and the use of all parts is well known, from the use
as fodder for animals to water purification, including their use for treating
infant malnutrition (Oyeyinka and Oyeyinka, 2018; Terefe et al., 2017). Due
to the number of uses and properties, it has been called the “miraculous
tree” among cultures that commonly use it (Badmos et al., 2014). M. oleifera
has been reported to have many beneficial compounds, from vitamins such
as vitamins A, C, and E, to minerals such as calcium, magnesium, potas-
sium, copper, iron, and manganese. Also, M oleifera have carotenoid, acid
ascorbic, and tocopherols among others (Oyeyinka and Oyeyinka, 2018). For
its nutritional, medicinal, and technological properties, M. oleifera is used
in different areas such as food, food additives, pharmaceuticals, wastewater
treatment, and more (Table 5.1).
TABLE 5.1 Uses of M. oleifera in Different Areas.

Study area Application
Food All parts of the plant are edible, seeds oil is used to cook, and the leaves
are used as livestock feed
Food additive Thickener in yogurt, milk coagulant for cheese elaboration, clarifier in
fruit juices, and food preservative
Medicine Treatment of disease such as inflammation, hepatorenal disorders,
hematological, cardiovascular, gastrointestinal, and as wound healing
Treatment of Coagulant and clarifier
wastewater Adsorbent of organic pollutants
Others Polysaccharides from trunk for gum formation, biofilm inhibition caused
by S. aureus
M. oleifera is used in a variety of foods because all its parts are edible
from the pods, leaves, flowers, seeds, and even the roots (Benitez and
Benítez Wilberth, 2012). Among important nutritional uses include its use
as food for humans mainly alleviating malnutrition for babies and children,
and livestock due is considered a great food to increase weight and milk
production in animals due to its high fiber content, proteins, vitamins, and
minerals, leaves being the main sources of forage for animals even more
than other leaves (Oyeyinka and Oyeyinka, 2018; Terefe et al., 2017). On
the other hand, studies highlight the great importance of its edible use in
humans, which is something very interesting, because important studies for
application in food start from there; M. oleifera has been used in various
countries over the years as a source of the human diet, as in the case of the
Philippines, in which the use of moringa flowers stands out consuming the
leaves of various firms, including cooked, fresh leaves, even as a dry powder
(Lee et al., 2016; Pontual et al., 2012). M. oleifera flour has been evaluated as
a food supplement in snacks, broiler breast meat, rice crackers, yogurt, and
cheese. Moreover, moringa seed flour has also been used in soups, bread, and
cookies to increase the fiber dietary and protein content and to reduce levels
of lipid oxidation as well as improve fatty acid profiles (Feitosa et al., 2020).
Also, moringa seeds have the power to compete against other oils for use in
the kitchen, as well as for the lubrication of mechanisms, as well as for the
manufacture of soaps and cosmetics (Benitez and Benítez Wilberth, 2012;
Rodríguez-Pérez et al., 2016). Due to its great antioxidant potential with
extracts and oils from all parts of M. oleifera, it has been able to demonstrate
its antioxidant effect in food matrices such as in meats. Moringa leaves have
demonstrated their effectiveness in the inhibition of lipid oxidation in meats
through packaging technologies (Shah et al., 2015).
Moringa oleifera has been used as food additive, highlighting the proper-
ties of the seed, since seed extracts of moringa seeds have been evaluated as
a thickener agent in yogurt, where moringa seeds demonstrated their effect as
a thickening agent and higher protein content (Cardines et al., 2018). Since
moringa leaves have shown their potential as a milk coagulant due to their
protease content and the action of proteins on the casein micelle (Pontual et
al., 2012). However, although moringa leaves are effective in coagulating
milk, the best properties for coagulation of milk have been seen in the seeds
of moringa, since this property has been low in other parts such as flowers
and leaves of this plant (Tajalsir et al., 2014; Sánchez-Muñoz et al.,2017).
This property of coagulation of milk has been verified for the elaboration of
food, being in the coagulation of milk for the formulation of cheeses with
different types of milk such as skim milk and soy milk (Sánchez-Muñoz et
al., 2017). Also, it was been used for clarification in honey as well as in the
clarification of sugar cane juice, both important for food production (Benitez
and Benítez Wilberth, 2012).
Its another important use is its application in cosmetics and pharmaceuti-
cals (Cardines et al., 2018), that lies in the bioactive compounds it contains,
from phenolic acids, alkaloids, phytosterols, flavonoids, as well as vitamins
and mineral content (Gautier et al., 2022). M. oleifera is also rich in phenolic
acids such as gallic acid, chlorogenic acid, ellagic acid, ferulic acid, and
flavanols, among which kaempferol and quercetin stand out (Feitosa et al.,
2020). Extracts from M. oleifera have been used in traditional medicine to
treat diseases such as inflammation, hepatorenal disorders, hematological,
cardiovascular, gastrointestinal diseases, treatment for gastric ulcers as well
as for the relief of fatigue and bronchitis, due to their high content of anti-
oxidants and anti-inflammatory compounds (Zhang et al., 2019; Vongsak et
al., 2013). Multiple in vitro and in vivo investigations in mice and humans
have studied the anti-obesity mechanisms of the different compounds from
moringa isolated or as crude extracts obtained from maceration, ultrasound-
assisted extraction, microwave-assisted extraction, supercritical fluid extrac-
tion, among others, in addition to the use of different solvents such as ethanol,
petroleum ether, acetone, and their aqueous mixtures. The main mechanisms
suggested are the improvement of the lipid profile, regulation of genes associ-
ated with adipogenesis, glucose uptake, insulin resistance, and satiety-related
hormones (Ali Redha et al. 2021). One of the properties of the hydroalcoholic
extracts of M. oleifera seeds recently studied is their potential for wound
healing. Seed extracts were incorporated into a 5% and 10% hydrogel and
applied to 1 cm wounds in male Swiss albino mice. After 4 days, a wound
closure rate of up to 81% was observed and total healing at 13 days, while
the control and standard (povidone) groups were 62% and 82% at the end of
treatment (14 days). In addition, mice to which the hydrogel with 10% M.
oleifera seed extract was applied showed higher resistance to skin breaking
of the healed wound, which was 243 g (Ali et al., 2022). Also, its beneficial
properties against asthma-related inflammatory responses have been studied
in Wistar rats. The ethanol extract of M. oleifera seeds was given to rats at
100 and 200 mg/kg body weight, a similar effect to that of dexamethasone
(2.5 mg/kg body weight) was found since asthmatic symptoms only occurred
in the control group (Mahajan et al., 2007). Oil-free M. oleifera seed extract
can alleviate arsenic toxicity, mitigate ROS generation, decrease Hcy, IL-6,
and TNF-α levels, and increase vitamin B9 and B12 levels (Jana et al., 2020).
Their use as a food source has already been demonstrated, but now indus-
trially the use of M. oleifera also stands out. One of the uses that have been
highlighted for M. oleifera is its potential as a coagulant for the treatment of
wastewater from the dairy industry and the slaughterhouse as a coagulant, to
decrease turbidity, and adsorbent of organic pollutants (Formentini-Schmitt
et al., 2013; del Real-Olvera et al., 2015). Among all the uses of M. oleifera
that has already been discussed about the leaves, seeds, and flowers, another
use is the trunk—this part of the plant has been used for the formation of
gums (Oyeyinka and Oyeyinka, 2018). One application, although in the food
or water industry, is for the inhibition of biofilms caused by S. aureus, being
the antibacterial property of extracts of M. oleifera the one that acts on this
microorganism on surfaces with S. aureus (de Oliveira et al., 2020). Also, it
has wide use both industrially, in wastewater treatment, as well as in food use
again both in nutritionally, fortifying foods, as well as acting as a potential
new food additive.
5.3 BIOACTIVE COMPOUNDS IN MORINGA
Plant extracts that have bioactive compounds can be obtained from different
parts of the plant such as roots, leaves, seeds, flowers, stems, and fruits. These
extracts are mainly composed of polyphenols and aromatic hydrocarbons
with antioxidant activity (Ong et al., 2021). In addition to plant extracts,
essential oils are another group of volatile compounds with a characteristic
aroma of the plant and possess antimicrobial and antioxidant properties,
consisting mainly of terpenes, terpenoids, phenylpropenes, and to a lesser
extent other compounds (Reis et al., 2022; El Khetabi et al., 2022). The pres-
ence of bioactive compounds in moringa extracts has been reported mainly
in leaves, seeds, and flowers (Fig. 5.1).
In a study for the optimization of the extraction of moringa leaves by
microwave and pressurized liquid-assisted extraction using aqueous ethanol,
a total of 62 compounds were identified by HPLC-ESI-QTOF-MS. These
include essential amino acids, organic acids, phenolic acids, flavonoids,
flavonoids, glucosinolates, and lignans. The most important bioactive
compounds in moringa leaves are the flavonoids kaempferol and quercetin
as well as all their derivatives, in addition to their antioxidant properties,
which are greater than those of traditional vitamins, they have anti-inflam-
matory effects (Rodríguez-Pérez et al. 2016). In addition to the conventional
flavonoids reported in M. oleifera leaves, four flavonoids (vitexin, robinetin,
FIGURE 5.1 Bioactive compounds in M. oleifera.
nictoflorin, and taxifolin) found in other plants such as Pigeonpea, Cayratia

trifolia, Lycium berries, and Pinus pinaster were recently reported for the
first time (Wang et al., 2022). Another type of bioactive compound identified
in the leaves of M. oleifera is carotenoids, with the highest concentrations
of β-carotene, lutein, violaxanthin, and neoxanthin, the concentrations found
were higher than those reported in other foods such as broccoli, chicory,
cabbage, and chard (Teixeira et al., 2014). This shows that in addition to their
nutritional value, the leaves of M. oleifera have a wide variety of bioactive
compounds that can have health benefits for people.
In the case of M. oleifera seeds, they have higher protein (~50.8%) and
oil (~11.2%) content than leaves and other vegetable sources such as yellow
peas, red lentils, and chickpeas. The essential amino acid content is also
higher in seeds. However, the contents of polyphenols and flavonoids are
considerably higher in moringa leaves than in seeds (Cattan et al., 2022).
In addition to phenolic compounds, in the seeds of M. oleifera, we can find
isothiocyanates with bioactive properties obtained by enzymatic conver-
sion from glucosinolates. These compounds can modulate the lipid profile
in diabetic patients, decreasing total cholesterol, triglycerides, and low and
very low-density lipoproteins, and at the same time increasing the levels of
high-density lipoproteins (Ali Redha et al., 2021).
Edible flowers are recognized for their nutritional, bioactive, and preser-
vative properties, including M. oleifera flowers (Purohit et al., 2021). M.
oleifera flowers extract containing polyphenols, flavonoids, terpenoids, and
tannins has a promising anti-arthritic effect on Wistar rats with Mycobac

terium tuberculosis-induced arthritis injected in the hind paw. The results
indicate that adverse physical changes such as weight loss, increased paw
volume, and arthritis index was reversed, and serum levels of rheumatoid
factor and proinflammatory cytokine levels were decreased (Mahajan et al.,
2009). In another study, one M. oleifera flower trypsin inhibitor (MoFTI)
was evaluated as a potential inflammatory agent in two rat models of acute
inflammation (paw edema and peritonitis) induced by carrageenan. The
results showed that a dose of 15–30 mg/kg decreased edema size, and
reduced leukocyte migration, nitric oxide levels, and proinflammatory cyto-
kines (Patriota et al., 2021).
Apolysaccharide isolated from the roots of M. oleifera MRP-1 is composed
mainly of rhamnose, arabinose, fructose, xylose, mannose, and galactose. It
possesses anti-inflammatory activity by preventing the LPS-induced increase
in nitric oxide and TNF-a production in RAW264.7 macrophage cells. In
another study, the methanol extract of M. oleifera roots was tested in two rat
models of acute and chronic inflammation induced by carrageenan (Cui et
al., 2019). Moringa root extract (660 mg/kg) showed inhibition of rat edema
(acute model) as well as cellular accumulation and fluid exudation (chronic
inflammation), while indomethacin (5 mg/kg) treatments only inhibited the
acute but not the chronic form (Ezeamuzie et al., 1996).
5.4 EXTRACTION METHODS

The extraction is the most important step in obtaining bioactive compounds
from vegetable sources (González‐González et al., 2022). The extraction of
M. oleifera has been used by different methods, such as percolation, ultra-
sound-assisted extraction (UAE), microwave-assisted extraction (MAE),
and maceration (Fig. 5.2) (Zhang et al., 2019). The percolation extraction
has been used for the extraction of bioactive compounds from M. oleifera
seeds, allowing them to heat to 100 °C and later filtered (Zhang et al., 2019).
Other alternatives to the extraction of bioactive compounds from M. oleifera
are solid-state fermentation (SSF) (Feitosa et al., 2020). The extraction for
SSF is an alternative method for the extraction of bioactive compounds,
which involves the action enzymatic of microorganisms to release the
compounds from the solid substrate. Moreover, other bioactive compounds
can be formed with a high capacity antioxidant as reported by Feitosa et
al. (2020), that showed a high content of total phenolic (134%) and an
increase in total flavonoid contents (783.1%) in extracts of M. oleifera seeds
of SSF compared with unfermented moringa seeds extracts compared with

flour unfermented. Furthermore, SSF showed an improved antimicrobial
activity on Bacillus subtilis. As result of fermentation, 14 new compounds
were obtained, such as, 2-hydroxycinnamic acid, kaempferol 3-glycoside,
p-coumaric acid, trans-cinnamic acid, luteolin, caffeic acid, isorhamnetin,
6,2 dihydroxyflavone, chlorogenic acid, epicatechin, epigallocatechin
gallate, quercetin, taxifolin, and ferulic acid, which they have been reported
with antimicrobial and antioxidant properties.
FIGURE 5.2 Extraction methods used for the bioactive compounds M. oleifera extraction.
Among solvents used for the extraction of bioactive compounds, ethanol

is more used, since it has submitted extractions of bioactive compounds on
best conditions of ethanol to 60%. However, extraction has been reported as
the best condition extract to 70% of ethanol in maceration (Vongsak et al.,
2013; Valdés-Hernández et al., 2015). The extraction of M. oleifera seeds
has been reported by the obtention of bioactive compounds with high-value
antioxidant, antimicrobial, and other properties through the use of solvents
such as ethanol, acetone, water, and/or a combination of these (Feitosa et
al., 2020; Zhang et al., 2019; Ali et al. 2022). The maceration with heating
has been used for the extraction of bioactive compounds for employed as
nanoparticles with antibacterial activity (Ao et al., 2022).
5.5 USE IN DAIRY FOODS

Due to consumer demands for dairy products that contain bioactive proper-
ties, fortified dairy products such as fortified yogurts, cheeses, and others
with preservation potential have been developed, such as yogurt fortified
with extracts of lyophilized M. oleifera seeds with various concentrations
from 0.05%, 0.1%, and 0.2% (Zhang et al., 2019). Likewise, the microbial
properties of yogurt during fermentation have been put under study, because
the microbiological analysis is the second important quality criterion in the
acceptability as well as the shelf-life of dairy products (Salem et al., 2015).
However, color, pH viscosity measurements, as well as determination of
radical scavenging activity and phenolic content are important parameters. It
has been shown that the addition of M. oleifera accelerates the fermentation
rate of yogurt and allows the elimination of free radicals and that it can even
be a food that provides health benefits, contributing to the development of
yogurt with bioactive properties (Zhang et al., 2019).
Some dairy products from other mammals besides cows have been the
subject of research to improve their conservation as sour cream from buffalo
milk due to M. oleifera as an oil or extract can improve microbial stability,
therefore an increase sensorial activity and they can improve the shelf life
of sour cream. In addition, the extracts and oils do not affect the punctua-
tion of the attributes such as flavor, texture, body, acidity, and appearance.
Furthermore, the M. oleifera extracts were permitted that the acidity
increased; however, the increase of M. oleifera oils decreased. M. oleifera
extracts and oils showed a decrease in the content of microorganisms such
as proteolytic bacteria, lipolytic bacteria, and mold and yeast to 4 weeks of
storage at 5 °C. (Salem et al., 2015). Badmos et al. (2014) applied extracts
from M. oleifera in soft cheese and evaluated the antibacterial effect. The
results showed that M. oleifera has antibacterial activity in soft cheese. The
extracts with a 3% ethanol condition had a decrease in bacterial content,
among them are enterococcus, Bacillus pasteuri, Staphylococcus spp., S
aureus, and Corynebacterium spp. Furthermore, was shown that moisture
content was low in the condition of extraction with 1 and 2% ether, which
helps to avoid contamination by microorganisms. Among the dairy products
which has presented positive effects of preservation with M. oleifera is the
cheese, as reported by Cui et al. (2020), who evaluated the preservative
effect of moringa oils applied by spray in mozzarella cheese, cheddar cheese,
parmesan cheese, and camembert cheese against L. monocytogenes and
evaluated a sensorial analysis. Moringa oils showed positive effects on the
inhibition of L. monocytogenes in cheese and demonstrated no changes in the
textural characteristics of the cheese during storage. The use of M. oleifera

has not only been oriented to preservation, since it has been used as an addi-
tive for the coagulation of milk in cheese processing (Abdeen et al., 2021).
Since the coagulation of milk is a very important step in the manufacture
of various types of cheese, a new source of enzymes for cheese production
has been sought, in addition to calf rennet, which has been widely used for
cheese production (Terefe et al., 2017). Among the application in cheeses,
M. oleifera is important in addition to the conservation against microorgan-
isms, it has lengthened the shelf life in gouda cheese through a film delaying
the oxidation of lipids in the cheese (Lee et al., 2016). Interestingly, two of
the main microorganisms among dairy products are L. monocytogenes and S.
aureus. A study has achieved to inhibit growing these two microorganisms in
cheeses through nanoparticles (Lin et al., 2019).
5.6 USE IN MEAT FOOS

Meat and meat by-products are very susceptible to lipid oxidation and micro-
bial spoilage resulting in undesirable physicochemical changes such as loss
of color, texture, and flavor. Bioactive compounds derived from plants have
been investigated for the preservation of meat products, in addition, they
provide functional properties (Munekata et al., 2020; Falowo et al., 2014).
M. oleifera extracts have been studied in beef preservation. A study by
Manzoor et al. evaluated the effect of aqueous extract of moringa leaves
on the preservation of raw beef packaged with modified atmospheres (80%
O2 and 20% CO2). The results showed significant effects on pH, thiobarbi-
turic acid reactive substances (TBARS) and color, but no effect on micro-
biological quality and water-holding capacity (Shah et al., 2015). Similarly,
direct application of moringa leaf powder at concentrations of 0.2–0.4% in
ground beef has the same effects on lipid oxidation without affecting the
sensory profile; in addition, the total microbial load, coliforms, fungi, and
yeasts were reduced (Mashau et al., 2021). On the other hand, the applica-
tion of moringa leaf extracts in combination with Bidens pilosa leaves in
veal minced meat showed a higher antioxidant activity, obtaining lower pH
and TBARS values. In addition, the antimicrobial properties of the extracts
showed inhibition against pathogens of interest such as S. aureus, E. coli,
and Shigella (Falowo et al., 2016, 2017). Furthermore, moringa seed meal
applied in beef burgers improves cooking properties as the concentration
of moringa seed meal increases, reduces thiobarbituric acid and pH values,
while higher concentration of moringa seed in meals reduces thiobarbituric
acid, pH values, and aerobic plate counts during storage. However, sensory
acceptability was lower in beef burger with moringa seed flour and reduced
at longer times, but that of beef burger without moringa seed flour decreased
significantly at longer storage times (Al-Juhaimi et al., 2015).
The addition of moringa leaf powder in the processing of chicken patties
decreases their moisture content, while increasing the fat, protein, and
ash content. As for the total microbial load, the patties with moringa leaf
powder remained at 3.78 Log CFU/g, while the control had 7.87 Log CFU/g,
MLP maintained the absence of E. coli and S. aureus at 2.52 Log CFU/g,
and the control showed 2.91 and 3.92 Log CFU/g, respectively. However,
sensory acceptability was significantly lower than the control (Elhadi et al.,
2016). Similarly, moringa flower extracts were added to the formulation of
nuggets, which were shown to improve oxidative stability decrease TBARS
values without demonstrating changes in sensory acceptability as occurs
with moringa leaves (Madane et al., 2019). On the other hand, the extract
of defatted moringa seeds applied in a chitosan-based coating was able to
inhibit the growth of B. cereus and E. coli until day 6 of refrigerated storage
in chicken sausages inoculated with both bacteria at a concentration of 4 Log
CFU/g, while after 15 days of storage the microbial load increased to 7.3
Log CFU/g but still lower than the control without treatment 8 Log CFU/g
(Sharma et al., 2020). In another study, it was reported that moringa leaf
powder in 2% added to irradiated chicken meat packaged under aerobic and
vacuum conditions decreased peroxide and TBARS values, and additionally
improved the amino acid and fatty acid content (Nisar et al., 2019). The same
treatment was evaluated in the preservation of chicken meatballs, reducing
the total volatile basic nitrogen values. As for the microbial load, M. oleifera
alone reduces more than 1 Log CFU/g of coliforms but in combination with
radiation it inhibits them completely (Nisar et al., 2020).
Sheep meat is very susceptible to damage by pathogenic microorganisms
and spoilage, having a very short shelf life. The use of natural antimicrobials
such as plant extracts and essential oils in combination with refrigeration
have been investigated for the preservation of mutton (Purgatorio et al. 2022).
In this sense, the influence of moringa leaf extracts on the nutritional, physi-
cochemical, and microbiological properties of sheep burgers was studied.
The values of protein, ash, total phenolics, and total flavonoids increased,
while moisture and fat decreased. The values of protein, ash, total phenolics
and total flavonoids increased, while moisture and fat decreased. Cooking
parameters such as yield, water and fat retention were also improved.
Regarding the microbial load, the additional hamburgers with 5% moringa
leaf extract had 4.30 Log CFU/g, while the control hamburgers had 6.63
Log CFU/g for the total plate count and 2.06 and 6.20 Log CFU/g for total
coliforms (Mashau et al., 2021).
These studies demonstrate the potential of M. oleifera as a preservative
in meat products. It has antimicrobial activity against pathogenic bacteria
such as E. coli and S. aureus that are regularly present in this type of food.
On the other hand, the antimicrobial properties of M. oleifera help against
the oxidation of lipids and proteins, decreasing the values of TBARS and
peroxide. In addition, they add functional characteristics due to bioactive
compounds, mainly polyphenols.
5.7 USE IN SEAFOOD AND FISH

Seafood and fish are one of the most perishable foods and are an important
part of the food supply in many countries, mainly on the coasts, and loss and
waste in value chains represent losses in the millions of dollars (Kruijssen et
al., 2020). One of the causes of spoilage of this type of food is proteases that
degrade the structural proteins of fish and shellfish. This process is generally
undesirable due to the changes in the physicochemical properties of fish as
loss of texture, and changes in color, moisture distribution, and water-holding
capability (Zhuang et al., 2021).
One solution to this problem is the use of protease inhibitors, which are
usually chemical preservatives and antibiotics that inhibit both the proteolytic
activity and the growth of the microorganisms that produce these enzymes.
However, the negative perception of the use of chemically synthesized anti-
microbials has increased the demand for natural preservatives, which would
also be beneficial by decreasing antibiotic resistance of pathogenic micro-
organisms and increasing shelf life (Cotabarren et al., 2020). The protease
inhibitor from M. oleifera leaves has been investigated and has shown
inhibition on serine proteases thrombin, elastase, chymotrypsin, and cysteine
proteases cathepsin B and papain, which are important in the industry. Their
application in shrimp (Penaeus monodon) as a natural preservative (0.2 mg/
mL) inhibited protein degradation and maintained a microbial load of 4×102
CFU/mL while the control registered 8×1010 CFU/mL, which shows that the
proteolytic activity is closely related to the microbial load and therefore the
deterioration of shrimp (Bijina et al., 2011). On the other hand, the applica-
tion of moringa and curry leaf extracts is effective in delaying melanosis
in Pacific white shrimp applied by immersion for 15 min, in addition to
delaying microbial deterioration (Soni et al., 2021).
Biological selenium nanoparticles (Bio-SeNPs) synthesized using selenite

and M. oleifera extracts are an effective antimicrobial to eliminate L. mono
cytogenes and Corynebacterium diphtheriae two pathogens of importance in
food safety, 0.7 mg/mL of Bio-SeNPs significantly inhibit the growth of these
two strains. the antimicrobial capacity of Bio-SeNPs was higher than when
using moringa extract or chemical-Se(0) (formed by the reduction of selenite
with cysteine). Furthermore, their application in raw salmon effectively
eliminates L. monocytogenes without causing cytotoxicity (Ao et al., 2022).
The incorporation of moringa seedmeal in a brine solution for the marinade
of smoked African mud catfish (Clarias gariepinus) at a concentration of
15 g/kg achieves a significant decrease in microbial load, especially fungi
and yeasts related to fish spoilage (Jimoh et al., 2021). In another study,
moringa leaf extract was added to an edible gelatin-based coating of gray
triggerfish (Balistes capriscus) skin and applied to smooth-hound shark
(Mulestus mulestus) fillets. The moringa leaf extract was mainly composed
of phenolic acids that presented high antioxidant and antimicrobial activity,
thus decreasing spoilage indices such as malondialdehyde formation, total
volatile basic nitrogen, microbial load, as well as physical parameters such
as weight loss and texture profile (Mezhoudi et al., 2022). However, the use
of moringa extracts does not always improve food preservation. Delfino et
al. (2021) applied extracts of M. oleifera leaves and Lavandula sp. foliage
to the preservation of tilapia fish burgers, which had an adverse effect by
increasing the production of TBARS and negatively affecting the overall
impression of tilapia fish burgers at the end of storage.
5.8 OXIDATIVE STABILIZER IN OILS
It is widely known that phenolic compounds have antioxidant properties and

that they are part of many foods in greater or lesser abundance, and their incor-
poration in cooking oils can increase their stability in frying foods without
affecting their sensory quality, through the elimination of free radicals and
the elimination of carbonyls (Wu et al., 2019). In addition, polyphenols from
Eucalyptus citriodora leaf extracts increase the stability of mixed vegetable
oils during storage (Ali et al., 2016). However, there are few studies related
to the stability of cooking oils during food frying. As reported by Dhibi et
al. (2022), methanolic extracts of M. oleifera and Zizipus Lotus contained
phenolic compounds (phenolic acids and flavonoids) which were able to
increase the antioxidant activity (DPPH radical inhibition), decrease the free
fatty acid content and peroxide value, and increase the oxidative stability of
corn oil, after being heated at 180 °C for 8 h. Moringa leaves extract has a
significant antioxidant effect in accelerated storage of soybean oil (12 days/60
°C), however, in the heating test its antioxidant activity decreased as the
temperature increased (105 to 180 °C) but always had lower values than the
control (Zhao et al., 2019). The mixture of moringa oil (20%) with sunflower
oil (80%) improves stability during potato frying, decreasing the formation
of polymers, and oxidized triglycerides reduces the value of p-anisidine and
total volatile aldehydes, and preserves vitamin E and antioxidant activity due
to a possible interaction with the components of moringa oil, which gives it
a protective effect (Boukandoul et al., 2019). Moreover, the incorporation of
moringa leaves extract in margarine increases oxidation resistance compared
to vitamin E, it also improves physical aspects such as water droplet size,
which is desirable since from a microbiological point of view it would help
to prolong shelf life. However, one characteristic that may not be acceptable
to the consumer is the change in color of the margarine for moringa leaves
extract addition (Ouahrani et al., 2022).
The use of natural antioxidants obtained from plants helps to extend the
shelf life of fish oils by retarding the oxidation of omega-3 unsaturated fatty
acids, known not only for their health benefits but also for their instability
and easy oxidation during storage, thus developing an undesirable flavor
and potentially toxic compounds (Hrebień‐Filisińska, 2021). Commercially
available flavonoids have been used to increase the stability of fish oil
by decreasing the formation of peroxides and thiobarbituric acid reactive
substances (Nieto et al., 1993). These compounds are present in moringa
extracts. The application of moringa leaf, flower, and seed extracts in fish oil
retarded the oxidation of fish oil; moringa extracts showed lower peroxide
and hydroperoxide values compared to the synthetic antioxidants tertiary
butylated hydroquinone (TBHQ) and butylated hydroxytoluene (BHT) in
fish oil in accelerated storage for 16 days at 60 °C and showed a greater
protective effect on polyunsaturated fatty acids (Nascimento et al., 2016).
5.9 USE IN OTHER FOODS
Another microorganism against which plants have antimicrobial effects is

Aspergillus flavus, allowing its application in nuts such as peanuts, which are
often attacked by this aflatoxigenic fungus (Razzaghi-Abyaneh et al., 2010).
The extracts of the leaves of the medicinal plants Cymbopogon citratus, M.
oleifera, Ocimum gratissimum, and Clerodendrum volubile have antifungal

activity against the toxigenic fungus A. flavus, phytochemical screening
showed the presence of saponins, tannins, glycoside, and volatile oils (Jeff-
Agboola et al., 2016). A study by Abu-Sree et al. (2021) demonstrated the
ability of M. oleifera leaves extracts in combination with oil of seeds to inhibit
A. flavus and aflatoxin production in peanuts. An emulsion consisting of the
methanol extract of moringa leaves, seed oil, and a commercial emulsifier
was applied. After three months, the results show a reduction from 6.8×104
to 9 CFU/kg for peanuts with red skin, while for peanuts without red skin
the reduction was 9.4×105 to 14 CFU/kg. As for aflatoxin B1 production,
it remained undetectable for peanuts with the moringa extract coating and
controls with a content of 480 and 945 ng/kg for peanuts with and without
the red skin, respectively.
In active films based on chitosan, guar gum, and polyvinyl alcohol, M.
oleifera extract improves characteristics such as tensile strength, thermal
stability, decrease in glass transition temperature and melting temperature,
and effectively inhibits the growth of E. coli and S. aureus. These character-
istics make possible its incorporation in this type of film that can be used in
food preservation due to its antimicrobial capacity (Bhat et al., 2022). The
application of M. oleifera coatings with chitosan nanoparticles increases the
shelf life of bananas during storage. This is achieved by reducing respira-
tion rate, ethylene production, weight loss, firmness, and delaying color
changes, thus increasing the sensory acceptability of the fruit after 30 days
of storage (Odetayo et al., 2022). On the other hand, papaya-based coatings
with moringa extract and/or ascorbic acid increased antioxidant activity and
shelf life of cut pears for more than 9 days; however, sensory acceptability
was affected by moringa extract (Rodríguez et al., 2020).
5.10 CONCLUSIONS
Food preservatives are one of the most widely used additives in the food
industry in order to keep food quality for a longer period. However, they are
closely related to different pathologies, so the demand for natural preserva-
tives is increasing and medicinal plants such as M. oleifera are an important
source of bioactive compounds with antioxidant and antimicrobial activity
which can be used to extend shelf life. The use of emerging technologies for
the extraction of bioactive compounds helps to improve yields, recovering
most of the phytochemicals of interest and preserving their properties. The
bioactive compounds of moringa have been identified in leaves, seeds,

flowers, and even roots, among which there are phenolic acids, polyphe-
nols, flavonoids, isothiocyanates, terpenoids, and others. Currently, many
studies support their potential use in the preservation of foods such as dairy
products, meats, seafood, and edible oils, among others. It is important to
establish clear mechanisms of action and application of moringa compounds
in food preservation so that they can be used safely and efficiently to ensure
safe food for consumers.
KEYWORDS
• antioxidant
• food preservation
• antimicrobial capacity
• healthy foods
• M. Oleifera extracts
• bioactive compounds
REFERENCES
Abd El-Hack, M. E.; Alqhtani, A. H.; Swelum, A. A.; El-Saadony, M. T.; Salem, H. M.;
Babalghith, A. O.; Taha, A. E.; Ahmed, O.; Abdo, M.; El-Tarabily, K. A. Pharmacological,
Nutritional and Antimicrobial Uses of Moringa Oleifera Lam. Leaves in Poultry Nutrition:
An Updated Knowledge. Poult. Sci. 2022, 101 (9), 102031. https://doi.org/10.1016/j.
psj.2022.102031.
Abdeen, E.-S. M. M.; Ibrahim, O. A.; Kholif, A. M. M. Utility of Moringa Oleifera Waste
as a Coagulant in Goat Soft Cheese Production. Heliyon 2021, 7 (7), e07536. https://doi.
org/10.1016/j.heliyon.2021.e07536.
Abu-Sree, Y. H.; Abdel-Fattah, S. M.; Abdel-Razek, A. G.; Badr, A. N. Neoteric Approach for
Peanuts Biofilm Using the Merits of Moringa Extracts to Control Aflatoxin Contamination.
Toxicol. Rep. 2021, 8, 1685–1692. https://doi.org/10.1016/j.toxrep.2021.08.006.
Ali Redha, A.; Perna, S.; Riva, A.; Petrangolini, G.; Peroni, G.; Nichetti, M.; Iannello, G.;
Naso, M.; Faliva, M. A.; Rondanelli, M. Novel Insights on Anti-Obesity Potential of the
Miracle Tree, Moringa Oleifera: A Systematic Review. J. Funct. Foods 2021, 84, 104600.
https://doi.org/10.1016/j.jff.2021.104600.
Ali, A.; Garg, P.; Goyal, R.; Khan, A.; Negi, P.; Li, X.; Kulshrestha, S. An Efficient Wound
Healing Hydrogel Based on a Hydroalcoholic Extract of Moringa Oleifera Seeds. SA J. Bot.
2022, 145, 192–198. https://doi.org/10.1016/j.sajb.2021.05.003.
Ali, S.; Chatha, S. A. S.; Ali, Q.; Hussain, A. I.; Hussain, S. M.; Perveen, R. Oxidative
Stability of Cooking Oil Blend Stabilized with Leaf Extract of Eucalyptus Citriodora. Int.
J. Food Prop. 2016, 19 (7), 1556–1565. https://doi.org/10.1080/10942912.2015.1047514.
Al-Juhaimi, F.; Ghafoor, K.; Hawashin, M. D.; Alsawmahi, O. N.; Babiker, E. E. Effects of
Different Levels of Moringa (Moringa Oleifera) Seed Flour on Quality Attributes of Beef
Burgers. CyTA–J Food 2015, 14 (1), 1–9. https://doi.org/10.1080/19476337.2015.1034784.
Ao, B.; Lv, J.; Yang, H.; He, F.; Hu, Y.; Hu, B.; Jiang, H.; Huo, X.; Tu, J.; Xia, X. Moringa
Oleifera Extract Mediated the Synthesis of Bio-SeNPs with Antibacterial Activity against
Listeria monocytogenes and Corynebacterium Diphtheriae. LWT 2022, 165, 113751.
https://doi.org/10.1016/j.lwt.2022.113751.
Badmos, A. H. A. et al. The Effect of Crude Leaf Extracts of Moringa Oleifera on the.
Wayamba J. Anim. Sci. 2014, No. 1400002484, 939–946.
Benitez, M.; Benítez Wilberth, M. Aprovechamiento Poscosecha De La Moringa (Moringa
Oleífera). Rev. Iber. Tecnol. Postcosecha 2012, 13 (2), 171–174.
Bhat, V. G.; Narasagoudr, S. S.; Masti, S. P.; Chougale, R. B.; Vantamuri, A. B.; Kasai, D.
Development and Evaluation of Moringa Extract Incorporated Chitosan/Guar Gum/Poly
(Vinyl Alcohol) Active Films for Food Packaging Applications. Int. J. Biol. Macromol.
2022, 200, 50–60. https://doi.org/10.1016/j.ijbiomac.2021.12.116.
Bijina, B.; Chellappan, S.; Krishna, J. G.; Basheer, S. M.; Elyas, K. K.; Bahkali, A. H.;
Chandrasekaran, M. Protease Inhibitor from Moringa Oleifera with Potential for Use as
Therapeutic Drug and as Seafood Preservative. Saudi J. Biol. Sci. 2011, 18 (3), 273–281.
https://doi.org/10.1016/j.sjbs.2011.04.002.
Boukandoul, S.; Santos, C. S. P.; Casal, S.; Zaidi, F. Oxidation Delay of Sunflower Oil under
Frying by Moringa Oil Addition: More Than Just a Blend. J. Sci. Food Agric. 2019, 99 (12),
5483–5490. https://doi.org/10.1002/jsfa.9809.
Cardines, P. H. F.; Baptista, A. T. A.; Gomes, R. G.; Bergamasco, R.; Vieira, A. M. S. Moringa
Oleifera Seed Extracts as Promising Natural Thickening Agents for Food Industry:
Study of the Thickening Action in Yogurt Production. LWT 2018, 97, 39–44. https://doi.
org/10.1016/j.lwt.2018.06.028.
Cattan, Y.; Patil, D.; Vaknin, Y.; Rytwo, G.; Lakemond, C.; Benjamin, O. Characterization
of Moringa Oleifera Leaf and Seed Protein Extract Functionality in Emulsion Model
System. Innov. Food Sci. Emerg. Technol. 2022, 75, 102903. https://doi.org/10.1016/j.
ifset.2021.102903.
Cotabarren, J.; Lufrano, D.; Parisi, M. G.; Obregón, W. D. Biotechnological, Biomedical, and
Agronomical Applications of Plant Protease Inhibitors with High Stability: A Systematic
Review. Plant Sci. 2020, 292, 110398. https://doi.org/10.1016/j.plantsci.2019.110398.
Cui, C.; Chen, S.; Wang, X.; Yuan, G.; Jiang, F.; Chen, X.; Wang, L. Characterization of
Moringa Oleifera Roots Polysaccharide MRP-1 with Anti-Inflammatory Effect. Int. J. Biol.
Macromol. 2019, 132, 844–851. https://doi.org/10.1016/j.ijbiomac.2019.03.210.
Cui, H.; Li, H.; Li, C.; Abdel-Samie, M. A.; Lin, L. Inhibition Effect of Moringa Oil on the
Cheese Preservation and Its Impact on the Viability, Virulence and Genes Expression of
Listeria monocytogenes. LWT 2020, 134 (September), 110163. https://doi.org/10.1016/j.
lwt.2020.110163.
de Oliveira, A. M.; de Abreu Filho, B. A.; de Jesus Bassetti, F.; Bergamasco, R.; Gomes, R. G.
Natural Extract of Moringa Oleifera Leaves Promoting Control of Staphylococcus aureus
Strains Biofilm on PVC Surface. Food Bioprocess Technol. 2020, 13 (10), 1817–1832.
https://doi.org/10.1007/s11947-020-02521-x.
Debonne, E.; Van Bockstaele, F.; Samapundo, S.; Eeckhout, M.; Devlieghere, F. The Use of
Essential Oils as Natural Antifungal Preservatives in Bread Products. J. Essent. Oil Res.
2018, 30 (5), 309–318. https://doi.org/10.1080/10412905.2018.1486239.
del Real-Olvera, J.; Rustrian-Portilla, E.; Houbron, E.; Landa-Huerta, F. J. Adsorption of
Organic Pollutants from Slaughterhouse Wastewater Using Powder of Moringa Oleifera
Seeds as a Natural Coagulant. New pub Balaban 2015, 57 (21), 9971–9981. https://doi.org/
10.1080/19443994.2015.1033479.
Delfino, L. A.; Mattje, L. G. B.; Silva, M.; Araujo, M. C.; Tormen, L.; Bainy, E. M. Evaluation
of Moringa and Lavandula Extracts as Natural Antioxidants in Tilapia Fish Burger. J. Culin.
Sci. Technol. 2021, 1–16. https://doi.org/10.1080/15428052.2021.1883494.
Dhibi, M.; Amri, Z.; Bhouri, A. M.; Hammami, S.; Hammami, M. Comparative Study of
the Phenolic Profile and Antioxidant Activities of Moringa (Moringa Oleifera Lam.) and
Jujube (Ziziphus Lotus Linn.) Leaf Extracts and Their Protective Effects in Frying Stability
of Corn Oil. Meas. Food 2022, 7, 100045. https://doi.org/10.1016/j.meafoo.2022.100045.
Efenberger-Szmechtyk, M.; Nowak, A.; Czyzowska, A. Plant Extracts Rich in Polyphenols:
Antibacterial Agents and Natural Preservatives for Meat and Meat Products. Crit. Rev.
Food Sci. Nutr. 2021, 61 (1), 149–178. https://doi.org/10.1080/10408398.2020.1722060.
El Khetabi, A.; Lahlali, R.; Ezrari, S.; Radouane, N.; Lyousfi, N.; Banani, H.; Askarne,
L.; Tahiri, A.; El Ghadraoui, L.; Belmalha, S.; Barka, E. A. Role of Plant Extracts and
Essential Oils in Fighting against Postharvest Fruit Pathogens and Extending Fruit Shelf
Life: A Review. Trends Food Sci. Technol. 2022, 120, 402–417. https://doi.org/10.1016/j.
tifs.2022.01.009.
Elhadi, D. A. E.; Elgasim, E. A.; Mohamed Ahmed, I. A. Microbial and Oxidation
Characteristics of Refrigerated Chicken Patty Incorporated with Moringa (Moringa
Oleifera) Leaf Powder. CyTA–J Food 2016, 15 (2), 234–240. https://doi.org/10.1080/1947
6337.2016.1242157.
El-Saber Batiha, G.; Hussein, D. E.; Algammal, A. M.; George, T. T.; Jeandet, P.; Al-Snafi,
A. E.; Tiwari, A.; Pagnossa, J. P.; Lima, C. M.; Thorat, N. D.; Zahoor, M.; El-Esawi, M.;
Dey, A.; Alghamdi, S.; Hetta, H. F.; Cruz-Martins, N. Application of Natural Antimicrobials
in Food Preservation: Recent Views. Food Control 2021, 126, 108066. https://doi.
org/10.1016/j.foodcont.2021.108066.
Ezeamuzie, I. C.; Ambakederemo, A. W.; Shode, F. O.; Ekwebelem, S. C. Antiinflammatory
Effects of Moringa Oleifera Root Extract. Int. J. Pharmacogn. 1996, 34 (3), 207–212.
https://doi.org/10.1076/phbi.34.3.207.13211.
Falowo, A. B.; Fayemi, P. O.; Muchenje, V. Natural Antioxidants against Lipid–Protein
Oxidative Deterioration in Meat and Meat Products: A Review. Food Res. Int. 2014, 64,
171–181. https://doi.org/10.1016/J.FOODRES.2014.06.022.
Falowo, A. B.; Muchenje, V.; Hugo, A.; Aiyegoro, O. A.; Fayemi, P. O. Antioxidant Activities
of Moringa Oleifera L. and Bidens Pilosa L. Leaf Extracts and Their Effects on Oxidative
Stability of Ground Raw Beef during Refrigeration Storage. CyTA - J. Food 2017, 15 (2),
249–256. https://doi.org/10.1080/19476337.2016.1243587.
Falowo, A. B.; Muchenje, V.; Hugo, C. J.; Charimba, G. In Vitro Antimicrobial Activities
of Bidens Pilosa and Moringa Oleifera Leaf Extracts and Their Effects on Ground Beef
Quality during Cold Storage. CyTA - J. Food 2016, 14 (4), 541–546. https://doi.org/10.108
0/19476337.2016.1162847.
Feitosa, P. R. B.; Santos, T. R. J.; Gualberto, N. C.; Narain, N.; de Aquino Santana, L. C.
L. Solid-State Fermentation with Aspergillus Niger for the Bio-Enrichment of Bioactive
Compounds in Moringa Oleifera (Moringa) Leaves. Biocatal. Agric. Biotechnol. 2020, 27,
101709. https://doi.org/10.1016/j.bcab.2020.101709.
Formentini-Schmitt, D. M.; Alves, Á. C. D.; Veit, M. T.; Bergamasco, R.; Vieira, A. M.
S.; Fagundes-Klen, M. R. Ultrafiltration Combined with Coagulation/Flocculation/
Sedimentation Using Moringa Oleifera as Coagulant to Treat Dairy Industry Wastewater.
Water, Air, Soil Pollut. 2013, 224 (9), 1682. https://doi.org/10.1007/s11270-013-1682-2.
Gautier, A.; Duarte, C. M.; Sousa, I. Moringa Oleifera Seeds Characterization and Potential
Uses as Food. Foods 2022, 11 (11), 1629. https://doi.org/10.3390/foods11111629.
González‐González, G. M.; Palomo‐Ligas, L.; Nery‐Flores, S. D.; Ascacio‐Valdés, J. A.;
Sáenz‐Galindo, A.; Flores‐Gallegos, A. C.; Zakaria, Z. A.; Aguilar, C. N.; Rodríguez‐
Herrera, R. Coffee Pulp as a Source for Polyphenols Extraction Using Ultrasound,
Microwave, and Green Solvents. Environ. Qual. Manag. 2022, 32 (1), 451–461. https://doi.
org/10.1002/tqem.21903.
Gutiérrez-del-Río, I.; Fernández, J.; Lombó, F. Plant Nutraceuticals as Antimicrobial Agents
in Food Preservation: Terpenoids, Polyphenols and Thiols. Int. J. Antimicrob. Agents 2018,
52 (3), 309–315. https://doi.org/10.1016/j.ijantimicag.2018.04.024.
Hrebień‐Filisińska, A. Application of Natural Antioxidants in the Oxidative Stabilization
of Fish Oils: A Mini‐review. J. Food Process. Preserv. 2021, 45 (4), e15342. https://doi.
org/10.1111/jfpp.15342.
Jana, S.; Chattopadhyay, S.; Dey, A.; Perveen, H.; Dolai, D. Involvement of Metallothionein,
Homocysteine and B-Vitamins in the Attenuation of Arsenic-Induced Uterine Disorders in
Response to the Oral Application of Hydro-Ethanolic Extract of Moringa Oleifera Seed: A
Preliminary Study. Drug Chem. Toxicol. 2020, 43 (1), 1–12. https://doi.org/10.1080/0148
0545.2018.1508296.
Jeff-Agboola, Y. A.; Awe, L. B. Antifungal and Phytochemical Screening of Some Nigerian
Medicinal Plant Extracts against Toxigenic Aspergillus Flavus. Cogent Food Agric. 2016, 2
(1). https://doi.org/10.1080/23311932.2016.1210556.
Jimoh, W. A.; Ayeloja, A. A.; Badmus, G. O.; Olateju, K. O. Antibacterial and Antifungal
Effect of Moringa (Moringa Oleifera) Seedmeal on Marinated Smoked African Mud
Catfish (Clarias Gariepinus). J. Food Saf. 2020, 40 (3), e12772. https://doi.org/10.1111/
jfs.12772.
Kruijssen, F.; Tedesco, I.; Ward, A.; Pincus, L.; Love, D.; Thorne-Lyman, A. L. Loss and
Waste in Fish Value Chains: A Review of the Evidence from Low and Middle-Income
Countries. Glob. Food Sec. 2020, 26, 100434. https://doi.org/10.1016/j.gfs.2020.100434.
Kumar, M.; Selvasekaran, P.; Kapoor, S.; Barbhai, M. D.; Lorenzo, J. M.; Saurabh, V.; Potkule,
J.; Changan, S.; ElKelish, A.; Selim, S.; Sayed, A. A. S.; Radha; Singh, S.; Senapathy,
M.; Pandiselvam, R.; Dey, A.; Dhumal, S.; Natta, S.; Amarowicz, R.; Kennedy, J. F.
Moringa Oleifera Lam. Seed Proteins: Extraction, Preparation of Protein Hydrolysates,
Bioactivities, Functional Food Properties, and Industrial Application. Food Hydrocoll.
2022, 131, 107791. https://doi.org/10.1016/j.foodhyd.2022.107791.
Lee, K.-Y.; Yang, H.-J.; Song, K. Bin. Application of a Puffer Fish Skin Gelatin Film
Containing Moringa Oleifera Lam. Leaf Extract to the Packaging of Gouda Cheese. J.
Food Sci. Technol. 2016, 53 (11), 3876–3883. https://doi.org/10.1007/s13197-016-2367-9.
Lin, L.; Gu, Y.; Cui, H. Moringa Oil/Chitosan Nanoparticles Embedded Gelatin Nanofibers
for Food Packaging against Listeria monocytogenes and Staphylococcus aureus on
Cheese. Food Packag. Shelf Life 2019, 19 (Dec 2018), 86–93. https://doi.org/10.1016/j.
fpsl.2018.12.005.
Madane, P.; Das, A. K.; Pateiro, M.; Nanda, P. K.; Bandyopadhyay, S.; Jagtap, P.; Barba, F.
J.; Shewalkar, A.; Maity, B.; Lorenzo, J. M. Drumstick (Moringa Oleifera) Flower as an
Antioxidant Dietary Fibre in Chicken Meat Nuggets. Foods 2019, 8 (8), 307. https://doi.
org/10.3390/FOODS8080307.
Mahajan, S. G.; Mali, R. G.; Mehta, A. A. Effect of Moringa Oleifera Lam. Seed Extract
on Toluene Diisocyanate-Induced Immune-Mediated Inflammatory Responses in Rats. J.
Immunotoxicol. 2007, 4 (2), 85–96. https://doi.org/10.1080/15476910701337472.
Mahajan, S. G.; Mehta, A. A. Anti-Arthritic Activity of Hydroalcoholic Extract of Flowers of
Moringa Oleifera Lam. in Wistar Rats. J. Herbs. Spices Med. Plants 2009, 15 (2), 149–163.
https://doi.org/10.1080/10496470903139363.
Mashau, M. E.; Munandi, M.; Ramashia, S. E. Exploring the Influence of Moringa Oleifera
Leaves Extract on the Nutritional Properties and Shelf Life of Mutton Patties during
Refrigerated Storage. CyTA–J. Food 2021, 19 (1), 389–398. https://doi.org/10.1080/19476
337.2021.1910732.
Mashau, M. E.; Ramatsetse, K. E.; Ramashia, S. E. Effects of Adding Moringa Oleifera
Leaves Powder on the Nutritional Properties, Lipid Oxidation and Microbial Growth in
Ground Beef during Cold Storage. Appl. Sci. 2021, Vol. 11, Page 2944 2021, 11 (7), 2944.
https://doi.org/10.3390/APP11072944.
Mezhoudi, M.; Salem, A.; Abdelhedi, O.; Fakhfakh, N.; Mabrouk, M.; Khorchani, T.;
Debeaufort, F.; Jridi, M.; Zouari, N. Development of Active Edible Coatings Based on Fish
Gelatin Enriched with Moringa Oleifera Extract: Application in Fish ( Mustelus Mustelus
) Fillet Preservation. Food Sci. Nutr. 2022, 10 (11), 3979–3992. https://doi.org/10.1002/
fsn3.2993.
Munekata, P. E. S.; Rocchetti, G.; Pateiro, M.; Lucini, L.; Domínguez, R.; Lorenzo, J. M.
Addition of Plant Extracts to Meat and Meat Products to Extend Shelf-Life and Health-
Promoting Attributes: An Overview. Curr. Opin. Food Sci. 2020, 31, 81–87. https://doi.
org/10.1016/J.COFS.2020.03.003.
Nascimento, J. A.; Magnani, M.; Sousa, J. M. B.; Araújo, K. L. G. V.; Epaminondas, P. S.;
Souza, A. S.; Souza, A. L.; Silva, M. C. D.; Souza, A. G. Assessment of the Antioxidant
Effects of M Oringa Oleifera Lam. Extracts in Fish Oil during Storage. J. Food Process.
Preserv. 2016, 40 (1), 29–36. https://doi.org/10.1111/jfpp.12580.
Nieto, S.; Garrido, A.; Sanhueza, J.; Loyola, L. A.; Morales, G.; Leighton, F.; Valenzuela, A.
Flavonoids as Stabilizers of Fish Oil: An Alternative to Synthetic Antioxidants. J. Am. Oil
Chem. Soc. 1993, 70 (8), 773–778. https://doi.org/10.1007/BF02542599.
Nisar, M. F.; Arshad, M. S.; Yasin, M.; Arshad, M. U.; Nadeem, M. T. Influence of Irradiation
and Moringa Leaf Powder on the Amino Acid and Fatty Acid Profiles of Chicken Meat
Stored under Various Packaging Materials. J. Food Process. Preserv. 2019, 43 (9), e14166.
https://doi.org/10.1111/jfpp.14166.
Nisar, M. F.; Arshad, M. S.; Yasin, M.; Khan, M. K.; Afzaal, M.; Sattar, S.; Suleria, H. A. R.
Evaluation of Gamma Irradiation and Moringa Leaf Powder on Quality Characteristics of
Meat Balls under Different Packaging Materials. J. Food Process. Preserv. 2020, 44 (10),
e14748. https://doi.org/10.1111/JFPP.14748.
Odetayo, T.; Sithole, L.; Shezi, S.; Nomngongo, P.; Tesfay, S.; Ngobese, N. Z. Effect of
Nanoparticle-Enriched Coatings on the Shelf Life of Cavendish Bananas. Sci. Hortic.
(Amsterdam). 2022, 304, 111312. https://doi.org/10.1016/j.scienta.2022.111312.
Ong, G.; Kasi, R.; Subramaniam, R. A Review on Plant Extracts as Natural Additives in
Coating Applications. Prog. Org. Coatings 2021, 151, 106091. https://doi.org/10.1016/j.
porgcoat.2020.106091.
Ouahrani, S.; Tzompa‐Sosa, D. A.; Dewettinck, K.; Zaidi, F. Oxidative Stability, Structural,
and Textural Properties of Margarine Enriched with Moringa Oleifera Leaves Extract. J.
Am. Oil Chem. Soc. 2022, 99 (6), 485–499. https://doi.org/10.1002/aocs.12586.
Oyeyinka, A. T.; Oyeyinka, S. A. Moringa Oleifera as a Food Fortificant: Recent Trends
and Prospects. J. Saudi Soc. Agric. Sci. 2018, 17 (2), 127–136. https://doi.org/10.1016/j.
jssas.2016.02.002.
Patriota, L. L. de S.; Ramos, D. de B. M.; Gama e Silva, M.; dos Santos, A. C. L. A.; Silva,
Y. A.; Marinho, A. de O.; Coelho, L. C. B. B.; Paiva, P. M. G.; Pontual, E. V.; Mendes, R.
L.; Napoleão, T. H. The Trypsin Inhibitor from Moringa Oleifera Flowers (MoFTI) Inhibits
Acute Inflammation in Mice by Reducing Cytokine and Nitric Oxide Levels. South African
J. Bot. 2021, 143, 474–481. https://doi.org/10.1016/j.sajb.2021.09.032.
Pontual, E. V.; Carvalho, B. E. A.; Bezerra, R. S.; Coelho, L. C. B. B.; Napoleão, T. H.; Paiva,
P. M. G. Caseinolytic and Milk-Clotting Activities from Moringa Oleifera Flowers. Food
Chem. 2012, 135 (3), 1848–1854. https://doi.org/10.1016/j.foodchem.2012.06.087.
Purgatorio, C.; Serio, A.; Chaves-López, C.; Rossi, C.; Paparella, A. An Overview of the
Natural Antimicrobial Alternatives for Sheep Meat Preservation. Compr. Rev. Food Sci.
Food Saf. 2022, 21 (5), 4210–4250. https://doi.org/10.1111/1541-4337.13004.
Purohit, S. R.; Rana, S. S.; Idrishi, R.; Sharma, V.; Ghosh, P. A Review on Nutritional,
Bioactive, Toxicological Properties and Preservation of Edible Flowers. Futur. Foods 2021,
4, 100078. https://doi.org/10.1016/j.fufo.2021.100078.
Razzaghi-Abyaneh, M.; Shams-Ghahfarokhi, M.; Rezaee, M. B.; Sakuda, S. Natural Aflatoxin
Inhibitors from Medicinal Plants. Mycotoxins Food, Feed Bioweapons 2010, 329–352.
https://doi.org/10.1007/978-3-642-00725-5_19/COVER.
Reis, D. R.; Ambrosi, A.; Luccio, M. Di. Encapsulated Essential Oils: A Perspective in Food
Preservation. Futur. Foods 2022, 5, 100126. https://doi.org/10.1016/j.fufo.2022.100126.
Rodríguez, G. M.; Sibaja, J. C.; Espitia, P. J. P.; Otoni, C. G. Antioxidant Active Packaging
Based on Papaya Edible Films Incorporated with Moringa Oleifera and Ascorbic Acid
for Food Preservation. Food Hydrocoll. 2020, 103, 105630. https://doi.org/10.1016/j.
foodhyd.2019.105630.
Rodríguez-Pérez, C.; Gilbert-López, B.; Mendiola, J. A.; Quirantes-Piné, R.; Segura-
Carretero, A.; Ibáñez, E. Optimization of Microwave-Assisted Extraction and Pressurized
Liquid Extraction of Phenolic Compounds from Moringa Oleifera Leaves by Multiresponse
Surface Methodology. Electrophoresis 2016, 37 (13), 1938–1946. https://doi.org/10.1002/
elps.201600071.
Salem, A. S.; Salama, W. M.; Ragab, W. A. Prolonged Shelf Life of Sour Cream by Adding
Moringa Oleifera Leaves Extract (MOLE) or Moringa Oleifera Oil (MOO). Am. J. Food
Technol. 2015, 10 (2), 58–67. https://doi.org/10.3923/ajft.2015.58.67.
Sánchez-Muñoz, M.; Valdez-Solana, M.; Avitia-Domínguez, C.; Ramírez-Baca, P.; Candelas-
Cadillo, M.; Aguilera-Ortíz, M.; Meza-Velázquez, J.; Téllez-Valencia, A.; Sierra-Campos,
E. Utility of Milk Coagulant Enzyme of Moringa Oleifera Seed in Cheese Production from
Soy and Skim Milks. Foods 2017, 6 (8), 62. https://doi.org/10.3390/foods6080062.
Saucedo-Pompa, S.; Torres-Castillo, J. A.; Castro-López, C.; Rojas, R.; Sánchez-Alejo, E. J.;
Ngangyo-Heya, M.; Martínez-Ávila, G. C. G. Moringa Plants: Bioactive Compounds and
Promising Applications in Food Products. Food Res. Int. 2018, 111, 438–450. https://doi.
org/10.1016/j.foodres.2018.05.062.
Shah, M. A.; Bosco, S. J. D.; Mir, S. A. Effect of Moringa Oleifera Leaf Extract on the
Physicochemical Properties of Modified Atmosphere Packaged Raw Beef. Food Packag.
Shelf Life 2015, 3, 31–38. https://doi.org/10.1016/j.fpsl.2014.10.001.
Sharma, K.; Kumar, M.; Waghmare, R.; Suhag, R.; Gupta, O. P.; Lorenzo, J. M.; Prakash, S.;
Radha; Rais, N.; Sampathrajan, V.; Thappa, C.; Anitha, T.; Sayed, A. A. S.; Abdel-Wahab,
B. A.; Senapathy, M.; Pandiselvam, R.; Dey, A.; Dhumal, S.; Amarowicz, R.; Kennedy,
J. F. Moringa (Moringa Oleifera Lam.) Polysaccharides: Extraction, Characterization,
Bioactivities, and Industrial Application. Int. J. Biol. Macromol. 2022, 209, 763–778.
https://doi.org/10.1016/j.ijbiomac.2022.04.047.
Sharma, P.; Wichaphon, J.; Klangpetch, W. Antimicrobial and Antioxidant Activities of
Defatted Moringa Oleifera Seed Meal Extract Obtained by Ultrasound-Assisted Extraction
and Application as a Natural Antimicrobial Coating for Raw Chicken Sausages. Int. J. Food
Microbiol. 2020, 332, 108770. https://doi.org/10.1016/J.IJFOODMICRO.2020.108770.
Soni, A.; Pandiyan, P.; Elumalai, P. Effective Treatment of Curry ( Murraya Koenigii ) and
Moringa ( Moringa Oleifera ) Leaves Extracts on Melanosis of Pacific White Shrimp
(Litopenaeus Vannamei) During Chilled Storage. J. Aquat. Food Prod. Technol. 2021, 30
(10), 1304–1314. https://doi.org/10.1080/10498850.2021.1988792.
Sultana, S. Nutritional and Functional Properties of Moringa Oleifera. Metab. Open 2020, 8,
100061. https://doi.org/10.1016/j.metop.2020.100061.
Tajalsir, A. E.; Ebraheem, A. S. A.; Abdallah, A. M.; Khider, F. J.; Elsamani, M. O.; Mohamed
Ahmed, I. A. Partial Purification of Milk-Clotting Enzyme from the Seeds of Moringa
Oleifera. J. Microbiol. Biotechnol. Food Sci. 2014, 4 (1), 58–62. https://doi.org/10.15414/
jmbfs.2014.4.1.58-62.
Teixeira, E. M. B.; Carvalho, M. R. B.; Neves, V. A.; Silva, M. A.; Arantes-Pereira, L.
Chemical Characteristics and Fractionation of Proteins from Moringa Oleifera Lam.
Leaves. Food Chem. 2014, 147, 51–54. https://doi.org/10.1016/j.foodchem.2013.09.135.
Terefe, M. A.; Kebede, A.; Kebede, M.; Sciences, C. Clotting Activities of Partially Purified
Extracts of Moringa Oleifera L. on Dromedary Camel Milk. East African J. Sci. 2017, 11
(2), 117–128.
Valdés-Hernández, Geidy Vivian; Cruz-Viera, Luis; Comet-Rodríguez, R. Influencia de Las
Condiciones de Operación En La Extracción de Polifenoles a Partir de Hojas de Moringa
Oleifera Lam. CENIC. Ciencias Químicas ISSN 2015, 46, 135–145.
Vongsak, B.; Sithisarn, P.; Mangmool, S.; Thongpraditchote, S.; Wongkrajang, Y.; Gritsanapan,
W. Maximizing Total Phenolics, Total Flavonoids Contents and Antioxidant Activity of
Moringa Oleifera Leaf Extract by the Appropriate Extraction Method. Ind. Crops Prod.
2013, 44 (November 2017), 566–571. https://doi.org/10.1016/j.indcrop.2012.09.021.
Wang, Y.-Y.; Peng, C.; Zhang, Y.; Wang, Z.-R.; Chen, Y.-M.; Dong, J.-F.; Xiao, M.-L.; Li,
D.-L.; Li, W.; Zou, Q.-J.; Zhang, K.; Wei, P. Optimization, Identification and Bioactivity
of Flavonoids Extracted from Moringa Oleifera Leaves by Deep Eutectic Solvent. Food
Biosci. 2022, 47, 101687. https://doi.org/10.1016/j.fbio.2022.101687.
Wu, G.; Chang, C.; Hong, C.; Zhang, H.; Huang, J.; Jin, Q.; Wang, X. Phenolic Compounds
as Stabilizers of Oils and Antioxidative Mechanisms under Frying Conditions: A
Comprehensive Review. Trends Food Sci. Technol. 2019, 92, 33–45. https://doi.
org/10.1016/j.tifs.2019.07.043.
Zhang, T.; Jeong, C. H.; Cheng, W. N.; Bae, H.; Seo, H. G.; Petriello, M. C.; Han, S. G.
Moringa Extract Enhances the Fermentative, Textural, and Bioactive Properties of Yogurt.
LWT 2019, 101 (June 2018), 276–284. https://doi.org/10.1016/j.lwt.2018.11.010.
Zhao, B.; Lan, T.; Li, H.; He, Y.; Wu, D.; Chen, Z. Antioxidation Activity of Moringa Oleifera
Lam. Leaves Extract on Soybean Oil during Both Storage and Thermal Treatment. J. Food
Process. Preserv. 2019, 43 (8), e13975. https://doi.org/10.1111/jfpp.13975.
Zhuang, S.; Hong, H.; Zhang, L.; Luo, Y. Spoilage‐related Microbiota in Fish and Crustaceans
during Storage: Research Progress and Future Trends. Compr. Rev. Food Sci. Food Saf.
2021, 20 (1), 252–288. https://doi.org/10.1111/1541-4337.12659.
CHAPTER 6
Storage of Fruits and Vegetables

in Controlled Atmospheres and Its
Application in the Preservation of White
Onion Quality
CELIA CHÁVEZ-MENDOZA and ALEXANDRO GUEVARA-AGUILAR
Centro de Investigación en Alimentación y Desarrollo A C,
Coordinación Tecnología de Productos Hortofrutícolas y Lácteos,
Delicias, Chihuahua, México
ABSTRACT
Controlled atmosphere (CA) technology consists of storing fruits and
vegetables in a refrigeration chamber in which the initial or normal atmo-
sphere is replaced by another one that is poorer in oxygen and richer in
CO2. Through this technology, it is possible to control various metabolic
processes of fruits and vegetables, thus delaying the process of maturation
and senescence and maintaining their nutritional quality and optimal state
of maturation until consumption. However, if it is not applied correctly, it
can cause various quality problems. Changing the normal atmosphere to a
controlled one requires the use of gases such as oxygen, carbon dioxide, and
nitrogen, which are mixed to achieve the desired storage concentration. There
are several types of AC systems, which depend on the way of generating the
atmosphere and the concentrations of O2 and CO2 used. CAs are currently
used commercially almost exclusively for the storage of apple, pear, kiwi,
and cabbage. In Mexico, they are mainly used to preserve apples and pears in
the states of Chihuahua and Coahuila, but they have not been used in onions
despite being a seasonal product and widely consumed. Experimental studies
have shown that this vegetable, being a nonclimacteric product, responds
well to this technology, managing to preserve its quality for up to 7 months.

The use of CA, its characteristics, and its application, with emphasis on the
conservation of onion quality, is briefly described in this chapter.
6.1 INTRODUCTION
The recent concern of consumers and health organizations to maintain a
healthy diet has increased the demand for fresh fruits and vegetables to be
consumed on a daily basis; this is because they represent an important source
of antioxidant compounds that are beneficial against chronic diseases,
including cancer and cardiovascular diseases. This scenario has created new
opportunities for the horticultural industry to improve the quality of fruits
and vegetables through their nutritional value (López et al., 2011). However,
in many of these products, the storage period is short, damaging their quality
and limiting their commercialization, thus missing out on market niches.
An alternative to this drawback is the use of controlled atmospheres (CAs),
a technology that involves storage in low O2 and/or high CO2 concentra-
tions to delay the rate of respiration, ethylene production, and weight loss,
and therefore the fruit deterioration (Crisosto et al., 2002) The use of CAs
represents one of the most important advances in postharvest technology, as
the gas composition in storage has an impact on the shelf life of the stored
product, and is used to prevent damage to harvested fruits and vegetables
during storage, transport and distribution (Arshad et al., 2023).
There are many variables that affect the fruit degradation during postharvest
storage. Some of these variables are damage by microorganisms, the storage
environment, and the maturity at which they were harvested, among others.
Therefore, storage requirements may be different for each type of fruit, and they
tend to behave differently after harvest; for this reason, they are classified as
climacteric or nonclimacteric, and there are fruits with intermediate behavior.
Climacteric fruits show a growth in respiration rate during their decomposition
stage that is associated with ethylene production, and in nonclimacteric fruits,
their chemical composition is maintained over time, except for sugars that are
decomposed for metabolic processes. This leads to a greater number of physi-
ological changes in climacteric fruits that can be reduced by manipulating the
atmosphere in which they are stored (Thompson, 2016).
The scientific basis for the application of CA in fruit and vegetable storage
has been the subject of several research studies and has evolved tremendously,
some of the science dates back 200 years, but it was first refined and commercially
applied in the mid-20th century (Thompson, 2010). Commercially, it has been
applied in apples and pears due to its special adaptation to CA conditions. But
Storage of Fruits and Vegetables in Controlled Atmospheres 137
its effect has been evaluated in other fruits such as pitahaya (Magaña-Benitez et
al., 2010); papaya (Martins and De Resende, 2013); blackberry (Brackmann et
al., 2016); lychee (Ali et al., 2021); avocado (Chirinos et al., 2021; Fuentealba et
al., 2022); banana, mango, pineapple, cherimoya and rambutan (Kader, 1994).
In vegetables, it has been applied in tomato storage (Dilmaçünal et al., 2022);
in the preservation and shelf-life extension of spinach (Martínez-Damina and
Cantwell de Trejo, 2002), and in a more limited way in onion (Rios-Gonzalez
et al., 2018; Smittle, 1988; Yamashita et al., 2009).
Onion is one of the oldest and most widely consumed crops in the world,
is highly valued for its characteristic pungent flavor, and is an essential ingre-
dient for cooking worldwide. It is a seasonal vegetable and is sometimes
overproduced (Nega et al., 2015). Produced in many countries around the
world, it is the second most important crop after tomato. China is the leading
producer worldwide, followed by India and the United States. It is also the
most important income-generating crop in small, medium, and large-scale
production in many developing countries (Muhie, 2022).
On the contrary, one of the characteristics of this vegetable crop is the
susceptibility to postharvest losses due to its high water and low dry matter
content, although it is less perishable than other crops, losses are unacceptable
during medium and long storage periods; it has been reported that 40–50%
of the product never reaches the ultimate user/consumer due to postharvest
losses (Gorreapti et al., 2017).
Quality losses of this vegetable are due to several factors because, during
their collection, handling, transport, packaging, and storage, they are exposed
to several treatments and environmental conditions which not only can affect
their quality attributes and physiological characteristics but could also be
responsible for several reactions and stresses, causing important biochemical
changes in the bulb tissues (Benkeblia and Varoquaux, 2003).
Onion is classified as a low perishability product because it maintains a low
respiration rate (Kadder, 2002). In addition, after harvest, it enters a natural
state of dormancy that favors its conservation. This phenomenon is controlled
by several endogenous hormones that vary depending on the genetic compo-
sition of each variety, environmental factors, and in particular temperature,
which can extend or shorten it (Sharma et al., 2016). However, during storage,
especially for prolonged periods, there are losses of marketable bulbs mainly
due to sprouting, rooting, decay, weight loss, or phytopathological damage
(González et al., 2005). Prolonged storage can also significantly affect the
appearance and chemical composition of bulbs (Sharma et al., 2014).
CA has been proven that it could be used as an environmentally friendly
technology for prolonged storage of fruits and vegetables including onions
(Poldma et al., 2012). This technology helps to preserve its quality allowing
to offer quality products to the market in times of low supply and high prices,
satisfying the demand that exists throughout the year, some researchers have
used this technology and obtained positive results (Chávez-Mendoza et al.,
2018; Garba et al., 2014; Poldma et al., 2012).
Therefore, characteristics and importance of the technology of crop
storage in CAs are discussed in this chapter, as well as its application in
onion preservation.
6.2 CONTROLLED ATMOSPHERE TECHNOLOGY

Controlled atmosphere technology consists of storing fruits and vegetables in
a refrigeration chamber in which the initial or normal atmosphere is character-
ized by a concentration of 20.95% oxygen (O2), 0.03% carbon dioxide (CO2),
and 78.09% nitrogen (N2) is replaced by an atmosphere poorer in oxygen
and richer in CO2, maintaining precise control of the concentrations of these
gases during storage or transport (Graell and Ortiz, 2003). Air composition
is adjusted based on product requirements and the change of the atmosphere
is achieved by certain devices such as nitrogen generators, CO2, ethylene
absorbers, etc. Inside the chamber, the environmental physical conditions
such as temperature, humidity, and air circulation must also be controlled.
CA is successful in fruit and vegetable storage only when combined with
refrigeration or low temperatures. Therefore, standard refrigeration units are
integral components of CA storage systems.
This technique associated with cold, reiterates the effect of refrigeration on
the vital activity of the tissues, avoiding certain physiological problems and
reducing losses due to rotting. The action of the atmosphere on fruit respira-
tion is much more important than the action of low temperatures. A controlled
atmosphere delays biochemical reactions causing slower respiration and a
delay in ripening, the fruit being in a dormant condition, with the possibility
of vegetative reactivation once the fruit is placed in normal atmospheric air.
It is desirable that the created atmosphere composition remains constant
over time. However, internal changes (metabolic reactions) of certain products
consume some gases (oxygen) and generate others (carbon dioxide, ethylene)
that alter this initial composition. These variations are detected by monitoring
devices and compensated by various gas production/removal mechanisms.
These systems cannot be implemented in small packages for retail sales.
CA technology is relatively expensive compared to regular storage, but it
adds value to the crop by extending the marketability of the product, which
is important for those fruits or varieties that have a short shelf life such as
sweet onions (Poldma et al 2012).
This technique has been improved and is currently very important in the
preservation of foods of both vegetable and animal origin. Currently, large
quantities of fruits and vegetables, such as apples, pears, kiwis, sweet onions,
and cabbage, are stored in CA. Furthermore, this storage technology is also
widely used in several countries during the maritime transport of various
fruits, such as bananas, mangoes, avocados, and apples (Yahia et al., 2019).
6.3 GASES EMPLOYED IN CONTROLLED ATMOSPHERES

Changing the normal atmosphere to a CA basically requires three types of
gases: oxygen, carbon dioxide, and nitrogen, which are mixed to achieve the
desired concentration.
6.3.1 OXYGEN
A decrease in the amount of oxygen in the atmosphere results in reduced respira-
tory intensity and lower ethylene biosynthesis in fruits and vegetables (Graell and
Ortiz, 2003). It is a necessary compound for the metabolism of living organisms
and is involved in oxidation processes. It is usually reduced in CA chambers,
which may favor the growth of anaerobic pathogens (Yahia et al., 2019).
6.3.2 NITROGEN
It is employed to displace oxygen, thereby decreasing the growth of aerobic
spoilage microorganisms and protecting against food oxidation (Parry,
1993). It is also used to balance the atmosphere and achieve the desired
concentration inside the chamber.
6.3.3 CARBON DIOXIDE

The increase in the level of CO2 decreases the respiratory intensity and inhibits
the action of ethylene, this reduces the metabolic activity of plant tissues (in
medium concentrations) which enter a state of “dormancy.” Also inhibits the
activity of microorganisms (in high concentrations) (Graell and Ortiz, 2003;
Yahia, 1995b). CO2 dissolves in water to form carbonic acid and thus lowers
the cellular pH. It also modifies the enzymatic activity of several metabolic
pathways. Greater solubility at low temperatures and therefore more effective
in controlling microorganisms under these conditions (Yahia et al., 2019).
6.4 CA STORAGE SYSTEMS

There are several types of CA storage systems, some of the most commonly
used are described in the following:
a. Conventional CA (CCA)
This system stores the fruit under hermetic conditions and allows
the fruit to change the surrounding atmosphere due to the process
of respiration, which causes the amount of oxygen to decrease
and the amount of CO2 to increase until the necessary atmosphere
is established. Gas levels should be monitored daily and when the
desired concentrations are achieved, gas control begins. O2 levels
are maintained by the introduction of air from outside the room and
CO2 levels are maintained by removal. In this system, the time in
which the desired concentrations are achieved can be very long. In
apples, for example, it takes about 10 days and during this time the
fruit can continue to mature (Yahia, 1995b). In apples, a concentra-
tion of 3kPa O2 and 3kPa CO2 has been used (Yahia et al., 2019).
b. Fast conventional CA (FCCA)
In this system, the atmosphere is generated immediately after the
chamber is sealed. Oxygen concentration is reduced to desired levels
in less than two days using nitrogen or CA generators. A precise
control of the gas concentration, temperature, and relative humidity
of the chamber is carried out by means of certain analyzing devices.
It is currently the most commercially used system for the storage of
apples, pears, and cabbage (Yahia, 1995c).
c. Ultra-low oxygen CA (ULOCA)
In this system, the oxygen concentration is reduced to the minimum
levels tolerable by the fruit. It is a type of FCCA that uses very low O2
concentrations (less than 1%) and without using high levels of CO2.
Such low O2 levels are very critical, and a highly efficient monitoring
and control system is needed to prevent the amount of oxygen from
dropping to dangerous levels for the fruit (fermentation).
Preservation under such O2 conditions is called ultralow oxygen
(ULO). ULO preservation is carried out in airtight preservation
chambers and is used for the long-term preservation of apples, pears,
blueberries, and kiwi.
In Golden Delicious apples, for example, it was observed that the

use of this system preserved firmness and achieved good acidity
after storage for 6–10 months, especially when ULOCA was applied
in a rapid manner. It also decreased some important physiological
disorders such as darkening of the pulp core, reduced ethylene
biosynthesis, decreased respiration rate, and susceptibility to decay.
Special care must be taken when using this atmosphere as it may
favor the development of anaerobic processes in the fruit causing the
accumulation of fermentation-derived compounds such as ethanol,
ethyl acetate, and acetaldehyde (Yahia, 1995b).
The main advantages achieved with low oxygen atmospheres,
compared to standard CA, are longer shelf life and subsequent product
shelf life, better quality retention during storage and upon release, and
a reduction of certain physiological alterations (Graell et al., 1998).
d. High carbon dioxide CA
In this system, high amounts of CO2 (10–15% in the case of apple)
are used before starting with the ideal atmosphere. This CA has been
used for two decades in apples, but its commercial use is limited to
the northwestern United States for Golden Delicious apples. When
applied to this fruit, it was observed that using a CO2 concentration
of 10–15% for 2–4 weeks and a temperature between 0 and 5°C, it
was possible to reduce the texture and quality loss and prolong the
shelf life of the fruit (Yahia,1995b).
e. Low-ethylene CA (LECA)
This system began to be used commercially in 1983 in the Northwest
USA and consists of a fast CA with low O2 levels, high CO2 levels,
and strict control of ethylene levels in pre-harvest and postharvest
(Yahia, 1995c).
Levels of this hormone in this system are maintained at a concentra-
tion of 1 ppm or less. By storing “Mclntosh” apples in this system
for 8 months, the fruit only lost 3.5% of its initial firmness compared
to a 24.2% loss of firmness of apples stored in CCA (Yahia, 1995b).
f. Insecticidal CA
In this system, very low amounts of O2 and high amounts of CO2 are
used to control insects as an alternative to the use of chemicals.
Atmosphere levels needed for insect control in very short periods

are 0.5% O2 or less and 50% CO2 or more. These can control insects
and have the potential for use in postharvest, especially as quarantine
treatments. Insect response depends on their stage of development, but
many insects can be controlled within 2–4 days at room temperature.
CA use for insect control is very advantageous because it does
not leave toxic residues in the food and the cost is competitive
compared to the application of agrochemicals. However, insecticidal
atmospheres are very extreme and can cause fruit problems such as
anaerobiosis and fermentation. Therefore, it is very important to use
them only on those fruits that can tolerate these levels of atmosphere
and only for a period shorter than the tolerated period (Yahia, 1995a).
g. Dynamic control system CA (DCS)
This system consists of maintaining oxygen levels within the
minimum limits tolerated by the fruit, for which it is monitored by
sensors, and its levels are periodically adjusted.
CA conditions are dynamically regulated during storage (instead of
being set in advance). The CA dynamic conservation system equip-
ment reacts to the physiological circumstances of the fruit inside the
chamber, adjusting the conservation conditions to them automati-
cally. Achieving oxygen concentration percentages below 1% is not
a problem, as the oxygen concentration is adjusted directly as soon
as there is a risk of it being too low.
So far, the commercial use of DCS has only been evaluated in
apples, in which it favors the maintenance of firmness, and acidity,
and reduces the development of some physiopathologies without
affecting the sensory quality of the fruit (Calvo and Caldan, 2012).
6.5 CONTROLLED ATMOSPHERE APPLICATION

The atmosphere to be used for a given product is based on recommendations
derived from research carried out by many fruit research centers around the
world; it is necessary to contrast these recommendations in each specific area
given the influence of environmental factors on the behavior of the species
and/or varieties (Graell and Ortiz, 2003).
It is recommended that the desired product temperature and atmosphere
composition be reached within a maximum of 7–10 days after the food is
placed in storage before ripening begins, that is, when the ethylene in the
fruit is less than 1 ppm (Bishop, 1990).
CAs are currently used commercially almost exclusively for apple, pear,
kiwi, and cabbage storage. In Mexico, they are mainly used to store apples
and pears in the states of Chihuahua and Coahuila (Yahia, 1995b). Table 6.1
shows gas concentrations and temperatures for CA storage of various fruits
and vegetables.
TABLE 6.1 Recommended Gas Concentrations for the Equilibrium Atmosphere and
Temperature Range for the Preservation of Fruit and Vegetable Products.
Product Temperature (°C) %O2 %CO2
Apricot 0–5 2–3 2–3
Avocado 5–13 2–5 3–10
Broccoli 0–5 1–2 5–10
Pumpkin 0–5 3–5 5–7
Cherry 0–5 3–10 10–12
Brussels sprouts 0–5 1–2 5–7
Cauliflower 0–5 2–5 2–5
Mushroom 0–5 Air 10–15
Asparagus 0–5 Air 5–10
Spinach 0–5 Air 10–20
Strawberry 0–5 10 15–20
Pomegranate 4–6 5–10 0–2
Kiwi 0–5 2 5
Lettuce 0–2 2–5 0
Lemon 12–14 5–10 0–2
Tangerine 4–5 10–12 0–2
Mango 10–15 5 5
Apple 0–5 2–3 1–2
Cantaloupe melon 3–7 3–5 10–15
Peach 0–5 1–2 5
Orange 2–4 10–12 0–2
Papaya 10–15 5 10
Pear 0–5 2–3 0–1
Cucumber 8–12 3–5 0
Semi-ripe tomato 8–12 3–5 0
Pineapple 10–15 5 10
Olive 8–12 2–5 2–10
Source: Artés (1987); Kader (1990), Marcellin (1992), IIR (1995), Yahia et al. (2019).
6.6 DISADVANTAGES OF USING CONTROLLED ATMOSPHERES

Improper use of the CA and MA may cause some problems and result in
product loss (Yahia, 1995b). Some of the potential problems are discussed in
the following subsections.
6.6.1 ANAEROBIOSIS
Exposure of fresh products to very low O2 and/or very high CO2 concentra-
tions that are outside the tolerance limits for a particular product may cause
respiration to change from aerobic (with oxygen) to anaerobic (without
oxygen). Anaerobiosis refers to the fermentation of a fruit or vegetable,
which causes bad odors and flavors (Day, 1993; Farber, 1991).
6.6.2 DEVELOPMENT OF BAD FLAVORS AND ODORS

Long-term storage of fruits in CA may decrease their ability to develop their
characteristic flavor and aroma (Yahia, 1995b). Bad tastes and odors may also
occur at very low O2 concentrations as a result of anaerobic respiration and
very high CO2 concentrations due to fermentative metabolism (Kader, 2004).
6.6.3 PHYSIOLOGICAL DISORDERS

CA can stimulate or aggravate some physiological disorders of fruit and
vegetable products, for example, internal browning in pears and apples,
brown spots in lettuce, chilling injury of some products (Kader, 2004), and
freezing damage to texture.
6.6.4 INCREASED SUSCEPTIBILITY TO FUNGAL ATTACK
Inadequate gas levels can facilitate the penetration of fungi and other micro-
organisms through plant tissues (Yahia, 1995b).
6.6.5 BACTERIOLOGICAL ISSUES
Very low O2 and/or very high CO2 concentrations can stimulate the growth of
certain pathogenic bacteria such as Clostridium and Campylobacter, which
is a potential risk in products stored in modified atmospheres (Farber, 1991).
6.6.6 ABNORMAL RIPENING OF FRUITS AND VEGETABLES
Inadequate atmospheres can cause abnormal or irregular ripening of products

(Kader et al., 1989). This can occur in banana, mango, pear, and tomato with
O2 levels below 2% and CO2 levels above 5% (Kader, 2004).
6.6.7 HIGH INITIAL COST
A relatively high initial investment is required for equipment acquisition and

chamber construction with adequate conditions for CA storage, as well as
hiring trained personnel to operate the system. Furthermore, gas concentra-
tions are different for each particular product (Day, 1993; Farber, 1991).
6.7 ONION PRESERVATION
In onions, the organ of consumption is generally the bulb, which, unlike

other vegetable species, has some characteristics that favor conservation
for a relatively long time. Under photoperiod conditions suitable for bulb
formation, the plant initiates reserve accumulation at the base of the leaves,
forms cataphylls without laminae, and ends with the leaf primordia initia-
tion, before entering a dormancy period. During this period the bulbs have
a reduced metabolism and do not have the capacity to emit leaves and roots,
which facilitates conservation. Depending on the variety, growth restarts after
a variable number of days. Inner cataphylls are thick and firm, while outer
cataphylls are formed by dead cells, which form a barrier to gas exchange
and a protective element against mechanical damage and fungal attack.
Bulbs have low respiration rates, which can be considered as an indicator
of low metabolic activity, a characteristic also favorable for conservation
(Furlani and Rivero, 2010; UCDavis, 2022).
The main purpose of onion storage in CA is to preserve bulbs with
commercial qualities for a prolonged period of time.
Postharvest quality of onion is strongly influenced by cultivar, cultural
practices, climatic conditions during the growing and development period,
as well as storage conditions. Well-preserved onions are generally long-day
onions with prolonged dormancy periods, while short-day cultivars gener-
ally have storability. Fertilization, mainly nitrogen fertilization, also affects
onion storage, as well as late harvests (Poldma et al., 2012).
6.8 ONION QUALITY CRITERIA
Onion quality indices can be objective or subjective and should serve to

accurately express the quality and postharvest life of the product in different
situations.
Some of the main characteristics to be taken into account to define onion
quality are color, absence of sprouting and roots, firmness, soluble solids,
Dry matter, size, weight, pungency, and phytopathological damage.
a. Color. Onion commercial value is strongly dictated by its aesthetic
appearance, related in an important way to skin color, due to the high
sensitivity of the consumer to this quality factor (Kiura et al., 2021).
Onion color is defined according to the coloration of the epidermis
or skin, this parameter is verified visually or by means of a colorim-
eter. Onions consumed in Mexico are white, purple, and yellow in
color. In white onions, the bulb must have a white skin covering at
least 80% of its surface at the time of harvesting, while purple and
yellow onions must be 100% covered by these colors at the time of
harvesting (Norma Mexicana NMX-FF-021-SCFI-2003).
Bulb color can be affected by handling. It has been observed that
on onions of white varieties or those with few protective cataphylls,
a greenish coloration can appear on the external scales, which is
caused by exposing the bulbs to direct sunlight for a few days or
when they are kept in brightly lit places. This disorder detracts from
the commercial quality and may impart a strange taste to the product.
Onion color has been related to its antioxidant capacity. It was
found that red onions have a higher antioxidant capacity than yellow
and white onions, although yellow onions are richer in phenolic
compounds (Gokce et al., 2010).
b. Root sprouting and emission. The term sprouting in onion
refers to the new leaves that begin to develop from the bulb
after having been at rest for a period of time (Mexican Standard
NMX-FF-021-SCFI-2003).
Sprouting rate during onion storage is closely related to different
agronomic parameters such as soil moisture, nitrogen fertilization,
bulb ripening at harvest time, water stress, and variety (Sorensen
and Grevsen, 2002; Miedema, 1994).
Root sprouting and emission from bulbs not only accelerates spoilage
but also determines the end of its shelf life since the consumer rejects
the presence of sprouts and/or roots emerging from the product. Root
emission affects the visual quality of the onion and may furthermore
predispose to further deterioration due to root rot. It is caused by
excessive humidity during storage.
Once they have completed their development, bulbs enter a state of
"rest" or dormancy, characterized by a very reduced physiological
activity that does not respond to environmental conditions, that is,
they do not sprout even under optimal humidity and temperature
conditions. Different studies have shown that sprouting inhibitory
compounds such as abscisic acid predominate in this stage over
promoters such as gibberellins, auxins, and others. This balance
changes with storage time leading to a state in which it sprouts
or emits roots if exposed to favorable environmental conditions.
There is no clear differentiation between one state and another, but
rather a slow transition. As time goes by, promoters predominate,
and sprouting irremediably takes place. Refrigerated storage and
CAs slow down sprouting and rooting (Lopez, 2003; Sharma et al.,
2016). Sprouting in onions is considered to be the main contributor
to postharvest losses (20–40%) of total storage losses (50–90%), and
as a result, sprouted onions are discarded and placed in containers,
so no attention has been paid to onions with this condition, however,
it has been found that these onions contain high phenolic content and
high antioxidant activity (Majid et al., 2016).
Another physiological disorder of onion is sun damage, which is
the result of prolonged exposure of the onion to solar radiation.
The problem is associated with high temperatures and/or ultraviolet
rays. It manifests itself in the form of blisters and rupture of the
peridermis, especially when the temperature of the product is close
to 50°C (Furlani and Rivero, 2010).
c. Firmness. Bulb consistency or firmness is a very important quality
characteristic. Bulbs should be firm and not yield to strong pressure.
Soft bulbs are the result of excessive dehydration, sun or mechanical
damage, or due to the presence of a physiological disorder known
as translucent cataphyll. Also, the neck of the bulb should be narrow
and dry. The presence of wide necks indicates poor curing, initiation
of bud break, flower scape production, or root growth (Furlani and

Rivero, 2010).
d. Soluble solids. Soluble solids values are considered a good approxi-
mation of the onion sugar content. This parameter is related to the
shelf life and sweetness of this vegetable. There is a direct relation-
ship between the content of soluble solids and the percentage of dry
matter in onion and high amounts of these components indicate a
better aptitude for conservation (Fondo Europeo Agrícola de Desar-
rollo Rural, 2008).
e. Pungency. This parameter refers to the level of onion pungency and
is established by determining the pyruvic acid content of the bulbs.
The secondary organosulfur compounds of onion are responsible for
the pungent flavor and aroma. Thiosulfinates are the predominant
flavor compounds in raw onion and are formed when unflavored
S-alk(en) yl-l cysteine sulfoxides (ACSO) are lysed by alliinase after
tissue breakdown. The initial products of this reaction are sulfenic
acid, pyruvate, and ammonia. Depending on the alkenyl substituent
present, sulfenic acids either condense in pairs to form thiosulfinate
or organize to form thioprapanal sulfoxides or lachrymatory factors
(Lee et al., 2010).
There are several scales to measure the level of pungency, a simpli-
fied scale is used by the sweet onion industry in Georgia in the
United States of America, where pyruvic acid levels vary from 0
to 18 µmol/g of fresh onion (µmol/g). When values are between 0
and 3 µmol/g, they are classified as low pungency, when they are
between 3 and 7 medium pungency and if they are higher than 7, the
onions are classified as high pungency. Another scale is the one used
by Vidalia Labs International, Georgia, USA, where the scale ranges
from 0 to 10 µmol/g of fresh onion. When pyruvic acid values are
less than 3, they are classified as very mild, between 3 and 4 they are
classified as mild, between 4 and 5.5 they are classified as slightly
pungent, between 5.5 and 6 as pungent and greater than 6 as very
pungent. Thus, onions with a pyruvic acid level below 3.5 are called
extra sweet onions, between 3.6 and 5.5 sweet onions, and with
values above 5.6 spicy onions. The pungency level is a parameter
directly related to the variety and other agronomic parameters such
as fertilization, irrigation, climatic conditions, and soil type (Raigón,
2006).
f. Size onion. Size is determined by the weight or by the equatorial

diameter of the vegetable. According to Boyhan et al. (2004), the
small onion should measure <6.4 cm; medium, from ≥6.4 to <7.6
cm; large from ≥7.6 to ≤8.9 cm; and jumbo, >8.9 cm. Meanwhile, in
Mexico, ball onions must comply with the specifications presented
in Table 6.2 (NMX-FF-021-SCFI-2003).
TABLE 6.2 Size Specifications of Mexican Ball Onions.

Size Equatorial diameter range (cm)
Small Less than 3.5 cm
Medium small 3.6–5.0 cm
Medium 5.1–7.0 cm
Large 7.1–9.0 cm
Extra Greater than 9.0
Source: NMX-FF-021-SCFI-2003.
g. Dry matter. Dry matter is one of the most important factors in

onion quality, about 80% of the dry matter of the bulb is made up of
nonstructural carbohydrates such as sucrose, glucose, fructose, and
low molecular weight fructans. Sugar metabolism is closely linked
to onion dormancy and sprouting (Lee et al., 2010). Bulbs with high
amounts of dry matter are more suitable for prolonged storage and
also tend to contain a higher concentration of fructans, which repre-
sent the main carbohydrate reserve in most bulbs, and are converted
to fructose as a source of energy for sprouting (Chope et al., 2012).
h. Phytopathological damage. Numerous pathogens can cause
diseases that shorten shelf life and affect product quality. Most
common are Aspergillus niger, Botrytis spp., Erwinia carotovora,
Fusarium spp., Penicillium spp., and Rhyzopus spp.
In some cases, disease development can be initiated as a result of
nematode attacks that produce deformations and cracks in some
bulbs, which are not always eliminated in the selection process.
Overall, proper temperature management should be considered the
main weapon of control. Low temperatures coupled with low rela-
tive humidity are generally more effective than any other treatment,
including chemicals, in controlling diseases.
6.8.1 FUNGAL DAMAGE
A. niger (black mold). It is the main microorganism that attacks onions

during storage, especially at high temperatures and relative humidity. It
affects the outer parchment of the onion, producing black powdery spots
(fungal spores), which significantly detract from the commercial quality of
the product. Moreover, it can attack the fleshy cataphylls, showing more or
less sunken rotting grooves of dry consistency, depending on the environ-
mental humidity. It can move from diseased to healthy bulbs. The onion
surface must be kept dry during and after harvesting (storage) (Fernandez et
al., 1999).
When the entire bulb surface turns black, the onion shrivels, wrinkles
becomes secondarily infected with bacteria, and rots. After harvest, black
mold is avoided by transporting and storing the bulbs below 15°C or at very
low humidity. Infection is minimized by avoiding hitting and cutting the
bulbs.
Basal rot (Fusarium spp.). Generally, infection occurs in the field and
manifests itself in storage. Dry or semi-wet rot appears at the bulb base and
can affect the entire bulb. Damaged tissue becomes aqueous. Infection can
spread from diseased to healthy onions. It is one of the main causes of post-
harvest losses. Onion cultivation should be avoided in fields where onion
disease attacks have occurred in previous years (Fernández et al., 1999).
Blue mold. Watery rot on the collar and external scales, followed by the
appearance of blue–green (occasionally yellow–green) spores, is caused by
the fungus Penicillium. Bruising and other mechanical damage, sun scald,
and freeze damage should be minimized (UCDavis, 2022).
6.8.2 BACTERIAL DISEASES
These are characterized by soft rotting and often have an unpleasant odor.
Bacteria enter through wounds produced before or after harvest. The main
genera are Erwinia and Pseudomonas. Low temperatures and careful
management to avoid injury are the best preventive control of these diseases.
Bacterial Rot/Soft Rot. Characterized by watery, foul-smelling, slimy
liquid areas, this rot is caused by the bacterium E. carotovora subsp. Caroto-
vora (UCDavis, 2022). Erwinia can infect plants in the field. Leaves wilt and
eventually dry up. If a longitudinal cut of an infected plant is made, it will be
observed that the middle part of the new bulb appears completely loose and
very sticky. Coats become soft and watery in appearance. Later on, a light
yellow to light brown gelatinous substance will form. Affected bulbs are soft
and watery and, when squeezed, a sticky fluid or pulp comes out of the bulbs
(Bejo, 2011).
The bacteria that cause these problems in onion crops are only found in
soil, irrigation water, and in the remains of previous crops. Infection almost
always starts through wounds caused by insect damage, hail, heavy rain, and
strong wind. On the contrary, if it rains gently for a prolonged period, the
leaves may become soggy, either because they are held together or because
the leaf axils fill with water and begin to rot. Bacteria mostly enter the bulb
through wounds in the neck, but this is not always the case. The higher the
temperature, the faster the infection progresses. Bacteria are inactivated at
temperatures below 3°C (Bejo, 2011).
6.9 FACTORS AFFECTING THE CONSERVATION OF ONIONS
a. Variety. Generally, short-day varieties are less suitable for prolonged

storage than long-day varieties, as the latter have a higher dry
matter content and therefore have lower water losses and are less
susceptible to sprouting and postharvest diseases. However, signifi-
cant differences in dry bulb storability have been reported among
cultivars of the same photoperiod type, which may differ up to 3–3.5
months of storage period. Cultivar selection and pre-harvest condi-
tions, such as plant establishment methods (transplanting rather than
direct seeding) and harvest stage (early harvest when 20–50% of the
top of the plant has fallen off) can reduce the incidence of sprouting
after long-term storage. Other important crop characteristics that
can affect bulb storage capacity include the number and thickness
of coat layers and the pungency of inner layers (Petropoulos et al.,
2016; Grevsen et al., 2004).
b. Curing. Immediately after harvest, bulbs must undergo a drying
(“curing”) process to harden their outer scales, reduce skin cracks,
and allow the necks to narrow, thus inhibiting pathogen infections.
The curing method (field or forced air curing) and conditions
(temperature and relative humidity), as well as growing conditions
and harvest stage, can be of great importance to achieve maximum
quality of dried bulbs and minimum losses due to water losses and
pathogen infections (Petropoulos et al., 2016).
c. Temperature. It is one of the factors that have the greatest influ-

ence on the metabolic activity of fruit and vegetable products. If
the temperature increases, the rate of respiration will also increase,
generating a greater amount of heat, which decreases the postharvest
life of the products.
Temperature acts directly on physiological processes related to
sprouting inhibition and to a lesser degree on bulb root formation.
In turn, high temperature favors the growth of disease-causing
microbes.
The ideal temperature for a good conservation of onion is 0–5°C or
28–30°C. Freezing temperature is − 0.4 to − 0.8°C. Low temperatures
cause softening and blackening of tissues. When stored at low tempera-
tures, 1–2°C is recommended, followed by a period of reconditioning
before sale to avoid water condensation. Moreover, the maximum
temperature tolerated by the bulbs is 38–41°C, but at this temperature,
there is a risk of microorganism growth (Brice et al., 1997).
High temperatures cause damage, and softening, leaving the appear-
ance of cooked tissues, as well as promoting the development of
pathogens (fungi and bacteria) at a higher rate. Sprouting is later at
0°C than at storage temperatures of 5–10°C. Temperatures between
16 and 22°C promote sprouting, the optimum being 15°C, which is
inhibited at 30°C. A similar response to temperature changes is seen in
root formation, except that the optimum temperature is found in lower
ranges (9–11°C) ( Brice et al., 1997).
Onion can be affected by freezing, its freezing point is approxi-
mately -1°C depending on the variety and solids content. Bulbs
may be affected, showing a yellow-grayish coloration and a watery
consistency. This disorder progresses from the surface to the bulb
center and from the periphery to the central part of the vegetable.
Some measures to alleviate damage in affected bulbs consist of
unfreezing the product slowly to about 4°C and marketing it imme-
diately (Furlani and Rivero, 2010).
Another problem that can occur in onions is that of translucent cata-
phylls, a problem that occurs more frequently after 3 or 4 months
of cold storage, being its incidence more common in large bulbs.
It usually starts in the most curved portion of the cataphylls and
may involve the base of the outermost leaves. Cataphylls become
partially transparent, in contrast to the opaque appearance of healthy

tissue, and conducting vessels become more noticeable. In severe
cases, the bulb softens and is invaded by various pathogens. Its
incidence is significantly reduced by cooling the product as soon as
possible after curing (Furlani and Rivero, 2010).
d. Relative humidity (RH). High RH in the environment (above 80%)
is one of the most important enemies for onion conservation since it
stimulates the formation of new roots and sprouting in the bulbs and
promotes the development of pathogens (fungi and bacteria) in a wide
range of temperatures.
High relative humidity, with water condensation on the peel, causes
fermentation in the external cataphylls and consequently, the pres-
ence of stains, deteriorating the presentation and external quality of
the bulbs.
Low relative humidity (less than 60%) increases weight loss due to
bulb dehydration, resulting in wrinkling, which is undesirable for
marketing. An optimum RH for storage is 65–75%. Overall, the loss
of as little as 5% of a fruit's weight results in an unattractive appear-
ance, decreasing its market value (Brice et al., 1997).
6.10 ONION STORAGE IN CONTROLLED ATMOSPHERES
Onions to be stored in CAs must have the correct maturity, be very well
cured, dry, and healthy, so they must be rigorously selected, and they must
not be hit during the whole postharvest process so that they are free of any
damage. It should be remembered that storage will only preserve the quality
with which the onion enters.
A good quality onion should be undamaged, flawless, clean-skinned, free
of pathogens, and have the expected color for the variety (Muhie, 2022).
Controlled atmospheres have been found to give better results when
stored early in the season just after bulb curing. Factors such as irrigation,
harvest date, environmental conditions, variety, use of growth regulators,
fertilization, and type of curing have an effect on onion storage in CA.
The following are some general recommendations for onion storage in
CA (Sumner, 2000).
• The chamber must be airtight and moisture resistant.
Construction must provide the following:
An absolute vapor barrier to prevent moisture migration in the CA

chamber.
Adequate wall, ceiling, and floor insulation.
High tightness (chamber must contain a pressure medium, pressure
change 0.8–0.4 or 0.6–0.3 in of water for at least 30 min). A leaking
chamber will require excessive addition of nitrogen and carbon
dioxide. Leaks around the door and improper sealing are major
contributors to the loss of chamber tightness.
The CA chamber must be protected from wind and direct contact
with sunlight and rain to avoid sudden temperature changes that can
cause leakage.
Each chamber should have a vacuum–pressure release valve and
should be equipped with an airbag to prevent atmosphere loss when
small fluctuations in pressure occur.
A large air pocket may be necessary for chambers in onion storage
due to the large increase in air volume during the evaporator defrost
cycle.
Chambers should be interconnected (with PVC piping and valves) to
allow more efficient atmosphere establishment and pressure normal-
ization. This type of interconnection is especially important in small
chambers.
• Storage temperature.
Some portions of onions may freeze at temperatures of about -0.6°C
(30.8°F).
Each chamber must have at least two evaporator units with a
condensing unit (sized to meet the heat gain of the chamber)
equipped with reheat. Evaporator defrost cycles should be staggered
with PVC pipe interconnections between chambers to serve as atmo-
sphere expansion/contraction regulators. A backup cooling system
may be advisable.
• Air circulation.
Storage air circulation should be forced to 100–200 m3/ton of onion.
Forced air circulation is needed to maintain uniform temperature
and relative humidity throughout the CA chamber. Fans should be
placed at the rear of the room (at the opposite end of the entrance
door). The stacking of bins should allow air circulation through them.
The fan speed should be variable and controlled to achieve relative
humidity conditions as uniform as possible throughout the chamber.
Fan operation should start when the first onions are placed inside the
chamber and continue to run until the last onions are removed from
storage.
• Relative humidity.
Onions may lose a certain amount of moisture during CA storage.
For example, the Vidalia onion could lose 1–1.5% weight per month.
The dehumidification system must be capable of removing this
amount of water without significantly affecting the storage tempera-
ture. The operation of the dehumidification units is controlled by the
hygrostat. Improper control of relative humidity, air circulation, and
temperature can cause complete loss of onions during storage.
• Low oxygen concentration in CA chamber.
A low oxygen level, for example, 3%, reduces respiration, and
sugar loss and prolongs the storage life of onions. In this environ-
ment, humans can lose consciousness in less than 30 seconds, they
cannot breathe and their heartbeat stops within minutes. Therefore,
do not enter a CA chamber with low oxygen concentration without
breathing apparatus.
• High CO2 concentration in CA chamber.
A high carbon dioxide atmosphere, for example, 5%, inhibits respi-
ration and pungency development and prolongs storage and shelf
life after onion storage. The CO2 produced by the onion is probably
not enough to develop and maintain a high level of this gas.
• CA chamber monitoring and control system.
It is essential to control the temperature, humidity, and atmosphere
in the CA chambers without entering them.
This control includes the measurement of at least twice a day of the

following:
a. Temperature in the warmest and coldest areas of the chamber with
an alarm for high and low temperatures.
b. Oxygen and carbon dioxide concentrations. Adjustments can be

made manually or automatically with a computerized system.
c. Relative humidity. Relative humidity sensors located at the front and
rear of the chamber are used to monitor RH differences on which the
airflow rate is based, when this difference is too large the fan speed
should be increased. This fan speed adjustment can also be done
manually or automatically with a computerized system. A record of
all measurements and adjustments should be kept. Sensors should
operate for at least 6 months without recalibration.
• Heating the onion before removing it from the CA chamber.

Onions should be “warmed” before leaving the CA chamber to avoid
moisture condensation on the bulbs. Fans should operate during
the entire heating period. Additional heat may be needed for this
process, but the difference between air temperature and bulb surface
temperature should not exceed the dew point (approximately 15°F
or -9°C). The chamber must be ventilated before opening the door.
• Chamber size should reflect the sales rate of onions.
Overall, each CA chamber should contain the volume of onions to
be sold in 1 week. If weekly sales are less than the capacity of a
chamber, onions should be conditioned in another area outside the
chamber.
It should be noted that the removal of a portion of the onions from a
CA chamber will reduce the shelf life and quality of the remaining
onions left in the chamber if storage conditions are not restored.
Furthermore, regeneration of the CA atmosphere can be prohibi-
tively expensive if there are very large empty spaces in the chamber
(partially filled chambers).
Onions should be sold before starting the heating procedure. Shelf
life loss due to sprouting can increase rapidly after about 3 weeks.
Onions should have good air circulation through marketing to prevent
deterioration by root development and surface fungus growth.
a. Onion storage conditions in CAs.
Commercial storage of onion bulbs in CA is limited in part due to
variable success and inconsistent effects on bulb quality. Response
to CA varies among cultivars, therefore, experiments should be
conducted under local conditions to determine the appropriate levels

and composition of gases for each specific variety or local cultivar.
It has been found that the onion preservation in CA has significantly
improved its postharvest quality with respect to bulbs stored both refriger-
ated and at room temperature in several varieties. Conditions such as 1%
oxygen and 5% CO2 at a temperature of 1–2°C and a RH of 58–75% reduced
storage losses, and showed higher dry matter and total soluble solids content.
With this technology, onions with high pungency maintained good market-
able quality for more than 8 months (Poldman et al., 2012). According to
Tanaka et al. (1996), a mixture of 1% O2 and 1% CO2 at 1°C and 65–70%
humidity inhibited sprouting and preserved the vegetable for up to 1 year.
Chávez-Mendoza et al. (2016) and Ríos-González et al. (2018) on the
contrary, worked with storage conditions of 1% O2 + 1% CO2 and 3% O2 +
5% CO2, 2.5°C and 60–75% humidity, reporting minimal quality changes
in onion kept in CA for 7 months. With both CA treatments, sprouting was
inhibited by 100%, as well as microbial growth and there were no significant
changes in color, but a slight increase in pungency was observed.
Chope et al. (2007) reported that an atmosphere consisting of 5 kPa CO2
and 3 kPa O2 has been recommended for soft onions of the Granex variety,
low oxygen concentration inhibited bulb sprouting and reduced weight
losses, however, this may have a detrimental effect on sweet onion quality
through increased pungency, although the effect is not as great as in onions
stored in normal atmosphere or air, furthermore it may limit the subsequent
bulb shelf life with accelerated sprouting 3 weeks after removal from CA
storage.
Uddin et al. (2003) stored Hysam variety bulbs in regular atmosphere
and CA with two treatments 2% O2 + 2% CO2 and 2% O2 and 8% CO2, 0.5
± 0.5°C, 80% RH for 9 weeks, finding that pungency and flavor precursors
increased by 9–11% in onion stored in regular atmosphere and were signifi-
cantly reduced by the CA effect.
Smittle (1989) reported that the combination of CA storage (5% CO2, 3%
O2) and refrigerated storage (1°C) resulted in 99% of bulbs being considered
marketable after 7 months of storage; however, there was a 9% weight loss.
This author also recommended a CA of 3% O2, 5% CO2, 92% N2, 1 °C,
70–80% RH, and a ventilation rate of 5.m3.h-1.m3 of onions for Vidalia
sweet onions, which are known for their sweetness and low pungency.
Finally, Table 6.3 shows various CA compositions recommended for
onion storage (Thompson, 1996).
TABLE 6.3 Recommended CA Composition for Storage of Onion Bulbs.

Carbon dioxide (%) Oxygen (%) Temperature (°C)
0 1–2 –
0 1–2 0–5
0–5 1–2 0–5
0–5 0–1 0–5
5 3 1
5 5 4–5
10 3 4–5
Source: Thompson (1996).
6.11 CONCLUSION
AC storage technology is a very old technique that has evolved and expanded
its application in the conservation of more products of both animal and vege-
table origin. Its use allows preserving the quality of these for long periods.
However, its effect on each particular fruit must be previously researched to
obtain the desired results.
Onion is a widely cultivated and consumed crop in the world, it is a
seasonal product and although it is not very perishable like other crops, it
does suffer from storage losses, so AC storage technology is a good alter-
native for its conservation, also taking advantage of the fact that it is not
climacteric, which favors its conservation for longer periods.
The quality of the onion during storage is influenced by a large number
of factors, both pre and postharvest, which must be considered for a correct
application of CA. Up to now, atmospheres with 1% O2 + 1% CO2 and 3%
O2 + 5% CO2, in combination with low-temperature storage (0.5–2.5°C)
and relative humidity of 58–75%, are the most researched and the most
employed, which have allowed to maintain the quality of the onion from 7 to
12 months. The most important impact of AC technology is in the pungency
levels which tended to increase slightly during prolonged storage.
KEYWORDS
• controlled atmospheres
• conservation
• quality index
• onion storage
REFERENCES
Ali, S.; Khan, A. S.; Malik, A. U.; Anwar, R.; Anjum, M. A.; Nawaz, A.; Naz, S. Combined
Application of Ascorbic and Oxalic Acids Delays Postharvest Browning of Litchi Fruits
Under Controlled Atmosphere Conditions. Food Chem. 2021, 350, 129277. https://doi.
org/10.1016/j.foodchem.2021.129277
Arshad, M. O.; Chauhan, Y.; Singh, P.; Srivastav, P.; Gupta, M.; Patwa, N. Advancements
in Controlled Atmosphere Storage Technology—A Review. In Proceedings of Second
International Conference in Mechanical and Energy Technology; Yadav, S., Haleem, A.,
Arora, P. K., Kumar, A., Eds.; Springer, 2023; pp 399–410. https://link.springer.com/
chapter/10.1007/978-981-19-0108-9_42
Artés, F. Refrigeración y comercialización hortofrutícolas en la Región de Murcia [Fruit and
Vegetable Refrigeration and Marketing in the Region of Murcia]. II Edición. Ed. CEBAS-
CSIC, 1987; 150 p.
Bejo Z. B. V. Enfermedades y plagas importantes en cebollas [Important Diseases and
Pests in Onions], 2011; 17 pp. https://baixardoc.com/preview/20130126104951-bejo-
enfermedades-y-plagas-cebollas-5cba3001bfd7a
Benkeblia, N.; Varoquaux, P. Effects of Gamma Irradiation, Temperature and Storage Time
on the Status of the Glucose, Fructose and Sucrose in Onion Bulbs Allium cepa L. Int.
Agrophys. 2003, 17, 1–5.
Bishop, D. Controlled Atmosphere Storage. In: Cold and Chilled Storage Tecnology; Dellino,
C. V. J., Ed.; Blackie and Son Ltd.: Glasgow, 1990.
Bohyan, G. E.; Purvis, A. C.; Hurst, W. C.; Torrance, R. L.; Paulk, J. T. Harvest Date Effect
on Yield and Controlled-Atmosphere Storability of Short-Day Onions. HortScience 2004,
39 (7), 1623–1629. https://doi.org/10.21273/HORTSCI.39.7.1623
Brackmann, A.; Anese, R. D. O.; Thewes, F. R.; Fronza, D.; Hamann, J. J. Storability of
‘Tupy’and ‘Guarani’ Blackberries in Controlled Atmosphere. Bragantia 2016, 75, 240–246.
https://doi.org/10.1590/1678-4499.321
Brice, J. L.; Currah, L.; Malins, A.; Bancroft, R. Onion Storage in the Tropics: A Practical
Guide to Methods of Storage and Their Selection; Natural Resourxes Institute. The
University of Greenwich, 1997; 116 p.
Calvo, G.; Candan, A. P. Tendencias para la conservación de frutas de pepita [Trends for the
Preservation of Pome Fruits]. Revista Iberoamericana de Tecnología Postcosecha 2012, 13
(2), 153–159.
Chávez-Mendoza, C.; Vega-García, M. O.; Guevara-Aguilar, A.; Sánchez, E.; Alvarado-
González, M.; Flores-Córdova, M. A. Effect of Prolonged Storage in Controlled
Atmospheres on the Conservation of the Onion (Allium Cepa L.) Quality. Emirates J. Food
Agric. 2016, 28 (12), 842–852. DOI: 10.9755/ejfa.2016-06-643.
Chirinos, R.; Campos, D.; Martínez, S.; Llanos, S.; Betalleluz-Pallardel, I.; García-Ríos,
D.; Pedreschi, R. The Effect of Hydrothermal Treatment on Metabolite Composition of
Hass Avocados Stored in a Controlled Atmosphere. Plants 2021, 10 (11), 2427. https://doi.
org/10.3390/plants10112427
Chope, G. A.; Cools, K.; Hammond, J. P.; Thompson, A. J.; Terry, L. A. Physiological,
Biochemical and Transcriptional Analysis of Onion Bulbs During Storage. Ann. Bot. 2012,
109 (4), 819–831. https://doi.org/10.1093/aob/mcr318
Chope, G. A.; Terry, L. A.; White, P. J. The Effect of the Transition Between Controlled
Atmosphere and Regular Atmosphere Storage on Bulbs of Onion Cultivars SS1, Carlos
and Renate. Postharv. Biol. Technol. 2007, 44 (3), 228–239. https://doi.org/10.1016/j.

postharvbio.2006.12.018
Crisosto, C. H.; Garner, D.; Crisosto, G. High Carbon Dioxide Atmospheres Affect Stored
'Thompson Seedless' Table Grapes. HortScience 2002, 37 (7), 1074–1078. https://doi.
org/10.21273/HORTSCI.37.7.1074
Day, B. P. F. Fruit and Vegetables, Principles and Applications of Modified Atmosphere
Packaging, Ed. R. T. Parry; Blackie: Glaskow, 1993; pp 114–133.
Dilmaçünal, T.; Bayar, B.; Demirhan, Ö. Impact of Modified Atmosphere Packaging and
Controlled Atmosphere Applications on ‘Seval F1’ Tomato Fruit Quality and Marketability.
J. Inst. Sci. Technol. 2022, 12 (2), 527–538. https://doi.org/10.21597/jist.1037827
Farber, J. M. Microbiological Aspects of Modified Atmosphere Packaging—A Review. J.
Food Prot. 1991, 54, 58–70. https://doi.org/10.4315/0362-028X-54.1.58
Fernández, L. J.; Liverotti, O.; Wocca, F. Manejo postcosecha de cebolla, Control de calidad,
Inspección de Frutas y Hortalizas [Onion Postharvest Handling, Quality Control, Fruit and
Vegetable Inspection].Corporación del Mercado Central de Buenos Aires [Corporation of
the Central Market of Buenos Aires], 1999; 30 pp. https://www.yumpu.com/es/document/
read/14747439/cebolla-mercado-central-de-buenos-aires
Fondo Europeo Agrícola de Desarrollo Rural. Cebolla Fuentes de Ebro: Estudios sobre
la calidad del material vegetal en Aragón. Centro de Transferencia Agroalimentaria.
Informaciones técnicas No. 194, 2008. https://bibliotecavirtual.aragon.es/es/catalogo_
imagenes/grupo.do?path=3705355
Fuentealba, C.; Vidal, J.; Zulueta, C.; Ponce, E.; Uarrota, V.; Defilippi, B. G.; Pedreschi,
R. Controlled Atmosphere Storage Alleviates Hass Avocado Black Spot Disorder.
Horticulturae 2022, 8 (5), 369. https://doi.org/10.3390/horticulturae8050369
Furlani, M. A.; Rivero, M. L. Manual del Cultivo de la cebolla. Centro Regional Cuyo INTA,
2010. 10 p. https://inta.gob.ar/documentos/manual-del-cultivo-de-la-cebolla
Garba, U. A.; Mohammed, D.; Ja´afaru, A. Study and Analysis of a Refrigerated and
Controlled Atmosphere Onion Storage System. JORIND 2014, 12, 246–252.
Gökçe, A. F.; Kaya, C.; Serçe, S.; Özgen, M. Effect of Scale Color on the Antioxidant
Capacity of Onions. Sci. Hortic. 2010, 123 (4), 431–435. https://doi.org/10.1016/j.
scienta.2009.11.007
González, M. V.; Miravé, J. P.; Saluzzo, J. A. Relaciones entre el tamaño y la conservación
de bulbos de cebolla para diferentes cultivares y condiciones de almacenamiento. Revista
FAVE- Ciencias Agrarias 2005, 4, 21–30.
Gorreapti, K.; Thangasamy, A.; Bhagat, Y.; Murkute, A. A. Curing of Onion: A Review.
Indian Hortic. J. 2017, 7 (1), 8–14.
Graell, J.; López, M. L.; Larrigaudiere, C. Manzanas y peras. Avances técnicos en atmosfera
controlada [Apples and Pears: Technical Advances in Controlled Atmosphere]. Horticultura
Internacional Agosto 1998, 67–69. https://www.mapa.gob.es/ministerio/pags/biblioteca/
revistas/pdf_hortint/hortint_1998_21_66_69.pdf
Graell, J.; Ortiz, A. Recomendaciones para almacenamiento en atmosfera controlada
[Recommendations for Controlled Atmosphere Storage]. Revista Horticultura 2003, 172,
38–43. https://xdoc.mx/preview/recomendaciones-para-almacenamiento-en-atmosfera-
controlada-60348e2d61c42
Grevsen, K.; Sorensen, J. N. Sprouting and Yield in Bulb Onions (Allium cepa L.) as
Influenced by Cultivar, Plant Establishment Methods, Maturity at Harvest and Storage
Conditions. J. Hortic. Sci. Biotech. 2004, 79 (6), 877–884. https://doi.org/10.1080/146203

16.2004.11511860
International Institute of Refrigeration (IIR). Guide to Refrigerated Transport; Ed. Int. Inst.
Refrigerationm 1995; 150 pp.
Kader, A. A. A Summary of CA Requirements and Recommendations for Fruit Other Than
Pome Fruits. In: Proceedings of the Fifth International Controlled Atmosphere Research
Conference, Wenatchee, Washington, USA, Vol. 2; 14–16 June, 1989; pp 303–328.
Kader, A. A. Controlled Atmosphere Storage. In: Agricultural Handbook Number 66; Gross,
K. C., Wang, C. Y., Saltveit, M. E., Eds.; The Commercial Storage of Fruits, Vegetables and
Florist and Nursery Stocks; USDA, 2004.
Kader, A. A. Modified Atmospheres During Transport and Storage of Fresh Fruits and
Vegetables. I Int. Cong. Food Technol. Develop. Murcia. 1990, 1, 149–163.
Kader, A. A. Postharvest Technology of Horticultural Crops, 3rd ed.; UC ANR Publications,
Davis, 2002; p 40.
Kader. A. A. Fruit, Maturity, Ripening and Quality Relationships. Perishables Handling
Newslett. 1994, 80, 2.
Kiura, I. N.; Bernard M. Gichimu, B. M.; Rotich, F. Visual and Keeping Quality of Stored
Bulb Onions as Affected by Harvest and Postharvest Treatments. Int. J. Agron. 2021, 2021.
https://doi.org/10.1155/2021/9969571
Lee, E. J.; Jeon, J. K.; Suh, J. K. Pyruvic Acid and Sugar Contents during Storage Duration in
Onion (Allium cepa L.). J. Bio-Environ. Control. 2010, 19 (4), 377–381.
López, C. A. F. Manual Para la Preparación y Venta de Frutas y Hortalizas. Del campo al
mercado [Manual for the Preparation and Sale of Fruits and Vegetables: From the Field to
the Market]; FAO Agricultural Services Bulletin 15, 2003. https://www.fao.org/3/y4893s/
y4893s.pdf
López, V. J. Á.; Valverde Juárez, F. J.; Mejía Torres, S. L.; López Angulo, G.; Vega García,
M. O. Efecto del almacenamiento en atmósfera controlada sobre la calidad postcosecha y
nutricional del tomate. Revista Chapingo. Serie Horticultura 2011, 17 (2), 115–128.
Magaña-Benítez, W.; Balbín, A. M.; Corrales, J.; Saucedo, G. V. C.; Sauri, D. E. Frutas de
pitahaya (Hylocereus undatus) frigoconservadas a 4°C en atmósferas controladas. Revista
Iberoamericana de Tecnología Postcosecha 2010, 11 (2), 143–147.
Majid, I.; Dhatt, A. S.; Sharma, S.; Nayik, G. A.; Nanda, V. Effect of Sprouting on
Physicochemical, Antioxidant and Flavonoid Profile of Onion Varieties. Int. J. Food Sci.
Technol. 2016, 51 (2), 317–324. https://doi.org/10.1111/ijfs.12963
Marcellin, P. Les maladies physiologiques du froid. In: Les végétaux et le froid; Côme, D.,
Ed.; Hermann: París, 1992; pp 53–105.
Martínez-Damián M. T.; M. Cantwell de Trejo. Cambios de calidad en espinaca almacenada
en atmósferas controladas [Quality Changes in Spinach Stored in Controlled Atmospheres].
Revista Chapingo Serie Horticultura 2002, 8 (1), 49–62.
Martins, D. R.; de Resende, E. D. Quality of Golden Papaya Stored Under Controlled
Atmosphere Conditions. Food Sci. Technol. Int. 2013, 19 (5), 473–481. https://doi.
org/10.1177/10820132124622
Miedema, P.; Bulb Dormancy in Onion. I: The Effects of Temperature and Cultivar on
Sprouting and Rooting. J. Hortic. Sci. 1994, 69, 29–39. https://doi.org/10.1080/1462031
6.1994.11515245
Muhie, S. H. Preharvest Production Practices, and Postharvest Treatment and Handling

Methods for Best Quality Onion Bulbs. J. Hortic. Sci. Biotechnol. 2022, 97 (5), 552–559.
https://doi.org/10.1080/14620316.2022.2041493
Nega, G.; Mohammed, A.; Menamo, T. Effect of Curing and Top Removal Time on Quality
and Shelf Life of Onions (Allium cepa L.). GJSFR 2015, 15, 26–36.
NMX-FF-021-SCFI-2003. Productos alimenticios no industrializados para Consumo
humano–bulbos–cebolla (Allium cepa L.) [Non-Industrialized Food Products for Human
Consumption–Bulbs–Onion (Allium cepa L.)-] Especificaciones (cancela a la NMX-FF-
021-1986) CDU: 635.25.
Parry, R. T. Introduction. In: Principles and Applications of Modified Atmosphere Packaging
of Food; Parry, R. T., Ed.; Blackie Academic & Professional: Glasgow, 1993; pp 1–18.
Petropoulos, S. A.; Ntatsi, G.; Ferreira, I. C. F. R. Long-Term Storage of Onion and the Factors
That Affect Its Quality: A Critical Review. Food Rev. Int. 2017, 33 (1), 62–83. https://doi.
org/10.1080/87559129.2015.1137312
Põldma, P.; Moor, U.; Merivee, A.; Tõnutare, T. Effect of Controlled Atmosphere Storage on
Storage Life of Onion and Garlic Cultivars. In: IV International Conference Postharvest
Unlimited, 2011, 945; pp 63–69. https://www.actahort.org/books/945/945_7.htm
Raigón, M. D. El nivel de pungencia en las cebollas [The Level of Pungency in Onions].
Revista Horticultura Internacional 2006, 51, 48–51.
Ríos-González, I. J.; Guevara-Aguilar, A.; Sida-Arreola, J. P.; Sánchez, E.; Ronquillo-Aboite,
J. O.; Alvarado-González, M.; Chávez-Mendoza, C. Effect of Controlled Atmospheres on
the Quality of White Onion (Allium cepa L.). J. Food Sci. Technol. 2018, 55 (9), 3564–3574.
Sharma, K.; Assefa, A. D.; Kim, S.; Ko, E. Y.; Park, S. W. Change in Chemical Composition
of Onion (Allium cepa L0. cv. Sunpower) During Post-Storage Under Ambient Conditions.
New Zealand J. Crop Hortic. Sci. 2014, 42 (2), 87–98. https://doi.org/10.1080/01140671.
2013.860039
Sharma, K.; Rok Lee, Y.; Park, S. W.; Nile, S. H. Importance of Growth Hormones and
Temperature for Physiological Regulation of Dormancy and Sprouting in Onions. Food
Rev. Int. 2016, 32 (3), 233–255.
Smittle, D. A. Controlled Atmosphere Storage of Vidalia Onions. In: International Controlled
Atmosphere Conference 5th Proceedings, Wenatchee, Washington, USA, Vol. 2, Other
Commodities and Storage Recommendations, 1989; pp 171–177.
Smittle, D. A. Evaluation methods for ‘Granex’ onions. J. Am. Soc. Hortic. Sci. 1988, 113,
877–880.
Sorensen, J. N.; Grevsen, K. Nitrogen and Water Stress Affects Sprouting in Bulb Onions
Stored Over Winter. Acta Hortic. 2002, 571, 98–86.
Sumner, P. E. Vidalia onion CA Storage Requirements. Cooperative Extension Service.
The University of Georgia College of Agricultural and Environmental Sciences.
USA, 2000. http://www.caes.uga.edu/commodities/fruits/veg/pubs/documents/
VIDALIAONIONCASTORAGEREQUIREMENTS.pdf.
Thompson, A. K. Controlled Atmosphere Storage of Fruits and Vegetables, 2nd ed.; CAB
International Edition, MPG Books Group, 2010; pp 1–2.
Thompson, A. K. Fruit and Vegetable Storage Hypobaric, Hyperbaric and Controlled
Atmosphere; Springer Briefs in Food, Health, and Nutrition, Springer: Huddersfield, 2016;
p 8.
Thompson, A. K. Postharvest Technology of Fruits and Vegetables; Blackwell Science Ltd.:
London, 1996.
UCDavis. Cebollas de Guarda: Recomendaciones para Mantener la Calidad Postcosecha.

Postharvest Center. University of California, 2022. https://postharvest.ucdavis.edu/
Commodity_Resources/Fact_Sheets/Datastores/Vegetables_Spanish/?uid=10&ds=803
Uddin, M. M.; MacTavish, H. S. Controlled Atmosphere and Regular Storage-Induced
Changes in S-alk (en) yl-l-cysteine Sulfoxides and Alliinase Activity in Onion Bulbs
(Allium cepa L. cv. Hysam). Postharv. Biol. Technol. 2003, 28 (2), 239–245. https://doi.
org/10.1016/S0925-5214(02)00195-3
Yahia, E. Uso de atmósferas modificadas y controladas como medio de conservación de frutas
y hortalizas. Memorias del simposio internacional de manejo postcosecha y perspectivas
de comercialización de frutas y hortalizas [Use of Modified and Controlled Atmospheres as
a Means of Preserving Fruit and Vegetables. Proceedings of the International Symposium
on Post-Harvest Handling and Marketing Prospects for Fruits and vegetables], Universidad
Nacional de Colombia [National university of Colombia], 27–30 June, 1995a.
Yahia, E. La tecnología de las atmosferas modificadas y controladas parte I. [The Technology
of Modified and Controlled Atmospheres Part I]. Horticultura Internacional 1995b, 7,
37–39.
Yahia, E. La tecnología de las atmosferas modificadas y controladas parte II [The Technology
of Modified and Controlled Atmospheres Part II]. Horticultura Internacional 1995c, 8,
20–25.
Yahia, E. M.; Fadanelli, L.; Mattè, P.; Brecht, J. K. Controlled Atmosphere Storage. In:
Postharvest Technology of Perishable Horticultural Commodities; Woodhead Publishing,
2019; pp 439-479.
Yamashita, K.; Sasaki, W.; Fujisaki, K.; Haga, H. Effects of Controlled Atmosphere Storage
on Storage Life and Shelf Life of the Japanese Onion Cultivar Super-Kitamomiji. In:
Proceedings of the 10th International Controlled & Modified Atmosphere Research
Conference; Turkey, 2009; p 53.
SECTION 2
FOOD BIOTECHNOLOGY
CHAPTER 7
Larrea tridentata Extracts and the

Capacity of Its Biocompounds in
Medicine and Agriculture
JOLANTA E. MARSZALEK1, DANIELA M. SÁNCHEZ-PÉREZ2,
ERIKA FLORES LOYOLA1, AGUSTINA RAMÍREZ-MORENO1, and
LUCIO RODRÍGUEZ-SIFUENTES1
1
Facultad de Ciencias Biológicas, Universidad Autónoma de Coahuila,
Coahuila, México
2
Instituto Tecnológico de Torreón (ITT)-DEPI, Carretera Torreón-San
Pedro, Torreón, Coahuila, México
ABSTRACT
The use of plants is of great interest in traditional medicine and when

looking for new pharmacological materials for drug development. Creo-
sote bush, Larrea tridentata (LT), is a perennial shrub growing in southern
parts of the USA and northern semideserts of Mexico. The extracts of that
plant contain a variety of metabolites, among which lignans are the most
prevalent. Nordihydroguaiaretic acid (NDGA) is an extensively studied
lignan with antibacterial, antiviral, anthelmintic, antifungal, and anticancer
properties. However, other lignans and phenolic compounds found in LT
extracts demonstrate similar activities. Even though extended exposure to
LT extracts is believed to be toxic to the organism, it is valid to exploit the
use of its isolated biocompounds. This review aims to give a good overview
of the research into possible applications of these metabolites in medicine
and agriculture.
7.1 INTRODUCTION
Research always directs its interests in improving human life; recently, novel
treatments for diverse human diseases have been sought. The reasons can be
found in increased bacterial resistance to antibiotics (Andersson et al., 2017),
developed resistance to antifungal treatments (Perlin et al., 2017), and a high
number of difficult-to-treat cancers (Holohan et al., 2013; Tsimberidou et al.,
2015). Consequently, alternatives that would have a strong activity with no
toxicity and preferably low cost are wanted. In that aspect, more researchers
explore natural sources, such as compounds from plants, bacteria, marine
plants, and others, that could fulfill that need (Firenzuoli and Gori, 2007).
Diverse studies demonstrate the presence of phytocompounds in plants that
have activity against several human illnesses (Wen et al., 2012; Thanga-
pazham et al., 2016, Hosseini and Ghorbani, 2015; Aboody and Mickymaray
, 2009). In different regions of the World, many of these plants have been
used in traditional medicine for hundreds of years. Some plants native to
North America possess many of these qualities. They are found in arid zones,
and because of their environmental conditioning, these plants contain many
phytocompounds with exciting properties. One such plant is the creosote
bush, Larrea tridentata (Zygophyllaceae) (LT), also locally referred to as
greasewood and chaparral in the USA, and gobernadora in Mexico (Fig.
7.1), which is a common shrub in North American deserts. It is abundant
in the semidesert areas of San Luis Potosí, Coahuila, Chihuahua, Durango,
Sonora, Zacatecas, Baja California, and Baja California Sur in Mexico,
as well as in the Southwest United States in Arizona, California, Nevada,
Texas, and New Mexico (Rzedowski and Huerta, 1994). These plant extracts
have been commonly used in broad applications in traditional Mexican
medicine (Arteaga et al., 2005). In this chapter, we examine information
about L. tridentata and its biocompounds that found applications in the field
of medicine and agriculture.
7.2 PLANT COMPOSITION AND ANTIOXIDANT CAPACITY
The leaves of L. tridentata contain around 5% moisture and within the solids:
reducing sugars (1.9%), nonreducing sugars (12%), fat (3.4%), crude fiber
(12.3%), and ash (10.7%) (Treviño-Cueto et al., 2007; Mendez et al., 2012).
As a result, the plant is an outstanding source of natural compounds, with
approximately 50% of the dry weight of leaves being extractable compounds
(Arteaga et al., 2005).
Larrea tridentata Extracts and the Capacity of Its Biocompounds 169
FIGURE 7.1 Image of the plant L. tridentata taken in the area of Torreon, Coahuila, México.
It has been reported that up to 62% of the plants’ phytocompounds

belong to the tannins (Treviño-Cueto et al., 2007). Besides tannins, saponins,
sterols, monoterpenes, sesquiterpenes, lignans, flavonoids, and terpenes were
detected in the aqueous and organic extracts of L. tridentata (Mendez et al.,
2012; Gnabre et al., 2015). These varied phytocompounds are accompanied
by essential oils (0.1% of the dry weight) and 67 nonvolatile compounds
(Arteaga et al., 2005).
Among bioactive compounds present in this plant, nordihydroguaiaretic

acid (NDGA), kaempferol, and quercetin are found at considerably high
concentrations (Hyder et al., 2002). Phytoestrogens, including flavonoids
comprising isoflavonoids and flavanols, derivates, lignans, and coumestans,
are secondary plant metabolites. All of these have attracted sizable atten-
tion due to their protective action against several health disorders. They
prevent and treat menopausal symptoms, skin aging, osteoporosis, cancer,
and cardiovascular, neurodegenerative, immune, and metabolic diseases
(Sirotkin and Halim Harrath, 2014).
LT’s principal compounds are phenolics, which are present in high
concentrations in leaves (36.2 mg/g), green steams (40.8 mg/g), and roots
(28.6 mg/g) (Fig. 7.2A). Condensed tannins also are present in all tissues
with high concentrations, specifically in flowers (1.7 mg/g), seeds (1.1
mg/g), and roots less than 5 mm in diameter (1.1 mg/g) (Fig. 7.2B). The
most studied lignan, NDGA, is found in flowers, leaves, green stems, and
small woody stems (<5 mm in diameter), with the highest concentrations
in leaves (38.3 mg/g) and green steams (32.5 mg/g) (Fig. 7.2C) (Hyder et
al., 2002). In the aerial parts of LT, about 25 main phytocompounds are
found. Their structure was mostly triterpene glycoside (Jitsuno and Mimaki,
2010). In 2011 two new lignans glycosides were described, larreatricin A
and larreatricin B (Yokosuka et al., 2011). Recently, thymol (5.38 mg/mL)
and carvacrol (4.18 mg/mL) were detected in an extract exhibiting strong
antibacterial properties (Morales-Ubaldo et al., 2022). A list of some of the
described phytocompounds is presented in Table 7.1, with an indication of
what properties they exhibited.
FIGURE 7.2 Percent distribution of (A) total phenolics, (B) condensed tannins, and (C)
NDGA in different plant parts of L. tridentata adapted from Hyder et al. (2002).
Source: Adapted from Hyder et al., 2002
TABLE 7.1 NMR-Identified Compounds from L. tridentate and Their Cytotoxic, Antiprotozoal, and Antioxidant Activity.
Larrea tridentata Extracts and the Capacity of Its Biocompounds

Compound Antiprotozoal Antibacterial Cytotoxic activity against (µg/mL) References
Activity Activity HL-60 cells ACC-375 L-6 MCF-7 SW-480
IC50 cells cells cells cells
(+)-Dihydroisorhamnetin >50.0 Abou-Gazar et al. (2004)
(7r,7′r)-4,4′-Dihydroxy-8,8’-dehydro-  >90 Schmidt et al. (2012)
7,7′-epoxylignan
(7s,7′s,8s,8′s)-3,3′,4,4′-Tetrahydroxy-  154 Schmidt et al. (2012)
7,7′-epoxylignan
(7s,7′s,8s,8′s)-3,3′,4′-Trihydroxy-4-  188 Schmidt et al. (2012)
methoxy-7,7′-epoxylignan
(7s,8s,7′s,8′s)-3,3′,4′-Thrihydroxy-4- 17.3 Abou-Gazar et al. (2004)
methoxy-7′,7′-epoxy-lignan
(s)-4′,5-Dihydroxy-7-methoxyflavone >80 60 >80 Lambert et al. (2005)
3,3′-di-o-Demethylisoguaiacin  132 Schmidt et al. (2012)
3,4-Dehydrolarreatricin 27.6 Abou-Gazar et al. (2004)
3,4-Dihidroxi-3′,4′-dimetoxi-6,7′-  51 Morales-Ubaldo et al. (2022)
ciclolignano
3,4′-Dihidroxy-3′,4-domethoxy-6,7′- 50 25 50 Lambert et al. (2005)
cyclolignan
3′-Demethoxy-6-o-demethylisoguaicin  13.6 Favela-Hernández et al. (2012);
Abou-Gazar et al. (2004)
3′-Demethoxy-6-demethylisoguaiacin >20 Yokosuka et al. (2021)
6-O-β-d-glucopyranoside
3′-Demethoxy-6-demethylisoguaiacin >20 Morales-Ubaldo et al. (2022);
6,40-di-O-β-d-glucopyranoside Yokosuka et al. (2021)
171
172
Engineering Principles for Food Processing Technology and Product Realization

6,30-Didemethylisoguaiacin >20 Yokosuka et al. (2021)
6,30,40-tri-O-β-d-glucopyranoside
3′′-hydroxy-4-epi-larreatricin 13.6 Abou-Gazar et al. (2004)
3’-o-Methyltaxifolin  27 Schmidt et al. (2012)
4,4′-Dihydroxy-7,7′-dioxolign-8(8′)-ene >50 Abou-Gazar et al. (2004)
4,4′-Dihidroxi-3-metoxi-6,7′-ciclolignano  56 Morales-Ubaldo et al. (2022)
1,2,4-Trihydro-1-(3,4-dihydroxyphenyl)- 16 Yokosuka et al. (2021)
2,3-dimethyl-3,4,6,7-naphthalenetetrol.
5,7,4′-Trihydroxy-3,8,3′- >50.0 Abou-Gazar et al. (2004)
trimethoxyflavone
5,4′-Dihydroxy-3,7,8,3′-  Favela-Hernández et al. (2012)
tetramethoxyflavone
5,4′-Dihydroxy-7-methoxyflavone  Favela-Hernández et al. (2012)
5,4′-Dihydroxy-3,7,8-trimethoxyflavone  Favela-Hernández et al. (2012)
2-[1-(3,4-Dihydroxyphenyl)-2-methyl-3- 17 Yokosuka et al. (2021)
oxobutyl]-4,5-dihydroxybenzaldehyde
(2R,3R)-2,3-Dihydro-2-(3,4- 4.1 Yokosuka et al. (2021)
dihydroxyphenyl)-3- methyl-5-(E)-
propenylbenzofuran.
(2R,3R)-2,3-Dihydro-2-(3-hydroxy- 15 Yokosuka et al. (2021)
4-methoxyphenyl)-3- methyl-5-(E)-
propenylbenzofuran
Larrea tridentata Extracts and the Capacity of Its Biocompounds

Butane-type di-o-methylated lignan 10 12 31 Lambert et al. (2005)
Butane-type di-o-methylated lignan 7 10 30 Lambert et al. (2005)
Butane-type tri-o-methyl lignan 7 5.3 55 Lambert et al. (2005)
Carvacrol  Morales-Ubaldo et al. (2022)
Cyclolignan-3′-demethoxyisoguaiacin 22 9.6 55 Lambert et al. (2005)
Herbacetin-3,8,4′-trimethyl ether >50.0 Abou-Gazar et al. (2004)
Herbacetin-3,7′-dimethylether  8 Schmidt et al. (2012)
Kaempferol-3,7-dimethylether  15 Schmidt et al. (2012)
Kaempferol-3-methyl1 ether 28.3 Abou-Gazar et al. (2004)
(isokempferide)
Lignoids-4-epi-larreatricin  >50 Abou-Gazar et al. (2004);
Favela-Hernández et al. (2012)
meso-(rel 7s,8s,7′s,8′s)-3,4,3′,4′- >50.0 Abou-Gazar et al. (2004)
Tetrahydroxy-7,7′-epoxylignan
meso-Dihydroguaiaretic acid  Schmidt et al. (2012)
meso-Nordihydroguaiaretic acid   33 Schmidt et al. (2012)
Naringenin  >90 Schmidt et al. (2012)
NDGA   2.6 Bashyal et al. (2017); Abou-
Gazar et al. (2004); Favela-
Hernández et al. (2012)
Thymol  Morales-Ubaldo et al. (2022)
173
7.2.1 LIGNANS
Lignans, a class of secondary plant metabolites, are produced by oxida-
tive dimerization of two phenylpropanoid units. They are optically active
phenolic dimers possessing a 2,3-dibenzylbutane structure (Zitterman, 2003;
Saleem et al., 2005). Although their molecular backbone consists only of
two phenylpropane (C6–C3) units, lignans show an enormous structural
diversity. In rye wheat, for example, these compounds appear mainly in the
glycosidic form. In many plants, the lignans are the building blocks of the
plant cell wall (Zitterman, 2003). Thus, they are widely distributed in the
plant kingdom and have been found in species belonging to more than 70
families. Lignans are found in roots, rhizomes, stems, leaves, seeds, and
fruits. Unfortunately, with some exceptions, the plants studied do not provide
commercially practical quantities of lignans. Flaxseed has been reported to
have the highest concentrations of dietary lignans in the form of secoisolar-
iciresinol diglucoside, and these are available for purchase as a supplement
(Source Naturals Lignan Extract Caps, n.d.).
Regarding biological interactions, lignans are phytoestrogens with
estrogenic or antiestrogenic activity (Barakat et al., 2018). In addition, they
respond to various biological issues in the form of anticancer, antioxidant,
antihypertensive, antiviral, and even insecticidal properties (Simpson and
Amos, 2017). These varied activities increased the interest in lignans and
their synthetic derivatives and proposed their applications as therapeutics;
for example, in cancer treatment.
7.2.1.1 NORDIHYDROGUAIARETIC ACID

Nordihydroguaiaretic acid [l,4-bis-(3,4-dihydroxyphenyl)-2,3-dimethyl-
butan] (NDGA) is a simple lignan that is linear with phenylpropane groups
connected via a b–b′ linkage. NDGA could be called the dominant lignan of
LT because it is present in all parts of the plant, representing approximately
5–10% of the leaves’ dry weight and 80% of all phenolics. This phytocom-
pound is of great interest due to its biological properties as a potent antioxi-
dant (Floriano-Sánchez et al., 2006).
7.2.2 ANTIOXIDANT ACTIVITY OF LIGNANS

The efficiency of lignans as antioxidants is related to their structure and
solubility. Antioxidants are defined as any molecule that inhibits oxida-
tion, that is, loss of electrons and increase in the oxidative state of another
molecule. Oxidation forms free radicals that are unstable atoms and molecules
possessing an insufficient number of electrons. Antioxidants usually termi-
nate many reactions by removing free radical intermediates and inhibiting
other oxidation reactions. Thus, antioxidants often serve as reducing agents
(e.g., thiols, ascorbic acid, and polyphenols). Antioxidants work through
single or combined mechanisms: free radical scavenging, complexing of
pro-oxidant, reducing activity, quenching of singlet oxygen, and scavenging
lipid peroxyl radicals. Preventive actions are represented by the antioxidants
that serve as inhibitors of free radical oxidation reactions (Kasote, 2013;
Mehta and Gowder, 2015).
During the normal function of the human body, free radicals, reactive
oxygen species (ROS), and nitrogen species are continuously generated.
They are important for energy supply, chemical signaling, detoxification,
and immune functions (Dimitrios, 2006). However, overproduction of
these species due to exposure to external oxidant substances or a failure in
the body’s defense mechanisms such as endogenous enzymes (superoxide
dismutase, glutathione peroxidase, and catalase), leads to damage of valu-
able biomolecules such as DNA, lipids, and proteins (Mehta and Gowder,
2015; Aruoma, 1998; Dimitrios, 2006).
It has been established that these damages are associated with an increased
risk of various degenerative diseases such as cardiovascular disease, cancer,
and others. On the contrary, phenolic compounds are excellent natural
antioxidants. They are known to counteract the excess ROS and erase their
pathological effects. The antioxidant capacity of phenolic compounds is
mainly due to their redox properties, which allow them to act as reducing
agents, hydrogen donors, singlet oxygen quenchers, or metal chelators
(Mehta and Gowder, 2015).
In LT, the antioxidants are represented by lignans, flavonoids, condensed
tannins, and other phenolics. As aforementioned, the main constituent of
the lignans in LT is NDGA. Its polyphenol-bearing o-dihydroxy structure,
with four phenolic hydroxyl groups, promotes radical scavenging ability
(Floriano-Sánchez et al., 2006; Choi and Jung, 2016). The antioxidant
activity of NDGA has been studied extensively over the years. Interest-
ingly, NDGA’s antioxidant ability was explored as a food protection agent
in the 1950s (Bickoff and Williams, 1945; Lundberg et al., 1944; Stull et
al., 1948). Recently, the characteristics of NDGA have been well-reviewed
(Manda et al., 2020); thus, here we summarize shortly only its action. The
potent scavenging activity of NDGA against multiple types of ROS comes
from donating one electron and one proton from each of its hydroxyl
groups contained in the catechol rings, which converts NDGA structure

into an oxidized catechol-quinone. The quinone is, firstly, a semi-quinone
radical, which is further converted into an ortho-quinone and superoxide
anion. Ortho-quinone may be changed back to the reactive semi-quinone by
cellular NADPH-dependent reductases, but this redox cycle is highly toxic,
especially at high NDGA concentrations. As a consequence, NDGA at a high
dose might lead to cytotoxic effects in the organism, as discussed later in the
chapter.
Free radical scavenging activity and stabilization of newly formed tran-
sient radicals are facilitated in compounds that do not possess steric hindrance
(Kitts et al., 1999; Foti et al., 1996) but have an ortho-hydroxyl group (Lu
and Liu, 1992). In the case of lignans found in LT, due to tetrahydrofuran
moiety, the number and the position of the phenyl moieties in epoxylignans
may contribute to robust antioxidant effects.
7.3 MEDICINAL APPLICATION OF L. TRIDENTATA
Creosote bush has been used in traditional medicine as a treatment for around
50 different illnesses, as reported by Arteaga et al. (2005). These include skin
problems (acne, dandruff, psoriasis); blood problems (altered blood pres-
sure, anemia, blood purifier, bruises, and hemorrhoids); digestive system
issues (bowel cramps and inflammation, stomach pain and diarrhea, diseases
of the liver, kidney, and gallbladder stones, kidneys pain and cystitis, ulcer,
indigestion, urinary tract infections, and diuretic); bacterial and varial infec-
tions (cold, influenza, bronchitis, chickenpox, tuberculosis); women issues
(menstrual pains, inflammation after delivery, cramping, contraceptive, and
sterility) as well as allergic treatment, diabetes, headache and toothache,
and snakebite pain. In addition, tonics and extracts from LT are believed to
have anti-inflammatory, antiamoebic, antibiotic, antifungal, antineoplastic,
antiseptic, and antiviral properties (Arteaga et al., 2005; Heron and Yarnell,
2001).
7.3.1 CYTOTOXICITY OF LIGNANS, ANTICANCER APPLICATION
Cytotoxicity study using human umbilical vein endothelial cells (HUVEC)

and isolated compounds from LT extracts showed to inhibit HUVEC cell
viability with half maximal inhibitory concentration (IC50) values of 86
and 59 μM for NDGA and 3′-O-methyl-NDGA, respectively. These two
compounds were correspondingly 2.4- and 1.6-fold less toxic to human cells
than to Naegleria fowleri (the parasite studied in the research) (Bashyal et
al., 2017).
For eight lignans and cyclolignans, including NDGA, isolated from the
flowering tops of LT (Table 7.1), the median inhibition concentration, IC50,
was evaluated against three cancerous cell lines (Lambert et al., 2005). The
results showed IC50 values of 7–80 µM, with the linear butane-type lignans
being the most potent and colon cancer cells being the least sensitive cell
type. The relative capacity of linear butane-type lignans against human
breast cancer appears to be aligned with the number of O-methyl groups in
the NDGA molecule. The potency of O-methylation is seen in the increased
lipophilicity, which allows the compounds to easier cross the cell’s plasma
membrane, or in the molecule’s ability to bind to a lipophilic target, such as
the estrogen receptor expressed in MCF-7 cells (Lambert et al., 2005)40.
Two meso-nordihydroguaiaretic acid glycoside derivatives, larrealignans
A and B, tetra- and tri-desmosides lignans, respectively, did not exhibit
cytotoxicity against HL-60 human leukemia cells even at a concentration of
20 µM. Other compounds isolated from LT showed moderate cytotoxicity
(two aglycone compounds had IC50 values of 7.0 and 4.9 mM) (Yokosuka
et al., 2011). The same group isolated 17 lignan compounds from LT leaves
and determined 10 as described for the first time. The cytotoxicity of these
compounds was evaluated against promyeoloblasts HL-60 cells, and the
majority of these lignans presented IC50 values in the range of 2.7–17 µM.
Compound (2R,3R)-2,3-dihydro-2-(3,4-dihydroxyphenyl)-3- methyl-5-(E)-
propenylbenzofuran showed the highest cytotoxicity and induced apoptotic
cell death (Yokosuka et al., 2021).
NDGA exhibits anticancer activity in numerous in vivo and in vitro
studies (Rowe et al., 2008; Leon et al., 2016; Park et al., 2005; Wang et
al., 2011). The ability of NDGA to hinder the proliferation of breast cancer
cells is directly related to the inhibition of the function of two receptor tyro-
sine kinase (RTKs), the insulin-like growth factor receptor (IGF-1R) and
the c-erbB2/HER2/neu (HER/neu) receptor (Youngren et al., 2005). Other
aspects of NDGA’s anticancer activity were established as an inhibitor of
lipoxygenase isoenzymes and showed the strongest growth inhibition with
an IC50 of 1.9 ± 0.5 µg (Hausott et al., 2003).
To improve the anticancer action of NDGA, a new derivate NDGA-
P21 was synthesized by methylation of -OH groups and linearization of
the molecule. With these changes, the new compound exhibited a higher
suppression of glioma (GBM) cells by arresting the cell cycle of GBM cells
in G0/G1 phase, blocking cell proliferation sequentially, and suppressing the

stemness of glioma stem-like cells (GSLCs) (Zhao et al., 2017).
NDGA has been tested as a potential treatment for skin cancer and restored
reduced glutathione levels and activities of antioxidant enzymes. As a result,
elevated myeloperoxidase activities, xanthine oxidase, and skin edema forma-
tion in TPA-treated mice (treated with 12-O-tetradecanoylphorbol-13-acetate)
were also lowered by NDGA, indicating a restrained inflammatory response.
At the same time, the histological results demonstrated an inhibitory effect of
NDGA on cellular inflammatory responses (Raisuddin et al., 2011).
Other reports (Lambert et al., 2002) established that intraperitoneal
administration of NDGA is lethal to the mouse at LD50 of 75 mg/kg, showing
an increase in serum alanine aminotransferase levels, which indicates liver
damage. At the same, freshly isolated mouse hepatocytes were more sensitive
to NDGA than human melanoma cells. Furthermore, glucuronidation was
identified as a potential detoxification mechanism for NDGA. Both mono and
diglucuronide conjugates of NDGA were formed after intravenous dosing.
The monoglucuronide was also formed after the cultivation of NDGA with
human hepatic microsomes. This suggested that glucuronide conjugation is
essential in the metabolism of NDGA by humans (Lambert et al., 2002).
Additionally, NDGA has been tested for animal medicine as an effective
anti-angiogenic agent in ovalbumin-sensitized guinea pigs (Bergren and
Valentine, 2016).
Luo et al. (1988) determined the reduction in cell viability of a methanol
LT extract on P388, a murine leukemia line with an EC50 of 0.57 µg/mL
(Luo et al. 1988). Gil Salido et al. (2016) studied the antiproliferative effect
of various extracts from Mexican plants against various cancer lines. In
murine cell lines, LT had one of the highest hindering effects and showed a
dominant selective index, which is a measure of the selectivity of the extract
for eliminating cancer cells line rather than normal cells. The cancer cells
were eliminated 3.17 times more often than normal cells suggesting that LT
would be an excellent candidate for cancer treatment. In the human cell lines,
the extracts of LT showed IC50 of below 200 µg/mL (range, 64.28–163.73
µg/mL) (Gil Salido et al., 2016).
7.3.2 ANTIMICROBIAL ACTIVITY
It has been proven that the extracts from LT show antibacterial activity
against a wide range of bacterial strains (Morales-Ubaldo et al., 2021, 2022).
In addition, solutions of purified compounds such as NDGA, methylated-

NDGA (Reyes-Melo et al., 2017), 3′-demethoxy-6-O-demethyl-isoguaiacin
(Favela-Hernández et al., 2015), and others (Favela-Hernández et al., 2012),
also have antibacterial effects.
When testing for antibacterial activity, the most straightforward and
well-established method is the agar-substrate diffusion method (Das et
al., 2010). Martins et al. (2013) screened various LT extracts (methanol,
dichloromethane, and ethyl acetate as solvents) against Gram-positive and
Gram-negative bacteria. In general, the crude extracts hindered Gram-
positive bacteria’s growth more effectively; however, all of the extracts had
antibacterial properties and showed MIC between 62.5–125 µg/mL. The
ethyl acetate fractions from LT leaves were the most efficient in blocking the
growth of Staphylococcus aureus (Martins et al., 2013). When 95% ethanolic
extract (1:3 plant to solvent ratio) was used against methicillin-resistant S.
aureus the MIC was determined to be 60 µg/mL, and retardation activity
saw a 10−6-fold reduction in bacterial replication as compared to untreated
samples (Snowden et al., 2014).
Similarly, when crude aqueous and ethanolic extracts were tested against
food-borne pathogen bacteria Enterobacter aerogenes, Escherichia coli,
Salmonella typhi, and S. aureus, they showed high growth hindering activity
(80% growth suppression). In this case, however, the authors attributed
the LT extracts’ activity to the presence of tannins (Mendez et al., 2012).
Several mechanisms for tannin inhibition of bacterial growth can be found
in the literature, but they can be classified into the interaction of tannins with
bacterial and substrate proteins, interaction with bacterial cell wall plasma
membrane, and chelation of metal ions (Štumpf et al., 2020).
It also has been shown that ethanolic extracts (80%, 3 g/20 mL) of LT
were effective against several pathogens: Listeria monocytogenes (10 mg/
mL), Clostridium perfringens (12 mg/mL), Shigella dysenteriae (14 mg/
mL), Yersinia enterocolitica (19 mg/mL), Proteus vulgaris (10 mg/mL), the
actinomycetes, and some of the molds tested (Verástegui et al., 1996).
Looking more closely at specific lignans and flavonoids found in LT
extracts, Favela-Hernández et al. (2012) isolated seven compounds (NDGA;
4-epi-larreatricin; 3′-demethoxy-6-O-demethylisoguaiacin; 5,4′-dihydroxy-
3,7,8,3′-tetramethoxyflavone; 5,4′-dihydroxy-3,7,8-trimethoxyflavone;
5,4′-dihydroxy-7-methoxyflavone; and 5,8,4′-trihydroxy-3,7-dimethoxyfla-
vone). They showed that except for the last compound (5,8,4′-trihydroxy-
3,7-dimethoxyflavone), all compounds tested had some degree of activity
against five of the 12 bacteria tested. The main compound responsible for the
antimicrobial and antimycobacterial activity was the lignan 3′-demethoxy-

6-O-demethylisoguaiacin which displayed activity against resistant S.
aureus (MIC 25 mg/mL), Enterococcus faecalis (MIC 12.5 mg/mL), E. coli
(MIC 50 mg/mL), Enterobacter cloacae (MIC 12.5 mg/mL), and multidrug-
resistance strains of Mycobacterium tuberculosis (MIC 12.5–50 mg/mL). In
addition, the other lignans (NDGA and 4-epi-larreatricin) showed activity
against S. aureus MR (MIC 50 and >50 mg/mL) and E. cloacae (MIC 50 and
12.5 mg/mL), respectively. As the lignans responded to antimycobacterial
strains, the flavonoids had limited activity (Favela-Hernández et al., 2012).
Lignan meso-dihydroguaiaretic acid isolated from LT targets the α-subunit
of coenzyme A transferase of M. tuberculosis H37Rv present in both geraniol
and 1-and 2-methylnaphthalene degradation pathways (Clemente-Soto et al.,
2014). On the contrary, NDGA inhibits the growth of M. tuberculosis at low
concentrations (8 µg/mL) due to destabilization of the bacterial cell wall and
blocking the synthesis of fatty acids and mycolic acids (Guzmán-Beltrán et
al., 2016; Tsuchiya et al., 1996).
Morales-Ubado et al. (2022) proved that hydroalcoholic crude extracts
of LT, as well as their various fractions, exhibit antibacterial activity against
various bacteria: S. aureus6538, L. monocytogenes19113, E. coli35218,
Pseudomonas aeruginosa9027, as well as against multidrug-resistants S.
aureus01, S. aureus02, Bacillus cereus, E. coli01, E. coli02, Klebsiella pneu-
moniae, and Pasteurella multocida. The crude extract was more sensitive to
inhibiting Gram-positive bacteria. In addition, the authors showed the pres-
ence of thymol and carvacrol in the extract, which are proven antibacterial
compounds. Their mechanism of action is related to the inhibition of biofilm
formation and cell membrane damage. The presence of other compounds
(coumarin, caffeic acid derivatives, rutin, quercetin glucoside, kaempferol,
and nor 3′-demethoxyisoguaiacin) was also shown, and all of these have
been reported to have antibacterial activity, especially nor 3′-demethoxy-
isoguaiacin displays bactericidal activity against multidrug-resistant bacteria
associated with bovine mastitis (Morales-Ubado et al., 2022). In a similar
study, antibacterial activity against phytopathogenic multidrug-resistant
bacteria: Clavibacter michiganensis subsp. michiganensis, Pseudomona
syringae, and Xanthomonas campestris (strains causing plant diseases: bacte-
rial canker, bacterial speck, and bacterial spot, respectively) were shown in
ethanolic crude extract and its subfractions (Morales-Ubaldo et al., 2021).
Gram-negative bacteria, including E. coli and P. aeruginosa, show resistance
to 40% ethanoic LT extract treatment (MIC >1000 μg/mL). However,
Gram-positive bacteria are more sensitive to the treatments. Further analysis
suggested that the LT extracts act similar to β-lactam antibiotics; thus, they
inhibit the synthesis of the peptidoglycan layer of bacterial cell walls (Turner
et al., 2021).
7.3.3 ANTHELMINTIC
As mentioned above, tea from LT has been used in traditional medicine

against many different illnesses and within them as a treatment for intestinal
parasites (Arteaga et al., 2005; Mabry et al., 1978). Treatments against para-
sites were tested in an in vitro study of LT methanolic extracts on sheathed
and exsheathed larvae of Homonchus contortus. The anthelmintic activity at
low concentrations exhibited a low mortality rate (16–34%). At the highest
concentration tested (200 mg/mL), LT extracts displayed some cytotoxicity
for the sheathed larvae (30%), but a higher mortality rate was recorded in
the exsheathed larvae (about 70%). The extract of LT damaged the cuticle of
the larvae, with the degree of mortality dependent on the dose of the hydro-
methanolic extracts but not on the length of exposure time, which proves that
these extracts possess anti-H. contortus properties, predominantly during the
exsheathed stage of this nematode (García et al., 2018).
Compounds isolated from LT were studied against three parasites:
Entamoeba histolytica, Giardia lamblia, and N. fowleri. The ethyl acetate
fraction showed antiparasitic activity at 50 μg/mL. In this fraction,
nine compounds were identified; lignans: NDGA, 3′-O-methylnordihy-
droguairetic acid, nor-3′-demethoxyisoguaiacin, two analogs of nor-3′-
demethoxyisoguaiacin, ((7R, 7′R)-7, 7′-bis(4′,3,4-trihydroxyphenyl)-(8R,
8′S)-8,8′-dimethyltetrahydrofuran), and a flavonoid (3-Methoxy-6,7,4′-
trihydroxyflavonol). The compounds displayed a dose–response antiparasitic
activity against all three pathogens by reducing the culture density by 50%
compared to untreated trophozoite cultures. Looking at structure–activity
relationships, it has been suggested that the more flexible straight chain
structure offers more results in higher anthelmintic activity. On the contrary,
the difference in hydrophilicity may be a physical property of two compounds
preventing diffusion into the parasite trophozoites (Bashyal et al., 2017).
7.3.4 ANTIPROTOZOA ACTIVITY
Protozoa-caused diseases are grave health problems that have a profound

global impact on the human and animal populations (Nash, 2014). Protozoan
parasites that cause the majority of organ-damaging and often fatal diseases
are Giardia intestinalis, Trypanosoma cruzi, Plasmodium falciparum, L.
mexicana, and Trichomonas vaginalis (Lee et al., 2019). Unfortunately, the
treatment and medications available for these parasites are often toxic, induce
resistance, and present many adverse effects in the patients (Lee et al., 2019;
Gelb and Hol, 2002). In traditional Mexican medicine, LT tea has been used
as a treatment for parasites such as amoebiasis (Arteaga et al., 2005; Heron
and Yarnell, 2001); however, only a few proper investigations have been
carried out to evaluate these extracts as antiprotozoal agents. Nevertheless,
as early as 1978 (Segura, 1978), there was a report of ethanoic LT extract
containing NDGA (10−6 to 10−8 M) inhibitory activity against Entamoeba.
Among biocompounds in LT, meso-nordhydroguariaretic acid (m-NDGA)
shows the highest antiprotozoal activity (Schmidt et al., 2012). Regardless
of its composition, a crude dichloromethane extract of LT acted as a natural
antiprotozoal agent against Trypanosoma brucei rhodesiense, T. cruzi, Leish-
mania donovani, and P. falciparum. The major detected lignan, m-NDGA,
was responsible for the anti-inflammatory effects and was found to have the
IC50 values: of 4.5, 33.1, 12, and 7.7 µM, respectively, against the listed
above parasites (Schmidt et al., 2012). Bashyal et al. (2017) evaluated nine
lignans separated from ethanolic LT extract against E. histolytica, G. lamblia,
and N. fowleri. Six compounds exhibited a good antiparasitic response against
the pathogens by reducing the culture density by 50% (EC50 83–236 µM for
E. histolytica, 36–188 µM for G. lamblia, and 37–155 µM for N. fowleri)
compared to untreated trophozoite cultures. The authors draw a connection
between the structures of the compounds and their antiparasitic activity due
to flexible chains and additional methoxy groups in the structure. Two hydro-
phobic compounds did not display antiparasitic activity, most likely due to
difficulty diffusing into the parasite. The only isolated hydrophilic flavonol
had activity against G. lamblia (EC50 = 153 µM) and N. fowleri (EC50 =
235 µM) and was believed to diffuse actively through organism membranes.
NDGA and m-NDGA were also indicated to have the highest potential for
future study as novel drugs against N. fowleri since they are probably inhibi-
tors of cysteine protease activity in that parasite (Bashyal et al., 2017).
7.3.5 ANTIVIRAL ACTIVITY

Viruses as intracellular parasites are composed of either DNA or RNA
genetic information enclosed in a lipid covering. They are nonautonomous
and require a host body or living cell to perform their living function, such
as replication (Behl et al., 2021). The treatment against viruses focuses on

controlling the development of high infections via immunization (vaccina-
tion) or suppression of varial replication after exposure to the viral vector.
Typically, antiviral drugs target a virus-encoded factor that can prevent
the virus’s genetic information’s adhesion, inclusion, or replication. More
novel approaches target the host cells’ factors the virus needs but are not
mandatory for host cell functions (Kumar et al., 2020). Biocompounds active
against viruses are found within three main groups: alkaloids, terpenes, and
polyphenols (Behl et al., 2021).
Within biocompounds described in LT lignans, specifically, NDGA and
mNDGA, show antiviral activity (Cui et al., 2020). It is speculated that NDGA
inhibits viral infection by targeting genome replication and viral assembly
in the dengue virus (Soto-Acosta et al., 2014). In vitro studies of NDGA
displayed its action against the hepatitis C virus (Syed et al., 2011), West
Nile virus, and Zika virus (Merino-Ramos et al., 1996). NDGA probably
reduces genome replication in these viruses and disturbs lipid metabolism
by interfering with the sterol regulatory element binding proteins pathway.
Compounds from methylated derivate NDGA, such as 3-O-methyl-
NDGA, and other lignans with various degrees of O-methylation, were
first shown to possess activity against HIV in the mid-1990s (Gnabre et al.,
1996). These compounds were effective at inhibiting HIV replication in cell
cultures. The creosote bush lignans have structures that promote binding to
DNA sections loaded in cytosine/guanine connections and Specificity Protein
One (Sp1). If bound to Sp1, the blockade by the plant molecules, as tetra-O-
methylnordihydroquaiaretic acid, prevents Sp1 factors from connecting to
their related site, and the virus replication is revoked (Gnabre et al., 2015).
In the primary research, anti-HIV lignans from LT were isolated by
multistep solvent fractionation. Each fraction was evaluated by secreted
alkaline phosphatase assay against Tat transactivation of HIV-1. Obtained
compounds representing O-methyl and O-acetyl-substituted NDGA were
recognized as a new class of anti-HIV agents with crucial clinical relevance
(Gnabre et al., 1996).
7.3.6 TOXICITY
As with other phytocompounds, when consuming plant extracts, there is

always a possibility of toxicity or even poisoning by long-term exposure
(Harutyunyan et al., 2019; Wojcikowski et al., 2004; Licata et al., 2013).
In the late 1990s, reports surfaced on renal and hepatotoxicity issues due
to chronic use of LT tea and NDGA supplementation (Arteaga et al., 2005),
triggering several studies on the safety of LT compounds to properly assess
the level of toxicity, especially when it relates to NDGA exposure.
Sheikh et al. (1997) described 18 patients with LT-associated toxicity,
72% of whom revealed liver injury, with different clinical presentations,
containing cholestatic hepatitis, mild hepatitis, cirrhosis, and even acute
liver failure (Sheikh et al., 1997). As mentioned in the previous section, the
redox cycle of NDGA at specific concentrations leads to cytotoxic effects in
an organism; thus, the evaluation of its toxicity has been studied by various
research groups. One of these reported that LT toxicity is caused by NDGA
activity which inhibits cyclooxygenase and cytochrome P-450-dependent
monooxygenase activity in epidermal and hepatic microsomal preparations
(Agarwal et al., 1991).
NDGA is an inhibitor of intracellular vesicular transport at concentra-
tions between 50 and 100 µM it interrupts the secretory vesicular route and
the endocytic pathway in human dendritic cells (Drecktrah et al., 1998). At
lower concentrations (below 12 µM), it has a reversible cholestatic effect
in hepatocyte couplets (Arteaga et al., 2005), which strongly suggests that
it needs to be used in a specific range of effective concentrations. In their
review, Arteaga et al. (2005) propose a range between 10–100 µM as safe
concentrations since higher concentrations cause irreversible system damage.
Aarland et al. (2015) demonstrated the low toxicity of an LT-methanolic
extract in an in vivo model of Artemia salina, with a lethal dose of 50%
(DL50) of 1592 ppm when toxicity is considered at 200 ppm (Aarland et
al., 2015). Noncancerous Hs27 fibroblast cells were treated with LT extracts
(ethanol, ethanol: water, and water) at different concentrations (3.75–120 µg/
mL) to evaluate their toxicity, and no considerable cell death was observed at
concentrations below 120 µg/mL compared to the negative control (Skouta
et al., 2018). Thus, regarding a whole extract of LT, if a small dosage is
administered to patients, no visible damage is detected. Additionally, medi-
cating with LT should be done cautiously in persons with a history of liver
disease (Heron and Yarnell, 2001).
7.4 THE USE OF L. TRIDENTATA IN AGRICULTURE
Other studies looked for utilization of LT extracts in other than medical

applications due to the previously mentioned unique set of properties. For
example, purified polyphenols from LT were mixed into a bioactive and

biodegradable coating deposited on avocados cv Hass to prolong their shelf
life. As a result, the bioactive film was able to significantly reduce loss of
weight (40% and 58% for room and refrigerated conditions, respectively),
preserve firmness (9 times under refrigerated conditions), maintain bright-
ness (50% more than the control), and inhibit the development of internal
grey pulp (100%) (Aguirre-Joya et al., 2017). The tannin-rich aqueous
extract of LT has been used as a carbon source in solid-state fermentation
inoculated with Aspergillus niger. In the fermentation, gallic and ellagic
acids were produced at a fair rate within 96 h (Ventura et al., 2008).
7.4.1 ANTIFUNGAL
Due to the composition of high polyphenolic content, extracts from LT have

been explored as potential antifungal treatments. The plant phytoanticipins,
as generally secondary metabolites, generate molecular interaction of the
organelles and tissues of the fungi through targeting biomembranes, proteins,
or nucleic acids (Engelmeier and Hadacek, 2006). Alcohols are enzyme
inhibitors due to their lipophilic characteristic, especially phenolic alcohols’
free OH groups (Asael et al., 2018). Isothiocyanates and phenolic compounds
inactivate fungal proteins by binding to the amino group or sulfhydryl group
of amino acids (Tiznado and Troncoso, 2008). The alkaloids get introduced
into the DNA and cause mutations in the microorganism. Quinones, flavones,
flavonoids, tannins, and flavonols form nucleophilic complexes with proteins
catalyzing inactivation (Cowan, 1999; Selitrennikoff, 2001).
Tequida et al. (2002) reported the fungicide activity of several plant
extracts, including LT, against various fungi. They saw higher antifungal
activity in ethanolic vs. methanolic LT extract for all tested fungi (Aspergillus
niger; 77 vs 41, Aspergillus flavus; 76 vs 72, Penicillium chrysogenum; 63
vs 52, Penicillium expansum; 72 vs 58, Fusarium poae; 100 for both, and
Fusarium moniliforme.; 100 vs 88) (Tequida-Meneses et al., 2002). The
polyphenolic extract prepared from dry LT material in 70% acetone was
determined to be a strong fungicide that inhibited 100% inhibition in a variety
of moderately sensible and very sensible fungi (Pythium sp., Colletotrichum
truncatum, Colletotrichum coccodes, Alternaria alternata, Fusarium verticil-
lioides, Fusarium solani, Fusarium sambucinum, and Rhizoctonia solani)
(Tequida et al., 2002).
Previously, it has been believed that tannins as poly and monomeric

phenols present in LT extracts exhibit a potent antifungal activity because
they can form complexes with polysaccharides and proteins of external
layers of the fungal cells interfering with the functions of cell walls and
membranes, eventually causing the death of microorganisms (Aguilar et al.,
2007).
Extracts from LT showed good activity against A. flavus and Aspergillus
parasiticus. Within the extract, two lignans, NDGA and methyl-NDGA,
were identified as the phytoanticipins. The 100% inhibition for both fungi
treated was achieved at 300 and 500 µg/mL for methyl-NDGA and NDGA,
respectively (Vargas-Arispuro et al., 2005).
At the same time, aqueous extracts of LT are effective against Botrytis
cinerea, C. coccodes, and Fusarium oxysporum (Lira-Saldivar et al., 2006).
Polyphenols from LT ethanol extracts can inhibit 100% Phytophthora
cinnamomi at the concentration of 1000 ppm but also showed high growth
inhibitions in other solvents (water, lanolin, and cocoa), and the minimum
inhibitory concentration value of 6.96 mg/L (Castillo Reyes et al., 2015).
Leaf dichloromethane (DCM) and methanolic extracts can inhibit 100%
F.oxysporum f. sp. radicis-lycopersici at 3000 and 4000 ppm, respectively,
both in vitro and in vivo. The LT extract with DCM exhibited inhibition of
98% at 750 ppm; 94% for methanolic at 2000 ppm, and 97% for ethanolic
at 500 ppm, which indicates that the compounds extracted with EtOH had
greater fungicidal power over the fungus in vitro. When directly applied to
tomato plants, it reduced the severity and incidence of disease caused by
phytopathogenic fungus without significantly affecting the plant biology,
such as chlorophyll synthesis, plant height, and the number of leaves
(Peñuelas-Rubio et al., 2017).
7.4.2 ANTIPROTOZOA ACTIVITY
LT can be used as an unconventional and functional fooder for livestock, even

though it might not be preferential for the animals in taste respect. Protozoa
(Eimeria spp. oocysts) count was measured in rumen fluid and feces of sheep
fed with 5 and 10% LT substituted fodder. 10% creosote bush as a source
of fodder in sheep diets did not increase daily weight gain (very similar
nutritive value to control) but reduced the count of Eimeria spp. oocysts in
rumen fluid (down by 55%) and feces (down by 40%) (Hernández-Baez et
al., 2019).
7.5 OTHER APPLICATIONS
Recently, aqueous extracts from fresh leaves of LT were used to synthesize

silver nanoparticles (Méndez‐Andrade et al., 2022). The authors used the
nanoparticles to control the growth of C. michiganensis subsp. michiganensis
in tomato plants and determined that their presence had a positive influence
on the antioxidant protection of the plants.
7.6 CONCLUSIONS
Creosote bush (L. tridentata) has been used in traditional medicine and
research for many years. The abundance of various extractable polyphenols
creates an opportunity for many applications of that plant in medicine and
agriculture. These applications are related to LT’s antiseptic, antiamoebic,
antibiotic, antifungal, and antiviral properties. Recently described lignans
with potent antibacterial activity against multidrug-resistant bacteria give
hope for the development of a new line of medical treatment against these
microbes. However, there is still much to research in this field. It is unfortu-
nate that, thus far, there is only limited research on the antiviral application
of LT extracts and phytocompounds.
Similarly, there is a high expectation of using these biomolecules as
novel treatments in fighting various cancer diseases. Thus, expanding and
deepening the applied research of LT materials as antiviral and anticancer
agents is suggested. On the contrary, the application of polyphenols as
antifungal treatment in agriculture could replace the use of toxic chemicals
on plants.
KEYWORDS
• Larrea tridentate
• lignans
• phytocompounds
REFERENCES
Aarland, R. C.; Peralta-Gómez, S.; Sanchéz, C. M.; Parra-Bustamante, F.; Villa-Hernández, J.

M.; de León-Sánchez, F. D.; Pérez-Flores, L. J.; Rivera-Cabrera, F.; Mendoza-Espinoza, J.
A. A Pharmacological and Phytochemical Study of Medicinal Plants Used in Mexican Folk
Medicine. Indian J. Tradit. Knowl. 2015, 14 (4), 550–557.
Aboody, M. S. Al; Mickymaray, S. Anti-Fungal Efficacy and Mechanisms of Flavonoids.
Antibiotics 2020, 9 (2). https://doi.org/10.3390/antibiotics9020045.
Abou-Gazar, H.; Bedir, E.; Takamatsu, S.; Ferreira, D.; Khan, I. A. Antioxidant Lignans from
Larrea tridentata. Phytochemistry 2004, 65 (17), 2499–2505. https://doi.org/10.1016/j.
phytochem.2004.07.009.
Agarwal, R.; Wang, Z.; Bik, D.; Mukhtar, H. Nordihydroguaiaretic Acid, an Inhibitor of
Lipoxygenase, Also Inhibits Cytochrome P-450-Mediated Monooxygenase Activity in Rat
Epidermal and Hepatic Microsomes. Drug Metab. Dispos. 1991, 19 (3), 620–624.
Aguilar, C. N.; Rodríguez, R.; Gutiérrez-Sánchez, G.; Augur, C.; Favela-Torres, E.; Prado-
Barragan, L. A.; Ramírez-Coronel, A.; Contreras-Esquivel, J. C. Microbial Tannases:
Advances and Perspectives. Appl. Microbiol. Biotechnol. 2007, 76 (1), 47–59. https://doi.
org/10.1007/s00253-007-1000-2.
Aguirre-Joya, J. A.; Ventura-Sobrevilla, J.; Martínez-Vazquez, G.; Ruelas-Chacón, X.; Rojas,
R.; Rodríguez-Herrera, R.; Aguilar, C. N. Effects of a Natural Bioactive Coating on the
Quality and Shelf Life Prolongation at Different Storage Conditions of Avocado (Persea
Americana Mill.) Cv. Hass. Food Packag. Shelf Life 2017, 14 (September), 102–107.
https://doi.org/10.1016/j.fpsl.2017.09.003.
Andersson, D. I.; Hughes, D. Selection and Transmission of Antibiotic-Resistant
Bacteria. Microbiol. Spectr. 2017, 5 (4), 1–17. https://doi.org/10.1128/microbiolspec.
mtbp-0013-2016.
Arteaga, S.; Andrade-Cetto, A.; Cárdenas, R. Larrea tridentata (Creosote Bush), an Abundant
Plant of Mexican and US-American Deserts and Its Metabolite Nordihydroguaiaretic Acid.
J. Ethnopharmacol. 2005, 98 (3), 231–239. https://doi.org/10.1016/j.jep.2005.02.002.
Aruoma, O. I. Free Radicals, Oxidative Stress, and Antioxidants in Human Health and Disease.
J. Am. Oil Chem. Soc. 1998, 75 (2), 199–212. https://doi.org/10.1007/s11746-998-0032-9.
Asael, R. G. H.; Guevara-Gonzalez, R. G.; De Jesus, R. G. S.; Angelica, F. P. A. Antifungal
Activity of Mexican Endemic Plants on Agricultural Phytopathogens: A Review.
2018 14th Int. Eng. Congr. CONIIN 2018 2018, No. October. https://doi.org/10.1109/
CONIIN.2018.8489793.
Barakat, R.; Park, C. J.; Perez, P. A.; Chiu, K.; Ko, C. Female Antiestrogens. In:
Encyclopedia of Reproduction; Elsevier, 2018; pp 740–747. https://doi.org/10.1016/
B978-0-12-801238-3.64414-8.
Bashyal, B.; Li, L.; Bains, T.; Debnath, A.; LaBarbera, D. V. Larrea tridentata: A Novel Source
for Anti-Parasitic Agents Active Against Entamoeba histolytica, Giardia Lamblia and
Naegleria fowleri. PLoS Negl. Trop. Dis. 2017, 11 (8), e0005832. https://doi.org/10.1371/
journal.pntd.0005832.
Behl, T.; Rocchetti, G.; Chadha, S.; Zengin, G.; Bungau, S.; Kumar, A.; Mehta, V.; Uddin,
M. S.; Khullar, G.; Setia, D. et al. Phytochemicals from Plant Foods as Potential Source
of Antiviral Agents: An Overview. Pharmaceuticals 2021, 14 (4), 1–46. https://doi.
org/10.3390/ph14040381.
Bergren, D. R.; Valentine, J. L. Anti-Anaphylactic Action of Nordihydroguaiaretic Acid in

Antigen Sensitized Guinea Pigs. In: Respiratory Physiology and Neurobiology; Elsevier
B.V., 2016; pp 26–31. https://doi.org/10.1016/j.resp.2016.09.003.
Bickoff, E.; Williams, K. T.; Sparks, M. Stabilization of Carotene with Nordihydrouaiaretic
Acid and Other Antioxidants. Oil Soap 1945, 5 (19), 128–131.
Castillo Reyes, F.; Hernandez Castillo, F. D.; Clemente Constantino, J. A.; Gallegos Morales,
G.; Rodriguez Herrera, R.; Aguilar, C. N. In Vitro Antifungal Activity of Polyphenols-Rich
Plant Extracts against Phytophthora cinnamomi Rands. African J. Agric. Res. 2015, 10 (50),
4554–4560. https://doi.org/10.5897/ajar2013.8072.
Choi, Y. S.; Jung, M. Y. Kinetic Study on the Singlet Oxygen Quenching Activity of
Nordihydroguaiaretic Acid (NDGA) Using Methylene Blue Sensitized Photooxidation
of α-Terpinene. Food Sci. Biotechnol. 2016, 25 (5), 1333–1336. https://doi.org/10.1007/
s10068-016-0209-1.
Clemente-Soto, A. F.; Balderas-Rentería, I.; Rivera, G.; Segura-Cabrera, A.; Garza-
González, E.; Del Rayo Camacho-Corona, M. Potential Mechanism of Action of meso-
Dihydroguaiaretic Acid on Mycobacterium tuberculosis H37Rv. Molecules 2014, 19 (12),
20170–20182. https://doi.org/10.3390/molecules191220170.
Cowan, M. M. Plant Products as Antimicrobial Agents. Clin. Microbiol. Rev. 1999, 12 (4),
564–582. https://doi.org/10.1128/CMR.12.4.564.
Cui, Q.; Du, R.; Liu, M.; Rong, L. Lignans and Their Derivatives from Plants as Antivirals.
Molecules 2020, 25 (1), 1–17. https://doi.org/10.3390/molecules25010183.
Das, K.; Tiwari, R. K. S.; Shrivastava, D. K. Techniques for Evaluation of Medicinal Plant
Products as Antimicrobial Agent: Current Methods and Future Trends. J. Med. Plants Res.
2010, 4 (2), 104–111. https://doi.org/10.5897/JMPR09.030.
Dimitrios, B. Sources of Natural Phenolic Antioxidants. Trends Food Sci. Technol. 2006, 17
(9), 505–512. https://doi.org/10.1016/j.tifs.2006.04.004.
Drecktrah, D.; De Figueiredo, P.; Mason, R. M.; Brown, W. J. Retrograde Trafficking of
Both Golgi Complex and TGN Markers to the ER Induced by Nordihydroguaiaretic Acid
and Cyclofenil Diphenol. J. Cell Sci. 1998, 111 (7), 951–965. https://doi.org/10.1242/
jcs.111.7.951.
Engelmeier, D.; Hadacek, F. Chapter 17 Antifungal Natural Products: Assays and Applications;
2006; pp 423–467. https://doi.org/10.1016/S1572-557X(06)03017-0.
Favela-Hernández, J. M. J.; Clemente-Soto, A. F.; Balderas-Rentería, I.; Garza-González,
E.; Camacho-Corona, M. D. R. Potential Mechanism of Action of 3′-Demethoxy-6-O-
Demethylisoguaiacin on Methicillin Resistant Staphylococcus aureus. Molecules 2015, 20
(7), 12450–12458. https://doi.org/10.3390/molecules200712450.
Favela-Hernández, J. M. J.; García, A.; Garza-González, E.; Rivas-Galindo, V. M.; Camacho-
Corona, M. R. Antibacterial and Antimycobacterial Lignans and Flavonoids from Larrea
tridentata. Phyther. Res. 2012, 26 (12), 1957–1960. https://doi.org/10.1002/ptr.4660.
Firenzuoli, F.; Gori, L. Herbal Medicine Today: Clinical and Research Issues. Evid. Based
Complement. Altern. Med. 2007, 4 (SUPPL. 1), 37–40. https://doi.org/10.1093/ecam/
nem096.
Floriano-Sánchez, E.; Villanueva, C.; Medina-Campos, O. N.; Rocha, D.; Sanchez-González,
D. J.; Cardenas-Rodríguez, N.; Pedraza-Chaverrí, J. Nordihydroguaiaretic Acid Is a Potent
in Vitro Scavenger of Peroxynitrite, Singlet Oxygen, Hydroxyl Radical, Superoxide Anion
and Hypochlorous Acid and Prevents in Vivo Ozone-Induced Tyrosine Nitration in Lungs.
Free Radic. Res. 2006, 40 (5), 523–533. https://doi.org/10.1080/10715760500419365.
Foti, M.; Piattelli, M.; Baratta, M. T.; Ruberto, G. Flavonids, Coumarins, and Cinnamic Acids
as Antioxidants in a Micellar System. Structure–Activity Relationship. J. Agric. Food
Chem. 1996, 44 (2), 497–501. https://doi.org/10.1021/jf950378u.
García, J. E.; Gómez, L.; Mendoza-de-Gives, P.; Rivera-Corona, J. L.; Millán-Orozco, J.;
Ascacio, J. A.; Medina, M. A.; Mellado, M. Anthelmintic Efficacy of Hydro-Methanolic
Extracts of Larrea tridentata against Larvae of Haemonchus contortus. Trop. Anim. Health
Prod. 2018, 50 (5), 1099–1105. https://doi.org/10.1007/s11250-018-1535-5.
Gelb, M. H.; Hol, W. G. J. Tropical Protozoan Parasites. Science 2002, 297, 343–344.
Gil Salido, A. A.; Iloki Assanga, S. B.; Lewis Luján, Lidianys Maria Fernández Ángulo, D.;
Lara Espinoza, C. L.; Acosta Silva, A. L.; Rubio Pino, J. L. Composition of Secondary
Metabolites in Mexican Plant Extracts and Their Antiproliferative Activity towards Cancer
Cell Lines. Int. J. Sci. 2016, 2 (03), 63–77. https://doi.org/10.18483/ijsci.971.
Gnabre, J. N.; Ito, Y.; Ma, Y.; Huang, R. C. Isolation of Anti-HIV-1 Lignans from Larrea
tridentata by Counter-Current Chromatography. J. Chromatogr. A 1996, 719 (2), 353–364.
https://doi.org/10.1016/0021-9673(95)00727-X.
Gnabre, J.; Bates, R.; Huang, R. C. Creosote Bush Lignans for Human Disease Treatment and
Prevention: Perspectives on Combination Therapy. J. Tradit. Complement. Med. 2015, 5
(3), 119–126. https://doi.org/10.1016/j.jtcme.2014.11.024.
Guzmán-Beltrán, S.; Rubio-Badillo, M. Á.; Juárez, E.; Hernández-Sánchez, F.; Torres, M.
Nordihydroguaiaretic Acid (NDGA) and α-Mangostin Inhibit the Growth of Mycobacterium
tuberculosis by Inducing Autophagy. Int. Immunopharmacol. 2016, 31, 149–157. https://
doi.org/10.1016/j.intimp.2015.12.027.
Harutyunyan, K.; Balayan, K.; Tadevosyan, G.; Hayrapetyan, M.; Musayelyan, R.; Grigoryan,
R.; Khondkaryan, L.; Sarkisyan, N.; Babayan, N. Genotoxic Potential of Selected Medicinal
Plant Extracts in Human Whole Blood Cultures. J. HerbMed. Pharmacol. 2019, 8 (2),
160–162. https://doi.org/10.15171/jhp.2019.25.
Hausott, B.; Greger, H.; Marian, B. Naturally Occurring Lignans Efficiently Induce Apoptosis
in Colorectal Tumor Cells. J. Cancer Res. Clin. Oncol. 2003, 129 (10), 569–576. https://
doi.org/10.1007/s00432-003-0461-7.
Hernández-Baez, I.; García-López, J. C.; Espinosa-Reyes, G.; Lee-Rangel, H. A.;
Faz-Colunga, D. Á.; Pinos-Rodríguez, J. M. Creosote Bush (Larrea tridentata) Biomass as
Fodder for Sheep. Rev. Chapingo Ser. Zo. Áridas 2019, 18 (2), 1–9. https://doi.org/10.5154/r.
rchsza.2019.09.023.
Heron, S.; Yarnell, E. The Safety of Low-Dose Larrea tridentata (DC) Coville (Creosote
Bush or Chaparral): A Retrospective Clinical Study. J. Altern. Complement. Med. 2001, 7
(2), 175–185. https://doi.org/10.1089/107555301750164262.
Holohan, C.; Van Schaeybroeck, S.; Longley, D. B.; Johnston, P. G. Cancer Drug Resistance:
An Evolving Paradigm. Nat. Rev. Cancer 2013, 13 (10), 714–726. https://doi.org/10.1038/
nrc3599.
Hosseini, A.; Ghorbani, A. Cancer Therapy with Phytochemicals: Evidence from Clinical
Studies. Avicenna J. Phytomed. 2015, 5 (2), 84–97. https://doi.org/10.22038/ajp.2015.3872.
Hyder, P. W.; Fredrickson, E. L.; Estell, R. E.; Tellez, M.; Gibbens, R. P. Distribution and
Concentration of Total Phenolics, Condensed Tannins, and Nordihydroguaiaretic Acid
(NDGA) in Creosotebush (Larrea tridentata). Biochem. Syst. Ecol. 2002, 30 (10), 905–912.
https://doi.org/10.1016/S0305-1978(02)00050-9.
Jitsuno, M.; Mimaki, Y. Triterpene Glycosides from the Aerial Parts of Larrea tridentata.
Phytochemistry 2010, 71 (17–18), 2157–2167. https://doi.org/10.1016/j.phytochem.2010.
09.012.
Kasote, D. M. Flaxseed Phenolics as Natural Antioxidants. Int. Food Res. J. 2013, 20 (1),
27–34.
Kitts, D. D.; Yuan, Y. V.; Wijewickreme, A. N.; Thompson, L. U. Antioxidant Activity of the
Flaxseed Lignan Secoisolariciresinol Diglycoside and Its Mammalian Lignan Metabolites
Enterodiol and Enterolactone. Mol. Cell. Biochem. 1999, 202 (1–2), 91–100. https://doi.
org/10.1023/a:1007022329660.
Kumar, N.; Sharma, S.; Kumar, R.; Tripathi, B. N.; Barua, S.; Ly, H.; Rouse, B. T. Host-
Directed Antiviral Therapy. Clin. Microbiol. Rev. 2020, 33 (3), e00168-19 1-36. https://doi.
org/10.1128/CMR.00168-19.
Lambert, J. D.; Sang, S.; Dougherty, A.; Caldwell, C. G.; Meyers, R. O.; Dorr, R. T.;
Timmermann, B. N. Cytotoxic Lignans from Larrea tridentata. Phytochemistry 2005, 66
(7), 811–815. https://doi.org/10.1016/j.phytochem.2005.02.007.
Lambert, J. D.; Zhao, D.; Meyers, R. O.; Kuester, R. K.; Timmermann, B. N.; Dorr, R. T.
Nordihydroguaiaretic Acid: Hepatotoxicity and Detoxification in the Mouse. Toxicon 2002,
40 (12), 1701–1708. https://doi.org/10.1016/S0041-0101(02)00203-9.
Lee; Kim; Hayat; Shin. Recent Advances in the Discovery of Novel Antiprotozoal Agents.
Molecules 2019, 24 (21), 3886. https://doi.org/10.3390/molecules24213886.
Leon, D.; Parada, D.; Vargas-Uribe, M.; Perez, A. A.; Ojeda, L.; Zambrano, A.; Reyes, A.
M.; Salas, M. Effect of Nordihydroguaiaretic Acid on Cell Viability and Glucose Transport
in Human Leukemic Cell Lines. FEBS Open Bio. 2016, 6 (10), 1000–1007. https://doi.
org/10.1002/2211-5463.12106.
Licata, A.; Macaluso, F. S.; Craxì, A. Herbal Hepatotoxicity: A Hidden Epidemic. Intern.
Emerg. Med. 2013, 8 (1), 13–22. https://doi.org/10.1007/s11739-012-0777-x.
Lira-Saldivar, R. H.; Hernández-Suárez, M.; Hernández-Suárez, F. D. Activity of Larrea
tridentata (D .C .) Coville L . Extracts and Chitosan against Fungi That Affect Horticultural
Crops. Chapingo Ser. Hortic. 2006, 12 (2), 211–216.
Lu, H.; Liu, G.-T. Anti-Oxidant Activity of Dibenzocyclooctene Lignans Isolated from
Schisandraceae.Planta Med.1992,58(04), 311–313. https://doi.org/10.1055/s-2006-961473.
Lundberg, W. O.; Halvorson, H. O.; Burr, G. O. The Antioxidant Properties of
Nordihydroguaiaretic Acid. Oil Soap 1944, 21 (2), 33–35. https://doi.org/10.1007/
BF02593156.
Luo, Z.; Meksuriyen, D.; Erdelmeier, C. A. J.; Fong, H. H. S.; Cordell, G. A. Larreantin, a
Novel, Cytotoxic Naphthoquinone from Larrea tridentata. J. Org. Chem. 1988, 53 (10),
2183–2185. https://doi.org/10.1021/jo00245a011.
Mabry, T. J.; Hunziker, J. H.; Jr., D. R. D. Creosote Bush: Biology and Chemistry of Larrea in
New World Deserts; Dowden, Hutchinson & Ross, Inc., USA, 1978.
Manda, G.; Rojo, A. I.; Martínez-Klimova, E.; Pedraza-Chaverri, J.; Cuadrado, A.
Nordihydroguaiaretic Acid: From Herbal Medicine to Clinical Development for Cancer
and Chronic Diseases. Front. Pharmacol. 2020, 11 (Feb), 1–21. https://doi.org/10.3389/
fphar.2020.00151.
Martins, S.; Amorim, E. L. C.; Sobrinho, T. J. S. P.; Saraiva, A. M.; Pisciottano, M. N. C.;
Aguilar, C. N.; Teixeira, J. A.; Mussatto, S. I. Antibacterial Activity of Crude Methanolic
Extract and Fractions Obtained from Larrea tridentata Leaves. Ind. Crops Prod. 2013, 41
(1), 306–311. https://doi.org/10.1016/j.indcrop.2012.04.037.
Mehta, S. K.; Gowder, S. J. T. Members of Antioxidant Machinery and Their Functions. In

Basic Principles and Clinical Significance of Oxidative Stress; Thatha Gowder, J. S., Ed.;
InTech, 2015; p 13. https://doi.org/10.5772/61884.
Mendez, M.; Rodríguez, R.; Ruiz, J.; Morales-Adame, D.; Castillo, F.; Hernández-Castillo,
F. D.; Aguilar, C. N. Antibacterial Activity of Plant Extracts Obtained with Alternative
Organics Solvents against Food-Borne Pathogen Bacteria. Ind. Crops Prod. 2012, 37 (1),
445–450. https://doi.org/10.1016/j.indcrop.2011.07.017.
Méndez‐Andrade, R.; Vallejo‐Perez, M. R.; Loera‐Alvarado, E.; los Santos‐Villarreal, G.;
García‐Cerda, L. A.; Vera‐Reyes, I. Efficacy of Biosynthesized Silver Nanoparticles from
Larrea tridentata against Clavibacter michiganensis. J. Phytopathol. 2022, 170 (2), 91–99.
https://doi.org/10.1111/jph.13058.
Merino-Ramos, T.; Jiménez De Oya, N.; Saiz, J. C.; Martín-Acebes, M. A. Antiviral Activity
of Nordihydroguaiaretic Acid and Its Derivative Tetra-O-Methyl Nordihydroguaiaretic
Acid against West Nile Virus and Zika Virus. Antimicrob. Agents Chemother. 2017, 61 (8).
https://doi.org/10.1128/AAC.00376-17.
Morales-Ubaldo, A. L.; Gonzalez-Cortazar, M.; Zaragoza-Bastida, A.; Meza-Nieto, M.
A.; Valladares-Carranza, B.; A. Alsayegh, A.; El-Saber Batiha, G.; Rivero-Perez, N.
Nor 3′-Demethoxyisoguaiacin from Larrea tridentata Is a Potential Alternative against
Multidrug-Resistant Bacteria Associated with Bovine Mastitis. Molecules 2022, 27 (11),
3620. https://doi.org/10.3390/molecules27113620.
Morales-Ubaldo, A. L.; Rivero-Perez, N.; Avila-Ramos, F.; Aquino-Torres, E.; Prieto-Méndez,
J.; Hetta, H. F.; El-Saber Batiha, G.; Zaragoza-Bastida, A. Bactericidal Activity of Larrea
tridentata Hydroalcoholic Extract against Phytopathogenic Bacteria. Agronomy 2021, 11
(5), 957. https://doi.org/10.3390/agronomy11050957.
Nash, T. E. Parasitic Diseases That Cause Seizures. Epilepsy Curr. 2014, 14 (2_suppl), 29–34.
https://doi.org/10.5698/1535-7511-14.s2.29.
Osorio, E.; Flores, M.; Hernández, D.; Ventura, J.; Rodríguez, R.; Aguilar, C. N. Biological
Efficiency of Polyphenolic Extracts from Pecan Nuts Shell (Carya Illinoensis), Pomegranate
Husk (Punica Granatum) and Creosote Bush Leaves (Larrea tridentata Cov.) against Plant
Pathogenic Fungi. Ind. Crops Prod. 2010, 31 (1), 153–157. https://doi.org/10.1016/j.
indcrop.2009.09.017.
Park, R.; Chang, C. C.; Liang, Y. C.; Chung, Y.; Henry, R. A.; Lin, E.; Mold, D. E.; Huang, R.
C. C. Systemic Treatment with Tetra-O-Methyl Nordihydroguaiaretic Acid Suppresses the
Growth of Human Xenograft Tumors. Clin. Cancer Res. 2005, 11 (12), 4601–4609. https://
doi.org/10.1158/1078-0432.CCR-04-2188.
Peñuelas-Rubio, O.; Arellano-Gil, M.; Verdugo-Fuentes, A. A.; Chaparro-Encinas, L. A.;
Hernández-Rodríguez, S. E.; Martínez-Carrillo, J. L.; Vargas-Arispuro, I. D. C. Extractos
de Larrea tridentata Como Una Estrategia Ecológica Contra Fusarium oxysporum Radicis-
Lycopersici En Plantas de Tomate Bajo Condiciones de Invernadero. Rev. Mex. Fitopatol.
Mex. J. Phytopathol. 2017, 35 (3), 360–376. https://doi.org/10.18781/r.mex.fit.1703-3.
Perlin, D. S.; Rautemaa-Richardson, R.; Alastruey-Izquierdo, A. The Global Problem of
Antifungal Resistance: Prevalence, Mechanisms, and Management. Lancet Infect. Dis.
2017, 17 (12), e383–e392. https://doi.org/10.1016/S1473-3099(17)30316-X.
Raisuddin, S.; Rahman, S.; Ansari, R. A.; Rehman, H.; Parvez, S. Nordihydroguaiaretic
Acid from Creosote Bush (Larrea tridentata) Mitigates 12- O -Tetradecanoylphorbol-
13-Acetate-Induced Inflammatory and Oxidative Stress Responses of Tumor Promotion
Cascade in Mouse Skin. Evid. Based Complement. Altern. Med. 2011, 2011. https://doi.
org/10.1093/ecam/nep076.
Reyes-Melo, K.; García, A.; Romo-Mancillas, A.; Garza-González, E.; Rivas-Galindo, V. M.;
Miranda, L. D.; Vargas-Villarreal, J.; Favela-Hernández, J. M. J.; Camacho-Corona, M.
del R. Meso-Dihydroguaiaretic Acid Derivatives with Antibacterial and Antimycobacterial
Activity. Bioorganic Med. Chem. 2017, 25 (20), 5247–5259. https://doi.org/10.1016/j.
bmc.2017.07.047.
Rowe, D. L.; Ozbay, T.; Bender, L. M.; Nahta, R. Nordihydroguaiaretic Acid, a Cytotoxic
Insulin-like Growth Factor-I Receptor/HER2 Inhibitor in Trastuzumab-Resistant Breast
Cancer. Mol. Cancer Ther. 2008, 7 (7), 1900–1908. https://doi.org/10.1158/1535-7163.
MCT-08-0012.
Rzedowski, J.; Huerta, L. Vegetación de México, 1st ed.; Editorial Limusa, S. A. Noriega
Editores, 1994.
Saleem, M.; Kim, H. J.; Ali, M. S.; Lee, Y. S. An Update on Bioactive Plant Lignans. Nat.
Prod. Rep. 2005, 22 (6), 696. https://doi.org/10.1039/b514045p.
Schmidt, T. J.; Rzeppa, S.; Kaiser, M.; Brun, R. Larrea tridentata—Absolute Configuration
of Its Epoxylignans and Investigations on Its Antiprotozoal Activity. Phytochem. Lett.
2012, 5 (3), 632–638. https://doi.org/10.1016/j.phytol.2012.06.011.
Segura, J. J. Effects of Nordihydroguairetic Acid and Ethanol on the Growth of Entamoeba
Invadens. Arch. Invest. Med. (Mex). 1978, 1, 157–162.
Selitrennikoff, C. P. Antifungal Proteins. Appl. Environ. Microbiol. 2001, 67 (7), 2883–2894.
https://doi.org/10.1128/AEM.67.7.2883-2894.2001.
Sheikh, N. M. Chaparral-Associated Hepatotoxicity. Arch. Intern. Med. 1997, 157 (8), 913.
https://doi.org/10.1001/archinte.1997.00440290099011.
Simpson, D.; Amos, S. Other Plant Metabolites. In Pharmacognosy; Elsevier, 2017; pp
267–280. https://doi.org/10.1016/B978-0-12-802104-0.00012-3.
Sirotkin, A. V.; Halim Harrath, A. Phytoestrogens and Their Effects. Eur. J. Pharmacol. 2014,
741, 230–236. https://doi.org/10.1016/j.ejphar.2014.07.057.
Skouta, R.; Morán-Santibañez, K.; Valenzuela, C. A.; Vasquez, A. H.; Fenelon, K.
Assessing the Antioxidant Properties of Larrea tridentata Extract as a Potential Molecular
Therapy against Oxidative Stress. Molecules 2018, 23 (7), 1–17. https://doi.org/10.3390/
molecules23071826.
Snowden, R.; Harrington, H.; Morrill, K.; Jeane, L. D.; Garrity, J.; Orian, M.; Lopez, E.;
Rezaie, S.; Hassberger, K.; Familoni, D.; et al. A Comparison of the Anti-Staphylococcus
aureus Activity of Extracts from Commonly Used Medicinal Plants. J. Altern. Complement.
Med. 2014, 20 (5), 375–382. https://doi.org/10.1089/acm.2013.0036.
Soto-Acosta, R.; Bautista-Carbajal, P.; Syed, G. H.; Siddiqui, A.; Del Angel, R. M.
Nordihydroguaiaretic Acid (NDGA) Inhibits Replication and Viral Morphogenesis
of Dengue Virus. Antiviral Res. 2014, 109 (1), 132–140. https://doi.org/10.1016/j.
antiviral.2014.07.002.
Source Naturals Lignan Extract Caps. https://www.swansonvitamins.com/p/source-naturals-
lignan-extract-70-mg-60-caps (accessed 18 Sept 2022).
Stull, J. W.; Herreid, E. O.; Tracy, P. H. AStudy of the Use of the Antioxidant Nordihydroguaiaretic
Acid in Dairy Products. II: Its Antioxygenic Properties in Unsweetened Frozen Cream. J.
Dairy Sci. 1948, 31 (12), 1024–1028. https://doi.org/10.3168/jds.S0022-0302(48)92292-9.
Štumpf, S.; Hostnik, G.; Primožič, M.; Leitgeb, M.; Salminen, J.-P.; Bren, U. The Effect
of Growth Medium Strength on Minimum Inhibitory Concentrations of Tannins and
Tannin Extracts against E. coli. Molecules 2020, 25 (12), 2947. https://doi.org/10.3390/

molecules25122947.
Syed, G. H.; Siddiqui, A. Effects of Hypolipidemic Agent Nordihydroguaiaretic Acid on
Lipid Droplets and Hepatitis C Virus. Hepatology 2011, 54 (6), 1936–1946. https://doi.
org/10.1002/hep.24619.
Tequida-Meneses, M.; Cortez-Rocha, M.; Rosas-Burgos, E. C.; López-Sandoval, S.; Corrales-
Maldonado, C. Efecto de Extractos Alcohólicos de Plantas Silvestres Sobre La Inhibición de
Crecimiento de Aspergillus flavus, Aspergillus niger, Penicillium chrysogenum, Penicillium
expansum, Fusarium moniliforme y Fusarium poae. Rev. Iberoam. Micol. 2002, 19, 84–88.
Thangapazham, R. L.; Sharad, S.; Maheshwari, R. K. Phytochemicals in Wound Healing. Adv.
Wound Care 2016, 5 (5), 230–241. https://doi.org/10.1089/wound.2013.0505.
Tiznado, E.; Troncoso, R. Control of Fungal Diseases with Isothiocyanates. Stewart
Postharvest Rev. 2008, 2 (1), 1–14. https://doi.org/10.2212/spr.2006.1.4.
Treviño-Cueto, B.; Luis, M.; Contreras-Esquivel, J. C.; Rodríguez, R.; Aguilera, A.; Aguilar,
C. N. Gallic Acid and Tannase Accumulation during Fungal Solid State Culture of a Tannin-
Rich Desert Plant (Larrea tridentata Cov.). Bioresour. Technol. 2007, 98 (3), 721–724.
https://doi.org/10.1016/j.biortech.2006.02.015.
Tsimberidou, A. M. Targeted Therapy in Cancer. Cancer Chemother. Pharmacol. 2015, 76
(6), 1113–1132. https://doi.org/10.1007/s00280-015-2861-1.
Tsuchiya, H.; Sato, M.; Miyazaki, T.; Fujiwara, S.; Tanigaki, S.; Ohyama, M.; Tanaka, T.;
Iinuma, M. Comparative Study on the Antibacterial Activity of Phytochemical Flavanones
against Methicillin-Resistant Staphylococcus aureus. J. Ethnopharmacol. 1996, 50 (1),
27–34. https://doi.org/10.1016/0378-8741(96)85514-0.
Turner, T.; Ruiz, G.; Gerstel, J.; Langland, J. Characterization of the Antibacterial Activity
from Ethanolic Extracts of the Botanical, Larrea tridentata. BMC Complement. Med. Ther.
2021, 21 (1), 177. https://doi.org/10.1186/s12906-021-03344-9.
Vargas-Arispuro, I.; Reyes-Báez, R.; Rivera-Castañeda, G.; Martínez-Téllez, M. A.; Rivero-
Espejel, I. Antifungal Lignans from the Creosotebush (Larrea tridentata). Ind. Crops Prod.
2005, 22 (2), 101–107. https://doi.org/10.1016/j.indcrop.2004.06.003.
Ventura, J.; Belmares, R.; Aguilera-Carbo, A.; Gutiérrez-Sanchez, G.; Rodríguez-Herrera, R.;
Aguilar, C. N. Fungal Biodegradation of Tannins from Creosote Bush (Larrea tridentata)
and Tar Bush (Fluorensia Cernua) for Gallic and Ellagic Acid Production. Food Technol.
Biotechnol. 2008, 46 (2), 213–217.
Verástegui, M. A.; Sánchez, C. A.; Heredia, N. L.; García-Alvarado, J. S. Antimicrobial
Activity of Extracts of Three Major Plants from the Chihuahuan Desert. J. Ethnopharmacol.
1996, 175–177. https://doi.org/10.1016/0378-8741(96)84802-1.
Wang, B.; Yu, S.-c.; Jiang, J.-y.; Porter, G. W.; Zhao, L.-t.; Wang, Z.; Tan, H.; Cui, Y.-h.;
Qian, C.; Ping, Y.-f. et al. An Inhibitor of Arachidonate 5-Lipoxygenase, Nordy, Induces
Differentiation and Inhibits Self-Renewal of Glioma Stem-Like Cells. Stem Cell Rev.
Reports 2011, 7 (2), 458–470. https://doi.org/10.1007/s12015-010-9175-9.
Wen, C. C.; Chen, H. M.; Yang, N. S. Developing Phytocompounds from Medicinal Plants as
Immunomodulators, Vol. 62; 2012. https://doi.org/10.1016/B978-0-12-394591-4.00004-0.
Wojcikowski, K.; Johnson, D. W.; Gobé, G. Medicinal Herbal Extracts—Renal Friend or Foe?
Part One: The Toxicities of Medicinal Herbs. Nephrology 2004, 9 (5), 313–318. https://doi.
org/10.1111/j.1440-1797.2004.00310.x.
Yokosuka, A.; Iguchi, T.; Jitsuno, M.; Mimaki, Y. Structure and Cytotoxicity of Novel Lignans
and Lignan Glycosides from the Aerial Parts of Larrea tridentata. Molecules 2021, 26 (20),
6186. https://doi.org/10.3390/molecules26206186.
Yokosuka, A.; Matsuo, Y.; Jitsuno, M.; Adachi, K.; Mimaki, Y. Larrealignans A and B, Novel
Lignan Glycosides from the Aerial Parts of Larrea tridentata. Chem. Pharm. Bull. 2011, 59
(12), 1467–1470. https://doi.org/10.1248/cpb.59.1467.
Youngren, J. F.; Gable, K.; Penaranda, C.; Maddux, B. A.; Zavodovskaya, M.; Lobo, M.;
Campbell, M.; Kerner, J.; Goldfine, I. D. Nordihydroguaiaretic Acid (NDGA) Inhibits the
IGF-1 and c-ErbB2/HER2/Neu Receptors and Suppresses Growth in Breast Cancer Cells.
Breast Cancer Res. Treat. 2005, 94 (1), 37–46. https://doi.org/10.1007/s10549-005-6939-z.
Zhao, Q. W.; Lin, Y.; Xu, C. R.; Yao, Y. L.; Cui, Y. H.; Zhang, X.; Bian, X. W. NDGA-P21,
a Novel Derivative of Nordihydroguaiaretic Acid, Inhibits Glioma Cell Proliferation and
Stemness. Laboratory Investigation. Nature Publishing Group 2017; pp 1180–1187. https://
doi.org/10.1038/labinvest.2017.46.
Zitterman, A. DIETARY FIBER | Bran. In Encyclopedia of Food Sciences and Nutrition;
Elsevier, 2003; pp 1844–1850. https://doi.org/10.1016/B0-12-227055-X/00346-1.
CHAPTER 8
Perspectives for Innovation of New Food

Preservatives from Euphorbiaceae
REBECA VILLANUEVA CHÁVEZ, LUIS ENRIQUE COBOS-PUC,
ELDA PATRICIA SEGURA-CENICEROS, ANNA ILINÁ, and
SONIA YESENIA SILVA BELMARES
Department of Food Research, School of Chemical Sciences,
Autonomous University of Coahuila, Saltillo, Coahuila, Mexico
ABSTRACT
The microbial deterioration of food and its intake represents a public health
problem due to bacterial resistance. According to this, the losses of food
products cause great economic losses in the food industry. Therefore, the
food industry incorporates preservatives to prolong the shelf life of foods.
Preservatives frequently added to food production are sorbates and benzo-
ates. However, they present undesirable effects on health that can be mild or
severe. The most common are bloating and changes in the gut microbiota, but
they can cause tachycardia, hives, and heart disease. Therefore, it is required
to innovate preservatives with fewer adverse health effects. Plants represent
a great alternative to obtaining natural preservatives that, when incorporated
into food, improve the quality of life. Since ancient times, plants have been
traditionally used as food preservatives because they contain antimicrobial
compounds such as essential oils. However, due to its volatility, the effect
may be lost. Euphorbiaceae represents a potential source of innovative new
preservatives since its species contain compounds with remarkable anti-
bacterial effects. These plants synthesize different compounds as a defense
mechanism to survive against pathogens. Therefore, the generation of
knowledge about the chemical structure of antimicrobial compounds could

lay the groundwork for innovating new preservatives.
8.1 INTRODUCTION
Food spoilage occurs frequently causing economic losses for the food
industry. The deterioration begins with a natural oxidative process triggered
by a series of chemical and enzymatic reactions that result in the senescence
of the product (Anwar et al., 2019). Therefore, the food gradually loses its
properties over time, reflected in physiological and organoleptic changes (De
Oliveira et al., 2021). For this reason, preservation methods have been used
for years to preserve foods of animal or plant origin to feed ever-growing
populations. Conservation allows the integrity of the food to be maintained
throughout the production process until the final product is marketed (Floros
et al., 2010). However, the most important thing is its conservation during
handling, transport, and storage (Thyberg and Tonjes, 2008). Consequently,
different preservation methods have been used but some modify the nutritional
composition of foods during the process. Additionally, synthetic additives
have been conventionally added into processed foods for years (Joardder
and Masud, 2019). Some preservatives maintain the integrity of nutrients
due to antioxidant properties mean prevention of oxidative deterioration,
while others inhibit microbial growth. However, they have side effects that
harm health because they exceed the recommended daily intake (Trasande
et al., 2018). They are generally recognized as safe (GRAS) substances, but
some cause serious adverse effects such as cancer, multiple sclerosis, and
heart disease (Mazdeh et al., 2014; Baran et al., 2021). Therefore, natural
preservatives in processed foods could reduce health risks from the intake
of synthetic additives (Sharif et al., 2017). Plant terpenoids and phenolic
compounds inhibit or retard the growth of microorganisms (Gonelimali et
al., 2018; Negi, 2012).
Essential oils are an example of terpenoids used since ancient times as
food preservatives. Thymol, eugenol, and carvacrol have an effect against
bacteria transmitted by contaminated food and are used to preserve meat,
dairy, and vegetable products (Pandey et al., 2017). The main mechanism
of the antibacterial effect is damage to the bacterial membrane (Faustino
et al., 2019). Consequently, plants represent a source to discover new food
preservatives that replace conventional additives with even higher activi-
ties in the short term (Pisoschi et al., 2019). The plants used since ancient
Perspectives for Innovation of New Food Preservatives from Euphorbiaceae 199
times to preserve food are aromatic due to terpenoids, but the Euphorbiaceae
contain them and have an antimicrobial effect. Euphorbiaceae grow widely
in Africa and America (Islam et al., 2019). Therefore, they easily adapt to
different habitats and produce a variety of bioactive compounds (Mwine and
van Damme, 2011). One of its most outstanding effects is the antimicrobial
due to monoterpenes, diterpenes, and triterpenes (Félix-Silva et al., 2018).
However, it has antioxidant activity due to the content of flavonoids, alka-
loids, and tannins (Greay and Hammer, 2015; Cavalcante et al., 2020). In
consequence, they can be incorporated as preservatives in foods to improve
shelf life and prevent their senescence due to environmental factors and
microorganisms (Pandey et al., 2017). Some species of Euphorbiaceae that
could be used as preservatives are described below since they have an anti-
oxidant or antimicrobial effect.
Euphorbia heterophylla has an antibacterial effect attributed to its
aqueous extract (Ughachukwu et al., 2014). Manihot multifida (L.) Crantz’s
antioxidant and antimicrobial effect is attributed to its extracts (Fabri et al.,
2015). Euphorbia ebracteolata Hayata has an antimicrobial effect attributed
to terpenoids, acetophenones, and flavonoids (Ting et al., 2020). Euphorbia
neriifolia L. contains β-amyrin recognized as responsible for the antibacte-
rial effect. However, it is also related to flavonoids, terpenoids, and alkaloids
(Sultana et al., 2022). Euphorbia tirucalli contains alkaloids, flavonoids,
tannins, and terpenoids. The stem has polyphenols with antioxidant activity
(Nguyen et al., 2021) and peroxidases used to synthesize aromatic chemicals
and remove peroxides from food products and industrial waste (Shukla et
al., 2017). According to this, Euphorbiaceae plants are of industrial interest
(Keneni and Marchetti, 2017); Jatropha curcas and Jatropha dioica are
examples employed to obtain biodiesel from their oils (Zahan and Kano,
2018). However, phorbol esters from J. curcas extracted from a by-product
of the biodiesel fuel industry affect phytopathogenic fungi (Saetae and
Suntornsuk, 2012). In addition, J. dioica has an antimicrobial effect attrib-
uted to β-sitosterol (Silva-Belmares et al., 2014). Hevea brasiliensis contains
antioxidant compounds that can be used in the food industry to inhibit the
formation of free radicals to preserve the existing properties of food matrices
(Oleinik et al., 2022). Therefore, extraction methods have been implemented
to recover bioactive extracts with environmentally friendly solvents (Oleinik
et al., 2022)].
Manihot esculenta is used to formulate industrial products (Okezie et
al., 1983), but it generates waste rich in bioactive compounds. Therefore,
the waste has the potential to produce products with added value (Cui et
al., 2017). On the contrary, starch has been incorporated into biocomposite
materials to preserve food since it includes antimicrobial compounds such as
essential oils (De Souza et al., 2014). Since these materials have advantages
over conventional ones formulated based on synthetic plastics, the M. escu
lenta films are edible (Bharti et al., 2020). Therefore, bioactive compounds
and plant polymers from Euphorbiaceae could be used to formulate food
preservatives, previously evaluating their toxicity (Saetae and Suntornsuk,
2012).
8.2 FOOD PRESERVATION AND ADVERSE EFFECTS OF SYNTHETIC

PRESERVATIVES
Preservation methods have been used for years to preserve hunted foods or
vegetables gathered from the field to feed ever-growing populations. There-
fore, the integrity of the food must be maintained throughout the production
process, from its cultivation at the time of planting food to the commercializa-
tion of the final product (Floros et al., 2010). Therefore, changes in a society
focused on nutritional value, taste, texture, and quality of food have emerged
throughout Consequently, food preservation focuses on reducing food waste
and maintaining accessibility, including in geographic areas where food is
not produced. However, the most important thing is its conservation during
handling, transport, and storage (Thyberg and Tonjes, 2008; Kumari et al.,
2019). Consequently, food preservation focuses on reducing food waste
and maintaining accessibility, including in geographic areas where food is
not produced. However, the most important thing is its conservation during
handling, transport, and storage (Thyberg and Tonjes, 2008). Therefore,
throughout history has been used conservation methods such as cooking
(50,000 BC), drying (12,000 BC), fermentation (10,000 BC), sautéing,
pickling (3600 BC), smoking (3500 BC), refrigeration (1784), and canning
(1809). Although these methods are effective, some of them modify the
nutritional composition of the food during the process due to temperature.
For this reason, since 1950 synthetic additives have been incorporated into
conventionally processed foods (Joardder and Masud, 2019).
Some preservatives focus on maintaining the nutrient integrity of the
food through the use of antioxidant compounds since they prevent oxidative
deterioration of the food. However, others are used to inhibit the growth of
bacteria, yeasts, and molds. Butylated hydroxyanisole (BHA) is a widely
used food preservative for its antioxidant effect. Additionally, it is used in
the pharmaceutical and cosmetic industries to prevent lipid oxidation. On

the contrary, A synthetic antimicrobial additive is a substance added to fruits,
vegetables, and prepared foods to increase their shelf life and maintain
quality and safety. Because it retards the growth of microbial contamination,
fermentative processes and decomposition. Benzoates; sorbates; sodium
or potassium nitrite and nitrate; sulfites; and glutamate are the most used.
However, it has been seen that they have secondary effects due to their
excessive use, which is why they harm the health of children and adults
(Trasande et al., 2018). Since they frequently exceed the recommended
daily intake (Chaleshtori et al., 2018). They are generally GRAS substances,
but some salts, such as benzoate and sorbate produce allergies, intolerance,
cancer, multiple sclerosis, nausea, heart disease, and hives at high concentra-
tions (Mazdeh et al., 2014). BHA has mutagenic and carcinogenic potential
(Baran et al., 2021). Sodium nitrate is frequently used to preserve cured meat
products but has a toxic effect on humans and animals. It also causes methe-
moglobinemia due to the N-nitrosamines that are produced during metabolic
conversion after ingestion (Özen et al., 2014).
Accordingly, the effect produced by the intake of preservatives that are
ingested regularly through the consumption of processed foods that exist in
the market is alarming. On the contrary, it is advisable to include scientific
information on adverse effects in the labeling to reduce excessive intake due
to misinformation towards the consumer. Therefore, to reduce health risks
due to the intake of synthetic additives proposed addition of natural preser-
vatives on food such as organic acids, vinegar, sugar, and spices (Sharif et
al., 2017), since these are more tolerable for humans.
8.3 NATURAL ALTERNATIVES FOR FOOD PRESERVATION
Currently, leaves, stems, roots, or the entire plant are known to contain essen-
tial oils, phenolic compounds, terpenes, alkaloids, aldehydes, ketones, organic
acids, saponins, thiosulfates, and sterols that inhibit or retard the growth of
bacteria, fungi, and yeasts (Gonelimali et al., 2018; Lyu et al., 2019). Terpe-
noids are a group of compounds derived from the mevalonic acid pathway
characterized by their antibacterial effects and include terpenes, carotenoids,
and steroidal compounds. Therefore, for years, humans have explored new
sources of natural preservatives in nature, focusing mainly on plants that
contain essential oils since their compounds have the main advantage of
having no harmful effects on health. Thymol, eugenol, and carvacrol are
obtained from leaves, stems, or roots through different extraction techniques

to improve their yield (Bulama et al., 2015). These compounds have an effect
against Listeria monocytogenes, Salmonella typhimurium, Escherichia coli
O157:H7, Shigella dysenteriae, Bacillus cereus, and Staphylococcus aureus.
Therefore, used to preserve fish, dairy products, vegetables, fruits, and
cooked rice (Burt et al., 2004). Carvacrol, thymol, and eugenol alter bacterial
membrane functions by inhibiting ATPase activity, followed by the excre-
tion of ATP and other cellular components. Carvacrol and thymol dissolve
the phospholipid bilayer through alignment between the fatty acid chains
(Moghaddam and Mehdizadeh, 2017).
The main mechanism of the antibacterial effect is the damage to the
bacterial membrane (Faustino et al., 2019). The cell permeability of the
microbial membrane is altered by the interaction of hydroxyl groups with
membrane proteins, causing a change in its structure and function due
to a loss of internal biomolecules such as ribose and sodium glutamate.
Therefore, electron transport, nutrient absorption, protein, and nucleic acid
synthesis are affected, and the enzymatic activity of the pathogen is affected
(Rahman and Sarker, 2020). Another mechanism of antibacterial activity
of phenolic compounds is alkyl substitution towards the aromatic nucleus
(Bajpai et al., 2008). It has been known for years that terpenoids, phenolic
compounds, and plant enzymes have an antibacterial effect (Bubonja-Šonje
et al., 2020). Bean protease is an example, given that it has more antioxidant
and antimicrobial effects than sorbate and benzoate salts (El-Saadony et al.,
2020). Consequently, plants represent a source for discovering new food
preservatives that replace conventional additives with even higher activities
in the short term. In addition, they are safer, increase the shelf life of foods,
and have more effect than conventional preservatives (Pisoschi et al., 2019)].
Table 8.1 describes some of the more common bioactive compounds of plant
origin used as food preservatives.
TABLE 8.1 Antimicrobial Compounds Isolated from Plants Bioactive Antimicrobial

Effects.
Plant Bioactive compound Microorganism Minimal References
inhibitory
concentration
(MIC) (mg/mL)
Syzygium Eugenol L. monocytogenes 0.5 Cui et al.
aromaticum (2018)

Plant Bioactive compound Microorganism Minimal References
inhibitory
concentration
(MIC) (mg/mL)
Origanum Carvacrol and thymol S. aureus, 0.25–0.5 Wijesundara
vulgare Staphylococcus and
epidermidis, Rupasinghe
Sthaphylococcus (2018)
sciuri, and E. coli
Kaffir lime d-Limonene, β-pinene, S. aureus, 1.0–8.0 Sreepian et al.
and sabinene Elizabethkingia (2019)
meningoseptica,
and L.
monocytogenes
Cuminum 1,8-cineole, limomene S. dysenteriae, L. 0.3–5 Sharafati
cyminum and pinene monocytogenes, Chaleshtori et
Salmonella typhi al. (2106)
y Streptococcus
pyogenes
Elletaria α-Terpinyl acetate, Candida albicans, 5–10 Abdullah et
cardamomum 1.8-cineole, linalool Streptococcus al. (2017)
acetate, sabinene, and mutans, S. aureus,
linalool L. monocytogenes,
B. cereus, and S.
typhimurium
Myristica Safrole, S. mutans 2 Ali et al.
fragrans β-phellandrene, (2021)
Houtt 3-p-menthene and
l-terpinen-4-ol
Zingiber Mono y di terpenos L. monocytogenes, 8.7 Beristain-
Officinale del aceite esceencial S. typhimurium, Bauza et al.
(cineole, eugenol y and S. aureus (2019)
geraniol).
Hibiscus Cyanidine-3- S. typhi and E. coli 0.20 Sutiany et al.
sabdariffa L. sambubiose, (2019)
cyanidine-3-glucoside,
flavonoids, tannins,
and saponins
As described, the plants used from ancient times to conserve food are
known as aromatic due to the terpenes they contain in essential oils that
give them an antibacterial effect. However, other groups of plants contain
terpenoids with antimicrobial potential, such as Euphorbiaceae.
8.4 ANTIMICROBIAL POTENTIAL OF EUPHORBIACEAE
Euphorbiaceae plants grow in tropical and subtropical areas of Africa and

America (Islam et al., 2019). Euphorbiaceae includes around 300 genera
and 8000 species known for their medicinal components (Govea-Salas et
al., 2017). Euphorbiaceae has a ubiquitous geographic distribution resulting
in the adaptation of this group of plants to multiple habitats. Therefore,
they produce an enormous variety of bioactive compounds (Mwinea and
van Damme, 2011). The effect of each Euphorbiaceae plant depends on its
chemical composition since it produces a wide variety of metabolites for their
growth, reproduction, and survival (Azmir et al., 2013). These plants produce
both primary and secondary metabolites. Primary metabolites include amino
acids, carbohydrates, lipids, and nucleic acids. These compounds form cell
structures and regulate their vital functions (Pagare et al., 2015), so they
are necessary for plant life, while secondary metabolites improve survival
against environmental stress (Ashraf et al., 2018).
Therefore, its species are used in the traditional medicine of different
countries. One of the most outstanding effects of Euforbiaceae is antimi-
crobial. Euphorbiaceae produces antimicrobial compounds, highlighting
terpenoids such as monoterpenes, diterpenes, and triterpenes (Felix-Silva et
al., 2018). These compounds are detected in different concentrations in the
oily phase of their extracts, and their concentration depends on each species
(Elshamy et al., 2019).
In addition, it synthesizes flavonoids, alkaloids, and tannins (Cavalcante
et al., 2020). On the contrary, compounds of Euforbiaceae present antioxidant
activity (Greay and Hammer, 2015). Therefore, they can be incorporated into
foods as additives to improve shelf life and prevent early senescence due to
environmental factors and the presence of microorganisms (Pandey et al.,
2017). Additionally, some of its compounds have cytotoxic, antispasmodic,
antitumor, analgesic, and gastroprotective effects (Cavalcante et al., 2020;
Sabandar et al., 2013). Some Euhorbiaceae species used as antimicrobials
in different countries are described below. Therefore, they could be used as
food preservatives.
8.4.1 EUPHORBIA HETEROPHYLLA
The plant is used in India and Nigeria for its many properties, but its most
prominent use is to treat gastrointestinal, respiratory, and skin infections
and to treat malaria (Okeniyi et al., 2012). Ethanol extracts have been used
since ancient times and were extracted in gin. However, the extraction and
efficiency of its components depend on the geographical location and soil
conditions (Moghaddam and Mehdizadeh, 2017). The aqueous extracts also
have an inhibitory effect on Klebsiella pneumoniae, E. coli, Pseudomonas
aeruginosa, and S. aureus. However, S. aureus is more sensitive to extracts
(Ughachukwu et al., 2014).
8.4.2 EUPHORBIA EBRACTEOLATA HAYATA
The dried root of the plant is used in China, Korea, and Japan to treat bloating
and cough. The chemical components are terpenoids, acetophenones, and
flavonoids. The extracts possess antiviral and antimicrobial effects (Ting et
al., 2020).
8.4.3 EUPHORBIA NERIIFOLIA L.
The plant contains compounds with antimicrobial properties. The extracts

have an effect against various pathogens. Consequently, used to treat
emerging infectious diseases, focusing on bacterial, viral, fungal, and parasitic
infections. This plant contains Euhol, monohydroxy triterpene, nerifoliol,
taraxerol, β-amyrin, glut-5-(10)-en-1-one, neriifolone, and cycloartenol. In
addition, quercetin, rutin, friedelin taraxerol, epitaraxerol, taraxeryl acetate,
3β-friedelanol, 3β-acetoxy friedelane, 3β-simiarenol, afzelin, 24-methylene
cycloarenol, ingenol triacetate, and β-amyrin. β-Amyrin is one of those
responsible for the antibacterial effect but to is associated with flavonoids,
monoterpenoids, diterpenoids, triterpenoids, and alkaloids (Sultana et al.,
2022).
8.4.4 MANIHOT MULTIFIDA (L.) CRANTZ
The plant is used in folk medicine to treat infected wounds. It has antioxidant
and antimicrobial potential against C. albicans. The methanolic extract of
fruits, and aqueous and hexane extract of leaves have activity with MIC of
39 µg/mL. In addition, the methanolic extract has the potential to purify
DPPH (1,1-diphenyl-2-picrylhydrazyl) with an IC50 of 46.9 μg/mL, while
the hexane extract has an IC50 of 59.2 μg/mL. However, the ethanol extract
has activity against brine shrimp at an LC50 of 15.6 µg/mL (Fabri et al.,
2015).
8.4.5 MALLOTUS OPPOSITIFOLIUS
El género Mallotus presenta alrededor de 150 especies, incluyendo árboles,

arbustos y algunas plantas trepadoras (Abban et al., 2013). In Africa, the
leaves of M. oppositifolius are used as an antiparasitic since they cause the
expulsion of tapeworms and the elimination of ameba. Consequently, it is
used to treat diarrhea and dysentery. Furthermore, it is used to treat urinary
and venereal infections (Agyare et al., 2014). The antimicrobial effect
is attributed to phloroglucinols, benzopyrans, cardenolides, flavonoids,
steroids, and triterpenoids (Abban et al., 2013). However, the leaves of M.
oppositifolius produce compounds derived from acylphloroglucinol with
effect against E. coli, S. aureus, S. typhi, P. aeruginosa (Tchangoue et al.,
2020).
8.4.6 CNIDOSCOLUS QUERCIFOLIUS
The plant, endemic to northeastern Brazil, is used in popular medicine as

an antibiotic. Leaf extract has low activity against S. aureus and coagulase-
negative Staphylococcus with MIC ≥ 2000 µg/mL. The plant contains
linamarin cyanoglycoside, cinnamic acid, and a mixture of steroids and
terpenoids. However, Linamarin is responsible for the effect on E. coli,
S. aureus, and P. aeruginosa with MIC of 1000, 2000, and 2000 µg /mL,
respectively (Félix-Silva et al., 2018).
8.4.7 CROTON BETACEUS
The Croton genus contains an enormous diversity of antimicrobial

compounds against pathogens. Therefore, their species have a high potential
to develop alternative pharmacological treatments to treat infections or to
use them as food preservatives (Wright, 2017). This plant contains prom-
ising antibacterial compounds since its extract with hexane has an effect
against Enterococcus faecalis. This effect is attributed to its fractions rich in
flavonoid-type phenolic compounds. In addition, the most effective fractions
contain tannins and free steroids (Das Rocha et al., 2021).
8.4.8 CROTON LUNDIANUS
Croton lundianus (Didr.) Müll. Arg. is native to Brazil, but not endemic. The
hexane extract also affects E. faecalis attributed to flavonoid-type phenolic
compounds, tannins, and free steroids (Das Rocha et al., 2021).
8.4.9 CROTON DICHOGAMOUS
This plant is a shrub used in East Africa to treat infectious diseases. Extracts
in acetone, ethanol (50%), and distilled water at 250 mg/mL are effective
against B. cereus, S. aureus, and C. albicans, but not on P. aeruginosa and E.
coli. The bacteria most sensitive to its extracts is B. cereus, with an MIC of
10.42 mg/mL (Matara et al., 2021).
8.4.10 JATROPHA CURCAS
The name comes from the Greek “jatros” meaning doctor and “trophe”
meaning food. This group of plants is distributed from the west to tropical
areas of Latin America due to the export and migration of ancient civiliza-
tions. J. curcas has been used in traditional medicine in Asia and Africa to
treat gastrointestinal infections such as diarrhea, typhoid fever, and cholera
(Dada et al., 2014). The effect of the plant is due to diterpenes with antimi-
crobial and anti-proliferative activity (Devappa et al., 2011). Antimicrobial
diterpenes such as rhamnofolan, daphnano, latirano, tigliano, and dinor-
diterpenes have been detected.
8.4.11 JATROPHA NEOPAUCIFLORA
The Jatropha genus has about 175 species and is native to tropical America.
However, it is distributed in the tropics and subtropics of Asia and Africa.
This genus contains compounds with antimicrobial and antioxidant effects.
In addition, lignans, flavonoids, coumarins, alkaloids, eudesmenoic acids,
but mainly terpenes have been detected (Cavalcante et al., 2020) In Mexico,
different species of the Europhobiaceae family grow, which includes around
48 genera. Most grow in semi-arid areas such as the state of Coahuila
(Devappa et al., 2011). Jatropha neopauciflora grows in northern Mexico
and has antimicrobial potential.
8.4.12 JATROPHA DIOICA
The plant grows like a bush from 50–150 cm it is known as Sangre de Drago
because it contains a colorless latex that turns red on contact with air. The plant
grows in dry climates and rocky soils on the mountains. Consequently, it is
easily found in semi-desert areas. Therefore, it grows in the northeast of Mexico
and infusion is used in Mexican traditional medicine for skin infections, vagi-
nitis, urethritis, and conjunctivitis (Govea-Salas et al., 2017; Sabandar et al.,
2013). Hexane extracts have an effect against bacteria that cause gastrointes-
tinal diseases and dermatophyte fungi such as S. aureus, E. coli, B. cereus,
Salmonella thypimurium, C. albicans, Candida parapsilosis, Cryptococcus
neoformans, and Sporotrix schenckii (Silva-Belmares et al., 2014).
Information on the chemical composition of the plant is scarce. However,
Jatrophona, riolozatrione, and β-sitosterol have been detected in extracts.
Additionally, the plant contains polyphenols, tannins, flavonoids, ellagitan-
nins, and terpenes as the natural defense of the plant. These compounds have
biological activities such as antimicrobials and antioxidants (Castro-Ríos et
al., 2020). Accordingly, J. dioica has the potential to design a food preser-
vative based on the standardization of its extract based on the content of
β-sitosterol or riolozatrione. To contribute to the decrease in the consump-
tion of synthetic preservatives (Pérez-Pérez et al., 2020)]. Table 8.2 shows
some antimicrobial compounds detected in extracts of different species of
Euphorbiaceae, their MIC, and the spectrum of action.
8.5 INDUSTRIAL APPLICATIONS OF EUPHORBIACEAE

The constant change in society and its demands over the years led to the
development of new technologies to replace the usual ones due to the
exponential demographic increase since it causes the overexploitation of
ecosystems and gives rise to problems such as climate change (Keneni and
Marchetti, 2017).
Therefore, in recent years, the use of natural resources has been resumed
to manufacture products for constant consumption, such as food. The demand
for conventional energy is scarce in some countries. Therefore, research is
focused on producing biofuels from natural sources such as Euphorbiaceae
because they synthesize terpenoids that can used to obtain biodiesel and
others to obtain preservatives for the food industry due to their antimicrobial
properties (Zahan and Kano, 2018). Therefore, obtaining biodiesel has been
proposed to correct the environmental impact of conventional energy sources
Perspectives for Innovation of New Food Preservatives from Euphorbiaceae
TABLE 8.2 The Spectrum of Action and MIC of Antimicrobial Compounds Detected in Plant Extracts of Euphorbiaceae.
Plant Antimicrobial compound Extraction type MIC (mg/mL) Microorganism References
J. curcas L. Δ-Cadinene, α-epi-cadinol, Hydrodistillation 0.3–0.6 P. aeruginosa Babahmad et al. (2018)
pulegone, thymol
Jatropha integerrima Cineole, p-cymene, α-pinene Hydrodistillation 625–1250 B. cereus, S. aureus Eshilokuna et al. (2007)
and P. aeruginosa
Jatropha podagrica Japodagrine, Japodagrone Methanol 5–20 Bacillus subtilis Minh et al. (2019)
Jatropha multifida Multifidanol, Jatrophen Ethanol 12.5 P. aeruginosa y S. Senou et al. (2022)
aureus
Jatropha gossypiifolia Jatrofeno, jatrofenone, jatrofatrione, Ethanol 0.025–0.05 E. coli, Enterococcus Wu et al. (2019)
L. curcusone, japogadrol, multifolone faecium, and S. aureus
J. neopauciflora β-Pinene Methanol/hexane 2 S. aureus Hernández-Hernández et al.
(2016)
Croton Lupeol Metanol/hexane 16–32 S. aureus, P. Tatsimo et al. (2020)
pseudopulchellus aeruginosa y E. coli
Croton ceanothifolius α-Pinene, (E)-caryophyllene, Hydrodistillation 12–16 S. aureus y E. coli De Araújo et al. (2020)
Baill β-elemene, sesquiterpenes
Croton adipatus α-Thujene, β-myrcene, d-limonene Hydrodistillation 286 B. subtilis Cucho-Medrano et al. (2021)
Croton thurifer β-Elemene, germacrene-D, Hydrodistillation 296 S. aureus Cucho-Medrano et al. (2021)
α-thujene
Croton collinus Β-Caryophyllene, γ-terpinene, Hydrodistillation 72 B. subtilis Cucho-Medrano et al. (2021)
α-thujene, d-limonene
209
FIGURE 8.1 Chemical structures with antimicrobial activity present in species of the
Euphorbiaceae family Wu et al. (2019).
from J. curcas and J. dioica. Biodiesel is obtained by transesterification of

the oil contained in the plant (Pérez-Pérez et al., 2020).
J. curcas to obtain biodiesel using lipase enzymes from Candida rugosa.
Liquid recombinant Candida rugosa lipase (CRL) isoenzymes have been
investigated to convert various non-edible oils to biodiesel. These enzymes
could be excellent biocatalysts to produce biodiesel from Jatropha crude
oil at a low cost (Kuo et al., 2015). Therefore, the economic value of J.
curcas largely depends on the lipid content in the seeds that are stored in
the oil bodies (Liu et al., 2015). Additionally, some studies have focused on
establishing in vitro cultures of J. curcas and J. gossypifolia to obtain novel
compounds for pharmaceutical or industrial applications (Solís-Ramos et al.,
2013). For this reason, cell cultures of J. curcas have been established to
increase lipid production in vitro. Because to know industrial applications
requires an understanding of growth conditions affecting lipid metabolism
and yield (Solís-Ramos et al., 2020).
On the contrary, phorbol esters with antifungal properties have been
extracted from J. curcas seed cake as a by-product of the biodiesel fuel industry.
These esters affect Fusarium oxysporum, Pythium aphanidermatum, Lasiodi
plodia theobromae, Curvularia lunata, Fusarium semitectum, Colletotrichum
capsici, and Colletotrichum gloeosporiodes. Therefore, they could be used
for agricultural applications (Saetae and and Suntornsuk, 2012). Castor oil is
extracted from the seed of Ricinus communis L. It has a high commercial value
for the manufacture of soaps, lubricants, and coatings. It is rich in hydroxy
fatty acids and widely used in the cosmetic, pharmaceutical, oleochemical,
and agricultural industries. The oil is a bactericidal, anti-inflammatory, and
antiherpetic agent due to ricinoleic acid (Nitbani et al., 2022). The Jatropha
genus has a wide range of bioactivities and it is expected that new avenues
will be established to exploit its chemicals by the pharmaceutical industry to
develop chemotherapeutic agents (Devappa et al., 2011).
Euphorbia cotinifolia contains enzymes that are easy to purify. Conse-
quently, it could have applications in biophysical studies, biotechnological
and industrial applications (Kumar et al., 2011). E. tirucalli has many bioac-
tive properties and contains alkaloids, flavonoids, tannins, and terpenoids.
In addition, the stem contains polyphenols with antioxidant activity. Scopo-
letin, gallic acid, and piperic acid isolated from the ethyl acetate fraction
have an effect against citrus bacterial canker (Nguyen et al., 2021). On the
contrary, the peroxidase E. tirucalli immobilized on chitosan beads with
magnetic properties has been recovered to provide an industrial application.
Because peroxidases can be used to treat wastewater containing phenolic
compounds. In addition, they are used to the synthesis of aromatic chemicals

and peroxide removal from food products, industrial waste, biosensors, and
in the development of diagnostic kits (Shukla et al., 2017).
On the contrary, Euphorbiaceae species are known to synthesize anti-
microbial compounds in response to invasion by phytopathogenic fungi. H.
brasiliensis is an example that produces bioactive proteins due to Phytophthora
infestation in tropical regions. These proteins protect the rubber tree from
any fungal invaders; this allows us to understand some defense mechanisms
and the improvement of plantations (Havanapan et al., 2016). However, the
plant also contains antioxidant flavonoids and phenols that can be used in the
food industry to inhibit the formation of free radicals to preserve the existing
properties of food matrices. Therefore, extraction methods such as soxhlet
have been implemented to recover bioactive extracts with environmentally
friendly solvents (Oleinik et al., 2022).
Manihot esculenta is known as cassava and is a source of food for
much of the world’s population, particularly in countries in South America,
Africa, and Asia (Okezie et al., 1983). It is used in cooking and is applied
in industrial products such as adhesive for laundry, manufacture of paper,
alcohol, butanol, dextrin, adhesive tape, textile finishing, and glue (Okezie et
al., 1983). Cassava-based industries generate large amounts of waste rich in
bioactive compounds, which offers the potential for obtaining value-added
products (Li et al., 2017). An application of cassava starch is the formulation
of biodegradable foams incorporated with grape stems because they have
properties to protect foods stored with low moisture content (Engel et al.,
2019). Additionally, some polymers of cassava starch, have been used to
design biocomposite materials for food packaging. This material has been
added with antimicrobial compounds such as cinnamaldehyde (De Souza et
al., 2014). On the contrary, there are films formulated based on M. esculenta
and carrageenan added with essential oils. These films have been used to
improve the shelf life of chicken nuggets during refrigerated storage. In
addition, they have advantages over conventional ones formulated based on
synthetic plastics since the M. esculenta films are edible (Bharti et al., 2020).
8.6 CONCLUSION
The deterioration of food causes economic losses for the food industry
and causes the loss of its nutritional and healthy properties. Consequently,
for years, they have used methods for their conservation. However, some
conventional preservation methods change the nutritional composition, while
others have detrimental side effects on health, causing adverse effects such as
cancer, multiple sclerosis, and heart disease. Therefore, natural preservatives
of a terpenic or polyphenolic nature could reduce health risks. The plants of
Euphorbiaceae are rich in this compound, and some produce polymers with
properties to develop materials for food preservation since the terpenoid and
polyphenolic compounds of this plant have antimicrobial and antioxidant prop-
erties. On the contrary, some Euphorbiace generate agro-industrial residues
that could be used for the production of new food preservatives. Consequently,
they represent a source to discover new food preservatives that replace conven-
tional additives with higher activities in the short term. However, the formula-
tion of new preservatives requires trials to recover bioactive compounds such
as optimization of extraction methods from Euphorbiaceae. In addition, it is
necessary to evaluate its effectiveness and safety.
KEYWORDS
• euphorbiaceae
• antimicrobial effect
• terpenoids
• polyphenolic compounds
• preservative
• foods
• phytochemicals
• natural sources
REFERENCES
Abban, G.; Ahiadu, B. K.; Barku, V. Y. A. Phytochemical Studies and Antioxidant Properties
of Methanolic and Aqueous Extracts of the Leaves of Mallotus oppositifolius. J. Basic Appl.
Sci. 2013, 20–31.
Abdullah, A.; Asghar, M. S.; Shahid, M.; Huang, Q. Evaluating the Antimicrobial
Potential of Green Cardamom Essential Oil Focusing on Quorum Sensing Inhibition of
Chromobacterium violaceum. J. Food Sci. Technol. 2017, 54, 2306–2315.
Agyare, C.; Amuah, E.; Adarkwa-Yiadom, M.; Osei-Asante, S.; Ossei, P. P. S. Medicinal
Plants Used for Treatment of wounds and Skin Infections: Assessment of Wound Healing
and Antimicrobial Properties of Mallotus oppositifolius and Momordica charantia. Int. J.
Phytomed. 2014, 6, 50.
Ali, A.; Yasir, M.; Jilani, M. I.; Shoaib, S.; Latif, M.; Ahmad, A.; Nazir, A. Chemical
Composition an In Vitro Evaluation of Cytotoxicity, Antioxidant and Antimicrobial
Activities of Essential Oil Extracted from Myristica Fragrans Houtt. Polish J. Environ.
Stud. 2021, 30, 1585–1590.
Anwar, R.; Mattoo, A. K.; Handa, A. K. Postharvest Biology and Nanotechnology. Ripening
and Senescence of Fleshy Fruits. John Wiley & Sons, Canada, Springer, 2019; pp 15–51.
Ashraf, M. A.; Iqbal, M.; Rasheed, R.; Hussain, I.; Riaz, M.; Arif, M. S. Environmental Stress
and Secondary Metabolites in Plants: An Overview; Elsevier Inc., 2018; pp 153–167.
Azmir, J.; Zaidul, I. S. M.; Rahman, M. M.; Sharif, K. M.; Mohamed, A.; Sahena, F.; Omar, A.
K. M. Techniques for Extraction of Bioactive Compounds from Plant Materials: A Review.
J. Food Eng. 2013, 117, 426–436.
Babahmad, R. A.; Aghraz, A.; Boutafda, A.; Papazoglou, E. G.; Tarantilis, P. A.; Kanakis, C.;
Ouhammou, A. Chemical Composition of Essential Oil of Jatropha curcas L. Leaves and
Its Antioxidant and Antimicrobial Activities. Ind. Crops Prod. 2018, 121, 405–410.
Bajpai, V. K.; Rahman, A.; Dung, N. T.; Huh, M. K.; Kang, S. C. In vitro inhibition of
Food Spoilage and Foodborne Pathogenic Bacteria by Essential Oil and Leaf Extracts of
Magnolia liliflora Desr. J. Food Sci. 2008, 73, 314–320.
Baran, A.; Yildirim, S.; Ghosigharehaghaji, A.; Bolat, I.; Sulukan, E.; Ceyhun, S. B. An
Approach to Evaluating the Potential Teratogenic and Neurotoxic Mechanism of BHA
Based on Apoptosis Induced by Oxidative Stress in Zebrafish Embryo (Danio rerio). Hum.
Exp. Toxicol. 2021, 40, 425–438.
Beristain-Bauza, S. D. C.; Hernández-Carranza, P.; Cid-Pérez, T. S.; Ávila-Sosa, R.; Ruiz-
López, I. I.; Ochoa-Velasco, C. E. Antimicrobial Activity of Ginger (Zingiber officinale)
and Its Application in Food Products. Food Rev. Int. 2019, 35, 407–426.
Bharti, S. K.; Pathak, V.; Alam, T.; Arya, A.; Singh, V. K.; Verma, A. K.; Rajkumar, V.
Materialization of Novel Composite Bio‐Based Active Edible Film Functionalized with
Essential Oils on Antimicrobial and Antioxidative Aspect of Chicken Nuggets During
Extended Storage. J. Food Sci. 2020, 85, 2857–2865.
Bubonja-Šonje, M.; Knezević, S.; Abram, M. Challenges to Antimicrobial Susceptibility
Testing of Plant-Derived Polyphenolic Compounds. Arhiv za Higijenu Rada i Toksikologiju
2020, 71, 300–311.
Bulama, J. S.; Dangoggo, S. M.; Mathias, S. N. Isolation and Characterization of Beta-
sitosterol from Ethyl Acetate Extract of Root Bark of Terminalia Glaucescens. Int. J. Sci.
Res. Pub. 2015, 5, 1–3.
Burt, S. Essential Oils: Their Antibacterial Properties and Potential Applications in Foods: A
Review. Int. J. Food Microbiol. 2004, 94, 223–253.
Castro-Ríos, R.; Melchor-Martínez, E. M.; Solís-Cruz, G. Y.; Rivas-Galindo, V. M.; Silva-
Mares, D. A.; Cavazos-Rocha, N. C. HPLC Method Validation for Jatropha dioica Extracts
Analysis. J. Chromatogr. Sci. 2020, 58 (5), 445–453.
Cavalcante, N. B.; da Conceição Santos, A. D.; da Silva Almeida, J. R. G. The Genus Jatropha
(Euphorbiaceae): A Review on Secondary Chemical Metabolites and Biological Aspects.
Chem. Biol. Interact. 2020, 318, 1–18.
Chaleshtori, F. S.; Arian, A.; Chaleshtori, R. S. Assessment of Sodium Benzoate and Potassium
Sorbate Preservatives in Some Products in Kashan, Iran with Estimation of Human Health
Risk. Food Chem. Toxicol. 2018, 120, 634–638.
Cucho-Medrano, J. L. L.; Mendoza-Beingolea, S. W.; Fuertes-Ruitón, C. M.; Salazar-
Salvatierra, M. E.; Herrera-Calderon, O. Chemical Profile of the Volatile Constituents and
Antimicrobial Activity of the Essential Oils from Croton adipatus, Croton thurifer, and
Croton collinus. Antibiotics 2021, 10, 1387.
Cui, H.; Zhang, C.; Li, C.; Lin, L. Antimicrobial Mechanism of Clove Oil on Listeria
monocytogenes. Food Control 2018, 94, 140–146.
Cui, H.; Zhang, C.; Li, C.; Lin, L. The Industrial Applications of Cassava: Current Status,
Opportunities and Prospects. J. Org. Behav. 2017, 28, 303–325.
Dada, E. O.; Ekundayo, F. O.; Makanjuola, O. O. Antibacterial Activities of Jatropha curcas
(LINN) on Coliforms Isolated from Surface Waters in Akure, Nigeria. Int. J. Biomed. Sci.
2014, 10, 25–30.
Das Rocha, A. R.; Sousa, H. G.; do Vale Júnior, E. P.; De Lima, F. L.; Costa, A. S.; De
Araújo, A. R.; Lago, E. C. Extracts and Fractions of Croton L.(Euphorbiaceae) Species
with Antimicrobial Activity and Antioxidant Potential. LWT—Food Sci. Technol. 2021,
139, 110521.
De Araújo, A. C.; Freitas, P.; Rodrigues dos Santos Barbosa, C.; Muniz, D. F.; Esmeraldo
Rocha, J.; Neto, J. B. D. A.; Melo Coutinho, H. D. Essential Oil of Croton ceanothifolius
Baill. Potentiates the Effect of Antibiotics Against Multiresistant Bacteria. Antibiotics
2020, 9, 1–9.
De Oliveira, M. M.; Lago, A.; Dal’ Magro, G. P. Food Loss and Waste in the Context of the
Circular Economy: A Systematic Review. J. Clean. Prod. 2021, 294, 126–284.
De Souza, A. C.; Dias, A. M.; Sousa, H. C.; Tadini, C. C. Impregnation of Cinnamaldehyde
into Cassava Starch Biocomposite Films Using Supercritical Fluid Technology for the
Development of Food Active Packaging. Carbohydr. Polym. 2014, 102, 830–837.
Devappa, R. K.; Makkar, H. P.; Becker, K. Jatropha diterpenes: A Review. J. Am. Oil Chem
Soc. 2011, 88, 301–322.
El-Saadony, M. T.; Elsadek, M. F.; Mohamed, A. S.; Taha, S. A.; Ahmed, B. M.; Saad, A. M.
Effects of Chemical and Natural Additives on Cucumber Juice’s Quality, Shelf Life, and
Safety. Foods. 2020, 9, 1–17.
Elshamy, A. I.; Abd‐ElGawad, A. M.; El Gendy, A. E. N. G.; Assaeed, A. M. Chemical
Characterization of Euphorbia heterophylla L. Essential Oils and Their Antioxidant Activity
and Allelopathic Potential on Cenchrus echinatus L. Chem. Biodivers. 2019, 16, 1–18.
Engel, J. B.; Ambrosi, A.; Tessaro, I. C. Development of Biodegradable Starch-Based Foams
Incorporated with Grape Stalks for Food Packaging. Carbohydr. Polym. 2019, 225, 115234.
Eshilokuna, A. O.; Kasalia, A. A.; Ogunwandea, I. A.; Walkerb, T. M.; Setzerb, W. N.;
Chemical Composition and Antimicrobial Studies of the Essential Oils of Jatropha
integerrima Jacq (Leaf and Seeds). Nat. Prod. Commun. 2007, 2, 853–855.
Fabri, R. L.; De Sá, D. S.; Pereira, A. P. O.; Scio, E.; Pimenta, D. S.; Chedier, L. M.
Antimicrobial, Antioxidant and Cytotoxicity Potential of Manihot multifida (L.) Crantz
(Euphorbiaceae). An. Acad. Bras. Cienc. 2015, 87, 303–311.
Faustino, M.; Veiga, M.; Sousa, P.; Costa, E. M.; Silva, S.; Pintado, M. Agro-Food Byproducts
as a New Source of Natural Food Additives. Molecules 2019, 24, 1–23.
Félix-Silva, J.; Gomes, J. A. S.; Fernandes, J. M.; Moura, A. K. C.; Menezes, Y. A. S.; Santos,
E. C. G.; Tambourgi, D. V.; Silva-Junior, A. A.; Zucolotto, S. M.; Fernandes-Pedrosa, M. F.
Comparison of Two Jatropha species (Euphorbiaceae) used popularly to Treat Snakebites
in Northeastern Brazil: Chemical Profile, Inhibitory Activity Against Bothrops erythromelas
Venom and Antibacterial Activity. J. Ethnopharmacol. 2018, 213, 12–20.
Floros, J. D.; Newsome, R.; Fisher, W.; Barbosa-Cánovas, G. V.; Chen, H.; Dunne, C. P.;
German, J. B.; Hall, R. L.; Heldman, D. R.; Karwe, M. V.; Knabel, S. J.; Labuza, T. P.;
Lund, D. B.; Newell-McGloughlin, M.; Robinson, J. L.; Sebranek, J. G.; Shewfelt, R. L.;
Tracy, W. F.; Weaver, C. M.; Ziegler, G. R. Feeding the World Today and Tomorrow: The
Importance of Food Science and Technology. Compr. Rev. Food Sci. Food Saf. 2010, 9,
572–599.
Gonelimali, F. D.; Lin, J.; Miao, W.; Xuan, J.; Charles, F.; Chen, M.; Hatab, S. R. Antimicrobial
Properties and Mechanism of Action of Some Plant Extracts Against Food Pathogens and
Spoilage Microorganisms. Front. Microbiol. 2018, 9, 1–9.
Govea-Salas, M.; Morlett-Chávez, J.; Rodriguez-Herrera, R.; Ascacio-Valdés, J. Some Mexican
Plants Used in Traditional Medicine. Aromatic Med. Plants—Back Nat. 2017, 192–200.
Greay, S. J.; Hammer, K. A. Recent Developments in the Bioactivity of Mono and Diterpenes:
Anticancer and Antimicrobial Activity. Phytochem. Rev. 2015, 14, 1–6.
Havanapan, P. O.; Bourchookarn, A.; Ketterman, A. J.; Krittanai, C. Comparative Proteome
Analysis of Rubber Latex Serum from Pathogenic Fungi Tolerant and Susceptible Rubber
Tree (Hevea brasiliensis). J. Proteomics 2016, 131, 82–92.
Hernández-Hernández, A. B.; Alarcón-Aguilar, F. J.; Jiménez-Estrada, M.; Hernández-
Portilla, L. B.; Flores-Ortiz, C. M.; Rodríguez-Monroy, M. A.; Canales-Martínez, M.
Biological Properties and Chemical Composition of Jatropha Neopauciflora Pax. Afr. J.
Tradit. Complement. Altern. Med. 2016, 14, 32–42.
Islam, M. S.; Ara, H.; Ahmad, K. I.; Uddin, M. M. A Review on Medicinal Uses of Different
Plants of Euphorbiaceae Family. Univers. J. Pharm. Res. 2019, 4, 47–51.
Joardder, M. U. H.; Masud, M. H. Food Preservation in Developing Countries: Challenges
and Solutions. A Brief History of Food Preservation; Springer International Publishing,
2019; pp 57–66.
Keneni,Y. G.; Marchetti, J. M. Oil Extraction from Plant Seeds for Biodiesel Production.
AIMS Energy. 2017, 5, 316–340.
Kumar, R.; Singh, K. A.; Tomar, R.; Jagannadham, M. V. Biochemical and Spectroscopic
Characterization of a Novel Metalloprotease, Cotinifolin from an Antiviral Plant Shrub:
Euphorbia cotinifolia. Plant Physiol. Biochem. 2011, 49, 721–728.
Kumari, P. K.; Akhila, S.; Rao, Y. S.; Devi, B. R. Alternative to Artificial Preservatives. Syst.
Rev. Pharm. 2019, 10, 99–102.
Kuo, T. C.; Shaw, J. F.; Lee, G. C. Conversion of Crude Jatropha curcas Seed Oil into Biodiesel
Using Liquid Recombinant Candida rugosa Lipase Isozymes. Bioresour. Technol. 2015,
192, 54–59.
Liu, H.; Wang, C.; Chen, F.; Shen, S. Proteomic Analysis of Oil Bodies in Mature Jatropha
curcas Seeds with Different Lipid Content. J. Proteomics 2015, 113, 403–414.
Lyu, X.; Lee, J.; Chen, W. N. Potential Natural Food Preservatives and Their Sustainable
Production in Yeast: Terpenoids and Polyphenols. J. Agric. Food Chem. 2019, 67,
4397–4417.
Matara, D. N.; Nguta, J. M.; Musila, F. M.; Mapenay, I. Phytochemical Analysis and
Investigation of the Antimicrobial and Cytotoxic Activities of Croton dichogamus pax
Crude Root Extracts. Evid. Based Complement. Altern. Med. 2021, 2021.
Mazdeh, F. Z.; Aftadbari, F. E.; Moradi-Khatoonabadi, Z.; Shaneshin, M.; Torabi, P.; Ardekani,
M. R. S.; Hajimahmoodi, M. Sodium Benzoate and Potassium Sorbate Preservatives in
Iranian Doogh. Food Addit. Contam. Part B Surveill. 2014, 7, 115–119.
Minh, T. N.; Xuan, T. D.; Tran, H. D.; Van, T. M.; Andriana, Y.; Khanh, T. D.; Ahmad,
A. Isolation and Purification of Bioactive Compounds from the Stem Bark of Jatropha
podagrica. Molecules 2019, 24, 889.
Moghaddam, M.; Mehdizadeh, L. Chemistry of Essential Oils and Factors Influencing Their
Constituents. Soft Chem. Food Fermentation 2017, 379–419.
Mwine, J. T.; van Damme, P. Why Do Euphorbiaceae Tick as Medicinal Plants? A Review of
Euphorbiaceae Family and Its Medicinal Features. J. Med. Plants Res. 2011, 5, 652–662.
Negi, P. S. Plant Extracts for the Control of Bacterial Growth: Efficacy, Stability and Safety
Issues for Food Application. Int. J. Food Microbiol. 2012, 156, 7–17.
Nguyen, T. M. L.; Dang, X. C.; Pham, V. T.; Truong, N. M.; Tran, D. M.; Thuc-Huy, D.;
Tran, Thi, L. M.; Vo, T. T. O. Phytochemical Screening and Evaluation of Antioxidant
Properties and Antimicrobial Activity Against Xanthomonas axonopodis of Euphorbia
tirucalli Extracts in Binh Thuan Province, Vietnam. Molecules 2021, 26, 2–11.
Nitbani, F. O.; Tjitda, P. J. P.; Wogo, H. E.; Detha, A. I. R. Preparation of Ricinoleic Acid from
Castor Oil: A Review. J. Oleo Sci. 2022, 71, 781–793.
Okeniyi, S. O.; Adedoyin, B. J.; Garba, S. Phytochemical Screening, Cytotoxicity, Antioxidant
and Antimicrobial Activities of Stem and Leave Extracts of Euphorbia heterophylla. J.
Biol. Life Sci. 2012, 4, 24–31.
Okezie, B. O.; Kosikowski, F. V.; Markakis, P. Cassava as a Food. Crit. Rev. Food Sci. Nutr.
1983, 17, 259–275.
Oleinik, G.; Dario, P. P.; De Morais Gasperin, K.; Benvegnú, D. M.; Lima, F. O.; Soares, L.
C.; Gallina, A. L. In Vitro Antioxidant Extracts Evaluation from the Residue of the Hevea
brasiliensis Seed. Sci. Rep. 2022, 12, 1–14.
Özen, H.; Kamber, U.; Karaman, M.; Gül, S.; Atakişi, E.; Özcan, K.; Atakişi, O.
Histopathologic, Biochemical and Genotoxic Investigations on Chronic Sodium Nitrite
Toxicity in Mice. Exp. Toxicol. Pathol. 2014, 66, 367–375.
Pagare, S.; Bhatia, M.; Tripathi, N.; Pagare, S.; Bansal, Y. K. Secondary Metabolites of Plants
and Their Role: Overview. Curr. Trends Biotechnol. Pharm. 2015, 9, 293–304.
Pandey, A. K.; Kumar,P.; Singh, P.; Tripathi, N. N.; Bajpai, V. K. Essential Oils: Sources of
Antimicrobials and Food Preservatives. Front. Microbiol. 2017, 7, 1–14.
Pérez-Pérez, J. U.; Guerra-Ramírez, D.; Reyes-Trejo, B.; Cuevas-Sánchez, J. A.; Guerra-
Ramírez, P. Actividad Antimicrobiana In Vitro de extractos de Jatropha dioica seseé contra
bacterias fitopatógenas de tomate. Polibotánica 2020, 125–133.
Pisoschi, A. M.; Pop, A.; Georgescu, C.; Turcuş, V.; Olah, N. K.; Mathe, E. An Overview of
Natural Antimicrobials Role in Food. Eur. J. Med. Chem. 2019, 143, 922–935.
Rahman, M.; Sarker, S. D. Antimicrobial Natural Products. Annu. Rep. Med. Chem. 2020,
55, 77–113.
Sabandar, C. W.; Ahmat, N.; Jaafar, F. M.; Sahidin, I. Medicinal Property, Phytochemistry and
Pharmacology of Several Jatropha Species (Euphorbiaceae): A Review. Phytochemistry
2013, 85, 7–29.
Saetae, D.; Suntornsuk, W. Antifungal Activities of Ethanolic Extract from Jatropha curcas
Seed Cake. J. Microbiol. Biotechnol. 2012, 20, 319–324.
Senou, M.; Lokonon, J. E.; Abissi, E. O. T. R.; Agbogba, F.; Dehou, R. J.; Medoatinsa, E.;
Baba-Moussa, L. Antibacterial Activity and Toxicity of the Sap and Aqueous Extract of the
Leaves of Jatropha multifida Linn. J. Biosci. Med. 2022, 10, 171–182.
Sharafati Chaleshtori, F.; Taghizadeh, M.; Rafieian-kopaei, M.; Sharafati-chaleshtori, R.
Effect of Chitosan Incorporated with Cumin and Eucalyptus Essential Oils As Antimicrobial
Agents on Fresh Chicken Meat. J. Food Process. Preserv. 2016, 40, 396–404.
Sharif, Z. I. M.; Mustapha, F. A.; Jai, J.; Yusof, N. M.; Zaki, N. A. M. Review on Methods
for Preservation and Natural Preservatives for Extending the Food Longevity. Ing. Química
2017, 19, 145–153.
Shukla, A.; Gundampati, R. K.; Jagannadham, M. V. Immobilization of Euphorbia tirucalli
Peroxidase onto Chitosan-Cobalt Oxide Magnetic Nanoparticles and Optimization Using
Response Surface Methodology. Int. J. Biol. Macromol. 2017, 102, 384–395.
Silva-Belmares, Y.; Rivas-Morales, C.; Viveros-Valdez, E.; de la Cruz-Galicia, M. G.;
Carranza-Rosales, P. Antimicrobial and Cytotoxic Activities from Jatropha dioica Roots.
Pak. J. Biol. Sci. 2014, 17, 748–750.
Solís-Ramos, L. Y.; Carballo, L. M.; Valdez-Melara, M. “Establishment of Cell Suspension
Cultures of Two Costa Rican Jatropha Species (Euphorbiaceae). Revista de Biologia
Tropical 2013, 67, 1095–1107.
Solís-Ramos, L. Y.; Carballo, L. M.; Valdez-Melara, M. Model-Assisted Identification of
Metabolic Engineering Strategies for Jatropha curcas Lipid Pathways. Plant J. 2020, 104,
76–95.
Sreepian, A.; Sreepian, P. M.; Chanthong, C.; Mingkhwancheep, T.; Prathit, P. Antibacterial
Activity of Essential Oil Extracted from Citrus Hystrix (Kaffir lime) Peels: An In Vitro
Study. Trop. Biomed. 2019, 36, 531–541.
Sultana,A.; Hossain, M. J.; Kuddus, M. R.; Rashid, M. A.; Zahan, M. S.; Mitra, S.; Roy,
A.; Alam, S.; Sarker, M. M. R.; Mohamed, I. N. Ethnobotanical Uses, Phytochemistry,
Toxicology, and Pharmacological Properties of Euphorbia neriifolia Linn. Against
Infectious Diseases: A Comprehensive Review. Molecules 2022, 27, 2–31.
Sutiany, A.; Dwiyanti, R. D.; Oktiyani, N. Inhibition Strength of Rosella (Hibiscus sabdariffa
L.) Boiled Water on Salmonella typhi In Vitro. Trop. Health Med. Res. 2019, 1, 62–67.
Tatsimo, J. S. N.; Tamokou, J. D. D.; Havyarimana, L.; Kengne, I. C.; Ekom, S. E.; Lannang,
A. M.; Sewald, N. Antimicrobial and Antioxidant Activities of Isolated Compounds
from the Root Bark of Croton pseudopulchellus (Euphorbiaceae). Investig. Med. Chem.
Pharmacol. 2020, 13, 1–6.
Tchangoue, Y. A. N.; Tchamgoue, J.; Lunga, P. K.; Knepper, J.; Paltinean, R.; Ibrom, K.;
Schulz, S. Antibacterial Phloroglucinols Derivatives from the Leaves of Mallotus
oppositifolius (Geisler) Müll. Arg.(Euphorbiaceae). Fitoterapia 2020, 142, 104527.
Thyberg, K. L.; Tonjes, D. J. Drivers of Food Waste and Their Implications for Sustainable
Policy Development. Resour. Conserv. Recycl. 2008, 106, 110–123.
Ting, Y.; Jun, H.; Yan,Y.; Wen-Wen, L.; Cong-Yuan, X.; Jie-Kun, X.; Wei-Ku, Z. Euphorbia
ebracteolata Hayata (Euphorbiaceae): A Systematic Review of Its Traditional Uses,
Botany, Phytochemistry, Pharmacology, Toxicology, and Quality Control. Phytochemistry
2020, 186, 1–25.
Trasande, L.; Shaffer, R. M.; Sathyanarayana, S. Food Additives and Child Health. Pediatrics
2018, 142, 1–8.
Ughachukwu, P. O.; Ezenyeaku, C. C. T.; Ochiogu, B. C.; Ezeagwuna, D. A.; Anahalu, C.
Evaluation of Antibacterial Activities of Euphorbia heterophylla. IOSR J. Dent. Med. Sci.
2014, 13, 69–75.
Wijesundara, N. M.; Rupasinghe, H. P. V. Essential Oils from Origanum vulgare and Salvia
Officinalis Exhibit Antibacterial and Anti-Biofilm Activities Against Streptococcus
pyogenes. Microb. Pathogen. 2018, 117, 118–127.
Wright, G. D. Unlocking the Potential of Natural Products in Drug Discovery. Microb.
Biotechnol. 2019, 12, 55–57.
Wu, Q.; Patocka, J.; Nepovimova, E.; Kuca, K. Jatropha gossypiifolia L. and Its Biologically
Active Metabolites: A Mini Review. J. Ethnopharmacol. 2019, 234, 197–203.
Zahan, K. A.; Kano, M. Biodiesel Production from Palm Oil, Its By-Products, and Mill
Effluent: A Review. Energies 2018, 11, 2132.
CHAPTER 9
Biotechnology and Food Safety: Zoonotic

Perspectives
A. ARUN PRINCE MILTON, G. BHUVANA PRIYA, SAMIR DAS,
SANDEEP GHATAK, and K. SRINIVAS
ABSTRACT
Pork (meat of pig) is the most commonly consumed animal meat world-
wide and there is increasing global demand for pork according to future
projections. The pork production system has witnessed a drastic change
from pasture-based rearing to the industrialized intensive system of rearing.
Though pork is less attributed to foodborne diseases in comparison to other
meat (poultry, beef), it is imperative to achieve pork safety owing to its large
consumption. Pathogens enter the pork value chain through infected pigs,
unhygienic slaughter, postharvest handling, poor processing methods, etc.
Consumers acquire the infection due to the consumption of raw or improp-
erly cooked pork and other kitchen cross-contaminations. The important
challenges in achieving microbial safety in pork include cross-contamination
of other foodstuffs, contaminated water, traceability, rapid on-site pathogen
testing techniques, foodborne disease surveillance, implementation of hazard
analysis and critical control points (HACCP), awareness of pork handlers,
and consumers, etc. For effective pork safety interventions, it is decisive
to understand the zoonotic pathogens transmitted via pork including their
nature, control options, and prevalence in different stages of production and
processing. In this chapter, pork-associated zoonotic agents discoursed are
Salmonella, Yersinia enterocolitica, Campylobacter jejuni/Campylobacter
coli, Listeria monocytogenes, Clostridium perfringens, Staphylococcus
aureus, shigatoxigenic Escherichia coli, Streptococcus suis, Taenia solium,

Toxoplasma gondii, Trichinella spiralis, hepatitis E virus, rotavirus, and
norovirus. Attention has been given to the nature of zoonotic agents, preva-
lence, diseases caused in human beings, and various intervention strategies
used against the pathogens in on-farm, slaughter, and postprocessing stages.
9.1 INTRODUCTION
Food safety has become an increasingly significant public health

subject that has led all governments throughout the world to allocate
millions of currencies to strengthen their efforts to ensure and improve
food safety (Kaferstein et al., 1997). The efforts toward improving
food safety can be perceived as a response to a growing number of
food safety issues and rising concerns of the consumer (WHO, 2009).
The Centers for Disease Control and Prevention (CDC) estimates
that foodborne pathogens cause 47.8 million illness events and 3000
deaths every year (CDC, 2011). Most importantly, the foodborne
disease outbreaks reported in the last decades underscore the signifi-
cance of the formulation and application of preventive measures and
programs relating to food safety. Particularly, a farm-to-fork tactic
has been followed in all food production sectors to reduce probable
biological hazards and improve hygiene (Schirone et al., 2019).
Many factors associated with the increase in foodborne diseases have
been described, such as changes in consumer preferences and eating
habits, changes in food processing methods, inadequate sanitation
and public health services including vector control and awareness
building, increased global travel, the adaptation of pathogen to newer
environments including the acquisition of antimicrobial resistance and
virulence factors and lastly advancement in pathogen detection tools
(Smith and Fratamico, 2018).
Globally, over the past decade, the growth rate of pork consumption has
enhanced by 10.71% (Wu et al., 2015). World meat production is forecasted
to increase by 2.2% (pork: 4.2%) to 346 million tonnes (pork: 114.4 million
tonnes) (FAO, 2021). Pork (meat of pig) is the most commonly consumed
animal meat worldwide and there is increasing global demand for pork
according to future projections (Davies, 2010; Delgado et al., 2001). The
Biotechnology and Food Safety: Zoonotic Perspectives 223
pork production system has witnessed a drastic change from pasture-based

rearing to the industrialized intensive system of rearing (Enemark et al.,
2015). Although pork is less attributed to foodborne diseases in comparison
to other meat (poultry, beef), it is imperative to achieve pork safety owing to
its large consumption (Baer et al., 2013). As pork and pork products are exten-
sively consumed throughout the world, it is crucial for producers (farmers),
processors and other stakeholders in the value chain to be conscious of the
risk factors for contamination of pork with foodborne pathogens (Baer et al.,
2013). Diverse farm and husbandry systems (large/small/backyard) are asso-
ciated with specific microbiological hazards, which are spread to the next
stage in the pork value chain, and the subsequent stage in the value chain
could also be the point of contamination (Buncic et al., 2019; Karabasil et
al., 2021). The meat sector is a sector with inherent risks, so meat safety must
be risk-oriented to reduce or eliminate food safety risks, with integrated and
longitudinal approaches along the value chain, and incessant improvement
(Karabasil et al., 2021).
9.2 ZOONOTIC PATHOGENS OF PIG AND PORK
Pigs act as a reservoir for several zoonotic pathogens of viral, bacterial,

and parasitic origin (Fig. 9.1). Further, the presence of other reservoirs and
vectors in the environment of pigs can increase the prevalence of zoonotic
diseases and the risk for transmission to human beings (Salajpal et al., 2013;
Delsart et al., 2020). Pathogens enter the pork value chain through infected
pigs, unhygienic slaughter, postharvest handling, poor processing methods,
etc (Fig. 9.1). The majority of pork products are prepared by smallholders
and traded in classical wet markets. Nevertheless, fresh pork is exceedingly
susceptible to contamination, posing health and financial burdens (Lam et
al., 2020). Consumers acquire the infection due to the consumption of raw or
improperly cooked pork and other kitchen cross-contamination. For effective
pork safety interventions, it is decisive to understand the zoonotic pathogens
transmitted via pork including their nature, control options, and prevalence
in different stages of production and processing. Recent changes and inten-
sification in the pig production system have nearly eliminated the risk of
tissue-dwelling foodborne parasites such as Trichinella spiralis, Taenia
solium, and Toxoplasma gondii. This represents a significant public health
accomplishment that has gone largely underappreciated (Davies, 2011).
This could have become possible as these parasites involve carnivorous
transmission and effective prevention can be achieved by some preharvest

interventions (Davies, 2011). However, important bacterial pathogens of
concern to pork safety such as Salmonella, Listeria, Y. enterocolitica, Campy
lobacter, etc. have their basic ecological niche in the gastrointestinal tracts
of healthy animals as commensals, and thus the point of contamination can
be at preharvest, during harvest, postharvest, and any stages till reaching the
consumers’ plate. Consequently, such complex epidemiology makes them
less amenable to controlling such bacterial pathogens in the pork production
chain (Davies et al., 2004; Alban and Stark, 2005; Fosse et al., 2009; Oliver
et al., 2009; Davies, 2011).
FIGURE 9.1 Zoonotic pathogens in pig value chain.
9.2.1 SALMONELLA
Salmonella is a Gram-negative bacteria under the family Enterobacte
riaceae, that can survive in temperature and pH ranges of 5–45°C and 4–9,
respectively (Doyle and Cliver, 1990). Some strains of Salmonella can also
survive with a 2% NaCl concentration and low-moisture foods like dried
pork products (Montville and Matthews, 2008; Baer et al., 2013). Presently,
there are only 2 known species of Salmonella (S. enterica and S. bongori),
6 subspecies of S. enterica, and more than 2600 serovars of S. enterica

subsp. enterica including notable serovars such as Salmonella serovars
Typhimurium and Salmonella Enteriditis causing nontyphoidal salmonellosis
(Guibourdenche et al., 2010). Salmonella infections in human beings are the
foremost cause of hospitalization and deaths due to foodborne illnesses. It
causes illnesses ranging from mild gastroenteritis and diarrhea to serious
systemic infections (Scallan et al., 2011; Gupta et al., 2019). Adult pigs are
frequently asymptomatic carriers of wide-ranging Salmonella serovars and
can represent a reservoir for human infections (Gupta et al., 2019). Several
studies have reported that as the growth of pigs’ advances from weaning
to growing and finishing stages, the prevalence of Salmonella is enhanced
(Korsak et al., 2003; Dorr et al., 2009; Wilkins et al., 2010). Nevertheless,
the prevalence of Salmonella was also reported to decrease as pigs advance
through the production chain (Kranker et al., 2003; Rowe et al., 2003).
Notwithstanding, the prevalence of Salmonella in pigs is not unusual in any
of the stages of production and processing, and the prevalence rates vary
from 0.7 to 77.8 (Baer et al., 2013). In a study conducted by USDA in two
commercial slaughter establishments, it was found that 91% of prescald
carcasses were carrying Salmonella (Schmidt et al., 2012). In another study,
it was reported that 37% of carcasses harboring Salmonella contamination
contained Salmonella counts between 1 and 3.9 log CFU per 100 cm2.
Hence, there is a chance of cross-contamination with other carcasses and
processing equipment (Dickson et al., 2002). Several studies have indicated
the prevalence of Salmonella in retail fresh pork, pork products, and ready-
to-eat pork products and the reasons noticed were contaminated slaughter
environment, complacence in hygiene, under processing, contamination
from food handlers, raw materials like spices, and pests inside the facility
such as rodents, insects, and birds (Baer et al., 2013).
9.2.2 YERSINIA ENTEROCOLITICA
Y. enterocolitica is a Gram-negative psychrotrophic salt-tolerant bacteria

that is extensively distributed in nature, in animal and aquatic reservoirs,
with pigs representing the main known reservoir for zoonotic strains of Y.
enterocolitica (Bottone, 1997). The pathogenicity of Y. enterocolitica is
usually linked with its bio-serotype. The biotypes, 1B, 2, 3, 4, and 5 and
serogroups, O:3, O:5,27, 42 O:8, and O:9, are pathogenic to human beings.
The bio-serotype 4/O:3 is predominant in human and pork samples (Martins
et al., 2020). The usual symptoms of Y. enterocolitica infection in human

beings are fever, diarrhea, and abdominal pain. However, reactive arthritis
and nodular erythema are some uncommon symptoms (Wang et al., 2021).
Yersiniosis in humans is the fourth and fifth most common foodborne
zoonosis generally transmitted through pork and water in the European
Union and the United States, respectively (EFSA, 2019; Tack et al., 2019).
In the European Union, from 2005 to 2018, pork and pork products were
among the most commonly described food types causing yersiniosis of
foodborne origin (EFSA and ECDC, 2019). Fosse et al. (2008) assessed that
77% of human yersiniosis cases are linked to the consumption of pork and
cold storage might guard Y. enterocolitica as it is psychrotrophic in nature
(Martins et al., 2020). Pork chitterlings were also linked to a yersiniosis
outbreak (Anonymous, 1990). It is assessed that 35–70% of pig herds and
45–100% of individual pigs carry Y. enterocolitica (Jakobsen et al., 2019). A
higher prevalence of Y. enterocolitica was reported in conventional intensive
pig farms (Laukkanen et al., 2009; Fondrevez et al., 2014; Zdolec and Kiš,
2021). In Norway, a considerable (25%) reduction in the incidence of Y.
enterocolitica prevalence after the sealing-off of the rectum with a plastic
bag indicates that the fecal material of pigs acts as an important source of
contamination for pork and pork products (Nesbakken and Skjerve, 1996).
However, the tonsils of pigs are considered as an important source of Y.
enterocolitica than fecal and intestinal contents (Fondrevez et al., 2014). The
reasons for the contamination of pig carcasses and products are unhygienic
slaughtering and processing, and poor handling of possibly contaminated
sites including tonsils and mesenteric lymph nodes (Martins et al., 2020).
9.2.3 CAMPYLOBACTER
Campylobacter is a Gram-negative microaerophilic bacteria that can survive

in temperature ranges and pH of 30–47°C and 5.8, respectively (Doyle,
1990). The C. jejuni/C. coli group of the pathogen has come out from obscu-
rity to rank among the foremost causes of diarrhea of bacterial origin in
humans (Butzler and Skirrow, 1979). C. jejuni is predominantly recovered
from gastroenteritis cases and is widely distributed in the animal kingdom
with the majority isolations from poultry, wild birds, exotic zoo animals,
and domestic animals like pigs, dogs, cats, cattle, sheep, etc (Milton et al.,
2020a). C. coli is the second most frequent and neglected cause of human
campylobacteriosis (Kelley et al., 2020). C. coli cause an illness in humans
that is identical to the one caused by C. jejuni (Karmali et al., 1983). Despite
pig being its major reservoir, C. coli is also found in other animals, but with
an occurrence considerably less than that of C. jejuni. Retail raw pork and
products are one of the important food vehicles (Noormohamed and Fakhr,
2013; Quintana-Hayashi and Thakur 2012; Beier et al. 2018). The widely
held Campylobacter infections in humans are self-limiting, which set hurdles
for the determination of its true incidence (Hanninen et al., 2000). Human
campylobacteriosis caused by C. jejuni/C. coli is typically manifested by
diarrhea (often bloody), vomiting, abdominal cramps, and fever and after
acute illness, it has been associated with long-term complications such as
Guillain–Barre syndrome, irritable bowel syndrome, and reactive arthritis
(Milton et al., 2020a). Swine get colonized with Campylobacter immediately
(<1 week) after birth and as they advance through various production phases,
the prevalence of Campylobacter usually increases (Schuppers et al., 2005).
A study reported a 15% increase in the prevalence of Campylobacter from
the growing to finishing stages of pigs (Farzan et al., 2010). In the finishing
stage, most of the pigs tested positive for Campylobacter, and across all the
stages of production, C. coli is found to be more prevalent than C. jejuni
(Schuppers et al., 2005). The occurrence of Campylobacter at the produc-
tion farm translates to the presence in the slaughter process and thereby in
pork. However, it can be reduced by following certain hygienic slaughter
procedures such asscalding, singeing, dehairing, polishing, etc. followed
by chilling as campylobacters are sensitive to low temperature and drying
(Pearce et al., 2003).
9.2.4 LISTERIA MONOCYTOGENES
Listeria is a Gram-positive, psychrotrophic, ubiquitously present bacteria

with common inhabitation in soil, water, and intestines of animals (Bahk
and Marth, 1990). Under the genus Listeria, L. monocytogenes is the only
zoonotic human pathogen representing the highest reasons for the hospital-
izations and death rates among infections caused by foodborne pathogens
(Scallan et al., 2011). The common clinical symptoms of L. monocytogenes
in humans are fever, chills, and back pain and mortality are usually due to
encephalitis, meningitis, and sepsis in 20–30% of infected cases, mostly
immunocompromised ones (Schlech, 2000). Listeriosis causes abortion and
fetal infection in pregnant women with around 1500 cases occurring in the
US alone and 99% are of foodborne origin (Scallan et al., 2011). Although
L. monocytogenes covers 13 serotypes, the serotypes 4b (28%), 1/2a (24%),

and 1/2c (22%) are commonly reported in slaughtered retail pork (Farber and
Peterkin, 1991; Kanuganti et al., 2002). Sows carry more L. monocytogenes
in feces than piglets (Fenlon et al., 1996). Apart from wet feed (or silage),
bedding or litter, and boots are all potential sources of Listeria contamination
to live pigs on the farm (Baer et al., 2013). As the environment acts as an
important source of L. monocytogenes to pigs, improper hygienic manage-
ment and biosecurity measures can lead to its enhanced prevalence on farms
(Hellstrom et al., 2010). Through serotype tracing, it was found that infected
live animals and cross-contamination of the carcass are the two important
routes of contamination to pork by L. monocytogenes (Hellstrom et al.,
2010). Higher levels of L. monocytogenes prevalence in tonsils and isolations
even after chilling of carcasses are the two noteworthy observations made
earlier (FSIS, 1996; Autio et al., 2000; Hellstrom et al., 2010). Ready-to-eat
pork products such as deli meats, frankfurters, etc., and home-stored meats
are known to harbor L. monocytogenes, and are associated with numerous
foodborne outbreaks (Schlech, 2000; de Valk et al., 2001; Baer et al., 2013).
These are also the indicators of cold insensitivity of L. monocytognes and
environmental contamination (Baer et al., 2013).
9.2.5 CLOSTRIDIUM PERFRINGENS AND CLOSTRIDIUM DIFFICILE
C. perfringens and C. difficile are Gram-positive anaerobic spore-forming

bacteria associated with economic and public health concerns in swine
production (Baker et al., 2010). These pathogens are normal colonizers of
the intestinal tract of various food animals mainly pigs (Songer and Uzal,
2005). Clostridial infections generally occur in piglets immediately (within
the first week) after birth and are associated with under-developed normal
microbiota, scouring, and antibiotic management (Yaeger et al., 2002; Baker
et al., 2010). Clinically, C. perfringens (type A and C) causes enteric and
systemic diseases in human beings including food poisoning, gas gangrene,
and enterocolitis (pigbel) (Milton et al., 2020b). C. difficile infections in
human beings can cause antibiotic-associated diarrhea to a life-threatening
infection such as severe pseudomembranous colitis (Milton et al., 2020b).
Both the pathogenic species of Clostridium have been reported from pork
and the environment of pig slaughterhouses (Rho et al., 2001; Metcalf et al.,
2010; Chan et al., 2012; Curry et al., 2012; Rodriguez et al., 2013; Mooyottu
et al., 2015; Wu et al., 2017). Nevertheless, although C. perfringens grades
among the most significant pathogens of food-borne diseases, the sorting of

C. difficile as a foodborne pathogen is still contentious (Milton et al., 2020b).
However, spotting identical strains of C. difficile in food animals, meat, and
humans put forward the possible transmission of C. difficile through foods as
well as pork products (Milton et al., 2020b).
9.2.6 STAPHYLOCOCCUS AUREUS
S. aureus is a Gram-positive, facultative anaerobic bacteria whose common

habitat is the upper respiratory tract and skin of humans and animals (Berg-
doll, 1990). As the primary reservoir of S. aureus seems to be skin, the pork
usually sold along with the skin makes them an important source if inad-
equately scalded and singed. The pathogen is zoonotic and the foodborne
illness caused by them involves clinical symptoms of vomiting, nausea,
diarrhea, and abdominal cramps after 1–6 h of ingestion (Bergdoll, 1990).
The pathogen releases enterotoxins that are the triggering factors of illness
and are accountable for the 14% hospitalization rate (Noskin et al., 2007).
However, for adequate enterotoxin levels to trigger illness, at least 105 S.
aureus cells/g must be present in the food (Bahtia and Zahoor, 2007). As S.
aureus is not a sturdy competitor compared to other bacterial flora present
in food, the contamination is typically linked to the postprocessing handling
of meat products when the competition has been removed (Argudin et al.,
2010). Despite the relatively low-risk attribute of S. aureus, the worldwide
emergence of methicillin-resistant S. aureus (MRSA) has become an
important public health concern (Baer et al., 2013). Varied prevalence of
S. aureus ranging from 42.2 to 76.7% has been reported in pork and prod-
ucts (Atanassova et al., 2001; van Loo et al., 2007; O’Brien et al., 2012;
Savariraj et al., 2018). Around 3.1–10.7% of fresh grounded and chopped
pork were described to be contaminated with MRSA (van Loo et al., 2007;
de Boer et al., 2009). Poor personal hygiene of slaughter and processing
personnel, unhygienic and unscientific abattoir, and the processing practices
contaminate the pork with a high count of S. aureus and consequent release
of enterotoxins (Savariraj et al., 2018).
9.2.7 SHIGATOXIGENIC ESCHERICHIA COLI (STEC)
E. coli is a Gram-negative, facultative anaerobic bacteria under the family

Enterobacteriaceae that is normally found in the intestines of warm-blooded
animals. E. coli is classified by its links with human disease, namely,

enterohemorrhagic or shiga toxigenic (EHEC/STEC), enterotoxigenic
(ETEC), enteropathogenic (EPEC), enteroinvasive (EIEC), enteroaggrega-
tive (EAEC), diffuse adhering (DAEC), and extraintestinal pathogenic E.
coli (ExPEC) (Logue et al., 2017). STEC possesses more zoonotic concerns
among all the pathogenic E. coli and pigs are important reservoirs of STEC
(Colello et al., 2016). STEC can cause severe foodborne diseases leading
to life-threatening complications like hemolytic uremic syndrome (HUS)
and bloody diarrhea due to colitis (Paton and Paton, 1998). The important
cause of acute kidney injury and acquired chronic kidney disease in children
are due to the damages caused by HUS through the cytotoxic actions of
Stx1 and Stx2. Normally, STEC does not cause disease in animals except
edema disease in pigs caused due to Stx2e subtype of STEC (Niewerth et
al., 2001). Pork and pork products have been incriminated in human STEC-
associated diarrhoeic and HUS infections (Sonntag et al., 2005; Kaufmann
et al., 2006; Trotz-Williams et al., 2012). A family outbreak of STEC (O157)
infection was reported to be due to the consumption of dry fermented pork
salami (Conedera et al., 2007). A varied prevalence of STEC in pigs ranging
from 2 to 31% was documented (Parma et al., 2000; Kaufmann et al., 2006;
Meng et al., 2014). A prevalence range of 0.2–26% of STEC was reported
earlier in pig carcasses during slaughtering (Leung et al., 2001; Bouvet et al.,
2002; Bohaychuk et al., 2011; Koláčková et al., 2014). In pig slaughtering
establishments, diverse STEC serogroups (O91, O121, and O157) that are
associated with human illnesses were detected earlier (Colello et al., 2016).
Slaughterhouse operations such as skinning, handling, and evisceration are
found to be the more likely reason for the contamination of pork (Koohma-
raie et al., 2005; Etcheverría et al., 2010).
9.2.8 STREPTOCOCCUS SUIS
S. suis is a Gram-positive facultative anaerobic bacteria whose common

habitat is the nasal cavities, tonsils, and alimentary and genital tracts of pigs
thus pig becomes the natural reservoir of the pathogen (Lun et al., 2007).
S. suis serotype 2 is a zoonotic pathogen largely of occupational origin and
most frequently associated with diseases in human beings and pigs (Lun et
al., 2007; Dutkiewicz et al., 2017). Human S. suis infections are clinically
manifested as pus-filled meningitis, the other described syndromes comprise
septicemia, streptococcal toxic shock-like syndrome with multiple organ
failure, cellulitis, endocarditis, rhabdomyolysis, arthritis, peritonitis, spon-

dylodiscitis, pneumonia, uveitis and endophthalmitis (Dutkiewicz et al.,
2017). The carriage rates of S. suis may vary between swine herds and can
range from 0% to up to 80–100% (Dutkiewicz et al., 2017). In the majority
of Western countries, the transmission of S. suis is considered occupational
affecting persons working in the pig or pork industries. But, in Southeast
Asian countries, the main risk is due to close contact with pigs or ingestion of
uncooked or partially cooked pork products (Wertheim et al., 2009; Segura,
2020). The percutaneous route of infection associated with handling pigs or
pork has also been reported multiple times (Chattopadhyay, 1979; Watkins et
al., 2001; Vilaichone et al., 2002; Wangkaew et al., 2006).
9.2.9 TAENIA SOLIUM
T. solium is a tapeworm (cestode) that dwells only in the intestines of human

beings (definitive host). Pigs acquire the infection by ingesting the eggs/
proglottids shed in human feces and play the role of intermediate hosts and
the parasite resides in the muscles and other tissues of pigs in the cyst form.
The human population acquires intestinal infections (taeniasis) of tapeworm
by consuming undercooked cyst-infested pork, that is, measly pork (Phiri et
al., 2003). In addition, human beings can also get infected with cystic form
infection (cysticercosis) if encounter infested human feces either directly or
indirectly. The cysts are formed throughout the body, but predominantly in
the eyes and brain leading to neurocysticercosis condition (Davies, 2011).
Neurocysticercosis accounts for 50% of epileptic cases in developing coun-
tries. Apart from seizures, it causes raised intracranial pressure, headaches,
focal neurologic deficits, and psychiatric and ocular symptoms (Rajshekhar
et al., 2006). It was estimated by the Food and Agriculture Organization
(FAO) that worldwide 50 million people harbor adult T. solium worm and up
to 50,000 mortalities annually are attributable to neurocysticercosis (Eddi et
al., 2003; Davies, 2011). T. solium prevails where there is poor sanitation and
traditional free-ranging or scavenging type of pig rearing is practiced (Phiri
et al., 2003; Davies, 2011). The elimination of T. solium from North America
and Europe was basically due to economic development and adaptation of
the modern intensive system of pig production (Gonzalez et al., 2003). The
United States’s pig industry has acknowledged that open defecation in swine
farms by the farmworkers is the sole potential concern and education efforts
can eliminate the risk (Davies, 2011). Presently, the T. solium infection is
endemic in South and Central America, Asia, sub-Saharan Africa, and

some parts of the Pacific regions. The prevalence of cysticercosis infection
in pigs in Africa, South/Central America, and Asia are 64%, 65.4%, and
26%, respectively (Djurković-Djaković et al., 2013). The prevalence of T.
solium infection in human beings in Africa, South/Central America, and
Asia are 10%, 22%, and 1%, respectively (Djurković-Djaković et al., 2013).
Nevertheless, increasing economic immigration from endemic countries to
developed countries and increased tourism in developing nations indicate T.
solium as a pathogen with global health implications and the ability to spread
to nonendemic zones (Pawlowski et al., 2005).
9.2.10 TOXOPLASMA GONDII
T. gondii is a coccidian parasite (protozoan) of felids which is the definitive

host. The life cycle of T. gondii comprises three infective stages, namely,
sporozoites, tachyzoites, and bradyzoites. Cats shed sporozoites (repro-
ducing stage) as oocysts in their feces and it remains in the environment
and contaminates soil and water, and infects other intermediate hosts
such as food animals (such as pigs), human beings, and wildlife. After
ingestion of oocysts by intermediate vertebrate hosts, the parasite assumes
tachyzoites and bradyzoites (nonreproducing) stages and develops cysts in
the tissues such as muscle, heart, and brain (Cliver 1990; Baer et al., 2013).
Human beings acquire infection mainly through the consumption of raw or
improperly cooked infested meat and through oocyst-contaminated water
(Foroutan-Rad et al., 2016; Sher et al., 2016). Severe clinical manifesta-
tions are usually noticed in immunosuppressed individuals such as AIDS
patients, and pregnant ladies. In pregnant women, it causes stillbirths and
abortion and if the fetus survives, T. gondii is passed to the neonate and
results in a congenital form of the disease with major abnormalities (Cliver,
1990; Carvalheiro et al., 2005). However, throughout the world, a large
number of people have antibodies to the parasite, indicating T. gondii is an
important public health concern. In the United States, it was reported that
out of 225000 cases, 50% were of foodborne origin (Mead et al., 1999). T.
gondii was described to be responsible for 8% of hospitalizations and 24%
of deceases from foodborne diseases (Scallan et al., 2011). The prevalence
of T. gondii in pigs in different countries has been well recognized (Baer
et al., 2013). The important risk factors for T. gondii infection in pigs were
described as free-range production system, increased contact with wild
animals or cats, improper discarding of pig carcasses, and contaminated

water or feed (Dubey et al., 2002; Gebreyes et al., 2008; Hill et al., 2010;
Baer et al., 2013). As T. gondii can endure infectivity in the tissues of
pigs for not less than 171 days, its presence in pork is a zoonotic concern
(Dubey et al., 1984). In a US-based study, it was found that pork was
the only retail meat reported to be positive for the parasite (Dubey et al.,
2005). The same study has estimated that people in the northeastern US
have a 78% chance of procuring pork contaminated with T. gondii (Dubey
et al., 2005). As T. gondii can be destructed by several preservation and
processing methods such as heating (min. 67°C), freezing (min. –12.4°C),
etc., improper processing or cooking remains a major risk factor (Dubey et
al., 1990; Lunden and Uggla, 1992).
9.2.11 TRICHINELLA SPIRALIS
T. spiralis is a nematode (roundworm) that causes trichinellosis in human

beings due to the consumption of undercooked meat containing encysted
larvae. In addition to the predominant and classical agent T. spiralis, a few
rare species of Trichinella implicated in human disease are Trichinella
pseudospiralis, Trichinella britovi, Trichinella murelli, Trichinella nativa,
Trichinella nelson, and Trichinella papuae (CDC, 2019). Human beings
who consume infested meat contract the disease with clinical manifestations
such as abdominal pain, fever, and periorbital edema. The larvae are released
from the cysts and invade the mucosa of the small intestine and develop
into adult worms and the adult female further releases larvae that migrate
to striated muscles and encyst. This leads to myalgia, joint pain, and muscle
contraction inefficiency (Cliver, 1990). It can also cause serious disease
in elderly individuals where serious complications such as encephalitis or
myocarditis can lead to death (Djurković-Djaković et al., 2013). Roughly,
150 trichinellosis cases are reported annually in the US and all of those cases
are of foodborne origin (Kennedy et al., 2009; Scallan et al., 2011). A survey
conducted in 198 countries revealed that 21.9% of the countries had Trichi
nella spp. in pig herds (Pozio, 2006). A CDC report on trichinellosis revealed
that during 2002–2007, 19% of trichinellosis in the US was caused by the
consumption of pork, whereas 50% were owing to the consumption of wild
game meat (Kennedy et al., 2009). The important risk factors for T. spiralis
infection in pigs were described as a free-range production system, feeding
of garbage and wildlife carcasses, and improper discarding of pig carcasses
(Cliver, 1990; Murrell and Pozio, 2000; van der Giessen, et al., 2007). A
study conducted between 1999 and 2004 in China revealed a prevalence of
1.6–3.7% of T. spiralis in retail meat (Cui et al., 2006). As thorough cooking
remains the best option to inactivate Trichinella, improper cooking becomes
the major risk factor in the transmission of T. spiralis to human beings via
the consumption of pork (Baer et al., 2013).
9.3 HEPATITIS E VIRUS

Hepatitis E virus (HEV) under the genus Hepevirus is an RNA virus recog-
nized as the leading cause of acute viral hepatitis. Among the diverse HEV
genotypes (GT) that affect human beings, GT3 and GT4 are zoonotic, have
been reported in various animal species (Zhou et al., 2019; Li et al., 2020).
Pigs serve as the main reservoir for the zoonotic HEV strains. Antibodies
against HEV have been extensively detected in both domestic and wild pigs
(Baechlein et al., 2010; Geng et al., 2010; Fredriksson-Ahomaa et al., 2020).
Though HEV infection is generally asymptomatic or self-limiting in healthy
people, GT3 and GT4 HEV infections in organ transplant recipients are more
likely to develop chronic hepatitis (Kamar et al., 2008). However, asymp-
tomatic individuals in endemic areas have been recognized to be potential
reservoirs shedding the virus, permitting its ecological persistence (Christou
and Kosmidou, 2013). Worldwide, it is estimated that around 939 million
equivalent to 1 in 8 persons have ever encountered HEV infection (Li et al.,
2020). The route of transmission of HEV from pigs to humans is frequently
through direct contact with pigs and indirectly through the consumption
of contaminated pork products and environmental exposure. Nevertheless,
the extensive consumption of pork products is certainly posing a great risk
of HEV transmission through food. HEV infections associated with the
consumption of undercooked pork and wild boar meat have been repeatedly
reported (Matsuda et al., 2003; Yazaki et al., 2003). It was estimated that
globally, close to 60% of domestic pigs and 27% of wild boars have ever
been HEV infected. Approximately 13% of domestic and 9.5% of wild pigs
are undergoing an active infection. In addition, globally, 10% of commercial
pork products are positive for HEV RNA (Li et al., 2022).
9.4 OTHER PATHOGENS IN THE CONTEXT OF PORK SAFETY

Shigella, Bacillus cereus, rotavirus, norovirus, and Sarcocystis, are other
pathogens associated with pork-associated foodborne diseases. Shigella
is a Gram-negative bacterial contaminant of food that is estimated to

cause 164.7 million foodborne illnesses (dysentery) and 1.1 million
deaths worldwide (Thiem et al., 2004). Foodborne outbreaks of shigella
are widely reported and 7.5% of Shigella outbreaks were attributed to
meat and meat products including pork (Pires et al., 2012). B. cereus
is a Gram-positive bacterial contaminant of food that is estimated to
cause 1.4–12% of foodborne outbreaks worldwide and 8.7% of B.
cereus outbreaks were attributed to pork products (Pires et al., 2012;
Grutsch et al., 2018). Rotavirus and norovirus are RNA viruses that are
predominantly responsible for diarrheal illnesses worldwide (Hakim et
al., 2018). Globally rotavirus accounts for 6% of diarrheic episodes and
is responsible for 35,2000–592,000 deaths in children below 5 years of
age (Parashar et al., 2003). Rotavirus has been previously detected in
retail pork and products (Soares et al., 2022). Norovirus is estimated
to be associated with 18% of diarrheal diseases and responsible for
212,000 deaths annually worldwide (Pires et al., 2015). Between 1988
and 2007, 2% of norovirus outbreaks were attributed to pork products
(Greig and Ravel, 2009). Sarcocystis is a coccidian parasite and three
of its species, namely, Sarcocystis suihominis, Sarcocystis miescheriana,
and Sarcocystis porcifelis have been recognized in pigs. However, only
S. suihominis can cause human infection upon consumption of raw pork
(Djurković-Djaković et al., 2013). A foodborne outbreak of S. suihominis
in children who consumed undercooked pork was reported in India
(Banerjee et al., 1994).
9.5 CONCLUSION
This chapter has thoroughly outlined all the zoonotic pathogens and their
significance in the pork industry. As the urge for intensive and alternative
(outdoor) pig production is equally increasing across the world, comprehen-
sive tailor-made preharvest plans specific to various situations or farms have
to be delineated and strictly followed to ensure the safety of pork. Further
hygienic slaughter, HACCP implementation, minimum handling, complete
processing, proper packaging, appropriate personal hygiene of handlers,
pest-proof food processing area, etc. are some of the postharvest strategies
that ensure safe pork on the consumer’s plate.
KEYWORDS
• pork
• safety
• Zoonoses
• pathogen
• foodborne diseases
REFERENCES
Alban, L.; Stark, K. D. Where Should the Effort be Put to Reduce the Salmonella Prevalence
in the Slaughtered Swine Carcass Effectively? Prev. Vet. Med. 2005, 68, 63–79.
Anonymous. Topics in Minority Health Yersinia enterocolitica Infections during the Holidays
in Black Families—Georgia. Morbid. Mortal. Weekly Rep. 1990, 39, 819–820.
Argudın, M. A.; Mendoza, M. C.; Rodicio, M. R. Food Poisoning and Staphylococcus aureus
Enterotoxins. Toxins. 2010, 2, 1751–1773.
Atanassova, V.; Meindl, A.; Ring, C. Prevalence of Staphylococcus aureus and Staphylococcal
Enterotoxins in Raw Pork and Uncooked Smoked Ham. Int. J. Food Microbiol. 2001, 68,
105–113.
Autio, T.; Sateri, T.; Fredriksson-Ahomaa, M.; Rahkio, M.; Lunden, J.; Korkeala, H. Listeria
monocytogenes Contamination Pattern in Pig Slaughterhouses. J Food Prot. 2000, 63,
1438–1442.
Baechlein, C.; Schielke, A.; Johne, R.; Ulrich, R. G.; Baumgaertner, W.; Grummer, B.
Prevalence of Hepatitis E Virus-Specific Antibodies in Sera of German Domestic Pigs
Estimated by Using Different Assays. Vet Microbiol. 2010, 144, 187–191.
Baer, A. A.; Miller, M. J.; Dilger, A. C. Pathogens of Interest to the Pork Industry: A Review
of Research on Interventions to Assure Food Safety. Compr. Rev. Food Sci. Food Saf. 2013,
12, 183–217.
Bahk, J.; Marth, E. H. Listeriosis and Listeria monocytogenes. Foodborne Diseases; Academic
Press Inc.: San Diego, California, 1990; pp 247–257.
Bahtia, A.; Zahoor, S. Staphylococcus aureus Enterotoxins: A Review. J. Clin. Diagn. Res.
2007, 3, 188–197.
Baker, A. A.; Davis, E.; Rehberger, T.; Rosener, D. Prevalence and Diversity of Toxigenic
Clostridium perfringens and Clostridium Difficile among Swine Herds in the Midwest.
Appl. Environ. Microbiol. 2010, 76, 2961–2967.
Banerjee, P. S.; Bhatia, B. B.; Pandit, B. A. Sarcocystis suihominis Infection in Human Beings
in India. Vet. Parasitol. 1994, 8, 57–58.
Beier, R. C.; Harvey, R. B.; Hernandez, C. A.; Hume, M. E.; Andrews, K.; Droleskey, R. E.
Interactions of Organic Acids with Campylobacter coli from Swine. PLoS One 2018, 13,
1–17.
Bergdoll, M. S. Staphylococcal Food Poisoning. Foodborne Diseases; Academic Press,

1990; pp 85–106.
Bohaychuk, V. M.; Gensler, G. E.; Barrios, P. R. Microbiological Baseline Study of Beef and
Pork Carcasses from Provincially Inspected Abattoirs in Alberta. Canada. Can. Vet. J. 2011,
52, 1095–1100.
Bottone, E. J. Yersinia enterocolitica: The Charisma Continues. Clin. Microbiol. Rev. 1997,
10, 257–276.
Bouvet, J.; Montet, M.; Rossel, R.; Le Roux, A.; Bavai, C.; Ray-Gueniot, S.; et al. Prevalence
of Verotoxin-Producing Escherichia coli (VTEC) and E. coli O157:H7 in French Pork. J.
Appl. Microbiol. 2002, 93, 7–14.
Buncic, S.; Alban, L; Blagojević, B. From Traditional Meat Inspection to Development of
Meat Safety Assurance Programs in Pig Abattoirs—the European Situation. Food Control.
2019, 106, 1–9.
Butzler, J. P.; Skirrow, M. B. Campylobacter enteritis. Clin. Gastroenterol. 1979, 8, 737–765.
Carvalheiro, C. G.; Mussi-Pinhata, M. M.; Yamamoto, A. Y.; Souza, C. B. S. D.; Maciel, L.
M. Z. Incidence of Congenital Toxoplasmosis Estimated by Neonatal Screening: Relevance
of Diagnostic Confirmation in Asymptomatic Newborn Infants. Epidemiol. Infect. 2005,
133, 485–491.
CDC, CDC Estimates of Foodborne Illness in the United States. Center for Disease Control
[Online] 2011. http://www.cdc.gov/foodborneburden/2011-foodborne-estimates.html
Centers for Disease Prevention and Control. Parasites—Trichinellosis (also known as
trichinosis) [Online], 2019. https://www.cdc.gov/parasites/trichinellosis/biology.html
(accessed May 17, 2022).
Chan, G.; Farzan, A.; Soltes, G.; Nicholson, V. M.; Pei, Y.; Friendship, R.; et al. The
Epidemiology of Clostridium perfringens Type A on Ontario Swine Farms, with Special
Reference to cpb2-Positive Isolates. BMC Vet Res. 2012, 8, 1–8.
Chattopadhyay, B. Group R Streptococcal Infection amongst Pig Meat Handlers. A Review.
Public Health. 1979, 93, 140–142.
Christou, L.; Kosmidou, M. Hepatitis E Virus in the Western World—A Pork-Related
Zoonosis. Clin. Microbiol. Infect. 2013, 19, 600–604.
Cliver, D. O. Parasites. Foodborne Diseases; Academic Press Inc.: San Diego, Calif., 1990;
PP 296–297.
Colello, R.; Cáceres, M. E.; Ruiz, M. J.; Sanz, M.; Etcheverría, A. I.; Padola, N. L. From
Farm to Table: Follow-up of Shiga Toxin-Producing Escherichia coli Throughout the Pork
Production Chain in Argentina. Front. Microbiol. 2016, 7, 1–7.
Conedera, G.; Mattiazzi, E.; Russo, F.; Chiesa, E.; Scorzato, I.; Grandesso, S.; et al. A
Family Outbreak of Escherichia coli O157 Haemorrhagic Colitis Caused by Pork Meat
Salami. Epidemiol Infect. 2007, 135, 311–314. doi: 10.1017/s0950268806006807. PMID:
17291366; PMCID: PMC2870571.
Cui, J.; Wang, Q.; Hu, D. S. The Epidemiology of Swine Trichinellosis in China during
1999–2004. Helminthologia. 2006, 43, 21–26.
Curry, S. R.; Marsh, J. W.; Schlackman, J. L.; Harrison, L. H. Prevalence of Clostridium
Difficile in Uncooked Ground Meat Products from Pittsburgh, Pennsylvania. Appl. Environ.
Microbiol. 2012, 78, 4183–4186.
Davies P. R. Pork Safety: Achievements and Challenges. Zoonoses Public Health 2010, 57,
1–5.
Davies, P. R. Intensive Swine Production and Pork Safety. Foodborne Pathog Dis. 2011
8,189–201. Doi: 10.1089/Fpd.2010.0717.
Davies, P. R.; Hurd, H. S.; Funk, J. A.; Fedorka-Cray, P. J.; Jones, F. T. The Role of
Contaminated Feed in the Epidemiology and Control of Salmonella enterica in Pork
Production. Foodborne Pathog. Dis. 2004, 1, 202–215.
De Boer, E.; Zwartkruis-Nahuis, J. T.; Wit, B.; Huijsdens, X. W.; De Neeling, A. J.; Bosch,
T.; et al. Prevalence of Methicillin-Resistant Staphylococcus aureus in Meat. Int. J. Food
Microb. 2009, 134, 52–56.
de Valk, H.; Vaillant, V.; Jacquet, C.; Le Querrec, F.; Stainer, F.; Quelquejeu, N.; Pierre,
O.; Pierre, V.; Desenclos, J. C.; Goulet, V. Two Consecutive Nationwide Outbreaks of
Listeriosis in France, October 1999–February 2000. Am. J. Epidemiol. 2001, 154, 944–995.
Delgado, C. L.; Rosegrant, M. W.; Meijer, S. Livestock to 2020: The Revolution Continues;
Intl. Trade Research Consortium (IATRC); Auckland, New Zealand, 2001.
Delsart, M.; Pol, F.; Dufour, B.; Rose, N.; Fablet, C. Pig Farming in Alternative Systems:
Strengths and Challenges in Terms of Animal Welfare, Biosecurity, Animal Health and Pork
Safety. Agriculture 2020, 10, 1–34. doi: 10.3390/agriculture10070261
Dickson, J.; Hurd, H. S.; Rostagno, M. H. Salmonella in the Pork Production Chain. Natl.
Pork Board/Pork Information Gateway Factsheet [Online], 2002. http://www.pork.org/
filelibrary/factsheets/porksafety/pork%20production%20chain.pdf
Djurković-Djaković, O.; Bobić, B.; Nikolić, A.; Klun, I, Dupouy-Camet J. Pork as a Source
of Human Parasitic Infection. Clin. Microbiol. Infect. 2013, 19, 586–594.
Dorr, P.; Tadesse, D. A.; Zewde, B.; Fry, P.; Thakur, S.; Gebreyes, W. A. Longitudinal Study
of Salmonella Dispersion and the Role of Environmental Contamination in Commercial
Swine Production Systems. Appl. Environ. Microbiol. 2009, 75, 1478–1486.
Doyle, M. P. Campylobacter jejuni. Foodborne Diseases; Academic Press Inc.; San Diego,
California, 1990; pp 217–228.
Doyle, M. P.; Cliver, D. O. Salmonella. Foodborne Diseases; Academic Press Inc.: San
Diego, California, 1990; pp 185–204.
Dubey, J. P.; Gamble, H. R.; Hill, D.; Sreekumar, C.; Romand, S.; Thulliez, P. High Prevalence
of Viable Toxoplasma gondii Infection in Market Weight Pigs from a Farm in Massachusetts.
J. Parasitol. 2002, 88, 1234–1238.
Dubey, J. P.; Hill, D. E.; Jones, J. L.; Hightower, A. W.; Kirkland, E.; Roberts, J. M.; et al.
Prevalence of Viable Toxoplasma gondii in Beef, Chicken and Pork from Retail Meat Stores
in the United States: Risk Assessment to Consumers. J. Parsitol. 2005, 91, 1082–1093.
Dubey, J. P.; Kotula, A. W.; Sharar, A.; Andrews, C. D.; Lindsay, D. S. Effect of High
Temperature on Infectivity of Toxoplasma gondii Tissue Cysts in Pork. J. Parsitol. 1990,
76, 201–204.
Dubey, J. P.; Murrell, K. D.; Fayer, R. Persistence of encysted Toxoplasma gondii in tissues of
pigs fed oocysts. Am. J. Vet. Res. 1984, 45, 1941–1943.
Dutkiewicz, J.; Sroka, J.; Zając, V.; Wasiński, B.; Cisak, E.; Sawczyn, A.; Kloc, A.; Wójcik-
Fatla, A. Streptococcus suis: A Re-emerging Pathogen Associated with Occupational
Exposure to Pigs or Pork Products. Part I—Epidemiology. Ann. Agric. Environ. Med. 2017,
24, 683–695.
Eddi, C.; Nari, A.; Amanfu, W. Taenia solium Cysticercosis/Taeniosis: Potential Linkage With
FAO Activities; FAO Support Possibilities. Acta Trop. 2003, 87, 145–148.
EFSA. The European Union One Health 2018 Zoonoses Report. EFSA J. 2019, 17, 1–275
Enemark, H. L.; Johansen, M. V.; Alban, L. Transmission Dynamics of Foodborne Parasites in

Pork (Pig and Wild Boar). In Parasite Transmission Dynamics in Food Sources; Gajadhar,
A., Ed.; Woodhead Publishing Ltd, 2015; pp 259–292.
Etcheverría, A.; Padola, N.; Sanz, M.; Polifroni, R.; Krüger, A.; Passucci, J.; et al. Occurrence
of Shiga Toxin-Producing E. coli (STEC) on Carcasses and Retail Beef Cuts in the
Marketing Chain of Beef in Argentina. Meat Sci. 2010, 86, 418–421.
European Food Safety Authority and European Centre for Disease Prevention and Control
(EFSA and ECDC). The European Union One Health 2018 Zoonoses Report. EFSA J.
2019, 17, E05926.
FAO. Food Outlook—Biannual Report on Global Food Markets (Rome: FAO) [Online] 2021.
http://www.fao.org/3/cb4479en/cb4479en.pdf
Farber, J. M.; Peterkin, P. I. Listeria monocytogenes, a Food-Borne Pathogen. Microbiol. Rev.
1991, 55, 476–511.
Fenlon, D. R.; Wilson, J.; Donachie, W. The Incidence and Level of Listeria monocytogenes
Contamination of Food Sources at Primary Production and Initial Processing. J. Appl.
Bacteriol. 1996, 81, 641–50.
Fondrevez, M.; Minvielle, B.; Labbé, A.; Houdayer, C.; Rose, N.; Esnault, E.; Denis, M.
Prevalence of Pathogenic Yersinia enterocolitica in Slaughter-Aged Pigs During a One-Year
Survey 2010 ̶ 2011 France. Int. J. Food Microbiol. 2014, 174, 56–62.
Foroutan-Rad, M.; Majidiani, H.; Dalvand, S.; Daryani, A.; Kooti, W.; Saki, J.; et al.
Toxoplasmosis in Blood Donors: A Systematic Review and Meta-Analysis. Transfus. Med.
Rev. 30, 116–122.
Fosse, J.; Seegers, H.; Magras, C. Foodborne Zoonoses Due to Meat: A Quantitative Approach
for A Comparative Risk Assessment Applied to Pig Slaughtering in Europe. Vet. Res. 2008,
39, 1–16.
Fosse, J.; Seegers, H.; Magras, C. Prevalence and Risk Factors for Bacterial Food-Borne
Zoonotic Hazards in Slaughter Pigs: A Review. Zoonoses Public Health 2009, 56, 429–454.
Fredriksson-Ahomaa, M.; London, L.; Skrzypczak, T.; Kantala, T.; Laamanen, I.; et al.
Foodborne Zoonoses Common in Hunted Wild Boars. Ecohealth 2020, 17, 512–522.
FSIS. Nationwide Pork Microbiological Baseline Data Collection Program: Market Hogs
(April 1995-March 1996). Food Safety and Inspection Service [Online] 1996. http://www.
fsis.usda.gov/science/baseline_data_market_hogs/index.asp
Gebreyes, W. A.; Bahnson, P. B.; Funk, J. A.; Mckean, J. D.; Patchanee, P. Seroprevalence of
Trichinella, Toxoplasma, and Salmonella in Antimicrobial-Free and Conventional Swine
Production Systems. Foodborne Pathog. Dis. 2008, 5, 199–203.
Geng, Y.; Wang, C.; Zhao, C.; Yu, X.; Harrison, T. J.; Tian, K.; et al. Serological Prevalence
of Hepatitis E Virus in Domestic Animals and Diversity of Genotype 4 Hepatitis E Virus in
China. Vector Borne Zoonotic Dis. 2010, 10, 765–770.
Gonzalez, A. E.; García, H. H.; Gilman, R. H.; Tsang, V. C. Control of Taenia solium. Acta
Trop. 2003, 87, 103–109.
Greig, J. D.; Ravel, A. Analysis of Foodborne Outbreak Data Reported Internationally for
Source Attribution. Int. J. Food Microbiol. 2009, 130, 77–87.
Grutsch, A. A.; Nimmer, P. S.; Pittsley, R. H.; Kornilow, K. G.; Mckillip, J. L. Molecular
Pathogenesis of Bacillus spp., with Emphasis on the Dairy Industry. Fine Focus. 2018, 4,
203–222.
Guibourdenche, M.; Roggentin, P.; Mikoleit, M.; Fields, P. I.; Bockemuhl, J.; Grimont, P. A.;
et al. Supplement 2003–2007 (No. 47) to the White-Kauffmann-Le Minor Scheme. Res.
Microbiol. 2010, 26–29. PMID:19840847.
Gupta, S. K.; Sharma, P.; Mcmillan, E. A.; Jackson, C. R.; Hiott, L. M.; Woodley, T.; et al.
Genomic Comparison of Diverse Salmonella Serovars Isolated From Swine. PLoS One
2019, 14, 1–24.
Hakim, M. S.; Nirwati, H.; Aman, A. T.; Soenarto, Y.; Pan, Q. Significance of Continuous
Rotavirus and Norovirus Surveillance in Indonesia. World J. Pediatr. 2018, 14, 4–12.
Hanninen, M. L.; Perko-Makela, P.; Pitkala, A; Rautelin, H. A Three Year Study of
Campylobacter jejuni Genotypes in Humans with Domestically Acquired Infections and in
Chicken Samples from the Helsinki Area. J. Clin. Microbiol. 2000, 38, 1998–2000.
Hellstrom, S.; Laukkanen, R.; Siekkinen, K. M.; Ranta, J.; Maijala, R.; Korkeala, H. Listeria
monocytogenes can Originate from Farms. J. Food Prot. 2010, 73, 641–648.
Hill, D. E.; Haley, C.; Wagner, B.; Gamble, H. R.; Dubey, J. P. Seroprevalence of and Risk
Factors for Toxoplasma gondii in the US Swine Herd Using Sera Collected During the
National Animal Health Monitoring Survey (Swine 2006). Zoonoses Public Health. 2010,
57, 53–59.
Jakobsen, A. M.; Bahl, M. I.; Buschhardt, T.; Hansen, T. B.; Al-Soud, W. A.; Brejnrod, A. D.;
Sørensen, S. J.; Nesbakken, T.; Aabo, S. Bacterial Community Analysis for Investigating
Bacterial Transfer from Tonsils to the Pig Carcass. Int. J. Food Microbiol. 2019, 295, 8–18.
Kaferstein, F. K.; Motarjemi, Y.; Bettcher, D.W. Foodborne Disease Control: A Transnational
Challenge. Emerg. Infect. Dis. 1997, 3, 503–510.
Kamar, N.; Selves, J.; Mansuy, J. M.; Quezzani, L.; Peron, J. M.; Guitard, J.; et al. Hepatitis
E Virus and Chronic Hepatitis in Organ-transplant Recipients. N. Engl. J. Med. 2008, 358,
811–817.
Kanuganti, S. R.; Wesley, I. V.; Reddy, P. G.; Mckean, J.; Hurd, H. S. Detection of Listeria
monocytogenes in Pigs and Pork. J. Food Prot. 2002, 65, 1470–1474.
Karabasil, N.; Boskovic, T.; Vasilev, D.; Betic, N.; Dimitrijevic, M. Pork Safety—Challenges
and Opportunities. IOP Conf. Ser.: Earth Environ. Sci. 2021, 854, 1–5.
Karmali, M. A.; Penner, J. L.; Fleming, P. C.; Williams, A.; Hennessy, J. N. The Serotype and
Biotype Distribution of Clinical Isolates of Campylobacter jejuni and Campylobacter coli
Over A Three-Year Period. J. Infect. Dis. 1983, 147, 243–246.
Kaufmann, M.; Zweifel, C.; Blanco, M.; Blanco, J.; Blanco, J.; Beutin, L.; et al. Escherichia
coli O157 and non-O157 Shiga Toxin–Producing Escherichia coli in Fecal Samples
of Finished Pigs at Slaughter in Switzerland. J. Food Prot. 2006, 69, 260–266. doi:
10.4315/0362–028x.jfp-10-579.
Kelley, B. R.; Ellis, J. C.; Large, A.; Schneider, L. G.; Jacobson, D.; Johnson, J. G. Whole-
Genome Sequencing and Bioinformatic Analysis of Environmental, Agricultural, and
Human Campylobacter jejuni Isolates From East Tennessee. Front. Microbiol. 2020, 11,
1–17.
Kennedy, E. D.; Hall, R. L.; Montgomery, S. P.; Pyburn, D. G.; Jones, J. L. Trichinellosis
Surveillance: United States, 2002–2007. MMWR Surveill Summ. 2009, 58, 1–7.
Kolackova, I.; Hazova, K.; Skockova, A.; Karpiskova, R. Occurrence of Shiga
Toxigenic Escherichia coli Strains in Pigs and Cattle at Slaughterhouses in the Czech
Republic in 2013. Klin. Mikrobiol. Infekc. Lek. 2014, 20, 36–39.
Koohmaraie, M.; Arthur, T.; Bosilevac, J.; Guerini, M.; Shackelford, S.; Wheeler, T. Post-
harvest Interventions to Reduce/Eliminate Pathogens in Beef. Meat Sci. 2005, 71, 79–91.
Korsak, N.; Jacob, B.; Groven, B.; Etienne, G.; China, B.; Ghafir, Y.; Daube, G. Salmonella
Contamination of Pigs and Pork in an Integrated Pig Production System. J. Food Prot.
2003, 66, 1126–1133.
Kranker, S.; Alban, L.; Boes, J.; Dahl, J. Longitudinal Study of Salmonella enterica Serotype
Typhimurium Infection in Three Danish Farrow-to-Finish Swine Herds. J. Clin. Microbiol.
2003, 41, 2282–2288.
Lam, S.; Nguyen, H. T. T.; Nguyen-Viet, H. Unger, F. Mapping Pathways toward Safer
Pork in Vietnam. ILRI Policy Brief [Online] 2020, 95. https://cgspace.cgiar.org/bitstream/
handle/10568/108768/research_brief_95.pdf?sequence=5&isallowed=y (accessed May 13,
2022).
Laukkanen, R.; Martínez, P. O.; Siekkinen, K. M.; Ranta, J.; Maijala, R.; Korkeala, H.
Contamination of Carcasses with Human Pathogenic Yersinia enterocolitica 4/O:3
Originates from Pigs Infected on Farms. Foodborne Pathog. Dis. 2009, 6, 681–688.
Leung, P.; Yam, W.; Ng, W.; Peiris, J. The Prevalence and Characterization of Verotoxin-
Producing Escherichia coli Isolated from Cattle and Pigs in an Abattoir in Hong
Kong. Epidemiol. Infect. 2001, 126, 173–179.
Li, P.; Ji, Y.; Li, Y.; Ma, Z.; Pan, Q. Estimating the Global Prevalence of Hepatitis E Virus in
Swine and Pork Products. One Health 2022; 14, 100362.
Li, P.; Liu, J.; Li, Y.; Su, J.; Ma, Z.; Bramer, W. M.; et al. The Global Epidemiology of
Hepatitis E Virus Infection: A Systematic Review and Meta-analysis, Liver Int. 2020, 40,
1516–1528.
Logue, C. M.; Barbieri, N. L.; Nielsen, D. W. Pathogens of Food Animals: Sources,
Characteristics, Human Risk, and Methods of Detection. Adv. Food Nutr. Res. 2017, 82,
277–365. doi: 10.1016/bs.afnr.2016.12.009.
Lun, Z. R.; Wang, Q. P.; Chen, X. G.; Li, A. X.; Zhu, X. Q. Streptococcus suis: An Emerging
Zoonotic Pathogen. Lancet. Infect. Dis. 2007, 7, 201–209.
Lunden, A.; Uggla, A. Infectivity of Toxoplasma gondii in Mutton Following Curing,
Smoking, Freezing or Microwave Cooking. Int. J. Food. Microbiol. 1992, 15, 357–363.
Martins, B. T. F.; De Azevedo, E. C.; Yamatogi, R. S.; Call, D. R.; Nero, L. A. Persistence of
Yersinia enterocolitica Bio-Serotype 4/O:3 in A Pork Production Chain in Minas Gerais,
Brazil. Food Microbiol. 2020, 94, 103660.
Matsuda, H.; Okada, K.; Takahashi, K.; Mishiro, S. Severe Hepatitis E Virus Infection after
Ingestion of Uncooked Liver from a Wild Boar. J. Infect. Dis. 2003, 188, 944.
Mead, P. S.; Slutsker, L.; Dietz, V.; Mccaig, L. F.; Bresee, J. S.; Shapiro, C.; et al. Food-
Related Illness and Death in the United States. Emerg. Infect. Dis. 1999, 5, 607–625.
Meng, Q.; Bai, X.; Zhao, A.; Lan, R.; Du, H.; Wang, T.; et al. Characterization of Shiga Toxin-
Producing Escherichia coli Isolated from Healthy Pigs in China. BMC Microbiol. 2014,
14, 1–14.
Metcalf, D.; Reid-Smith, R. J.; Avery, B. P.; Weese, J. S. Prevalence of Clostridium difficile in
Retail Pork. Can. Vet. J. 2010, 51, 873–876.
Milton, A. A. P.; Priya, G. B.; Angappan, M.; Momin, K. M.; Ghatak, S.; Kannan, P.
Campylobacteriosis: Emerging Foodborne Zoonosis. In Food Product Optimization For
Quality and Safety Control: Process, Monitoring, and Standards, 1st ed.; Contreras-
Esquivel, J. C., Badwaik, L. S., Kannan, P., Hagi, A. K., Eds.; CRC Press: United States,
2020a; pp165–187.
Milton, A. A. P.; Priya, G. B.; Momin, K. M.; Angappan, M.; Ghatak, S.; Kannan, P. Foodborne
Pathogenic Anaerobes. In Food Product Optimization For Quality and Safety Control:
Process, Monitoring, and Standards, 1st ed.; Contreras-Esquivel, J. C., Badwaik, L. S.,
Kannan, P., Hagi, A. K., Eds.; CRC Press: United States, 2020b; pp 31–52.
Montville, T. J.; Matthews, K. R. Food Microbiology: An Introduction, 2nd ed.; ASM Press:
Washington, D.C.; 2008.
Mooyottu, S.; Flock, G.; Kollanoor-Johny, A.; Upadhyaya, I.; Jayarao, B.; Venkitanarayanan,
K. Characterization of a Multidrug Resistant C. difficile Meat Isolate. Int. J. Food Microbiol.
2015, 192, 111–116.
Murrell, K. D.; Pozio, E. Trichinellosis: the Zoonosis that Won’t Go Quietly. Int. J. Parasitol.
2000, 30, 1339–1349.
Nesbakken, T.; Skjerve, E. Interruption of Microbial Cycles in Farm Animals from Farm to
Table. Meat Sci. 1996, 43, S47–S57.
Niewerth, U.; Frey, A.; Voss, T.; Le Bouguénec, C.; Baljer, G.; Franke, S.; et al. The AIDA
Autotransporter System is Associated With F18 and stx2e in Escherichia Coli Isolates
from Pigs Diagnosed with Edema Disease and Postweaning Diarrhea. Clin. Diagn. Lab.
Immunol. 2001, 8, 143–149.
Noormohamed, A.; Fakhr, M. K. A Higher Prevalence Rate of Campylobacter in Retail Beef
Livers Compared to Other Beef and Pork Meat Cuts. Int. J. Environ. Res. Public Health.
2013, 10, 2058–2068. doi: 10.3390/ijerph10052058
Noskin, G. A.; Rubin, R. J.; Schentag, J. J.; Kluytmans, J.; Hedblom, E. C.; Jacobson, C.;
Smulders, M.; Gemmen, E.; Bharmal, M. National Trends in Staphylococcus aureus
Infection Rates: Impact on Economic Burden and Mortality Over a 6-Year Period (1998–
2003). Clin. Infect. Dis. 2007, 45, 1132–1140.
O’Brien, A. M.; Hanson, B. M.; Farina, S. A.; Wu, J. Y.; Simmering, J. E., Wardyn, S. E.; et
al. MRSA in Conventional and Alternative Retail Pork Products. PLoS One. 2012, 7, 1–6.
Oliver, S. P.; Patel, D. A.; Callaway, T. R.; Torrence, M. E. Developments and Future Outlook
for Preharvest Food Safety. J. Anim. Sci. 2009, 87, 419–437.
Parashar, U. D.; Hummelman, E. G.; Bresee, J. S.; Miller, M. A.; Glass, R. I. Global Illness
and Deaths Caused by Rotavirus Disease in Children. Emerg. Infect. Dis. 2003, 9, 565–572.
Parma, A.; Sanz, M.; Blanco, J.; Blanco, J.; Viñas, M.; Blanco, M.; et al. Virulence Genotypes
and Serotypes of Verotoxigenic Escherichia coli Isolated from Cattle and Foods in
Argentina. Eur. J. Epidemiol. 2000, 16, 757–762.
Paton, J. C.; Paton, A. W. Pathogenesis and Diagnosis of Shiga Toxin-Producing Escherichia
coli Infections. Clin. Microbiol. Rev. 1998, 11, 450–479.
Pawlowski, Z.; Allan, J.; Sarti, E. Control of Taenia solium Taeniasis/Cysticercosis: from
Research Towards Implementation. Int. J. Parasitol. 2005, 35, 1221–1232.
Pearce, R. A.; Wallace, F. M.; Call, J. E.; Dudley, R. L, Oser, A.; Yoder, L.; Sheridan, J. J.;
Luchansky, J. B. Prevalence of Campylobacter Within a Swine Slaughter and Processing
Facility. J. Food Prot. 2003, 66, 1550–1556.
Phiri, I. K.; Sikasunge, C. S.; Siziya, S.; Gabriel, S.; Dorny, P.; Willingham, A. L. 3rd. The
Prevalence and Risk Factors of Porcine Cysticercosis in Zambia, Proceedings of the 5th
International Symposium On the Epidemiology and Control of Foodborne Pathogens in
Pork, Heraklion, Greece, 2003, pp 68–69.
Pires, S. M.; Fischer-Walker, C. L.; Lanata, C. F.; Devleesschauwer, B.; Hall, A. J.; Kirk, M.
D.; et al. Aetiology-Specific Estimates of the Global and Regional Incidence and Mortality
of Diarrhoeal Diseases Commonly Transmitted Through Food. PLoS One. 2015, 10, 1–17.
Pires, S. M.; Vieira, A. R.; Perez, E.; Lo Fo Wong, D.; Hald, T. Attributing Human Foodborne
Illness to Food Sources and Water in Latin America and the Caribbean Using Data from
Outbreak Investigations. Int. J. Food Microbiol. 2012, 152, 129–138.
Pozio, E.; Kapel, C. M.; Gajadhar, A. A.; Boireau, P.; Dupouy-Camet, J.; Gamble, H. R.
Trichinella in Pork: Current Knowledge on the Suitability of Freezing as a Public Health
Measure. Euro. Surveill. 2006, 11, 3079.
Quintana-Hayashi, M. P.; Thakur, S. Longitudinal Study of the Persistence of Antimicrobial-
Resistant Campylobacter Strains in Distinct Swine Production Systems on Farms, at
Slaughter, and in the Environment. Appl. Environ. Microbiol. 2012, 78, 2698–2705.
Rajshekhar, V.; Raghava, M. V.; Prabhakaran, V.; Oommen, A.; Muliyil, J. Active Epilepsy as
an Index of Burden of Neurocysticercosis in Vellore District, India. Neurology. 2006, 67,
2135–2139.
Rho, M. J.; Chung, M. S.; Lee, J. H.; Park, J. Monitoring of Microbial Hazards at Farms,
Slaughterhouses, and Processing Lines of Swine in Korea. J. Food Prot. 2001, 64,
1388–1391.
Rodriguez, C.; Avesani, V.; Van Broeck, J.; Taminiau, B.; Delmée, M.; Daube, G. Presence
of Clostridium difficile in Pigs and Cattle Intestinal Contents and Carcass Contamination at
the Slaughterhouse in Belgium. Int. J. Food Microbiol. 2013, 166, 256–262.
Rowe, T.; Leonard, F. C.; Kelly, G.; Lynch, P.; Egan, J.; Quirke, A.; Quinn P. Salmonella
Serotypes Present on a Sample of Irish Pig Farms. Vet. Rec. 2003, 153, 453–456.
Salajpal, K.; Karolyi, D.; Luković, Z. Sanitary Aspects of Outdoor Farming Systems. In
Proceedings of the Acta Argiculturae Slovenica; Slovenia: Ljubljana, 109–117.
Savariraj, W. R.; Ravindran, N. B.; Kannan, P.; Paramasivam, R.; Senthilkumar, T. M. A.;
Kumarasamy, P.; et al. Prevalence, Antimicrobial Susceptibility and Virulence Genes of
Staphylococcus aureus Isolated from Pork Meat in Retail Outlets in India. J. Food Saf.
2018, 39, 1–8.
Scallan, E.; Hoekstra, R. M.; Angulo, F. J.; Tauxe, R. V.; Widdowson, M. A.; Roy, S. L.; et
al. Foodborne Illness Acquired in the United States—Major Pathogens. Emerg. Infect. Dis.
2011, 17, 7–15.
Schirone, M.; Visciano, P.; Tofalo, R.; Suzzi, G. Editorial: Foodborne Pathogens: Hygiene
and Safety. Front. Microbiol. 2019, 10, 1974.
Schlech, W. F. Foodborne Listeriosis. Clin Infect Dis, 2000, 31, 770–775.
Schmidt, J.; Brichta-Harhay, D.; Kalchayanand, N.; Bosilevac, J.; Shackelford, S.; Wheeler,
T.; Koohmaraie, M. Prevalence, Enumeration, Serotypes, and Antimicrobial Resistance
Phenotypes of Salmonella enterica Isolates from Carcasses at Two Large United States
Pork Processing Plants. Appl. Environ. Microbiol. 2012, 78, 2716–2726.
Schuppers, M. E.; Stephan, R.; Ledergerber, U.; Danuser, J.; Bissig-Choisat, B.; Stark, K.
D. C.; Regula, G.. Clinical Herd Health, Farm Management and Antimicrobial Resistance
in Campylobacter coli on Finishing Pig Farms in Switzerland. Prev. Vet. Med. 2005,
69,189–202.
Segura, M. Streptococcus suis Research: Progress and Challenges. Pathogens. 2020, 9, 1–8.
Sher, A.; Tosh, K.; Jankovic, D. Innate Recognition of Toxoplasma gondii in Humans Involves
A Mechanism Distinct from that Utilized by Rodents. Cell. Mol. Immunol. 2016, 14, 36–42.
Smith, J. L.; Fratamico, P. M. Emerging and Re-emerging Foodborne Pathogens. Foodborne
Pathog. Dis. 2018, 15, 737–757.
Soares, V. M.; Dos Santos, E. A. R.; Tadielo, L. E.; Cerqueira-Cézar, C. K.; Da Cruz Encide
Sampaio, A. N.; Eisen, A. K. A.; et al. Detection of Adenovirus, Rotavirus, and Hepatitis E
Virus in Meat Cuts Marketed in Uruguaiana, Rio Grande Do Sul, Brazil. One Health. 2022,
14, 100377.
Songer, J. G.; Uzal, F. A. Clostridial Enteric Infections in Pigs. J. Vet. Diagn. Invest. 2005,
17, 528–536.
Sonntag, A.-K.; Bielaszewska, M.; Mellmann, A.; Dierksen, N.; Schierack, P.; Wieler, L. H.;
et al. Shiga Toxin 2e-Producing Escherichia coli Isolates From Humans and Pigs Differ in
Their Virulence Profiles and Interactions With Intestinal Epithelial Cells. Appl. Environ.
Microbiol. 2005, 71, 8855–8863.
Tack, D. M.; Marder, E. P.; Griffin, P. M.; Cieslak, P. R.; Dunn, J., Hurd, S.; et al. Preliminary
Incidence and Trends of Infections with Pathogens Transmitted Commonly Through
Food—Foodborne Diseases Active Surveillance Network, 10 U.S. Sites, 2015–2018.
Mortal. Week. Rep. 2019, 68, 369–373.
Thiem, V. D.; Sethabutr, O.; Von, S. L.; Van, T. B.; Chien, T.; Lee, H.; et al. Detection of
Shigella by A PCR Assay Targeting the IpaH Gene Suggests Increased Prevalence of
Shigellosis in Nha Trang, Vietnam. J. Clin. Microbiol. 2004, 42, 2031–2035.
Trotz-Williams, L. A.; Mercer, N. J.; Walters, J. M.; Maki, A. M.; Johnson, R. P. Pork
Implicated in A Shiga Toxin-Producing Escherichia coli O157: H7 Outbreak in Ontario,
Canada. Can. J. Public Health. 2012, 103, E322–E326.
Van Der Giessen, J.; Fonville, M.; Bouwknegt, M.; Langelaar, M.; Vollema, A. Seroprevalence
of Trichinella spiralis and Toxoplasma gondii in Pigs From Different Housing Systems in
the Netherlands. Vet. Parasitol. 2007, 148, 371–374.
Van Loo, I. H. M.; Diederen, B. M. W.; Savelkoul, P. H. M.; Woudenberg, J. H. C.; Roosendaal,
R.; Van Belkum, A. Methicillin-Resistant Staphylococcus aureus in Meat Products, the
Netherlands. Emerg. Infect. Dis. 2007, 13, 1753–1755.
Vilaichone, R. K.; Vilaichone, W.; Nunthapisud, P.; Wilde, H. Streptococcus suis Infection in
Thailand. J. Med. Assoc. Thai. 2002, 85, S109–S117.
Wang, J.; Liu, M.; Wang, H.; Wu, Q.; Xu, T.; Ma, G.; et al. Occurrence, Molecular
Characterization, and Antimicrobial Susceptibility of Yersinia enterocolitica Isolated from
Retail Food Samples in China. LWT 2021, 150, 111876.
Wangkaew, S.; Chaiwarith, R.; Tharavichitkul, P.; Supparatpinyo, K. Streptococcus suis
Infection: A Series of 41 Cases from Chiang Mai University Hospital. J. Infect. 2006, 52,
455–460.
Watkins, E. J.; Brooksby, P.; Schwelger, M. S.; Enright, S. M. Septicaemia in A Pig-Farm
Worker. Lancet, 2001, 357, 38.
Wertheim, H. F.; Nghia, H. D.; Taylor, W.; Schultsz, C. Streptococcus suis: An Emerging
Human Pathogen. Clin. Infect. Dis. 2009, 48, 617–625.
WHO. Food Safety and Foodborne Illnesses. Fact Sheet 237; WHO’s Health & Safety
Information Services: Geneva, Revised March 2007, 2009.
Wilkins, W.; Rajic, A.; Waldner, C.; Mcfall, M.; Chow, E.; Muckle, A.; Rosengren L.
Distribution of Salmonella Serovars in Breeding, Nursery, and Grow-to-Finish Pig, and Risk
Factors for Shedding in Ten Farrow-to-Finish Swine Farms in Alberta and Saskatchewan.
Can. J. Vet. Res. 2010, 74, 81–90.
Wu, L. H.; Wang, S. X.; Zhu, D.; Hu, W. Y.; Wang, H. S. Chinese Consumers’ Preferences
and Willingness to Pay for Traceable Food Quality and Safety Attributes: the Case of Pork.
China Econ. Rev. 2015, 35, 121–136.
Wu, Y. C.; Chen, C. M.; Kuo, C. J.; Lee, J. J.; Chen, P. C.; Chang, Y. C.; Chen, T. H. Prevalence
and Molecular Characterization of Clostridium difficile Isolates from a Pig Slaughterhouse,
Pork, and Humans in Taiwan. Int. J. Food Microbiol. 2017, 242, 37–44.
Yaeger, M.; Funk, N.; Hoffmann, L. A Survey of Agents Associated with Neonatal Diarrhea
in Iowa Swine Including Clostridium difficile and Porcine Reproductive and Respiratory
Syndrome Virus. J. Vet. Diagn. Invest. 2002, 14, 281–287.
Yazaki, Y.; Mizuo, H.; Takahashi, M.; Nishizawa, T.; Sasaki, N.; Gotanda, Y.; et al. Sporadic
Acute or Fulminant Hepatitis E in Hokkaido, Japan, may be Food-Borne, as Suggested by
the Presence of Hepatitis E Virus in Pig Liver as Food. J. Gen. Virol. 2003, 84, 2351–2357.
Zdolec, N.; Kiš, M. Meat Safety from Farm to Slaughter—Risk-Based Control of Yersinia
enterocolitica and Toxoplasma gondii. Processes 2021, 9, 1–13.
Zhou, J. H.; Li, X. R.; Lan, X.; Han, S. Y.; Wang, Y. N.; Hu, Y.; et al. The Genetic Divergences
of Codon Usage Shed New Lights on Transmission of Hepatitis E Virus from Swine to
Human. Infect. Genet. Evol. 2019, 68, 23–29.
SECTION 3
CASE STUDIES
CHAPTER 10
Functional Properties of Agroindustrial

Wastes from Pomegranate Peel (Punica
granatum)
ANA L. IZÁBAL-CARVAJAL1, CRISTIAN TORRES-LEÓN2,
LEONARDO SEPÚLVEDA1, MÓNICA L. CHÁVEZ GONZÁLEZ1,
CRISTÓBAL N. AGUILAR1, and J. A. ASCACIO-VALDÉS1
1
Bioprocesses and Bioproducts Group, Food Research Department,
School of Chemistry, Universidad Autónoma de Coahuila, Saltillo,
Coahuila, México
2
Research Center and Ethnobiological Garden, Universidad Autónoma de
Coahuila, Unidad Torreón, Viesca, Coahuila, México
ABSTRACT
Fruits are rich sources of bioactive components, which is mainly attributed

to various medicinal properties and beneficial effects on health (Kalaycıoğlu
and Erim, 2017). Pomegranate (Punica granatum L.) is being investigated
because it is a source of various beneficial phytochemicals for health since it
contains anthocyanins, ellagitannins, gallotannins, flavonoids, among other
polyphenolic compounds (Bonzanini et al., 2009; García-Villalba et al.,
2015; N. Seeram et al., 2005; Verotta et al., 2018).
The increase in food processing has greatly contributed to the increase
in agro-industrial waste, which is produced in all phases of the agricul-
tural production cycle, such as industrial manufacturing, processing, and
distribution (Kumar et al., 2017). In recent years, the use of agroindus-
trial wastes has become a global concern due to the scarcity of natural
resources and environmental protection; following this line of research,
some groups have carried out studies to value the residues of plant origin
(Devesa-Rey et al., 2011). The valorization of these wastes can be accom-
plished by extracting fibers, flavoring compounds, and phytochemicals
(Baiano, 2014).
For this reason, the use of pomegranate peel has been studied, due to
its high content of bioactive compounds with biological properties of great
relevance (antioxidant, antimicrobial, antiproliferative, etc.).
10.1 INTRODUCTION
According to the botanist N. I. Vavilov, pomegranate (Punica granatum

L.) is a very ancient plant and fruit first cultivated in Egypt, Italy, and
Greece, and then propagated to many regions as Asia, North Africa, and
especially grown in Mediterranean countries of Europe (Evreinoff, 1957).
Pomegranate (Punica granatum L.) is an important fruit crop adaptable to
a wide range of agro-climatic conditions. It is a fruit-bearing deciduous
shrub or a small tree that belongs to Punicaceae family (Singh et al.,
2018). The term Punica is the feminized Roman name for Carthage, an
ancient city in Tunisia where the best fruits came to Italy. The term Punica
granatum is derived from Latin word pōnum, which means apple, and the
word grānātus which means granulated. At the United States, the common
name means “seedy apple” (Stover and Mercure, 2007). This fruit can be
grown in arid areas due to its tolerance to drought conditions. However,
the main producers of pomegranate are India, Iran, Turkey China, United
States of America, Argentina, Brazil, Chile, Peru, Australia, and Italy
(Kahramanoglu, 2019a). Pomegranate has been considered for cultivating
in arid zones due to the high tolerance to drought (Kandylis and Kokkino-
magoulos, 2020).
Currently, the main producers and exporters of pomegranate in the world
are India, Iran, China, Turkey, the United States, South Africa, Spain, Peru,
Chile, and Argentina. The world production of this fruit is approximately
3.8 million tons (Kahramanoglu, 2019b). The pomegranate fruit is called
by various names depending on the region where it’s cultivated, because the
color of the rind and the aril can vary considerably depending on the growing
region. These differences mainly affect fruit size and peel color (red, yellow,
purple, and pink) (Pareek et al., 2015). Figure 10.1 shows the fruit’s different
parts, which can be divided into some anatomical origins like peel, seeds,
and arils (Viuda-Martos et al., 2010).
Functional Properties of Agroindustrial Wastes from Pomegranate Peel 251
FIGURE 10.1 Pomegranate (Punica granatum).
Pomegranate (Punica granatum L.) has been investigated for being a

dietary source with a high content of various phytochemicals beneficial to
health because it contains anthocyanins, ellagitannins, gallotanins, flavo-
noids, and some other polyphenolic compounds (Bonzanini et al., 2009;
García-Villalba et al., 2015; N. Seeram et al., 2005).
Ripe pomegranates are traditionally consumed fresh, because it’s difficult
to extract the arils from the fruit, for this reason processed fruits such as
jellies, syrups, juices, among others, are preferred (Ferrara et al., 2011).
Food processing has increased the production of agro-industrial residues or
by-products, which are produced throughout the food supply chain such as
agricultural production, industrial manufacturing, processing, and distribu-
tion (Kumar et al., 2017). In recent years, the use of agro-industrial wastes
has become a global concern due to environmental protection; following this,
some research groups have carried out studies with the aim to investigate the
value of the residues of plant origin (Devesa-Rey et al., 2011), which can be
achieved by extracting high-value compounds which can be reused as func-
tional and nutraceutical ingredients, for example, proteins, polysaccharides,
fibers, flavoring compounds, and phytochemicals (Baiano, 2014).
Bioactive compounds found in food waste can be extracted and used to
develop nutraceuticals and functional foods that (Baiano, 2014) can have
beneficial effects on health, such as the prevention of chronic diseases
(Kumar et al., 2017).
The aim of this chapter is to study the chemical composition of the pome-
granate and the use of agro-industrial wastes as a source of purification of
ellagitannins, biological activities, and bioactive compounds as nutraceutical

and functional foods.
10.2 CHEMICAL COMPOSITION OF POMEGRANATE

The interior of the pomegranate is composed of 40% arils and 10% seeds.
Aryls contain 85% water, 10% total sugars such as fructose and glucose,
1.5% pectin, phenolic compounds, and flavonoids, mainly anthocyanins;
organic acids like ascorbic acid, citric acid, and malic acid (Viuda-Martos
et al., 2010) (Fig. 10.2). Also, several studies have reported that the main
phenolic compounds in pomegranate juice are anthocyanins, while the main
phenolic compounds found in the mesocarp and pericarp of the fruit are
hydrolyzable tannins (Bonesi et al., 2019), which are classified as ellagitan-
nins and gallatonins, since their hydrolysis produces ellagic acid and gallic
acid (Arapitsas, 2012).
FIGURE 10.2 Major polyphenols found in pomegranate (Punica granatum). [Gallic acid
(1), ellagic acid (2), caffeic acid (3).]
10.3 PHENOLIC COMPOUNDS IN POMEGRANATE PEEL (PP)

The pomegranate peel (PP) represents approximately 50% of the total
weight of the fruit. It is a source of bioactive compounds such as phenols,
flavonoids, ellagitannins, proanthocyanidin compounds; minerals such as
potassium, nitrogen, calcium, phosphorus, magnesium, and sodium; and

complex polysaccharides (Sreekumar et al., 2014). The entire fruit is rich
in large polyphenolic compounds like punicalagin isomers, ellagic acid
derivatives, and anthocyanins (delphinidin, cyanidin, and pelargonidin
3-gluco-sides and 3,5-diglucosides); however, the peel contains a pool of
phenolics (hydrolyzable tannins) compared their concentration in another
part of the fruit.
PP is rich in hydrolyzable tannins, mainly punicalin, pedunculagin, and
punicalagin (N. Seeram et al., 2005). PP has a high antioxidant value. Several
studies have demonstrated the high concentration of phenolic compounds of
the peel (Akhtar et al., 2015).
The ellagitannins are one of the main groups of hydrolyzable tannins; they
are esters of hexahydroxydiphenic acid (HHDP) bound to a polyol, usually
glucose and quinic acid. When exposed to strong acids or bases, the ester
bonds hydrolyze, and HHDP spontaneously reorganizes into water-insoluble
dilactone, obtaining ellagic acid (2,3,7,8-tetrahydroxchromene [5,4,3-cde]
chromene-5,10-dione, according to IUPAC). Ellagic acid is a thermodynami-
cally stable molecule; it has four rings that perform the lipophilic domain,
four phenolic groups (hydroxyl), and two lactone groups that represent the
hydrophilic part, where they can proceed as donors or acceptors of hydrogen
bonds (Aguilera-Carbo et al., 2008a; Bala et al., 2006).
The ellagitannins are found mostly in the pomegranate’s pericarp, seeds,
and flowers, while the gallotanins are found at the leaves and seldom in other
parts of the pomegranate. The hydrolysis of ellagitannins is used to recognize
and quantify ellagitannins in food (Brighenti et al., 2017). The punicalagin is
the most abundant of the ellagitannins in the pomegranate (Figure 10.3), the
total antioxidant activity of the juice is attributed to this compound present
in the fruit peels (Tzulker et al., 2007).
10.4 AGRO-INDUSTRIAL WASTES AS A SOURCE OF BIOACTIVE

COMPOUNDS
After the extraction of sugar, oil, juice, and starch, there remains some
residues such as peelings, stems, trimmings, seeds, bran, and shells. The
recovered biocompounds and outgrowth can be applied to produce basic
foods (Baiano, 2014).
According to Kumar et al. (2017), the bioactive compounds present in
agro-industrial waste can be recovered using various techniques. According
FIGURE 10.3 Punicalagin chemical structure.
to the literature, the extraction methods for bioactive compounds are in basic
based on:
_ Subcritical water extraction (SWC),
_ Supercritical fluid extraction (SFE),
_ Solvent extraction (SE),
_ Microwaves,
_ Ultrasounds, and
_ Use pf anzymes
These technique’s availability provides an opportunity for the recovery
of specific compounds.
The extraction of bioactive compounds, mainly polyphenols, is an
alternative for recovering agro-industrial waste. However, the extraction
methods currently used have some disadvantages; for example, the use of
toxic solvents generates damage to the environment, and green technologies
are expensive, complex, and difficult to implement. Also, some traditional
techniques use high temperatures that can generate negative effects on the
biological activity of the extracts. Moreover, the techniques currently used
produce an incomplete release of phenolic compounds, due to the impos-
sibility of recovering the bound polyphenols (Bhanja Dey et al., 2016)
through the strong interactions of these compounds with the plant matrix
(Torres-León et al., 2019).
Recently emerging techniques for the extraction of ellagitannin have
been made since it seeks to reduce the use of polluting solvents and obtain
better extraction yields. Emerging technologies include pressurized liquid
extraction, ultrasound-assisted extraction, pressurized hot water extraction,
simultaneous distillation, microwave-assisted dual extraction, and super-
critical fluid extraction (Domínguez-Rodríguez et al., 2017).
Furthermore, solid state fermentation has great potential to obtain bioac-
tive compounds such as phenolic compounds (Martínez-Ávila et al., 2012)
being that microorganisms like fungi naturally produce enzymes that degrade
the cell wall, generating hydrolysis reactions (Jamal et al., 2011) and the
mobilization of compounds unto the extraction solvent (Torres-León et al.,
2019). Additionally, this bioprocess is economical and simple to perform;
it requires small equipment and a reduction in operating costs because it
doesn’t need sterilization, aeration, or agitation processes (Soccol et al.,
2017).
During these bioprocesses, it must be considered the choice of micro-
organisms and the substrates, since filamentous fungi are suitable solid
state fermentation as the technique simulates their natural habitat: in this
condition, they can synthesize considerable amounts of enzymes and some
other metabolites (Farinas, 2015). These extracellular enzymes are produced
and simultaneously utilized for the extraction/release of phenolics from
the matrix of substrates in a single process, along with the production of
new phenolic compounds by the secondary metabolism of microorganisms
(Bhanja Dey et al., 2016).
This bioprocess allows the use of solid agro-industrial waste as a substrate
that is considered a pollutant and produce less wastewater production. The
use of agro-industrial waste reduces production costs, and the extracts
high biological activities guarantee the bioprocess’s economic profitability
(Acosta-Estrada et al., 2014).
Ascacio-Valdés et al. (2013) reported that ellagitanase is directly related
to the degradation and accumulation of ellagitannins, since the ester bonds
between the HHDP group and the glycosides are degraded by this enzyme,
which has high specificity.
Vattem and Shetty (2003) mentioned that the β-glucosidase enzyme is
associated with the degradation of ellagitannins present in blueberry resi-
dues through solid state fermentation with the fungus Lentinus edodes to
produce ellagic acid. Huang et al. (2008) co-cultivated Aspergillus oryzae
and Trichoderma reesei using acorn crown extract with a content higher than
62% ellagitannins as a substrate to produce ellagic acid with high levels of
enzymes such as hydrolase, cellulase, and xylanase. The results indicated
that the mixed culture of T. reesei and A. oryzae is an efficient method to
produce an enzymatic system for the degradation of ellagitannins.
Aguilera-Carbo et al. (2008) reported on the production of ellagic acid
by cultivating fungi in the solid state using polyurethane foam (PUF) as
a support and an aqueous extract obtained from the peel of pomegranate
(Punica granatum) as a source of carbon and energy. In this study, Asper
gillus niger GH1 consumed ellagitannins during the first 36 h of culture with
a maximum concentration of ellagic acid reached at 48 h. The authors attrib-
uted the biodegradation of ellagitannins to a new tannase, which is probably
different from acyl hydrolase.
10.5 PURIFICATION OF ELLAGITANNINS
To purify ellagitannins, several steps must be followed—first, a column

chromatography must be performed using chromatographic fillers such
as Amberlite XAD-16, Sephadex LH-20 and macroporous resin H-103
(Aguilar-Zárate et al., 2017; Seeram et al., 2005). The most reported tech-
niques are high-performance liquid chromatography (HPLC), centrifugal
partition chromatography (CPC), high-speed countercurrent chromatography
(HSCC), and preparative HPLC (Aguilar-Zárate et al., 2017).
It is recommended to perform a HPLC analysis since they are commonly
used to detect and quantify compounds. Some methods have been reported
based on reverse phase C18 column technology, with solvents such as
methanol, ethanol, and acetonitrile, mixed with acidified water (acetic or
phosphoric acid) (Aguilera-Carbo et al., 2008b).
10.6 BIOLOGICAL ACTIVITIES
Fruits play a main part in the maintenance of a well-balanced diet. They

contain macro- and micronutrients, and also bioactive compounds that
promote health. Plenty of studies indicate the importance of fruit consump-
tion for the preventing of health-associated risks and diseases (Kandylis,
2020).
Pomegranate is a rich in polyphenols and have shown many health-related
properties, so it has become in the object of study in medicine. Various studies
have established that pomegranate has antibacterial, anti-inflammatory,

antiviral, and anticancer properties (Kalaycıoğlu and Erim, 2017). These
investigations have concluded that the regular consumption of pomegranate
fruit, juice, or even its added compounds in other food products, acts in a
beneficial way for health and can even protect or improve health against
various diseases such as obesity, diabetes, cardiovascular diseases, and even
some types of cancer (Kandylis et al., 2020).
The fruit’s health-promoting properties are mainly due to punicalagin’s
presence and, slightly, to other metabolites like anthocyanins and lavonols
(Syed et al., 2007).
The punicalagin, HHDP-hexosides, and ellagic acid are derived from the
partial or complete hydrolysis of punicalagin and have generated interest
because of the human health effects (Aguilar-Zárate et al., 2017). Punicalagin
is found in high amounts in PP. The powerful bioactivity of this compound
has been demonstrated in several studies.
10.6.1 ANTIOXIDANT ACTIVITY
Phenolic compounds have an aromatic ring bearing on one or more hydroxyl

groups and their structures may vary from a simple phenolic molecule to
a complex high-molecular-weight polymer. Flavonoids, have C6–C3–C6
structure, and account for more than half of the over 8000 different phenolic
compounds (Balasundram et al., 2006). Pomegranate’s phenolic content is
higher than those reported for cranberry, strawberry, red wine, orange juice,
and green tea (Gil et al., 2000).
The use of synthetic antioxidants has been increasingly restricted due to
the secondary effects they may produce; the determination of the antioxidant
capacity of pomegranate compounds and derivates it’s been studied for use
as natural additives to replace synthetic antioxidants (Viuda-Martos et al.,
2010).
Wang et al. (2013) studied a large-scale separation method of punicalin
from PP using medium pressure liquid chromatography (MLPC) and they
obtained large amounts of punicalin with high purity of 95.9%. Also, they
evaluated the antioxidant and protein precipitating capacities of the purified
punicalin; the results showed that punicalin has strong antioxidant potential
according to the DPPH and hydroxyl radical scavenging assay and exhibited
a low affinity for protein, suggested that would not be masked by tannin
protein precipitation.
10.6.2 ANTIMICROBIAL ACTIVITY/PREBIOTIC EFFECT
One of the oldest techniques to control the growth of microorganisms is

using chemical or synthetic agents as inhibitors, growth reducers, or even
inactivators. Several studies have demonstrated the antibacterial activity of
pomegranate against various pathogens and organisms resistant to antibiotics
(Viuda-Martos et al., 2010).
An important function of pomegranate and its derivatives is it’s effect
exerts on the intestinal microbiota and its antimicrobial activity (Acosta-
Estrada et al., 2014). Ellagitannins are hydrolyzed into ellagic acid in the
intestine before being metabolized by the colon microbiota to form urolithin
A and B (Finegold et al., 2014), which has been associated with the prebiotic
potential of pomegranate and its products (Kandylis, 2020).
Betanzos-Cabrera et al. (2015) evaluated the antimicrobial activity of
fresh pomegranate juice in clinical isolates of multi-resistant Staphylococcus
in epidermal strains. They observed inhibition in the growth kinetics of the
strain at different concentrations of the juice, which can be attributed to the
high content of polyphenols and the antioxidant capacity of the pomegranate.
Šavikin et al. (2018) studied PP as an antimicrobial agent and a reinforcing
agent was utilized in developing starch-based films like hydroxypropyl high-
amylose starch plasticized by water and glycerol. The results demonstrated
that PP inhibited gram-positive (S. aureus) and gram-negative (Salmonella)
bacteria growth. Meanwhile, PP also increased Young’s modulus, tensile
strength, and stiffness of the starch-based films.
Prebiotics can improve selected or randomized growth of colon micro-
biota. Pomegranate extracts inhibit pathogens and initiate the growth of
beneficial microbiota in human guts. Probiotic lactobacilli were relatively
unaffected by pomegranate chemical constituents: ellagic acid slightly
reduced the growth of lactobacilli (Lactobacillus pentosus, Lactobacillus
rhamnosus, and Lactobacillus acidophilus) to approximately 10–20% of
controls (Bialonska et al., 2009).
10.6.3 ANTI-INFLAMMATORY ACTIVITY
Various investigations have been reported in the literature on the anti-inflam-

matory activity of pomegranate extracts through in vitro and in vivo studies.
Macrophages function as immune system control switches maintaining the
balance between pro and anti-inflammatory activities; they can become
different subsets with positive inflammatory phenotypes, classical polariza-

tion (M1), and activation of macrophages in powerful pro-inflammatory cells
characterized by the generation of species reactive oxygen and nitrogen, and
the alternative activation profile (M2) characterized by the secretion of anti-
inflammatory cytokines (Bouhlel et al., 2007).
Hollebeeck et al. (2012) evaluated the extract’s anti-inflammatory
properties of the PP, rich in Punicalagin, using an in vitro model of the
epithelium of the human intestine. They concluded that the extracts showed
anti-inflammatory properties in intestinal cells, acting on the transcription
of the pro-inflammatory gene and protein levels, indicating that the latter
phenomenon is possibly due to a direct molecular capture of the ellagitan-
nins from the shell, helping to prevent inflammation in chronic intestinal.
Ellagic acid has been shown to be used to treat pain and inflammation
through central and peripheral sites. Mansouri et al. (2015) examined the
interaction between ellagic acid and venlafaxine (VLF) in intraperitoneal
acetic acid-induced pain in mice using an isobolographic analysis, which
indicated a synergistic antinociceptive and anti-inflammatory interaction
when these drugs were combined.
10.7 BIOACTIVE COMPOUNDS AS NUTRACEUTICALS AND

FUNCTIONAL FOODS
Bioactive compounds are known to lower the risk of developing diseases

like cancer, Alzheimer’s, cataracts, and Parkinson.
The idea of basic foods is complex and can refer to various possible
aspects, including foods obtained through any process, whose main char-
acteristic is that one or more of its components, even if that component is a
nutrient, affect the function of the organism of in a particular way, by helping
the physiological or psychological effect far away the nutritional. The
pomegranate fruit conforms to the definition of functional food, although
establishing any function implies identifying the bioactive components to
specify their possible beneficial effects on health (Viuda-Martos et al., 2010).
ACKNOWLEDGMENTS
Autor Izábal-Carvajal A.L. thanks the Mexican Council for Science and
Technology (CONACYT) for the financial support for the development of
the project (PhD) in Food Science and Technology offered by the Autono-
mous University of Coahuila.
KEYWORDS
• agroindustrial waste
• pomegranate
• bioactive compound
• environmental protection
• valorization
• phytochemicals
REFERENCES
Acosta-Estrada, B. A.; Gutiérrez-Uribe, J. A.; Serna-Saldívar, S. O. Bound Phenolics In Foods,
A Review. Food Chem. 2014, 152, 46–55. https://doi.org/10.1016/j.foodchem.2013.11.093
Aguilar-Zárate, P.; Wong-Paz, J. E.; Michel, M.; Buenrostro-Figueroa, J.; Díaz, H. R.; Ascacio,
J. A.; Contreras-Esquivel, J. C.; Gutiérrez-Sánchez, G.; Aguilar, C. N. Characterisation
of Pomegranate-Husk Polyphenols and Semi-Preparative Fractionation of Punicalagin.
Phytochem. Anal. 2017, 28 (5), 433–438. https://doi.org/10.1002/pca.2691
Aguilera-Carbo, A.; Augur, C.; Prado-Barragan, L. A.; Favela-Torres, E.; Aguilar, C. N.
Microbial Production of Ellagic Acid and Biodegradation of Ellagitannins. Appl. Microbiol.
Biotechnol. 2008a, 78 (2), 189–199. https://doi.org/10.1007/s00253-007-1276-2
Aguilera-Carbo, A. F.; Augur, C.; Prado-Barragan, L. A.; Aguilar, C. N.; Favela-Torres, E.
Extraction and Analysis of Ellagic Acid from Novel Complex Sources. Chem. Papers
2008b, 62 (4), 440–444. https://doi.org/10.2478/s11696-008-0042-y
Akhtar, S.; Ismail, T.; Fraternale, D.; Sestili, P. Pomegranate Peel and Peel Extracts:
Chemistry and Food Features. Food Chem. 2005, 174, 417–425. https://doi.org/10.1016/j.
foodchem.2014.11.035
Arapitsas, P. Hydrolyzable Tannin Analysis in Food. Food Chem. 2012, 135 (3), 1708–1717.
https://doi.org/10.1016/j.foodchem.2012.05.096
Ascacio-Valdés, J.; Burboa, E.; Aguilera-Carbo, A. F.; Aparicio, M.; Pérez-Schmidt,
R.; Rodríguez, R.; Aguilar, C. N. Antifungal Ellagitannin Isolated from Euphorbia
Antisyphilitica Zucc. Asian Pac. J. Trop. Biomed. 2013, 3 (1), 41–46. https://doi.
org/10.1016/S2221-1691(13)60021-0
Baiano, A. Recovery of Biomolecules from Food Wastes—A Review. Molecules 2014, 19 (9),
14821–14842. https://doi.org/10.3390/molecules190914821
Bala, I.; Bhardwaj, V.; Hariharan, S.; Kumar, M. N. V. R. Analytical Methods for Assay of
Ellagic Acid and Its Solubility Studies. Jo. Pharm. Biomed. Anal. 2006, 40 (1), 206–210.
https://doi.org/10.1016/j.jpba.2005.07.006
Balasundram, N.; Sundram, K.; Samman, S. Phenolic Compounds in Plants and Agri-
Industrial By-Products: Antioxidant Activity, Occurrence, and Potential Uses. Food Chem.
2006, 99 (1), 191–203. https://doi.org/10.1016/j.foodchem.2005.07.042
Betanzos-Cabrera, G.; Montes-Rubio, P. Y.; Fabela-Illescas, H. E.; Belefant-Miller, H.;
Cancino-Diaz, J. C. Antibacterial Activity of Fresh Pomegranate Juice Against Clinical
Strains of Staphylococcus epidermidis. Food Nut. Res. 2015, 59. https://doi.org/10.3402/
fnr.v59.27620
Bhanja Dey, T.; Chakraborty, S.; Jain, K. K.; Sharma, A.; Kuhad, R. C. Antioxidant
Phenolics and Their Microbial Production by Submerged and Solid-state Fermentation
Process: A Review. Trends Food Sci. Technol. 2016, 53, 60–74. https://doi.org/10.1016/j.
tifs.2016.04.007
Bialonska, D.; Kasimsetty, S. G.; Schrader, K. K.; Ferreira, D. The Effect of Pomegranate
(Punica Granatum l.) Byproducts and Ellagitannins on the Growth of Human Gut Bacteria.
J. Agric. Food Chem. 2009, 57 (18), 8344–8349. https://doi.org/10.1021/jf901931b
Bonesi, M.; Tundis, R.; Sicari, V.; Loizzo, M. R. The Juice of Pomegranate (Punica granatum
L.): Recent Studies on Its Bioactivities. In Quality Control in the Beverage Industry: Volume
17: The Science of Beverages; Elsevier, 2019; pp 459–489. . https://doi.org/10.1016/
B978-0-12-816681-9.00013-8
Bonzanini, F.; Bruni, R.; Palla, G.; Serlataite, N.; Caligiani, A. Identification and Distribution
of Lignans in Punica granatum L. Fruit Endocarp, Pulp, Seeds, Wood Knots and Commercial
Juices by GC-MS. Food Chem. 2009, 117 (4), 745–749. https://doi.org/10.1016/j.
Bouhlel, M. A.; Derudas, B.; Rigamonti, E.; Dièvart, R.; Brozek, J.; Haulon, S.; Zawadzki, C.;
Jude, B.; Torpier, G.; Marx, N.; Staels, B.;Chinetti-Gbaguidi, G. PPARγ Activation Primes
Human Monocytes into Alternative M2 Macrophages with Anti-inflammatory Properties.
Cell Metabolism. 2007, 6 (2), 137–143. https://doi.org/10.1016/j.cmet.2007.06.010
Brighenti, V.; Groothuis, S. F.; Prencipe, F. P.; Amir, R.; Benvenuti, S.; Pellati, F. Metabolite
Fingerprinting of Punica granatum L. (Pomegranate) Polyphenols by Means of High-
performance Liquid Chromatography with Diode Array and Electrospray Ionization-mass
Spectrometry Detection. J. Chromatogr. A 2017, 1480, 20–31. https://doi.org/10.1016/j.
chroma.2016.12.017
Devesa-Rey, R.; Vecino, X.; Varela-Alende, J. L.; Barral, M. T.; Cruz, J. M.; Moldes, A. B.
Valorization of Winery Waste vs. the Costs of Not Recycling. Waste Manag. 2011, 31 (11),
2327–2335. https://doi.org/10.1016/j.wasman.2011.06.001
Domínguez-Rodríguez, G.; Marina, M. L.; Plaza, M. Strategies for the Extraction and
Analysis of Non-extractable Polyphenols from Plants. J. Chromatograp. A 2017, 1514,
1–15. https://doi.org/10.1016/j.chroma.2017.07.066
Evreinoff, V. A. Contribution à l’étude du Grenadier. J. Agric. Trop. Bot. Appl. 1957, 4 (3),
124–138. https://doi.org/10.3406/jatba.1957.2380
Farinas, C. S. Developments in Solid-state Fermentation for the Production of Biomass-
degrading Enzymes for the Bioenergy Sector. Renew. Sustain. Ener. Rev. 2015, 52, 179–188.
https://doi.org/10.1016/j.rser.2015.07.092
Ferrara, G.; Cavoski, I.; Pacifico, A.; Tedone, L.; Mondelli, D. Morpho-pomological and
Chemical Characterization of Pomegranate (Punica granatum L.) Genotypes in Apulia
Region, Southeastern Italy. Sci. Hortic. 2011, 130 (3), 599–606. https://doi.org/10.1016/j.
scienta.2011.08.016
Finegold, S. M.; Summanen, P. H.; Corbett, K.; Downes, J.; Henning, S. M.; Li, Z. Pomegranate
Extract Exhibits Invitro Activity Against Clostridium Difficile. Nutrition 2014, 30 (10),
1210–1212. https://doi.org/10.1016/j.nut.2014.02.029
García-Villalba, R.; Espín, J. C.; Aaby, K.; Alasalvar, C.; Heinonen, M.; Jacobs, G.; Voorspoels,
S.; Koivumäki, T.; Kroon, P. A.; Pelvan, E.; Saha, S.; Tomás-Barberán, F. A. Validated
Method for the Characterization and Quantification of Extractable and Nonextractable
Ellagitannins after Acid Hydrolysis in Pomegranate Fruits, Juices, and Extracts. J. Agric.
Food Chemi. 2015, 63 (29), 6555–6566. https://doi.org/10.1021/acs.jafc.5b02062
Gil, M. I.; Tomas-Barberan, F. A.; Hess-Pierce, B.; Holcroft, D. M.; Kader, A. A. Antioxidant
Activity of Pomegranate Juice and Its Relationship with Phenolic Composition and
Processing. J. Agric. Food Chemi. 2000, 48 (10), 4581–4589. https://doi.org/10.1021/
jf000404a
Hollebeeck, S.; Winand, J.; Hérent, M. F.; During, A.; Leclercq, J.; Larondelle, Y.; Schneider,
Y. J. Anti-inflammatory Effects of Pomegranate (Punica granatum L.) Husk Ellagitannins
in Caco-2 Cells, an In Vitro Model of Human Intestine. Food Func. 2012, 3 (8), 875–885.
https://doi.org/10.1039/c2fo10258g
Huang, W.; Niu, H.; Li, Z.; He, Y.; Gong, W.; Gong, G. Optimization of Ellagic Acid
Production from Ellagitannins by Co-culture and Correlation Between Its Yield and
activities of Relevant Enzymes. Biores. Technol. 2008, 99 (4), 769–775. https://doi.
org/10.1016/j.biortech.2007.01.032
Jamal, P.; Idris, Z. M.; Alam, M. Z. Effects of Physicochemical Parameters on the Production
of Phenolic Acids from Palm Oil Mill Effluent Under Liquid-state Fermentation by
Aspergillus Niger IBS-103ZA. Food Chemi. 2011, 124 (4), 1595–1602. https://doi.
org/10.1016/j.foodchem.2010.08.022
Kahramanoglu, I. International Journal of Agriculture, Forestry and Life Sciences Trends in
Pomegranate Sector: Production, Postharvest Handling and Marketing; 2019a.
Kahramanoglu, I. Trends in Pomegranate Sector: Production, Postharvest Handling and
Marketing; 2019b.
Kalaycıoğlu, Z.; Erim, F. B. Total Phenolic Contents, Antioxidant Activities, and Bioactive
Ingredients of Juices from Pomegranate Cultivars Worldwide. Food Chem. 2017, 221,
496–507). https://doi.org/10.1016/j.foodchem.2016.10.084
Kandylis, P.; Kokkinomagoulos, E. Food Applications and Potential Health Benefits
of Pomegranate and Its Derivatives. Foods 2020, 9 (2), 122. https://doi.org/10.3390/
foods9020122
Kumar, K.; Yadav, A. N.; Kumar, V.; Vyas, P.; Dhaliwal, H. S. Food Waste: A Potential
Bioresource for Extraction of Nutraceuticals and Bioactive Compounds. Bioresour.
Bioprocess. 2017, 4 (1). https://doi.org/10.1186/s40643-017-0148-6
Mansouri, M. T.; Naghizadeh, B.; Ghorbanzadeh, B. Ellagic Acid Enhances the Antinociceptive
Action of Venlafaxine in Mouse Acetic Acid-induced Pain: An Isobolographic Analysis.
Pharmacol. Rep. 2015, 67 (3), 473–477. https://doi.org/10.1016/j.pharep.2014.11.004
Martínez-Ávila, G. C.; Aguilera-Carbó, A. F.; Rodríguez-Herrera, R.; Aguilar, C. N. Fungal
Enhancement of the Antioxidant Properties of Grape Waste. Ann. Microbiol. 2012, 62 (3),
923–930. https://doi.org/10.1007/s13213-011-0329-z
Pareek, S.; Valero, D.; Serrano, M. Postharvest Biology and Technology of Pomegranate. J.
Sci. Food Agric. 2015, 95 (12), 2360–2379. https://doi.org/10.1002/jsfa.7069
Šavikin, K.; Živković, J.; Alimpić, A.; Zdunić, G.; Janković, T.; Duletić-Laušević, S.;
Menković, N. Activity Guided Fractionation of Pomegranate Extract and Its Antioxidant,
Antidiabetic and Antineurodegenerative Properties. Ind. Crops Prod. 2018, 113, 142–149.
https://doi.org/10.1016/j.indcrop.2018.01.031
Seeram, N.; Lee, R.; Hardy, M.; Heber, D. Rapid Large-scale Purification of Ellagitannins
from Pomegranate Husk, A By-product of the Commercial Juice Industry. Sep. Purif.
Technol. 2005, 41 (1), 49–55. https://doi.org/10.1016/j.seppur.2004.04.003
Seeram, N. P.; Adams, L. S.; Henning, S. M.; Niu, Y.; Zhang, Y.; Nair, M. G.; Heber, D.
In Vitro Antiproliferative, Apoptotic and Antioxidant Activities of Punicalagin, Ellagic
Acid and a Total Pomegranate Tannin Extract are Enhanced in Combination with Other
Polyphenols as Found in Pomegranate Juice. J. Nutr. Biochem. 2005, 16 (6), 360–367.
https://doi.org/10.1016/j.jnutbio.2005.01.006
Singh, B.; Singh, J. P.; Kaur, A.; Singh, N. Phenolic Compounds as Beneficial Phytochemicals
in Pomegranate (Punica granatum L.) Peel: A Review. Food Chemi. 2018, 261, 75–86.
https://doi.org/10.1016/j.foodchem.2018.04.039
Soccol, C. R.; da Costa, E. S. F.; Letti, L. A. J.; Karp, S. G.; Woiciechowski, A. L.; de
Vandenberghe, L. P. S. Recent Developments and Innovations in Solid State Fermentation.
Biotechnol. Res. Innova. 2017, 1 (1), 52–71. https://doi.org/10.1016/j.biori.2017.01.002
Sreekumar, S.; Sithul, H.; Muraleedharan, P.; Azeez, J. M.; Sreeharshan, S. Pomegranate Fruit
as a Rich Source of Biologically Active Compounds. BioMed Res. Int. 2014, 2014, 686921.
https://doi.org/10.1155/2014/686921
Stover, E.; Mercure, E. W. The Pomegranate: A New Look at the Fruit of Paradise; n.d.
Syed, D. N.; Afaq, F.; Mukhtar, H. Pomegranate Derived Products for Cancer
Chemoprevention. Semin. Cancer Biol. 2007, 17 (5), 377–385. https://doi.org/10.1016/j.
semcancer.2007.05.004
Torres-León, C.; Ramírez-Guzmán, N.; Ascacio-Valdés, J.; Serna-Cock, L.; dos Santos
Correia, M. T.; Contreras-Esquivel, J. C.; Aguilar, C. N. Solid-state Fermentation with
Aspergillus Niger to Enhance the Phenolic Contents and Antioxidative Activity of Mexican
Mango Seed: A Promising Source of Natural Antioxidants. LWT 2019, 112. https://doi.
org/10.1016/j.lwt.2019.06.003
Tzulker, R.; Glazer, I.; Bar-Ilan, I.; Holland, D.; Aviram, M.; Amir, R. Antioxidant Activity,
Polyphenol Content, and Related Compounds in Different Fruit Juices and Homogenates
Prepared from 29 Different Pomegranate Accessions. J. Agric. Food Chemi. 2007, 55 (23),
9559–9570. https://doi.org/10.1021/jf071413n
Vattem, D. A.; Shetty, K. Ellagic Acid Production and Phenolic Antioxidant Activity in
Cranberry Pomace (Vaccinium macrocarpon) Mediated by Lentinus Edodes Using a
Solid-state System. Proc. Biochem. 2003, 39 (3), 367–379. https://doi.org/10.1016/
S0032-9592(03)00089-X
Verotta, L.; Panzella, L.; Antenucci, S.; Calvenzani, V.; Tomay, F.; Petroni, K.; Caneva, E.;
Napolitano, A. Fermented Pomegranate Wastes as Sustainable Source of Ellagic Acid:
Antioxidant Properties, Anti-inflammatory Action, and Controlled Release under Simulated
Digestion Conditions. Food Chemi. 2018, 246, 129–136. https://doi.org/10.1016/j.
Viuda-Martos, M.; Fernández-Lóaez, J.; Pérez-álvarez, J. A. Pomegranate and its Many
Functional Components as Related to Human Health: A Review. Compr. Rev. Food Sci.
Food Saf. 2010, 9 (6), 635–654. https://doi.org/10.1111/j.1541-4337.2010.00131.x
Wang, Y.; Zhang, H.; Liang, H.; Yuan, Q. Purification, Antioxidant Activity and Protein-
precipitating Capacity of Punicalin from Pomegranate Husk. Food Chemi. 2013, 138 (1),
437–443. https://doi.org/10.1016/j.foodchem.2012.10.092
CHAPTER 11
Foodborne Zoonoses: Current Status and

Control Strategies
M. SUMAN KUMAR1, HIMANI DHANZE1, S. S. REDDY VAKAMALLA1,
A. R. CHANDNI1, and S. WILFED RUBAN1,2
Division of Veterinary Public Health, ICAR-IVRI, Izatnagar,
1
Uttar Pradesh, India

2
Veterinary College, KVA&FSU, Hebbal, Bengaluru, Karnataka, India
ABSTRACT
The aim of this chapter is to review and report the latest control strategies
that is available on food borne diseases to public health considering the
socioeconomic development.
11.1 INTRODUCTION
The World Food Summit held at Rome in 1996 reaffirmed that gaining secure
and healthy food is the fundamental right of every person, in accordance with
the integrity to sufficient food and fair to be free from starvation. Foodborne
zoonotic diseases pose a continuous hazard to public well-being with impli-
cations on socioeconomic development and international trade worldwide.
Zoonoses refer to those illnesses and epidemic that is normally passed on
among vertebrate animals and human. More than 200 familiar illnesses are
transferred from food by pathogenic microorganisms such as bacteria and
their toxins, viruses, and parasites. The World Health Organization’s (WHO)
action to measure the world hardship of foodborne illness considered illness
caused by 31 foodborne hazards and reported over 600 million illnesses and
4,20,000 deaths in the year 2010 alone (WHO, 2015). The disease burden
was highest in Africa (43%), followed by South-East Asia (24%) and Eastern
Mediterranean (16%) sub regions. Around 40% of the foodborne diseases
(FBD) were reported from children less than 5 years. The highest rates of
under-5 mortality due to diarrhea were in sub-Saharan Africa and South Asia
with India considered for many loss of life in 2015 (Troeger et al., 2017).
Based on the CDC, 48 million people are affected by food-borne illness each
year, 128,000 are hospitalized, and around 3000 die as a result (CDC, 2021).
Norovirus is the most common cause of FBD, followed by Campylobacter,
Salmonella, and Listeria monocytogenes. The observed trend was mostly
comparable to that of the EU, the United States of America (USA), and
Australia. However, pathogenic E. coli caused the most cases in Korea, and L.
monocytogenes was not as lethal. The number of Campylobacter, norovirus,
and C. perfringens-related infections in Japan was quite high (Lee and Yoon,
2021). FBD cost India about $28 billion (Rs. 1,78,100 crore) or around 0.5%
of the country’s gross domestic product (GDP) every year (Kristkova et al.,
2017). It is estimated that the current FBD burden in India represents about
100 million cases per year. This corresponds to one in 12 people falling ill,
which might be well underestimated as not all cases are reported or recorded.
The risk of foodborne illness has been rising during the last two decades
due to several factors including pathogen behavior, rapid growth of popula-
tion, and an increased open global market trade in foods and farm animals
from countries where appropriate microbiological security strategy are not
followed or implemented. The improved transport logistics and conditions
facilitates some microorganisms to survive on food products for longer
periods and reach the consumer in a viable form. The change in eating habits
of consumer, such as the consumption of lightly cooked food and a shift
from low-to-high protein-based diets (e.g., meat and fish products) has also
increased the probability of occurrence of foodborne illnesses. The increased
movement of human population also facilitates easy spreading of pathogens
worldwide. These factors have impacted the severity of FBD and their
impacts on health of communities. Therefore, preventing illness and deaths
due to FBD remains a major public health challenge across the globe. Lack
of accurate epidemiological data on incidence and cost of FBD, particularly
in the developing countries, has delayed the setting up of suitable policies
and allocation of resources to mitigate the ills of FBD. Further, alarming
rise in antimicrobial resistance among bacterial pathogens particularly in the
food chain has also impacted the public health to a greater extent. Effec-
tive control involves a thorough understanding of the essential aspects of
Foodborne Zoonoses: Current Status and Control Strategies 267
epidemic dynamics, such as the spatiotemporal distribution and common

origins, as well as strong identification of outbreaks from baseline sporadic
individuals (Deng et al., 2016; Ronholm et al., 2016). Studies that gather
information regarding FBDs are vitally necessary to protect health. Greater
understanding of biological agents, foods, and the sources of contamina-
tion that are frequently implicated in FBD outbreaks allows agencies to
design effective preventative measures, thereby minimizing the incidence
of outbreaks.
11.2 MAJOR BACTERIAL FOODBORNE PATHOGENS
Human diarrheal disease are mostly caused by bacterial foodborne infec-

tions (64%) followed by viral (24%) and protozoan (12%) infections.
During the last two decades, Salmonella spp., Campylobacter spp., and
E. coli were reported to be the major foodborne bacterial agents. Recent
WHO estimates revealed that Campylobacter spp. (27%), enterotoxigenic E.
coli (25%), non-typhoidal Salmonella enterica (23%), Shigella spp. (15%),
and enteropathogenic E. coli (7%) are major diarrheal disease pathogens.
Campylobacter spp. has been estimated to cause 96 million cases of illness
in year 2010 alone. Globally, about 4.6 billion cases of infectious diarrhea
and 1.6 million deaths are reported annually due to contaminated food and
water (Pires et al., 2015).
11.2.1 CAMPYLOBACTERIOSES
Campylobacter is the leading cause of bacterial gastroenteritis world-

wide—one of four key global causes of FBD with an annual incidence of
4.4–9.3/1000 cases leading to 30,931 deaths (WHO, 2012). A report of Food
Borne Epidemiology Reference Group of World Health Organization (WHO,
2015) estimates global median of 95,613,970 cases of campylobacteriosis.
In Southeast Asian Region D (SEAR-D) (in which India is included), the
median 16,379,442 cases have been estimated. Global median deaths esti-
mated due to campylobacteriosis are 21,374 and in SEAR-D deaths are 5410.
Campylobacter foodborne illness is a concern and a monetary burden for
the human population, accounting for 8.4% of all diarrhea cases worldwide
(Connerton and Connerton, 2017). Campylobacteriosis is increasing at a rate
more than twice that of salmonellosis (Mace et al., 2015). Most campylo-
bacteriosis outbreaks have been endemic, with asymptomatic symptoms,
and C. jejuni, preceded by C. coli, has been found to be responsible for

nearly 80% of outbreaks in most EU countries and the USA (Kaakoush et
al., 2015). The public health impact of Campylobacter and its prevalence in
India is firmly established. However, the figures for actual prevalence and
economic losses incurred in India are sketchy. Sporadic studies conducted in
different parts of the country have revealed the presence of Campylobacter
organisms in foods of plant origin (Kumar et al., 2001), various species of
animals, birds, and foods of animal origin including milk and from cases
of human diarrhea (Kumar et al., 2015). Campylobacteriosis occurs in
people of all ages (HSE, 2017), with the highest incidence of notification
in children aged 0–5, and the largest percentage of patients hospitalized
and dying in adults aged 65 and older (Facciola et al., 2017). Troeger et al.
(2017) estimated nearly 10,211 mean number of deaths in children younger
than 5 years due to campylobacterioses in India alone. The complications
of infection include meningitis, urinary tract infections, reactive arthritis,
and Gullian Barré syndrome, a nerve disorder that causes muscle weakness
and paralysis of the limbs. Major sources of infection include undercooked
poultry meat, pets (especially young puppies), water, and raw milk. Several
studies in India have estimated the occurrence of C. jejuni and C. coli in
the range of 4.42–28% in chicken meat samples, 1.3–4.4% in cattle milk,
3.6–25% in meat samples, 13.52–22.64% in water samples, 2.35–25% in
human diarrheal cases, and 0.56–8% in healthy human fecal samples (OPZD
Annual Reports 2014–2019). Infections due to chicken eating account for
50–70% of all Campylobacter infections worldwide (Seliwiorsto et al.,
2015). Macrolides and fluoroquinolones are the drug of choice for treatment
of human diarrheal cases. However, increasing instances of antimicrobial
resistance have been reported of late against multiple drugs. Different
studies in India have shown an increasing trend of multidrug resistance of
2.2% (Prasad et al., 1994), 30.6% (Jain et al., 2005), and 51.33% (Kumar,
2016) in Campylobacter spp. isolates from diverse origin. Campylobacter
spp. also has the ability to form biofilms and survives in the environment. A
nation-wide surveillance of campylobacteriosis along with typing of strains
isolated from both human and animal cases is required. Preventive measures
such as rearing of campylobacter-free poultry flocks and the good hygienic
practices throughout the entire production chain can limit the transmission
of the pathogen. Withholding feed from chickens for around 12 h before
slaughter is another important preventive action that will help decrease the
amount of these bacteria (Hansson et al., 2018). Campylobacter infections
can be prevented in humans in a variety of ways, including improved sewage
sanitation, the availability of clean drinking water, public awareness of the

importance of pasteurization of milk, proper cooking of animal-sourced
foods, vaccination, and the use of therapeutics in cases of infection (Hansson
et al., 2018).
11.2.2 SALMONELLOSIS
Salmonella is the second leading causative agent of foodborne illness in

most developed countries and most common reason for foodborne zoonosis
in other developing countries. Salmonella enterica subsp. enterica causes
99% of the human and animal infections. It is estimated that approximately
78.7 million cases of foodborne illness cases occur due to non-typhoidal
Salmonella globally, leading to 59,000 deaths (WHO, 2015). Salmonella is
one of the most frequent foodborne infections in the world, responsible for
approximately 93.8 million foodborne illnesses and 155,000 fatalities per
year (Eng et al., 2015). In 2018, the European Union (EU) member states
reported 5146 outbreaks of FBD, giving rise in the infections of 48,365
persons. Salmonella was solely responsible for 33% of these outbreaks.
Salmonella spp. has wide range of livestock species (poultry, cattle, and
pigs) as host and the infected animals act as a reservoir of zoonotic salmo-
nellae. Salmonellosis is mostly a foodborne illness; however, reptiles are
responsible for 7% salmonellosis occurrences in humans (CFSPH, 2013).
Contaminated eggs, poultry meat, pork, and other meats and products are
common vehicles of salmonellosis to humans. The Salmonella epidemics
that resulted in 1581 cases in Poland, Slovakia, and Spain were directly
connected to infected eggs (Munck et al., 2020). Historically, plants have not
been regarded as hosts for human infections for Salmonella; nevertheless, in
the last several decades, the habitats for these species have shifted (Lenzi et
al., 2021). Devleesschauwer et al. (2017) observed that despite the fact that
salmonellosis outbreaks caused by fruits and vegetables have been widely
documented, their frequency remains intermittent. In India, foods of animal
origin are considered as the main source with contributing factors such as
inadequate cooking, lack of refrigeration, slow cooling of food, and inad-
equate reheating before serving. Various studies in India have documented
prevalence of Salmonella spp. in foods of animal origin including meat
(4–12.4%), carabeef (1.25%), pork (10%), water (3.4–14.89%), chicken
(14.89%), and in poultry litter (1.73–28.33%) (OPZD Annual Reports 2014–
2019). Serotypes Typhi and Paratyphi A of typhoidal Salmonella are more
frequent in Southeast Asia, while non-typhoidal Salmonella (NTS) is more

prominent in Africa (WHO, 2016). The non-typhoidal Salmonella serovars,
especially Typhimurium, Enteritidis, Heidelberg, and Newport have been
reported in many outbreaks of human salmonellosis around the globe which
have been linked with consumption of Salmonella-contaminated foods of
animal origins. Furthermore, the emergence of several clones of Salmonella
resistant to multiple antimicrobials worldwide underscores a significant food
safety hazard (Jajere, 2019). A detailed surveillance followed by risk assess-
ment studies is imperative to control salmonellosis.
11.2.3 E. COLI INFECTION
E. coli has wide range of host and so far at least seven different groups
of E. coli strains have been isolated from gastrointestinal diseases, out of
which foodborne infections are mostly associated with enterotoxigenic E.
coli (ETEC) and enteropathogenic E. coli (EPEC) strains although Shiga-
toxigenic E. coli (STEC) causes more severe disease leading to mortality.
Among the STEC strains, E. coli O157:H7 is widely recognized as an impor-
tant cause of foodborne illness. The clinical outcomes of STEC infections
would be mild-to-severe diarrhea and hemolytic uremic syndrome. Cattle
are the principal reservoirs of STEC (Bibbal et al., 2015) and beef has been
implicated in several global outbreaks. ETEC strains have also been isolated
from carabeef, dung, feces of diarrhea patients, and river water in India.
The presence of antimicrobial-resistant E. coli other than STECs in foods
of animal origin is also an emerging public health concern. Of particular,
public health concern among antimicrobial-resistant E. coli are strains
that produce extended-spectrum β-lactamases (ESBLs). ESBL-producing
E. coli have been documented in feces of healthy poultry and retail meat,
from several parts of the world including India (Kar et al., 2015). A pooled
prevalence of 15% was estimated for E.coli in the food products of livestock
origin in Ethiopia. The high prevalence may be due to unhygienic slaugh-
tering of food animals and consumption of raw meat in the area concerned
(Assefa and Bihon, 2018). Recently, Odo et al. (2021) reviewed different
prevalence studies of E. coli O157:H7 in Nigeria. Chicken and beef samples
from slaughterhouses of southwest Nigeria when tested showed an overall
prevalence of 17.1% for multidrug resistant E coli O157:H7. This bacterial
strain could also be detected in fermented milk products from Nasarawa state
of Nigeria. Spatial epidemiology study of E. coli O157:H7 in Netherlands
revealed the increased incidence being associated with exposure to small

ruminants rather than cattle (Mulder et al., 2020). The total number of
confirmed cases of foodborne ETEC infection in the EU were above 6000
in 2016 and 2017 and increased to 8167 in 2018 while the number reduced
to below 5000 during 2020 (EFSA-ECDC, 2020). The statistical analysis
to identify the major sources contributing to different foodborne pathogens
showed that STEC was largely transmitted through direct contact with
infected animals. Nevertheless, among the foodborne infection pathways,
beef/lamb accounted for 65% of the infections (Mughini-Gras et al., 2022).
In India, the occurrence of E. coli have been documented in cattle milk
(1–11%), water (2.6–33.3%), meat and meat products (24.10%), poultry
eggs (10%), carabeef (9%), and chicken meat and giblets (1.33%). In a study
conducted on street-vended foods, a higher presence of E. coli was observed
in raw food samples (60.62%), followed by environmental samples (46.73%)
and ready-to-eat foods (RTE) (42.99%), of which 31 (26.49%), 3 (6.97%),
and 7 (15.21%) samples, respectively, harbored ESBL producers. ESBL E.
coli were detected in finished food products like salad, chutney, and paneer
(Sivakumar, 2016).
Escherichia albertii, a close relative of E. coli, is considered as an
emerging zoonotic pathogen which is often misunderstood as other members
of Enterobacteriaceae. The organism has been isolated from meat and meat
products of chicken, pig, and duck and from the environmental sources.
Diarrhea outbreaks due to infection with E. albertii have been reported
in different areas across the globe including countries like China, Brazil,
Germany, Bangladesh, Alaska, USA, Poland, Japan, Iran, Mexico, and
Nigeria (Muchaamba et al., 2022). The presence of the organism could be
detected in a recent study in retail oysters with a prevalence of 1.9% which
may be accounted to the contamination of the water bodies with the bacteria
(Arai et al., 2022).
11.2.4 LISTERIOSIS
Listeriosis is a serious, but preventable and treatable zoonotic disease.

Listeria monocytogenes is one of the most virulent foodborne pathogens, with
20–30% case fatality. According to WHO (2015) estimates, annually 14,169
cases of foodborne illness and 3175 deaths occur globally due to listeriosis.
L. monocytogenes has been discovered to be a common pathogen in several
nations, including the United Kingdom, Mexico, Canada, Australia, and USA
(Yang et al., 2020). Although it is considered uncommon in comparison to

other foodborne infections, listeriosis has a fatality rate of 20–30% in its most
severe form (Li et al., 2020). The survival capability of L. monocytogenes in
wide range of pH, salt concentration, and other processing and refrigeration
conditions poses potential threat to public health. Processed, refrigerated,
and RTE of animal origin are the major sources often implicated in Listeria
outbreaks. Cooking or other antimicrobial treatments are not performed
between food production and consumer consumption in these circumstances,
which increases their risk (Jordan and McAuliffe, 2018; Gray et al., 2021).
Listeriosis is directly responsible for the deaths of 260 individuals per year
in the USA because of consuming contaminated food (Centers for Disease
Control and Prevention, 2015). A widespread outbreak of listeriosis resulted
to 937 cases and 216 fatalities in South Africa in 2018 due to the eating
of tainted RTE-processed beef (Thomas et al., 2020). Pregnant, elderly, or
immuno-compromised persons are at a greater risk of infection. L. monocy
togenes has also been reported from cases of spontaneous abortion in humans
(Kaur et al., 2007). Occurrence rates of 3.5–15.7% in water and 5.21–20.5%
in meat have been recorded for L. monocytogenes in different studies (OPZD
Annual Reports 2014–2019). Many studies have reported occurrence of L.
monocytogenes in animal origin foods such as milk and its products (1–23%),
chevon (0–17%) poultry meat (8–15%), and pork (1.3–2.8%) (Barbuddhe
et al. 2012; Shakuntala et al. 2019). Existence of multidrug-resistance and
biofilm-forming capacity of L. monocytogenes isolates is also a cause for
public health concern. The clinical outcomes of listeriosis include perinatal
and non-perinatal-related symptoms of septicaemia, meningitis, neurological
consequences, stillborn, and death. Listeria spp. isolates from food have
been found to be resistant to antibiotics currently used in the treatment of
human listeriosis, such as penicillin, ampicillin, tetracycline, and gentamicin
(Olaimat et al., 2018). Further research is needed on alternative therapies
to treat listeriosis and better understanding about bacterial persistence and
survival over a wide range of temperature outside mammalian hosts.
11.3 OTHER IMPORTANT BACTERIAL PATHOGENS CAUSING FBDS
11.3.1 VIBRIO SPP.

Among more than 100 species of Vibrio, Vibrio cholera (V. cholera), Vibrio
parahemolyticus (VP), and Vibrio Vulnificus (V. Vulnificus) are involved in
foodborne infection. VP is the most common kind of bacterial gastroenteritis
that is linked to seafood consumption all over the world (Baker-Austin, 2018).
The reservoirs include normal flora of brackish water, aquatic flora (macro-
phytes, microphytes), and fauna (zooplanktons, crustaceans), humans, and
animals. Raw oysters, crustaceans, crabs, and oysters or undercooked mussels
are common sources of the disease in man. V. cholera is found in oysters
for weeks under refrigeration and for more than 2 weeks in dairy products,
including soft deserts and cakes. Complications of infection include severe
dehydration, cardiac complications, and circulatory failure, which occur due
to loss of potassium, septicemia, wound infection, ear infection, cellulitis,
peritonitis, necrotizing fascitis, cholecystitis, endophthalmitis, and menin-
gitis. Various studies have documented occurrence of V. cholera from 3.45%
to 57.38% and that of V. vulnificus as 2.3–38.5% in marine fish, clams, shell
fish, water, brackish water fish, and aquatic environment. In a joint report
published by WHO and FAO, it was documented that there were between
500 and 800 outbreaks of VP gastroenteritis recorded in Japan each year,
affecting more than 10,000 persons. VP is one of the most common causes
of foodborne gastroenteritis in Japan (WHO and FAO, 2011). It is believed
that this bacterium is responsible for 50% of the infections that are caused by
consuming contaminated food in Asian nations (Chen et al., 2018). Occur-
rence of VP was found to be between 6.9% and 56.09% in crab, shrimps,
fish, and shell fish in coastal regions of India. Although the data regarding
burden of cholera in India is well documented (Ali et al., 2017), the food-
borne burden due to other Vibrio spp. is sketchy and focused studies are
required to estimate the true burden.
11.3.2 AEROMONAS
Aeromonas species, which are sometimes referred to as “jack of all trades,”

have drawn growing interest as human pathogens in recent years. Aeromonas
spp. are ubiquitous aquatic bacteria, recognized as increasingly important
human pathogen due to the frequent prevalence in all types of foods, particu-
larly in seafood. The reservoirs include wide range of fish, environment, and
water. This organism has been isolated from meat, raw milk, poultry, fish,
shellfish, and vegetables. Additionally, salt or sodium chloride (NaCl) is a
usual preservative for meat products and raw fish; nevertheless, Aeromonas
may thrive in NaCl concentration as high as 4%. pH is another factor that
regulates bacterial growth in food. Aeromonas has been shown to be able
to live at pH-5 (Figueras et al., 2014; John et al., 2019). In humans, the
symptoms range from acute gastroenteritis to life-threatening complications

such as septicemia and, in extremely rare instances, meningitis (Dallagassa
et al., 2018). Seafood products are more commonly contaminated because
of the widespread occurrence of these bacteria in the marine environment.
Aeromonas spp. can enter the food chain through cross-contamination during
processing from biofilms on food-contacted surfaces. Batra et al. (2016) esti-
mated 76 cases per million population in Asia. Occurrence rate of Aeromonas
spp. in various samples were 22.2–50% in fresh water fish, 18.34% in prawns,
46.67% in frozen fish, 10–22.5% in chicken, and 11–16.52% in different
water sources (Sharma et al., 2009; Smita and Brahmabhatt, 2011; Das et al.,
2012; Didugu et al., 2016).
11.3.3 BRUCELLOSIS
Brucellosis is a re-emerging foodborne bacterial zoonosis which is consid-

ered as a public health concern and is an important economic burden with
losses occurring in the human, livestock, and wildlife populations. Each year
around 5 lakhs human cases of brucellosis are reported globally. Countries
like Canada, Denmark, Germany, Austria, Australia, France, New Zealand,
Sweden, and USA are considered to be free from bovine brucellosis. Disin-
fectants like 70% ethanol, formaldehyde, hypochlorite, and iodophores
can easily destroy the organisms while pasteurization of milk remains the
important method to kill the organisms in milk. Consumption of unpasteur-
ized milk is the major route of acquiring the infection by humans, but direct
contact with infected animals, consumption of infected raw or undercooked
meat, and in rare cases blood transfusion may also result in infection.
A seroprevalence of 9% was recorded in hunted wild boars in Finland in
2016 (Fredriksson-Ahomaa et al., 2020), which hints to the probability
of acquiring brucellosis from wild animals. Disease outbreaks in humans
have been documented by different authors in countries including Mexico,
Turkey, Saudi Arabia, and South Korea of which most of them were due to
consumption of raw or unpasteurized milk and milk products (Pal, 2018).
Epidemiological study of human cases in Dalian and Liaoning of China
between 2014 and 2019 could report 571 total cases with no fatality (Yucun
et al., 2021). In India, brucellosis caused a median loss of USD 3.4 billion
to the livestock sector of which 96% was to the dairy sector alone (Singh et
al., 2015). In India, the annual median losses due to human brucellosis were
estimated to be INR 627.5 million (Singh et al., 2018). The main source
of infection in Indian population is dairy products. Cheese prepared from

unpasteurized milk from endemic areas is a particular risk for tourists. The
milk of infected sheep, goats, or cattle may contain large numbers of viable
organisms, which become concentrated in products such as yogurt, paneer,
and cheese (Ongor et al., 2006). In dairy animals, the organism localizes in
the supra-mammary lymph nodes and mammary glands of 80% of infected
animals, and these continue to excrete the pathogen in milk throughout their
lives acting as carriers but intermittently (OIE, 2016). Various studies have
reported occurrence rate of 4.35–55.6% in cattle milk samples in India. The
prevalence rate was found to be higher in unorganized farms. Recent studies
have indicated resistance toward commonly used drugs such as rifampicin
and co-trimoxazole, which is matter of concern and calls for judicious use
of antibiotics (Doimari et al., 2018). Although numerous studies have been
conducted on sero-prevalence of brucellosis in various livestock, very few
have focused on the occurrence of Brucella spp. in foods. Studies on esti-
mating risk of brucellosis through foods need to be carried out.
11.3.4 STAPHYLOCOCCUS AUREUS
Staphylococcus aureus (S. aureus) is the predominant agent associated with

food poisoning and is present in the soil, water, air, and on everyday objects
and surfaces, in humans and animals. Human hands and nasal tracts are
common areas of S. aureus inhabitation. S. aureus has also been identified
from a variety of animal-based foods and was described as the third-most
prevalent cause of food-borne disease worldwide (Hachemi et al., 2019).
S. aureus food poisoning occurs due to preformed heat stable enterotoxins
produced in foods. Among the 21 Staph enterotoxin (SE) or enterotoxin-like
proteins (SEls) identified, SEA is the most common toxin responsible for
food poisoning. Consuming food infected with toxins generated by S. aureus
can result in staphylococcal food poisoning, which can induce severe gastro-
enteritis, nausea, vomiting, diarrhea, and abdominal discomfort within 1–6
h (crago et al., 2012). Foods frequently associated include meat and meat
products; poultry and egg products; and salads, such as egg, tuna, chicken,
bakery products, and milk and dairy products. It has been reported that the
prevalence of S. aureus in milk and milk products produced in developing
nations is higher than in industrialized nations (EFSA, 2009), for instance, in
India wherein milk products such as sweets are a common source for infec-
tion. Methicillin-resistant S. aureus (MRSA), when associated with human
or animal infections, poses severe public health concern (Silva et al., 2013).
The transmission of MRSA occurred in clinical settings, but its prevalence
in the population, outside of hospitals, has been observed in recent decades
(Oniciuc et al., 2015). Recent research by Caggiano et al. (2016) evaluated
healthy persons who worked in the food sector and found that the presence
of S. aureus and MRSA posed a threat to public health. LA-MRSA strains
were detected in pork, poultry, and raw turkey products in the USA (Smith,
2015). Caggiano et al. (2016) conclude that the spread of S. aureus and
MRSA in non-hospital settings, such as communities and animals, require
ongoing and vigilant surveillance. Costa et al. (2015) performed S. aureus
isolation and MRSA identification in which 9.5% of the samples contained
MRSA. The high frequency of MRSA in meat, particularly in food prepared
for consumption, highlights the need for the best food handling techniques
in hospitals as well as the best animal handling methods.
Several reports on the presence of S. aureus in different foods such as
meat, poultry, milk, and their products with prevalence in different products
ranging from 10% to 100%. The description of several IDSP report outbreaks
indicates toward staph poisoning.
11.3.5 BACILLUS CEREUS
It is believed that Bacillus cereus (B. cereus) is responsible for between 1.4%
and 12% of foodborne outbreaks (FBOs) globally (Grutsch et al., 2018). In
general, the incidence of outbreaks attributed to bacterial toxins increased,
and in 2018, B. cereus was the fifth most common cause of FBOs (EFSA,
2019). After S. aureus , B. cereus is the second-most commonly detected
causal agent in FBOs in France (Glasset et al., 2016). It causes acute gastro-
enteritis due to B. cereus intoxication and is commonly transmitted through
consumption of rice, spices, and dairy products. A wide variety of foods such
as meats, milk, vegetables, and fish are implicated. In the Chinese market,
dairy products such as baby formula have produced a significant number of
B. cereus-induced FBOs (Liu et al., 2020). Moreover, diarrheal-type illness
has been recorded more often in Northern Europe, which includes Finland
and Norway, and emetic-type disease has been observed more frequently in
Japan and the United Kingdom, posing a significant issue in both industrial-
ized and emerging nations (Bennett et al., 2013; Jessberger et al., 2020). In
India, prevalence in meat and poultry products varies from 10% to 70%.
Sporadic reports are available from the country. Although several IDSP
outbreak descriptions from rice/biryani/noodles have been documented, no

further reports on aetiology are available. Even though B. cereus contributes
just a small percentage to the total number of FBOs, it is essential to highlight
that these figures are underestimations. This is because the majority of B.
cereus-caused food poisoning outbreaks are often minor and do not require
medical treatment. Consequently, the majority of instances are not reported
to health authorities (Jovanovic et al., 2021)
11.3.6 CLOSTRIDIUM PERFRINGENS
Foodborne illnesses caused by Clostridium perfringens (C. perfringens) are

always linked with meat and poultry products. During slaughtering, animal
products can get infected with this disease by contact with a contaminated
surface or faeces (Leung et al., 2017; EFSA, 2017). After Salmonella, C.
perfringens-type A food poisoning is the second-most widespread bacterial
food poisoning in the USA, with an estimated 1 million cases each year
(Scallan et al., 2011). The number of C. perfringens-related food poisoning
occurrences in EU member states was estimated to be roughly 5 million
per year (Felicio et al., 2015). C. perfringens is responsible for 5% of
FBD outbreaks, 10% of illnesses, and 4% of hospitalizations, according to
CDC epidemiology monitoring statistics (Dewey-Mattia et al., 2018). Heat
tolerance is connected with the production of spores that may germinate at
temperatures between 15 and 55 °C. Toxins of types A and C have been
proven to cause human illness. Type A causes the majority of food poisoning
and diarrheal diseases linked with C. perfringens. Important pathogen of
both humans and animals are present in soil, water, dust, and the intestinal
tracts of man and animals. Among the five types, type A is associated with
food poisoning and harbors enterotoxin genes. Meat and poultry products
are common sources. Inadequate heating, survival of spores, and post-
production temperature abuse especially in large-scale preparation of foods,
leads to human outbreaks. Prevalence in meats in India has been estimated to
be 40–85% in carabeef, 35–75% in chevon, 30–75% in poultry, and 20–85%
in RTE meat and poultry products.
11.3.7 CLOSTRIDIUM BOTULINUM
Foodborne botulism is a potentially lethal condition caused by the ingestion

of neurotoxins generated by Clostridium botulinum. Foodborne botulism
is due to preformed toxin botulinum neurotoxins (BoNTs), in food which

are secreted by Clostridium organisms known as BoNT-releasing clostridia.
Food sources include improperly canned foods, meat products, and seafoods.
Foodborne botulism usually develops 18–36 h (range 6–8 days) following
exposure (Peck et al., 2017). C. botulinum produces neurotoxins A, B, C,
D, E, F, and G among which toxin A is the most potent toxin known with
a toxic dose of 0.001 μg/kg BW. Inadequately heated and packed foods are
common sources. Honey is most commonly implicated in infant botulism.
During 2004−2020, a total of 80 foodborne botulism outbreaks occurred in
China, involving 386 illnesses and 55 deaths; most outbreaks were reported
between June and August, with a sharp peak in January (Li et al., 2022). In
Italy, botulism is a public health issue with 355 confirmed cases of botulism
involving 515 people from 1986 to 2019 (Benevenia et al., 2022). Between
2011 and 2015, an average of 162 yearly cases of botulism were recorded in
the USA. Foodborne botulism accounts for 1%–20% of all botulism cases as
documented by Jeffery and Karim (2017).
Lalitha and Surendran (2002) reported occurrence in foods such as cured
fishes and salt dried shrimps in retail market of Cochin with an overall preva-
lence of 19%.
11.3.8 MYCOBACTERIUM TUBERCULOSIS AND M. BOVIS (BOVINE

TUBERCULOSIS)
According to WHO, about 30% of total global tuberculosis (TB) cases are
found in India. M. bovis (bovine TB) plays a crucial role in India, where 70%
of the human population resides in rural areas. Consumption of unpasteurized
raw milk is the major risk factor for transmission. Srinivasan et al. (2018)
estimated a prevalence of 7.3% indicating that there may be approx. 21.8
million infected cattle in India affected with bovine TB. In another study
to estimate the occurrence of bovine TB in three high-risk populations, M.
bovis was detected in 11.4% of samples from farmers, dairy workers, and
livestock keepers; 8.9% zookeepers and animal handlers; and in 12.6% of
residents of TB endemic area (Bapat et al., 2017). In all these cases, previous
contact with active TB cases and raw milk consumption were found to
be important determinants of bovine TB. Though zoonotic TB organisms
are widely present in cattle herds of different regions across the globe, a
surveillance based on one health perspective is lacking in 89.9% of the WHO
signatory countries (De Macedo Couto et al., 2022). In 2020, EU recorded
88 cases of TB in humans of which 86 were caused by M. bovis while the
remaining two cases reported in Germany was due to M. caprae (EFSA-

ECDC, 2020). In the USA, 1.4% of the total TB cases in humans are due
to M. bovis infection. Some studies have proven that mortality is more for
M. bovis-infected patients than those infected with M. tuberculosis. Global
incidence of zoonotic TB estimated by WHO in 2010 came around 1.2 lakhs
and in countries like Africa, about 70,000 cases are being reported annually
(Olea-Popelka et al., 2017). Many studies have documented presence of M.
tuberculosis in milk at the occurrence rate of 4.8–52.17% depending on the
endemicity of region (OPZD Annual Reports 2014–2019). There is an urgent
need to study the contribution of M. bovis toward human TB burden in India.
11.3.9 PARATUBERCULOSIS
M. avium subspp. Paratuberculosis (MAP) is responsible for causing para-
tuberculosis and is a major pathogen of the livestock in India and globally.
The animals under sub-clinical to clinical stage continue to shed MAP bacilli
in their milk and feces. Among the 26,000 domestic livestock screened, the
bio-load of MAP in was found in 43% cattle, 36% buffaloes, 23% goats, and
41% sheep. Further, mass screening of 28,291 human samples from 2008
to 2016 also revealed a high bio-load of 33.7% (Chaubey et al., 2017). It is
believed that 90% of the herds in USA are possessing at least few animals
which are infected with MAP. A study conducted in disease-suspected
animals from selected abattoirs in Mexico revealed a seroprevalence of
2.08% (Osuna-Chávez et al., 2021). There are reports that MAP infection
in humans can trigger the development of many chronic and autoimmune
diseases including type 1 diabetes mellitus. Some studies could discuss that
the children who were seropositive for MAP antibodies were more prone to
develop the disease than the healthy controls who were otherwise considered
to be at risk (Ozana et al., 2022). The risk of MAP transmission exists even
in pasteurized milk and its products owing to the ability of the organism to
forms clumps in milk, which can lead to its survival during pasteurization.
When the risk assessment for acquiring infection from different sources of
milk and milk products were done using statistical models, it was found that
the risk was very high for the people who consume raw milk from infected
farms while pasteurization can decrease the risk substantially but not
completely (Knific et al., 2022). Live bacilli have also been recovered from
meat products and the environment thus illustrating the potential of MAP as
a pathogen of public health concern. The organism is known to survive for
3 months in soft cheese and 10 months in hard cheese. MAP causes chronic
inflammatory bowel disease in humans and is similar to Jhone’s disease in

ruminants. Due importance must be given toward the transmission of MAP
from milk and its products with research to ensure zero-survival during
pasteurization.
11.3.10 YERSINIOSIS
Yersinia enterocolitica is a widespread zoonotic pathogen found in soil,
water, animals, and numerous foods (Ahmed et al., 2019; Bari et al., 2013).
In some countries (including New Zealand), the intestinal condition yersini-
osis caused by Yersinia enterocolitica or Y. pseudotuberculosis is among
the most frequently reported foodborne infections, posing a significant
public health crisis (Pattis et al., 2019). Because of the effort required in pig
farming and the close interaction between dogs and humans, pigs and dogs
are a major source of Y. enterocolitica infections in humans (Duan et al.,
2014). Acute gastroenteritis is the most common word for Yersiniosis, and
the severity of the illness is mostly influenced by the age of the person who
becomes infected. (El Qouqa et al., 2011). Y. enterocolitica is an increasingly
diverse foodborne zoonotic pathogen that may cause significant morbidity
and death, particularly in newborns and young children (Pal, 2018). It is
also an important FBD, mostly spread by direct contact between humans
and animals or through the environment (Bancerz-Kisiel and Szweda, 2015).
Six biotypes of Y. enterocolitica exist: 1A, 1B, 2, 3, 4, and 5. Biotype 1B
was previously considered extremely pathogenic, whereas biotype 1A was
deemed to be non-pathogenic in humans; other biotypes are considered to be
of low pathogenicity in humans (Valentin-Weigand et al., 2015). As recently
calculated, the prevalence of Y. enterocolitica in instances of human gastro-
enteritis is 1.97% (95% confidence interval 1.32–2.74%), with serotype O:3
being the most prevalent (Riahi et al., 2021). Reports released by EFSA and
the ECDC Zoonoses Report for 2020 show the number of human cases of
this disease in Europe has increased to 5668, with very little data available
on the meat-producing supply chain (EFSA, 2021). Furthermore, just 0.2%
of the 2351 pigs examined in six European nations tested positive for Y.
enterocolitica. Y. enterocolitica infection is more prevalent in nations with
temperate climates than in tropical or subtropical locations. Europe had the
most severe epidemic of enteric yersiniosis in 2019, and it was the third-most
frequent zoonotic illness in Europe (EFSA, 2017). This foodborne bacteria is
thought to be the cause of around 87,000 cases per year throughout the world
(Pal et al., 2013).
11.3.11 FOODBORNE ZOONOTIC VIRUSES
Virology research is rapidly expanding leading to discovery of new viruses

at a comparatively higher pace. Out of the 22 families of viruses which are
found to infect humans, at least 10 are documented to cause foodborne trans-
mission. In the last decade, we have witnessed outbreaks due to consumption
of food contaminated with emerging viruses like Nipah virus, avian influenza
virus, and SARS coronavirus. Though those outbreaks were mainly associ-
ated with a particular food habit, they have raised the query if consump-
tion of food contaminated during preparation, handling, slaughtering, etc.,
could be one of the routes by which emerging viruses can infect the human
population. Foodborne viruses are mainly transmitted via the fecal-oral
route through ingestion of contaminated food and water, and/or by person-
to-person contact. Though burden of foodborne viruses is largely unknown,
according to WHO (2015) estimates, 15% of foodborne related deaths due to
diarrheal agents were because of Norovirus and 23.6% of foodborne-related
deaths due to invasive disease agents were due to Hepatitis A virus. FAO
and WHO have considered the noroviruses, hepatitis A virus, and the group
A rotaviruses as the priority viral foodborne hazards. The genomic studies
of these viruses have shown that they are chiefly transmitted through the
food handlers. Besides these, other viruses like enteroviruses, sapoviruses,
rotaviruses, astroviruses, and adenoviruses have also been implicated in
food- and/or water-borne illness. Several challenges including development
of reliable diagnostics to detect viruses in food and generation of reliable
surveillance data needs to be addressed for devising adequate preventive and
control measures.
11.3.12 NOROVIRUS
Worldwide, Norovirus is the leading causative agent for acute gastroenteritis,

particularly in countries where rotavirus vaccination is implemented. Noro-
virus infection can cause explosive vomiting, abdominal pain, and watery
diarrhea. The virus belongs to the family Caliciviridae and is reported to
cause infection mainly in humans, while its presence is detected in cattle,
sheep, dog, pig, and mice, too. Incubation period for the disease is usually
12–48 h and it usually lasts for 1–3 days (Velebit et al., 2019). Severe long-
drawn-out diarrhea can be observed in elderly and immuno-compromised
individuals, where they shed the virus for extended time period indicating
active viremia phase but slow viral clearance. The virus concentration in
the feces and vomitus (as high as 1010 viruses per gram) was found to be
independently associated with the protracted diarrhea (Lee et al., 2007).
Norovirus causes 124.8 million cases of foodborne illness and 34,929 deaths
(WHO, 2015). There are seven genogroups of norovirus, of which five are
responsible for human gastroenteritis. Infective dose of this virus is as low as
10 organisms. The low infective dose, the ability to survive in harsh environ-
ment, several transmission routes, and short-lived immunity against the virus
has hiked its impact in global population.
A meta-analysis report on a global scale regarding selected gastroenteritis
cases showed that 16% of the total cases were due to norovirus infection.
The prevalence of gastroenteritis produced by norovirus was found to be
maximum in South America with 22%, followed by Europe and Asia with
16% each while Africa showed a prevalence of 15% which was the least
(Liao et al., 2021). Globally, GII.4 and GII.3 were found to be the most
commonly occurring genotypes with highest frequency being reported in
children below 1 year of age (Farahmand et al., 2022). Asymptomatic infec-
tions with a prevalence of 7% was estimated by a meta-analysis and this
should be overlooked as those individuals can obviously contribute to the
disease transmission (Qi et al., 2018). In Southern India, out of 1856 diar-
rheal episodes, 207 (11.2%) were associated with norovirus (Menon et al.,
2016). NoV positivity varied between 6.3% and 12.6% in different cities of
Western India with the predominance of GII (96.6%) (Chhabra et al., 2009).
First instance of inter-genogroup recombination has been reported from
Kolkata, having polymerase sequence of GI and genotype 3 (GI.3), whereas
the capsid sequence belongs to GII and genotype 4 (GII.4) (Nayak et al.,
2008). To date, no animal norovirus have been detected in human stool,
but some serological evidence hints to possible transmission from animals
to humans (Villabruna et al., 2019). Some viruses that are categorized as
human norovirus today might have originated from animal source. Targeted
surveillance including samples of both humans and animals that are in close
contact is required to record a trans-species transmission event.
11.3.13 HEPATITIS A VIRUS AND HEPATITIS E VIRUS
These viruses are mainly transmitted via the fecal-oral route. Zoonotic infec-
tions by hepatitis E virus (HEV) can occur via contact with infected animals
or by consumption of contaminated foods of animal origin like meat, milk,
and eggs. HEV was identified initially from Indian state of Jammu and
Kashmir in the year 1978. The virus belongs to the family Hepeviridae. It
infects many animals including wild and domestic pigs, camel, chicken,
mongoose, rat, deer, and rabbits. Most acute hepatitis E infections in humans
have been linked to consumption of contaminated pork or pig liver. Extra-
hepatic diseases like neurological, hematological, cardiovascular, and renal
syndromes can also be caused by the virus (Cossaboom et al., 2016; Khuroo
et al., 2016). A systematic review of the global epidemiology of HEV
estimates that nearly 939 million individuals have got infected ever which
corresponds to 1 in 8 of the population and 15–110 million people are expe-
riencing ongoing infection (Li et al., 2020). Globally, the seroprevalence rate
was found to be 60% and 27% in domestic pigs and wild boars respectively
which had ever encountered the disease whereas the HEV RNA positivity
which indicates the active infection was 13% and 9.5% in domestic and wild
swine, respectively. The HEV RNA positivity in commercial pork products
was estimated to be 10% which puts forward the potential of its foodborne
zoonotic transmission (Li et al., 2022). In developed countries, HEV infec-
tion is considered as a leading cause of viral hepatitis which are zoonotic.
Out of the eight genotypes identified, genotypes 1, 2, 3 and 4 are frequently
infecting humans. The transmission of disease in developing nations of
Asia, Africa, and Latin America occurs mainly through contaminated water
wherein genotypes 1 and 2 are the most common in occurrence. Genotypes
3 and 4 are frequent when the transmission is via fecal contamination or
through consumption of contaminated or undercooked meat. While HEV1
and HEV2 shows endemic pattern of disease, HEV3 and HEV4 shows
sporadic occurrence. HEV1 and HEV2 are found to cause disease in humans
only while the other two produces disease both in humans as well as animals.
A single case of zoonotic infection with genotype HEV7 was reported in
UAE in a person who consumed meat and milk from an infected camel. Due
to lack of FDA-approved testing methods, the exact disease occurrence in
USA is undetermined (Damiris et al., 2022). Hepatitis A virus is typically
transmitted through undercooked or raw seafood or other contaminated raw
produce. WHO estimated 14 million HAV infection worldwide in the year
2010, but the occurrence of the disease is variable in different areas. India
and Sub-Saharan Africa are grouped as highly endemic areas while Australia,
USA, and Europe have low endemicity (Bachofen, 2018). Both these viruses
are highly endemic in India and according to one report, sero-prevalence for
HAV and HEV was found to be 92.68% and 17.05%, respectively, in individ-
uals of India (Kotwal et al., 2014). Further, they reported that hand washing
without soap, regular close contact with domestic animals, consumption of

unpasteurized milk, and regular consumption of food outside home were
risk factors for HAV (p < 0.05). While, for HEV, irregular hand washing,
consumption of unpasteurized milk, and irregular consumption of freshly
prepared food were risk factors (p < 0.05). Status of HEV in pig popula-
tion of India is largely unknown as only few studies have been conducted
till now. Although transmission through contact with infected animals have
been proven, status of butchers and farmers still needs to be assessed in
Indian context.
11.3.14 ROTAVIRUS
Rotavirus belongs to the family Reoviridae and it is the leading cause of

severe diarrhea in neonates and infants in developed and developing coun-
tries including India. Humans get exposed to the virus by direct contact with
the infected animals, through contaminated food and water and at times
through blood transfusion (Wilhelm et al., 2015). The virus is highly vari-
able in its genetics and thus classified into eight serogroups. Though reports
of zoonotic transmission exists, human-to-human transmission is more
frequent. A study in Southern Italy during 2015 recorded 16.87% prevalence
of rotavirus in mussels (Fusco et al., 2017). A study for the detection of
zoonotic viruses in pig fecal samples revealed 2.7% prevalence of rotavirus
on assessing the presence of viral RNA (Lyoo et al., 2020). In year 2013, an
estimated 47,100 rotavirus deaths occurred in India, 22% of all deaths due
to rotavirus that occurred globally that year (Tate et al., 2016). In year 2016,
Government of India launched vaccination program against rotavirus in
phased manner, but the data of diarrheal cases after vaccination against rota-
virus has not been analyzed yet. Though burden of rotavirus illness has been
extensively mapped across the world, very little attention has been paid to
identify the other potential modes of transmission of virus. The surveillance
of circulating rotaviruses in the human population has revealed the presence
of uncommon strains that are commonly found in domestic animals. Studies
on unusual rotaviruses from livestock and companion animals are limited
and the genomes of only a handful of these strains have been analyzed so far
through which rare interspecies transmission and reassortment events have
been proven. Surveillance for other groups of rotavirus besides Rotavirus A
is also required.
11.3.15 FOODBORNE PARASITES

Apart from bacteria and viruses, the parasitic organisms especially the
protozoa, nematodes, cestodes, and trematodes contribute widely to the
foodborne infections. Cryptosporidium, Giardia, Cyclospora, and Toxo
plasma are the major foodborne parasitic protozoa worldwide. Cryptospo-
ridium and Giardia are widespread in aquatic environment, and easily spread
via contaminated drinking water. 8.6 million cases of cryptosporidiosis and
3759 deaths were reported worldwide in 2010. It is the second major cause of
mortality in children younger than 5 years due to diarrheal diseases. Anthro-
ponotic and zoonotic transmission as well as through contaminated food and
water are common routes of acquiring infection. Waterborne outbreaks of
cryptosporidiosis had been reported in 1993 in USA wherein about four lakh
individuals encountered the disease (Ryan et al., 2018). Recent researches
has found that the disease can induce some long-term effects like lack of
cognitive skills in malnourished children and economic losses in terms of
livestock production. The parasite rapidly multiplies in the gut cells of the
host damaging its intestinal epithelium. The host can shed a huge number
of oocysts in feces leading to environmental contamination and spread.
Cryptosporidium parvum is zoonotic while most of the other species are not.
The parasite causes disease in many species of animals including cattle, pig,
sheep, goat, equines, and many wild species. Cryptosporidium is the major
cause of neonatal diarrhea in calves and at times in lambs and kids (Innes et
al., 2020). Seasonal pattern of disease occurrence was reported as a hike in
late spring and summer in some parts of Europe, and during spring in United
Kingdom which may be associated with the lambing season and subsequent
shedding of oocysts by young animals. The seasonality of disease occurrence
has also been discussed by various authors in countries such as Ireland, New
Zealand, Canada, and Netherlands. Epidemiological investigation of crypto-
sporidiosis in France during 2017–2019 revealed high occurrence of IIa and
IId subtypes of Cryptosporidium parvum suggestive of frequent zoonotic
transmission (Costa et al., 2020). Global prevalence of cryptosporidiosis was
estimated to be 7.6% with maximum frequency in Mexico (69.6%), followed
by Nigeria (34%) and Bangladesh (42.5%), according to a meta-analysis
study by Dong et al. (2020). It has been reported that Cryptosporidium is
responsible for 7% of children deaths due to diarrhea in India (Troeger et
al., 2017). In general, parasitic protozoa do not multiply in foods, but they
can survive in food stored under moist, cool, or damp conditions for months.
T. gondii is the potential foodborne pathogen which can be transmitted by
consumption of raw milk or undercooked meat where the infective tissue cyst
lodges. This parasite is responsible for causing abortion in pregnant women

and may also cause chorioretinitis and encephalitis. T. gondii is incriminated
as a fatal foodborne protozoan zoonotic disease in USA. Moreover, T. gondii
is reported to be chronically infecting one-third of the global human popula-
tion. A meta-analysis estimated a global prevalence of 33.8% latent infection
in pregnant women wherein South America reported the highest prevalence
of 56.2% (Almeria and Dubey, 2021). European Food Safety Authority
reported that 60% of all toxoplasma infections are meat borne wherein pork,
beef, and mutton are the major sources to contribute (Koutsoumanis, 2018).
The prevalence of T. gondii in poultry and livestock during the period from
2000 to 2019, when estimated using random-effect model, was estimated to
be 28.3%. The prevalence was highest in 2014 and lowest in 2013 in Asia
(Hajimohammadi et al., 2022).
Taenia solium cysticercosis and trichinellosis are assumed to be 100%
foodborne, but indirectly. Taeniasis/cysticercosis mainly affects the people
in developing countries of Africa, Asia, and Latin America. It is second
major cause of non-diarrheal foodborne deaths (WHO, 2015). Taeniasis and
neurocysticercosis (NCC) are common in areas where animal husbandry
practices are such that pigs and cattle come into contact with human feces.
The metacestode stage of the larva is infective while ingestion of eggs also
can cause infection with T. solium and T. saginata in humans. Cysticercosis is
placed under the WHO–FAO list of neglected zoonotic diseases. The disease
in endemic in developing nations of Asia, Latin America, and Africa while the
long-absent cases are re-emerging in Western Europe as a result of immigra-
tion and trade. The infection can result in mild or non-specific gastrointestinal
symptoms to fatal neurological manifestations (Symeonidou et al., 2018).
The disease is prevalent in all states of India, although the prevalence varies
between the states (NHP, 2019). The National Institute of Mental Health and
Neuro Sciences (NIMHANS), Bangalore, reported NCC in 2% of unselected
series of epilepsy patients. In a study from New Delhi, NCC accounted
for 2.5% of all intracranial space occupying lesions. An unusual feature
of the disease was seen that more than 95% of Indian patients with NCC
are vegetarians. The transmission of T. solium eggs through contaminated
water and salads might be one of the reasons for this finding. Consumption
of infected pork and wild animal meat results in trichinellosis. Trichinellosis
in animals is a notifiable disease and EC regulation 1375/2015 proposed
mandatory active surveillance during meat inspection (Van der Giessen et al.,
2021). Southwestern area of China is endemic to human Trichinellosis where
eight outbreaks of the disease were reported between the period of 2009 and
2020 with 0.4% case fatality rate (Zhang et al., 2022). Seropositivity of anti-
trichinella IgG antibodies in North Eastern Iran was calculated to be 2.6%.
Consumption of game meat by certain communities there could have exposed
the people to trichinellosis (Koohsar et al., 2021). Number of reported cases
of human Trichinellosis was 117 in the EU in 2020. The infection rate was
nil in the fattening pigs or breeding pigs while 0.0001% positive cases were
observed in pigs which were not kept under controlled housing conditions
which confirms that altering farming pattern can be a significant factor to
control the disease (EFSA-ECDC, 2020). The exact burden due to parasitic
contaminants in food is still largely unknown in India.
11.4 GLOBAL STATUS OF FBDs
World Health Organization estimates that 1 in 10 people are affected with

foodborne infections each year resulting in a death count of 420,000. The
highest burden of the diseases are found in South-East Asian and African
regions. The diarrheal diseases which are the most common form are
particularly caused by consumption of undercooked meat and dairy products,
contaminated by Norovirus, Campylobacter, Salmonella, and E. coli. While
pathogenic E. coli and Campylobacter are more frequent in low and high
income countries, non-typhoidal Salmonella is a concern over all regions
of the globe. Foodborne entero-pathogenic microorganisms are found to
produce 1.5 billion cases of diarrhea per year particularly in children below
3 years of age resulting in more than 3 million deaths (EFSA-ECDC, 2016).
The African region of WHO is with the highest rate of FBD with 91
million people being infected per year with death reports of 137,000 of which
32,000 people die of non-typhoidal Salmonella. Taenia solium is responsible
for 10% of the total foodborne infections in this region. FBD outbreak reports
in South Africa during the period between 2013 and 2017 was reviewed
and estimated to be 327 in number where above 11,000 reported cases with
0.4% case fatality were present. The etiology for most of the outbreaks were
foodborne bacterial pathogens (Shonhiwa et al., 2019).
The diseases like toxoplasmosis and taeniasis which are spread through
raw or undercooked meat are important foodborne zoonosis as far as Central
and South America are concerned. In these countries, 77 million people fall
ill each year due to foodborne infections.
FBD burden in Eastern Mediterranean region is estimated to be 100
million cases per year of which 32 million are children under 5 years of
age. Along with the diarrheal diseases which accounts to 70% of the total,
typhoid, brucellosis, and hepatitis A are reasons for death of about 37,000
people each year.
Lowest burden of FBDs are estimated in WHO European region with 23
million people getting diseased each year of which 15 million cases accounts
to Norovirus infection, 5 million Campylobacteriosis cases, and 1 million
foodborne Toxoplasmosis. Around 400 deaths are recorded annually in
European region due to Listeriosis caused by consumption of contaminated
meat and milk products, RTE meals as well as raw vegetables. According
to EFSA-ECDC annual report, the most reported cases of FBD in EU is
Campylobacteriosis followed by Salmonellosis in 2020 and the trend was
stable from 2016 to 2020. Yersiniosis, ETEC, and Listeriosis are among the
other infections which were less reported comparatively.
South-East Asian region is next to African region as far as foodborne
infections burden is considered. Above 150 million cases are reported each
year annually and deaths toll to 175,000. In the year 2020, China witnessed
7073 outbreaks of foodborne illnesses wherein 143 reported deaths occurred
among 37,454 affected individuals. About 41.7% accounted to the foodborne
microbial pathogens while poisonous mushrooms caused 58% outbreaks (Li
et al., 2021).
In Western Pacific Region, 125 million people fall ill annually due to
contaminated food and about 50,000 deaths occur. In this region, aflatoxin
is the major burden rather than diarrheal diseases. Annually, above 10,000
people are affected with hepatic cancer due to this (WHO, 2015).
11.5 INDIAN STATUS OF FBDs
The burden of FBD in India is largely unknown. Majority outbreaks of FBDs

go unreported, unrecognized, or un-investigated and may only be reported
after major health damage has occurred. Recent estimates state that at least
one out of 12 people fall ill due to food-related diseases in India, the number
of foodborne illnesses are projected to grow by as much as 70% resulting
in one out of 9 people falling ill by 2030, largely driven by population and
changing consumption patterns (Kristkova et al., 2017). It is important to
note that the expected FBDs increase might be still higher given the likely
underestimation of the current FBDs burden. The analysis of IDSP data
from the year 2011–2015 revealed that FBOs together with acute diarrheal
diseases constitute nearly half of all reported outbreaks. Moreover, for the
FBDs, passive surveillance systems represent only the tip of the iceberg
because most patients have mild symptoms of short duration and thus do
not visit doctor and many of those who seek medical care may not have
the laboratory test performed to determine a specific etiology. A review of
recorded FBD outbreaks in India from 1980 to 2016 revealed that S. aureus,
Vibrio spp., Salmonella spp., E. coli, Y. enteroclitica, and Norwalk-like virus
are some important microbial pathogens responsible for foodborne illnesses
(CD Alert, 2017). Different foods implicated in FBOs in India are milk and
milk products such as dahi, khoa, butter milk, sweets, kheer; meat, poultry,
fish, fowl, sea food such as prawns; cooked and uncooked rice; samosa,
tamarind; and cooked as well as uncooked vegetables.
The total number of cases and deaths due to acute diarrheal diseases in
India were 1,41,66,574 cases and 1555 deaths in year 2016 and 12,927,212
cases and 1331 deaths in year 2017 (NHP, 2018). Community studies revealed
that every child under 5 years of age had two or three episodes of diarrhea
every year, and 400,000–500,000 children under 5 years died from diarrhea.
Viral hepatitis was estimated about 12 cases per 100,000, although urban
communities had as high as 100 per 100,000. As per estimates by Kristkova
et al. (2017), if the current scenario of foodborne illnesses is not mitigated,
the number of people infected may increase from 100 million people today
to 170 million in 2030.
11.6 STRATEGIES AND PREVENTIVE APPROACHES
Farm to fork approach is required to ensure food safety which involves

series of steps to be taken by multidisciplinary team involving veterinarians,
microbiologists, environmentalists, industry people, and most importantly
consumers. Since the foodborne zoonotic diseases and their causative agents
differ between the regions, the preventive approach should be tailor-made
to each region at the national level. The existing difference in the burden of
FBD between low- and high-income regions clearly suggests that a major
fraction of the current burden is still avoidable through implementation of
general principles of food safety system. An enforcement of food safety
standards based on risk assessment in particular region along with effective
surveillance networks will aid in prevention of FBDs to a greater extent.
The following strategies may be implemented for effective prevention and
control of foodborne zoonotic diseases:
11.6.1 STRATEGY I—SURVEILLANCE AND RESPONSE
• Prioritization of important foodborne zoonotic diseases on the basis

of data available and interactions with planners, policy makers,
scientists, extension workers, NGOs, etc., from different profes-
sional and social organizations.
• Ensure that priority zoonotic diseases are agreed among all the
relevant sectors.
• Identification of network laboratories and working group for each
pathogen across the country.
• Use of validated diagnostics and common protocols for pathogen
detection and identification.
• Formulation of statistical plan for collection of samples from the
entire value chain during surveillance.
• Information sharing and interactions between the core groups, while
considering reliability of the data based on its sources.
• Strengthen public health systems to support surveillance and to
implement prevention and control programs.
• Improve ability to communicate electronically with state and local
health departments.
11.6.2 STRATEGY II—MICROBIOLOGICAL RISK ASSESSMENT AND

RISK MANAGEMENT
• Risk assessment studies to be carried out for important foodborne

zoonotic pathogens.
• Identification and implementation of different risk management
options throughout the food chain from farm to fork for different
food production systems.
• Establishing microbial standards based on the MRA studies in Indian
context.
• Consider the food supply chain as a whole and look at how contami-
nation might occur. Then, identify the individual control points where
contamination can be minimized or eliminated. The recent adoption
of WGS-based monitoring in several countries provides new pros-
pects for defining control objectives more precisely, reducing food
contamination, and reducing the enormous burden of disease.
11.6.3 STRATEGY III—APPLIED RESEARCH
• Reliable, rapid, simple, and affordable diagnostic tests need to be

developed.
• Rapid screening test may be devised for detection of food intoxica-
tion, especially to test foods which are prepared and served in mass
congregations.
• Alternatives to drugs and antimicrobials need to be explored.
• Genome analysis of foodborne pathogens to record interspecies
transmission and to gain insights in the field of pathogen behavior.
• Research on prevention and control throughout food chain.
11.6.4 STRATEGY IV—COORDINATED APPROACH AND AWARENESS

CAMPAIGNS
• Establish regional labs where experts from human health, animal

health and agricultural scientists work together to contain FBDs
with an integrated approach.
• Coordinated mechanisms to strengthen multisectoral collaboration.
• Awareness campaigns on zoonoses through interactions and train-
ings for professionals, high risk groups, common masses, and people
engaged in livestock industry.
• Need to instill true sense of sanitation and hygiene among various
stakeholders through motivation, education, and reward/recognition
using mass media.
• Kitchen hygiene is an area where basic food safety education is to be
provided so as to avoid risky food practices.
• Advocacy on the role of local welfare bodies, municipal corpora-
tion, and NGOs for prompt management and disposal of biohazard
waste and garbage.
11.7 CONCLUSION
FBDs cause huge morbidity and considerable mortality and pose significant
obstacle to socioeconomic development worldwide. However, information
regarding magnitude of burden due to specific pathogens is sketchy in our
country. On the other hand, precise information on the burden of foodborne

zoonotic diseases is essentially required for taking suitable decision by
policy-makers and to allocate appropriate resources for food safety control
and intervention efforts. The prevention and control measures should be initi-
ated by veterinarian’s from primary production level through good animal
husbandry practices (GAHP) and biosecurity measures. Further, during
production and processing level good manufacturing practices (GMP) and
good hygienic practices (GHP) and implementation of HACCP plan should
be followed for reducing FBDs. There is an urgent need for prudent use
of antibiotics in both human and veterinary medicine in order to mitigate
the rising antimicrobial resistance scenario. One health approach not only
improves the understanding of epidemiological nature of diseases, but also
aid in effective implementation of control measures. It is increasingly neces-
sary to consider holistically all aspects of FBD in the One Health Framework.
11.7.1 RECOMMENDATIONS/ACTIONABLE POINTS
• Formulate a “Network program on FBDs” with the mandate of

conducting surveillance studies on foodborne zoonotic diseases
followed by risk assessment studies for most prevalent/important
pathogens.
• Formation of Rapid Response Teams comprising multisubject
specialists for investigation of FBOs.
• Inclusion of veterinary public health specialist in all IDSP labs.
• Creation of a digital database like on the lines of “FoodNet” of CDC
USA.
• Integrating food safety into nutrition and food security policies and
programs.
• Incorporate zoonoses including foodborne infections and emerging
infectious diseases in the National Health Policy.
• Mandatory training of butchers on hygienic slaughter practices and
making it compulsory that only those personnel who have completed
this training can do dressing.
• Training of street food vendors in basics of hygiene and food safety
and issue of certificates so as to improve consumer confidence.
• A content on foodborne zoonotic disease and food safety may be
made part of course curriculum of school and for undergraduate
courses of home science and hospitality management.
KEYWORDS
• foodborne zoonoses
• control strategies
• public health
• socioeconomic development
• healthy food
REFERENCES
Ali, M.; Gupta, S. S.; Arora, N.; Khasnobis, P.; Venkatesh, S.; Sur, D.; Nair, G. B.; Sack,
D. A.; Ganguly, N. K.; Identification of Burden Hotspots and Risk Factors for Cholera in
India: An Observational Study. PLoS One 2017, 12 (8), e0183100.
Arai, S.; Yamaya, S.; Ohtsuka, K.; Konishi, N.; Obata, H.; Ooka, T.; ... Hara-Kudo, Y.
Detection of Escherichia Albertii in Retail Oysters. J. Food Prot. 2022, 85 (1), 173–179.
Assefa, A.; Bihon, A. A Systematic Review and Meta-Analysis of Prevalence of Escherichia
coli in Foods of Animal Origin in Ethiopia. Heliyon 2018, 4 (8), e00716.
Bachofen, C. Selected Viruses Detected on and in Our Food. Curr. Clin. Microbiol. Rep. 2018,
5 (2), 143–153.
Bancerz-Kisiel, A.; Szweda, W. Yersiniosis—A Zoonotic Foodborne Disease of Relevance to
Public Health. Annal. Agric. Environ. Med. 2015, 22 (3).
Bapat, P. R.; Dodkey, R. S.; Shekhawat, S. D.; Husain, A. A.; Nayak, A. R.; Kawle, A. P.; and
Kashyap, R. S. Prevalence of Zoonotic Tuberculosis and Associated Risk Factors in Central
Indian Populations. J. Epidemiol. Glob. Health 2017, 7 (4), 277–283.
Barbuddhe, S. B.; Malik, S. V. S.; Kumar, J. A.; Kalorey, D. R.; Chakraborty, T. Epidemiology
and Risk Management of Listeriosis in India. Int. Jo. Food Microb. 2012, 154 (3), 113–118.
Batra, P.; Mathur, P.; Misra, M. C. Aeromonas spp.: An Emerging Nosocomial Pathogen. J.
Lab. Phys. 2016, 8 (1), 1.
Benevenia, R.; Arnaboldi, S.; Dalzini, E.; Todeschi, S.; Bornati, L.; Saetti, F.; ... Losio, M. N.
Foodborne Botulism Survey in Northern Italy from 2013 to 2020: Emerging Risk or Stable
Situation?. Food Control. 2022, 132, 108520.
Bennett, S. D.; Walsh, K. A.; Gould, L. H. Foodborne Disease Outbreaks Caused by Bacillus
cereus, Clostridium perfringens, and Staphylococcus aureus—United States, 1998–
2008. Clin. Infect. Dis. 2013, 57, 425–433.
Brahmabhatt, M. N. Prevalence of Aeromonas spp. in Chicken Samples Collected from Retail
Shops of Anand (Gujarat). J. Vet. Public Health, 2011, 9 (2), 115–117.
Caggiano, G.; Dambrosio, A.; Ioanna, F.; Balbino, S.; Barbuti, G. de Giglio, O.; Diella,
G.; Lovero, G.; Rutigliano, S.; Scarafile, G.; Baldassarre, A.; Vimercati. L.; Musti, M.;
Montagna, M. T. Prevalence and Characterization of Methicillin-resistant Staphylococcus
aureus Isolates in Food Industry Workers. Ann Ig 2016, 28, 8–14.
Peck, M. W.; Smith, T. J.; Anniballi, F.; Austin, J. W.; Bano, L.; Bradshaw, M.; ... Stringer, S. C.
Historical Perspectives and Guidelines for Botulinum Neurotoxin Subtype Nomenclature.
Toxins 2017, 9 (1), 38.
CD Alert Foodborne Diseases and Food Safety in India. Monthly Newsletter of the National
Centre for Disease Control Directorate General of Health Services; Government of India,
2017.
CFSPH. Reptile-Associated Salmonellosis [Online] 2013. http://www.cfsph.iastate.edu/
Factsheets/pdfs/reptile_associated_salmonellosis.pdf
Chaubey, K. K.; Singh, S. V.; Gupta, S.; Singh, M.; Sohal, J. S.; Kumar, N.; ...; Dhama, K.
Mycobacterium Avium Subspecies Paratuberculosis—An Important Foodborne Pathogen
of High Public Health Significance with Special Reference to India: An Update. Vet.
Q. 2017, 37 (1), 282–299.
Chen, X.; Zhu, Q.; Yu, F.; Zhang, W.; Wang, R.; Ye, X.; Jin, L.; Liu, Y.; Li, S.; Chen, Y.
Serology, Virulence and Molecular Characteristics of Vibrio Parahaemolyticus Isolated
from Seafood in Zhejiang province. PLoS One 2018, 13, e0204892. [Google Scholar]
[crossref]
Chhabra, P.; Dhongade, R. K.; Kalrao, V. R.; Bavdekar, A. R.; Chitambar, S. D. Epidemiological,
Clinical, and Molecular Features of Norovirus Infections in Western INDIA. Journal of
medical virology, 2009, 81 (5), 922–932.
Cossaboom, C. M.; Heffron, C. L.; Cao, D.; Yugo, D. M.; Houk‐Miles, A. E.; Lindsay, D. S.;
Zajac, A. M.; Bertke, A. S.; Elvinger, F.; Meng, X.‐J. Risk Factors and Sources of Foodborne
Hepatitis E Virus Infection in the United States. J. Med. Virol. 2016, 88 (9), 1641–1645.
Costa, W. L. R.; Ferreira, J. D. S.; Carvalho, J. S.; Cerqueira, E. S.; Oliveira, L. C.; de Almeida,
R. C. Methicillin-resistant Staphylococcus aureus in Raw Meats and Prepared Foods in
Public Hospitals in Salvador, Bahia, Brazil. J. Food Sci. 2015, 80 (1), M147–M150. https://
doi.org/10.1111/1750-3841.12723
Costa, D.; Razakandrainibe, R.; Valot, S.; Vannier, M.; Sautour, M.; Basmaciyan,
L.; ... Favennec, L. Epidemiology of Cryptosporidiosis in France from 2017 to
2019. Microorganisms 2020, 8 (9), 1358.
Crago, B.; Ferrato, C.; Drews, S. J.; Svenson, L. W.; Tyrrell, G.; Louie, M. Prevalence
of Staphylococcus aureus and Methicillin-resistant S . Aureus (MRSA) in Food Samples
Associated with Foodborne Illness in Alberta, Canada from 2007 to 2010. Food Microbiol.
2012, 32, 202–205. https://doi.org/10.1016/j.fm.2012.04.012
Dallagassa, C. B.; Surek, M.; Vizzotto, B. S.; Prediger, K. C.; Moriel, B.; Wolf, S.; ... Fadel-
Picheth, C. M. Characteristics of an Aeromonas Trota Strain Isolated from Cerebrospinal
Fluid. Microb. Pathog. 2018, 116, 109–112.
Damiris, K.; Meybodi, M. A.; Niazi, M.; Pyrsopoulos, N. Hepatitis E in Immunocompromised
Individuals. World J. Hepatol. 2022, 14 (3), 482.
Das, A.; Vinayasree, V.; Santhosh, C. R.; Hari, S. S. Surveillance of Aeromonas sobria and
Aeromonas Hydrophila from Commercial Food Stuffs and Environmental Sources. J. Exp.
Sci. 2012.
De Macedo Couto, R.; Santana, G. O.; Ranzani, O. T.; Waldman, E. A. One Health and
Surveillance of Zoonotic Tuberculosis in Selected Low-income, Middle-income and High-
Income Countries: A Systematic Review. PLoS Neglect. Trop. Dis. 2022, 16 (6), e0010428.
Deng, X.; den Bakker, H. C.; Hendriksen, R. S. Genomic Epidemiology: Whole-Genome-
sequencing-powered Surveillance and Outbreak Investigation of Foodborne Bacterial
Pathogens. Annu. Rev. Food Sci. Technol. 2016, 7 (1), 353–374.
Dewey-Mattia, D.; Manikonda, K.; Hall, A. J.; Wise, M. E.; Crowe, S. J. Surveillance for
Foodborne Disease Outbreaks—United States, 2009–2015. MMWR Surveill Summ. 2018,
67 (10):, 1–11.
Dewey-Mattia, D.; Manikonda, K.; Hall, A. J.; Wise, M. E.; Crowe, S. J. Surveillance
for Foodborne Disease Outbreaks—United States, 2009–2015. MMWR Surveillance
Summaries 2018, 67 (10), 1.
Didugu, H.; Krishnaiah, N.; Rao, M. T.; Ramanipushpa, R. N. Antibiogram of Aeromonas
Species Isolated From Livestock Products. Int. J. Sci. Environ. Technol. 2016, 5 (2),
475–478.
Doimari, S.; Singh, V.; Kumari, R.; Kumar, M. S.; Singh, D. K. In Vitro Antimicrobial
Susceptibility of Brucella Species Isolated from Human and Animals in India. J. Antibiot.
Res. 2018, 3 (1), 101.
Dong, S.; Yang, Y.; Wang, Y.; Yang, D.; Yang, Y.; Shi, Y.; ... Zhou, Y. Prevalence of
Cryptosporidium Infection in the Global Population: A Systematic Review and Meta-
Analysis. Acta Parasitol. 2020, 65 (4), 882–889.
EFSA, (European Food Safety Authority) Assessment of the Public Health Significance of
Methicillin Resistant Staphylococcus Aureus (MRSA) in Animals and Foods. Scientific
Opinion of the Panel on Biological Hazards. EFSA J. 2009, 993, 1–73.
EFSA. European Food Safety Authority and European Centre for Disease Prevention and
Control. EFSA J. 2015, 13, 4329.
El Qouqa, I. A.; El Jarou, M. A.; Samaha, A. S. A.; Al Afifi, A. S.; Al Jarousha, A. M. K.
Yersinia Enterocolitica Infection among Children Aged Less than 12 Years: A Case–control
Study. Int. J. Infect. Dis. 2011, 15 (1), e48–e53.
European Food Safety Authority (EFSA). The European Union One Health 2018 Zoonoses
Report. EFSA J. 2019, 17 (12), e05926. Https://doi.org/10.2903/j.efsa.2019.5926
European Food Safety Authority (EFSA). European Centre for Disease Prevention and
Control (ECDC) The European Union Summary Report on Trends and Sources of Zoonoses;
Zoonotic Agents and Food-Borne Outbreaks in 2017.
European Food Safety Authority, and European Centre for Disease Prevention and Control.
The European Union one Health 2020 Zoonoses Report. EFSA J. 2021, 19 (12), e06971.
Farahmand, M.; Moghoofei, M.; Dorost, A.; Shoja, Z.; Ghorbani, S.; Kiani, S. J.; ... Tavakoli,
A. Global Prevalence and Genotype Distribution of Norovirus Infection in Children with
Gastroenteritis: A Meta‐analysis on 6 Years of Research from 2015 to 2020. Rev. Med.
Virol. 2022, 32 (1), e2237.
Felicio dasilva, et al. Risk Ranking of Pathogens in Ready-to-Eat Unprocessed Foods of
Non-animal Origin (FoNAO) in the EU: Initial Evaluation Using Outbreak Data (2007–
2011). Int. J. Food Microbiol. 2015, 195, 9–19. doi:10.1016/j.ijfoodmicro.2014.11.005.
Figueras, M. J.; Beaz-Hidalgo, R.; Hossain, M. J.; Liles, M. R. Taxonomic Affiliation of
New Genomes Should be Verified Using Average Nucleotide Identity and Multilocus
Phylogenetic Analysis. Genome Announc. 2014, 2 (6), e00927–e00914.
Food and Agriculture Organization of the United Nations (FAO) and World Health Organization
(WHO). Risk Assessment of Vibrio Parahaemolyticus in Seafood: Interpretative Summary
and Technical Report. Microbiological Risk Assessment Series No. 16. Rome, Italy. http://
www.fao.org/3/a-i2225e.pdf
Fredriksson-Ahomaa, M.; London, L.; Skrzypczak, T.; Kantala, T.; Laamanen, I.; Biström,
M.; ... Gadd, T. Foodborne Zoonoses Common in Hunted Wild Boars. Ecohealth, 2020, 17
(4), 512–522.
Fusco, G.; Di Bartolo, I.; Cioffi, B.; Ianiro, G.; Palermo, P.; Monini, M.; Amoroso, M. G.
Prevalence of Foodborne Viruses in Mussels in Southern Italy. Food Environ. Virol. 2017,
9 (2), 187–194.
Gray, J. A.; Chandry, P. S.; Kaur, M.; Kocharunchitt, C.; Bowman, J. P.; Fox, E. M.
Characterisation of Listeria Monocytogenes Food-associated Isolates to Assess
Environmental Fitness and Virulence Potential. Int. J. Food Microbiol. 2021, 350, 109247.
http://dx.doi.org/10.1016/j.ijfoodmicro.2021.109247. PMID:34023680
Grutsch, A. A.; Nimmer, P. S.; Pittsley, R. H.; mckillip, J. L. Bacillus spp. As Pathogens in the
Dairy Industry. In Foodborne Diseases; Academic Press, 2018; pp 193–211.
Hachemi, A.; Zenia, S.; Denia, M.F.; Guessoum, M.; Hachemi, M.M.; Ait-Oudhia, K.
Epidemiological Study of Sausage in Algeria: Prevalence, Quality Assessment, and
Antibiotic Resistance of Staphylococcus aureus Isolates and the Risk Factors Associated
with Consumer Habits Affecting Foodborne Poisoning, Vet. World 2019, 12 (8), 1240–1250.
Hajimohammadi, B.; Ahmadian, S.; Firoozi, Z.; Askari, M.; Mohammadi, M.; Eslami, G.; ...
Boozhmehrani, M. J. A Meta-Analysis of the Prevalence of Toxoplasmosis in Livestock
and Poultry Worldwide. Ecohealth 2022, 1–20.
Ingram, M.; John, J. S.; Applewhaite, T.; Gaskin, P.; Springer, K.; and Indar, L. Population-
based Estimates of Acute Gastrointestinal and Foodborne Illness in Barbados: A
Retrospective Cross-sectional Study. J. Health Popul. Nutr. 2013, 31 (4 Suppl 1), S81.
Innes, E. A.; Chalmers, R. M.; Wells, B.; Pawlowic, M. C. A One Health Approach to Tackle
Cryptosporidiosis. Trends Parasitol. 2020, 36 (3), 290–303.
Jain. D.; Sinha, S.; Prasad, N. K.; Pandey, M. C. Campylobacter Species and Drug Resistance
in a North Indian Rural Community. Trans. R. Soc. Trop. Med. Hyg. 2005, 99, 207–214.
Jajere, S. M. A Review of Salmonella enterica with Particular Focus on the Pathogenicity
and Virulence Factors, Host Specificity and Antimicrobial Resistance Including Multidrug
Resistance. Vet. World 2019, 12 (4), 504.
Jeffery, I. A.; Karim, S. Botulism. 2017.
Jessberger, N.; Dietrich, R.; Granum, P. E.; Märtlbauer, E. The Bacillus cereus Food Infection
as Multifactorial Process. Toxins 2020, 12, 701.
John, N.; Vidyalakshmi, V. B.; Hatha, A. M. Effect of ph and Salinity on the Production of
Extracellular Virulence Factors by Aeromonas from Food Sources. J. Food Sci. 2019, 84
(8), 2250–2255.
Jordan, K.; Mcauliffe, O. Listeria Monocytogenes in Foods. In Biological Emerging Risks
in Foods (Advances in Food and Nutrition Research), Vol. 86; Rodriguez-Lazaro, D., Ed.;
Cambridge: Elsevier, 2018; pp 181–213. http://dx.doi.org/10.1016/bs.afnr.2018.02.006
Kaur, S.; Malik, S. V. S.; Vaidya, V. M.; Barbuddhe, S. B. Listeria monocytogenes in
Spontaneous Abortions in Humans and Its Detection by Multiplex PCR. J. Appl.
Microbiol. 2007, 103 (5), 1889–1896.
Khuroo, M. S.; Khuroo, M. S.; Khuroo, N. S. Hepatitis E: Discovery, Global Impact, Control
and Cure. World J. Gastroenterol. 2016, 22 (31), 7030.
Knific, T.; Ocepek, M.; Kirbiš, A.; Krt, B.; Prezelj, J.; Gethmann, J. M. Quantitative Risk
Assessment of Exposure to Mycobacterium avium subsp. Paratuberculosis (MAP) via
Different Types of Milk for the Slovenian Consumer. Foods 2022, 11 (10), 1472.
Koohsar, F.; Naddaf, S. R.; Rokni, M. B.; Mirjalali, H.; Mohebali, M.; Shafiei, R.; Mowlavi,
G. Serological Detection of Trichinellosis among Suspected wild Boar Meat Consumers in
North and Northeast of Iran. Iran. J. Parasitol. 2021, 16 (2), 253.
Kotwal, A.; Singh, H.; Verma, A. K.; Gupta, R. M.; Jain, S.; Sinha, S.; Joshi, R. K.; et al. A
Study of Hepatitis A and E Virus Seropositivity Profile Amongst Young Healthy Adults in
India. Med. J. Armed Forces India 2014, 70 (3), 225–229.
Koutsoumanis, K. Public Health Risks Associated with Food-borne Parasites. EFSA J. 2018.
Kristkova, Z. S.; Grace, D.; Kuiper, M. The Economics of Food Safety in India: A Rapid
Assessment; Wageningen University and Research, 2017.
Kumar M. S. Molecular Studies on Antibiotic Resistance and Virulence Genes of
Campylobacter Isolates from Diverse Origin. Ph.D. Thesis, submitted to Deemed
University, Indian Veterinary Research Institute, Izatnagar, 2016.
Kumar, A.; Agarwal, R. K.; Bhilegaonkar, K. N.; Shome, B. R.; Bachhil, V. N. Occurrence
of Campylobacter jejuni in Vegetables. Int. J. Food Microbiol. 2001, 67 (1–2), 153–155.
Kumar, M.; Suman, T. P.; Ramees, H.; Dhanze, S.; Anjay, G.; Dubal, Z. B.; Sivakumar, M.;
Kumar, A. Occurrence of Thermophilic Campylobacters in Foods of Animal Origin, Animal
Faeces and Human Stool. J. Vet. Pub. Health 2015, 13 (2), 105–109.
Lalitha, K. V.; Surendran, P. K. Occurrence of Clostridium botulinum in Fresh and Cured Fish
in Retail Trade in Cochin (India). Int. J. Food Microbiol. 2002, 72 (1–2), 169–174.
Lee, H.; Yoon, Y. Etiological Agents Implicated in Foodborne Illness World Wide. Food Sci.
Anim. Resour. 2021, 41 (1), 1.
Lee, N.; Chan, M. C.; Wong, B.; Choi, K. W.; Sin, W.; Lui, G.; ... Leung, W. K. Fecal Viral
Concentration and Diarrhea in Norovirus Gastroenteritis. Emerg. Infect. Dis. 2007, 13 (9),
1399.
Lenzi, A.; Marvasi, M.; Baldi, A. Agronomic Practices to Limit Pre- and Post-harvest
Contamination and Proliferation of Human Pathogenic Enterobacteriaceae in Vegetable
Produce. Food Control 2021, 119, 107486.
Leung, V. H.; Phan, Q.; Costa, C. E.; Nishimura, C.; Pung, K.; Horn, L.; Sosa, L. Notes from
the Field: Clostridium Perfringens Outbreak at a Catered Lunch—Connecticut, September
2016. MMWR Morb. Morb. Mortal. Wkly. Rep. 2017, 66, 940–941.
Li, C.; Zeng, H.; Ding, X.; Chen, Y.; Liu, X.; Zhou, L.; Wang, X.; Cheng, Y.; Hu, S.; Cao,
Z.; Liu, R.; Yin, C. Perinatal Listeriosis Patients Treated at a Maternity Hospital in Beijing,
China, from 2013–2018. BMC Infect. Dis. 2020, 20 (1), 601. http://dx.doi. Org/10.1186/
s12879–020-05327-6. PMID:32799811.
Li, H.; Guo, Y.; Tian, T.; Guo, W.; Liu, C.; Liang, X.; ... Fu, P. Epidemiological Analysis of
Foodborne Botulism Outbreaks—China, 2004–2020. China CDC Week. 2022, 1–5.
Li, H.; Li, W.; Dai, Y.; Jiang, Y.; Liang, J.; Wang, S.; ... Guo, Y. Characteristics of Settings and
Etiologic Agents of Foodborne Disease Outbreaks—China, 2020. China CDC Week. 2021,
3 (42), 889.
Li, P.; Ji, Y.; Li, Y.; Ma, Z.; Pan, Q. Estimating the Global Prevalence of Hepatitis E Virus in
Swine and Pork Products. One Health 2022, 14, 100362.
Li, P.; Liu, J.; Li, Y.; Su, J.; Ma, Z.; Bramer, W. M.; ... Pan, Q. The Global Epidemiology of
Hepatitis E Virus Infection: A Systematic Review and Meta‐analysis. Liver Int. 2020, 40
(7), 1516–1528.
Liao, Y.; Hong, X.; Wu, A.; Jiang, Y.; Liang, Y.; Gao, J.; ... Kou, X. Global prevalence of
Norovirus in Cases of Acute Gastroenteritis from 1997 to 2021: An Updated Systematic
Review and Meta-analysis. Microb. Pathog. 2021, 161, 105259.
Liu, X.-Y.; Hu, Q.; Xu, F.; Ding, S.-Y.; Zhu, K. Characterization of Bacillus cereus in Dairy
Products in China. Toxins 2020, 12, 454.
Lyoo, E. L.; Park, B. J.; Ahn, H. S.; Han, S. H.; Go, H. J.; Kim, D. H.; ... Choi, I. S.
Detection and Genetic Analysis of Zoonotic Hepatitis E Virus, Rotavirus, and Sapovirus in
Pigs. Korean J. Vet. Res. 2020, 60 (2), 61–68.
Menon, V. K.; George, S.; Sarkar, R.; Giri, S.; Samuel, P.; Vivek, R.; . Gray, J. J. Norovirus
Gastroenteritis in a Birth Cohort in Southern India. PLoS One, 2016, 11 (6), e0157007.
Muchaamba, F.; Barmettler, K.; Treier, A.; Houf, K.; Stephan, R. Microbiology
and Epidemiology of Escherichia albertii—An Emerging Elusive Foodborne
Pathogen. Microorganisms 2022, 10 (5), 875.
Mughini‐Gras, L.; Benincà, E.; mcdonald, S. A.; de Jong, A.; Chardon, J.; Evers, E.; Bonačić
Marinović, A. A. A Statistical Modelling Approach for Source Attribution Meta‐analysis of
Sporadic Infection with Foodborne Pathogens; Zoonoses and Public Health, 2022.
Mulder, A. C.; van de Kassteele, J.; Heederik, D.; Pijnacker, R.; Mughini‐Gras, L.; Franz, E.
Spatial Effects of Livestock Farming on Human Infections with Shiga Toxin‐producing
Escherichia coli O157 in Small but Densely Populated Regions: The Case of the
Netherlands. Geohealth 2020, 4 (11), e2020gh000276.
Munck, N.; Smith, J.; Bates, J.; Glass, K.; Hald, T.; Kirk, M. D. Source Attribution of
Salmonella in Macadamia Nuts to Animal and Environmental Reservoirs in Queensland,
Australia. Foodborne Pathog. Dis. 2020, 17 (5), 357–364.
National Health Profile. 13th Issue. Central Bureau of Health Intelligence Directorate
General of Health Services Ministry of Health and Family Welfare; Government of India
WHO Collaborating Centre on Family of International Classifications, 2018.
National Health Profile. 14th Issue. Central Bureau of Health Intelligence Directorate
General of Health Services Ministry of Health and Family Welfare; Government of India
WHO Collaborating Centre on Family of International Classifications, 2019.
Nayak, M. K.; Balasubramanian, G.; Sahoo, G. C.; Bhattacharya, R.; Vinje, J.; Kobayashi,
N.; Sarkar, M. C.; Bhattacharya, M. K.; Krishnan, T. Detection of a Novel Intergenogroup
Recombinant Norovirus from Kolkata, India. Virology 2008, 377 (1), 117–123.
Odo, S. E.; Uchechukwu, C. F.; Ezemadu, U. R. Foodborne Diseases and Intoxication in
Nigeria: Prevalence of Escherichia coli 0157: H7, Salmonella, Shigella and Staphylococcus
aureus. J. Adv. Microbiol. 2021, 20 (12), 84–94.
OIE. Manual of Standards for Diagnostic Tests and Vaccines, 7th ed.; OIE Press; France,
2016; p 4.
Olaimat, A. N.; Al‐Holy, M. A.; Shahbaz, H. M.; Al‐Nabulsi, A. A.; Abu Ghoush, M. H.;
Osaili, T. M.; Ayyash, M. M.; Holley, R. A.; Emergence of Antibiotic Resistance in Listeria
Monocytogenes Isolated from Food Products: A Comprehensive Review. Compr. Rev. Food
Sci. Food Saf. 2018, 17 (5), 1277–1292.
Olea-Popelka, F.; Muwonge, A.; Perera, A.; Dean, A. S.; Mumford, E.; Erlacher-Vindel,
E.; ... Fujiwara, P. I. Zoonotic Tuberculosis in Human Beings Caused by Mycobacterium
Bovis—A Call for Action. Lancet Infect. Dis. 2017, 17 (1), e21–e25.
Ongor, H.; Cetinkaya, B.; Karahan, M.; Bulut, H. Evaluation of Immunomagnetic Separation-
Polymerase Chain Reaction in Direct Detection of Brucella abortus and Brucella melitensis
from Cheese Samples. Foodborne Pathog. Dis. 2006, 3, 245–250.
Oniciuc, E. A.; Ariza-Miguel, J.; Bolocan, A. S.; et al. Foods from Black Market at EU Border
as a Neglected Route of Potential Methicillin-resistant Staphylococcus aureus Transmission.
Int. J. Food Microbiol. 2015, 209, 34–38. https://doi.org/10.1016/j.ijfoodmicro.2014.11.015
Osuna-Chávez, R.; López-Robles, G.; Celaya-Michel, H.; Sosa-Castañeda, J.; Nieblas-
López, M.; Ibarra-Zazueta, C. Seroprevalence of Mycobacterium Avium Subspecie
Paratuberculosis in Suspect Cattle Slaughtered at Slaughterhouse from Sonora,

Mexico. Abanico veterinario 2021, 11.
Outreach Programme on Zoonotic Diseases (OPZD). Annual Report 2014–15, Submitted to
ICAR New Delhi.
ICAR New Delhi.
ICAR New Delhi.
ICAR New Delhi.
ICAR New Delhi.
Ozana, V.; Hruska, K.; Sechi, L. A. Neglected Facts on Mycobacterium Avium Subspecies
Paratuberculosis and Type 1 Diabetes. Int. J. Mol. Sci. 2022, 23 (7), 3657.
Pal, M. Brucellosis: A Highly Infectious Foodborne Zoonotic Disease of Public Health
Concern. Madridge J. Food Tech. 2018, S1 (1), 1–3.
Pal, M.; Beshatu, F.; Ritwick, D. Significance of Yersinia Entrocolitica as Emerging Zoonotic
Food Borne Pathogen. J. Natural Hist, 2013, 9, 36–45.
Pattis, I.; Cressey, P. J.; Lopez, L. D.; Roos, R.; Horn, B.; Soboleva, T. Annual Report
Concerning Foodborne Disease in New Zealand 2017; Ministry for Primary Industries:
Manatū Ahu Matua, 2019.
Pires, S. M.; Fischer-Walker, C. L.; Lanata, C. F.; Devleesschauwer, B.; Hall, A. J.; Kirk, M.
D.; et al. Aetiology-specific Estimates of the Global and Regional Incidence and Mortality
of Diarrhoeal Diseases Commonly Transmitted Through Food. PLoS One 2015,10 (12),
e0142927.
Prasad, K. N.; Mathur, S. K.; Dhole, T. N.; Ayyagari, A. Antimicrobial Susceptibility and
Plasmid Analysis of Campylobacter jejuni Isolated from Diarrheal Patients and Healthy
Chickens in Northern India. J. Diarrheal. Dis. Res. 1994, 12 (4), 270–273.
Qi, R.; Huang, Y. T.; Liu, J. W.; Sun, Y.; Sun, X. F.; Han, H. J.; ... Yu, X. J. Global Prevalence
of Asymptomatic Norovirus Infection: A Meta-analysis. Eclinicalmedicine 2018, 2, 50–58.
Riahi, S. M.; Ahmadi, E.; Zeinali, T. Global Prevalence of Yersinia Enterocolitica in Cases of
Gastroenteritis: A Systematic Review and Meta-analysis. Int. J. Microbiol. 2021.
Ronholm, J.; Nasheri, N.; Petronella, N.; Pagotto, F. Navigating Microbiological Food Safety
in the Era of Whole-genome Sequencing. Clin. Microbiol. Rev. 2016, 29 (4), 837–857.
Ryan, U.; Hijjawi, N.; Xiao, L. Foodborne Cryptosporidiosis. Int. J. Parasitol. 2018, 48 (1),
1–12.
Scallan, E.; et al. Foodborne Illness Acquired in the United States—Major Pathogens. Emerg.
Infect. Dis. 2011, 17, 7–15. doi: 10.3201/eid1701.p11101
Shakuntala, I.; Das, S.; Ghatak, S.; Milton, A. A. P.; Sanjukta, R.; Puro, K. U.; Aparajita,
D.; Sukhadeo, B. B.; Sen, A. Prevalence, Characterization, and Genetic Diversity of
Listeria monocytogenes Isolated from Foods of Animal Origin in North East India. Food
Biotechnol. 2019, 33 (3), 237–250.
Sharma, I.; Kumar, A.; Pramanik, A. K. Isolation and Identification of Mesophillic Aeromonas
Bacteria from Meat and Fish Foods of North East India. J. Pure Appl. Microbiol. 2009, 3
(2), 517–526.
Shonhiwa, A. M.; Ntshoe, G.; Essel, V.; Thomas, J.; Mccarthy, K. A Review of Foodborne
Diseases Outbreaks Reported to the Outbreak Response Unit, National Institute for
Communicable Diseases, South Africa, 2013–2017. Int. J. Infect. Dis. 2019, 79, 73.
Silva, N. C. C.; Guimarães, F. F.; Manzi, M. P.; Budri, P. E.; Gómez-Sanz, E.; Benito, D.;
Langoni, H.; Rall, V. L. M.; Torres, C. Molecular Characterization and Clonal Diversity of
Methicillin-Susceptible Staphylococcus aureus in Milk of Cows with Mastitis in Brazil. J.
Dairy Sci. 2013, 96 (11), 6856–6862.
Singh, B. B.; Dhand, N. K.; Gill, J. P. S. Economic Losses Occurring due to Brucellosis in
Indian Livestock Populations. Prev. Vet. Med. 2015, 119 (3–4), 211–215.
Singh, B. B.; Khatkar, M. S.; Aulakh, R. S.; Gill, J. P. S.; Dhand, N. K. Estimation of the
Health and Economic Burden of Human Brucellosis in India. Prev. Vet. Med. 2018, 154,
148–155.6
Sivakumar, M. Phenotypic and Genotypic Characterization of Methicillin Resistant
Staphylococcus aureus and ESBL Producing Escherichia coli Isolated from Street Vended
Foods of Animal Origin and Associated Environment. M.V.Sc. Thesis, submitted to Deemed
University, Indian Veterinary Research Institute, Izatnagar, 2016.
Smith, T. C. Livestock-associated Staphylococcus Aureus: The United States Experience.
PLoS Pathog. 2015, 11 (2), 1–8. https://doi.org/10.1371/journal.ppat.1004564
Srinivasan, S.; Easterling, L.; Rimal, B.; Niu, X. M.; Conlan, A. J.; Dudas, P.; Kapur,
V. Prevalence of Bovine Tuberculosis in India: A Systematic Review and Meta‐
analysis. Transbound. Emerg. Dis. 2018, 65 (6), 1627–1640.
Symeonidou, I.; Arsenopoulos, K.; Tzilves, D.; Soba, B.; Gabriël, S.; Papadopoulos, E.
Human Taeniasis/Cysticercosis: A Potentially Emerging Parasitic Disease in Europe. Ann.
Gastroenterol. 2018, 31 (4), 406.
Tate, J. E.; Burton, A. H.; Boschi-Pinto, C.; Parashar, U. D.; World Health Organization–
Coordinated Global Rotavirus Surveillance Network, Agocs, M.; Ranjan Wijesinghe, P.
Global, Regional, and National Estimates of Rotavirus Mortality in Children<5 Years of
Age, 2000–2013. Clin. Infec. Dis. 2016, 62 (Suppl 2), S96–S105.
Thomas, J.; Govender, N.; mccarthy, K. M.; Erasmus, L. K.; Doyle, T. J.; Allam, M.; Ismail,
A.; Ramalwa, N.; Sekwadi, P.; Ntshoe, G.; Shonhiwa, A.; Essel, V.; Tau, N.; Smouse,
S.; Ngomane, H. M.; Disenyeng, B.; Page, N. A.; Govender, N. P.; Duse, A. G.; Stewart,
R.; Thomas, T.; Mahoney, D.; Tourdjman, M.; Disson, O.; Thouvenot, P.; Maury, M. M.;
Leclercq, A.; Lecuit, M.; Smith, A. M.; Blumberg, L. H. Outbreak of Listeriosis in South
Africa Associated with Processed Meat. N. Engl. J. Med. 2020, 382 (7), 632–643. http://
dx.doi.org/10.1056/nejmoa1907462. PMID:32053299.
Troeger, C.; Forouzanfar, M.; Rao, P. C.; Khalil, I.; Brown, A.; Reiner Jr, R. C.; Alemayohu,
M. A. Estimates of Global, Regional, and National Morbidity, Mortality, and Aetiologies
of Diarrhoeal Diseases: A Systematic Analysis for the Global Burden of Disease Study
2015. Lancet Infec. Dis. 2017, 17 (9), 909–948.
Van der Giessen, J.; Deksne, G.; Gómez-Morales, M. A.; Troell, K.; Gomes, J.; Sotiraki, S.;
... Robertson, L. J. Surveillance of Foodborne Parasitic Diseases in Europe in a One Health
Approach. Parasite Epidemiol. Cont. 2021, 13, e00205.
Velebit, B.; Djordjevic, V.; Milojevic, L.; Babic, M.; Grkovic, N.; Jankovic, V.; Yushina,
Y. In The Common Foodborne Viruses: A Review. IOP Conference Series: Earth and
Environmental Science; IOP Publishing, September, 2019. Vol. 333, No. 1, p 012110.
Villabruna, N.; Koopmans, M. P.; de Graaf, M. Animals as Reservoir for human
Norovirus. Viruses 2019, 11 (5), 478.
World Health Organization. WHO Estimates of the Global Burden of Foodborne

Diseases: Foodborne Disease Burden Epidemiology Reference Group 2007–2015. No.
9789241565165. World Health Organization, 2015.
Yang, K.; Wang, A.; Fu, M.; Wang, A.; Chen, K.; Jia, Q.; Huang, Z. Investigation of Incidents
and Trends of Antimicrobial Resistance in Foodborne Pathogens in Eight Countries from
Historical Sample Data. Int. J. Environ. Res. Public Health 2020, 17 (2), 472.
Yucun, C.; Jin, H.; Shihong, Y.; Liyang, P.; Xiaohui, L.; Muyuan, M. Epidemiological
Characteristics of Human Brucellosis in Dalian, Liaoning, 2014–2019. Disease Surveillance
2021, 36 (1), 59–62. doi: 10.3784/jbjc.202008180279
Zhang, X. Z.; Wang, Z. Q.; Cui, J. Epidemiology of Trichinellosis in the People’s Republic of
China During 2009–2020. Acta Tropica 2022, 106388.
CHAPTER 12
Omics Technologies: An Overview

of Application of Proteomics in Meat
Quality and Safety Assurance
KIRAN MOHAN1, NAVEENA BASAPPA MAHESWARAPPA2,
S. WILFRED RUBAN3, and RITUPARNA BANERJEE2
1
Department of Livestock Products Technology, Veterinary College,
KVAFSU, Bidar, Karnataka, India
2
ICAR-National Research Centre on Meat, Chengicherla, Hyderabad,
Telangana, India
3
Department of Livestock Products Technology, Veterinary College
Bengaluru, KVAFSU, Bidar, Karnataka, India
ABSTRACT
Over the last two decades, sophisticated “-OMICs” technologies like

genomics, proteomics, peptidomics, transcriptomics, metabolomics, and
lipidomics have been explored successfully to address food quality chal-
lenges. Additionally, these strong methodologies have been extensively
employed to understand the biological mechanisms for phenotypic variance
in technical and sensory quality attributes of muscle foods. Omics-based
technologies allow researchers to investigate complex biological processes
quantitatively and integratively using a systems biology approach. Food
proteomics is a rapidly growing area that is gaining importance in product
innovation, food safety, and food quality. Proteomics is the application of
technology for the identification and quantification of a cell’s, tissue’s, or
organism’s total protein content. It complements other “omics” technolo-
gies such as genomics and transcriptomics in elucidating the identity of an
organism’s proteins and in comprehending their structure and function. The

complexity of proteomics arises from the need to examine and classify all of
a genome’s protein signatures. This chapter highlights the role of proteomics
in food research, the underlying assumptions, methodological consider-
ations, and bioinformatic techniques used in the field. Further, it summarizes
proteome data from the most important meat species used in food production
and examines significant food concerns from a proteomic perspective. This
chapter seeks to offer an overview of the various proteomics methods, their
recent advancements, and their application in research and analysis in order
to better highlight meat quality and safety problems.
12.1 INTRODUCTION
Proteomics may be characterized as a method of determining a cell’s

structure, sequence, and quantity of proteins, along with their interactions
with other proteins and the degree of modification (Campbell, 2003). Many
different variables are evaluated with respect to meat quality, including color
or appearance, palatability (texture/tenderness/juiciness), nutritional factors
such as composition (proteins/minerals/fatty acids), as well as meat safety
in broad sense (Leheska et al., 2008; Verbeke et al., 2009). In contrast to
genomes, the proteome is dynamic and changes in response to changes in
the body’s physiological state. Meat quality may be genetically influenced
by many quantitative factors, such as the fiber characteristics or metabolic
processes of postmortem or living muscle. Environmental factors and the
procedure of meat processing influence meat quality just as much as the
animal’s genetic make-up.
Meat authentication is a procedure used to make sure the product is as
described on the label. Biomarkers that can be used as indications of meat
quality and authenticity are now being studied following recent advances
in the fields of omics and proteomics. Food safety can be compromised
by economic-motivated adulteration, which also has a detrimental impact
on consumers’ trust. Food safety and quality regulators are increasingly
concerned about the authenticity of muscle foods because of their growing
global reputation as a source of negative practices. Authentication of foods
has been addressed by a variety of analytical techniques, including biochem-
ical, immunological, mass spectrometry-based proteomic, and metabolomic
approaches. There is no limit to the types of foods that may be studied
using mass spectrometry-based methods, which are reliable, credible, and
Omics Technologies: An Overview of Application of Proteomics 305
sensitive in their use. Protein fractionation techniques including OFFGEL

and GELFrEE fractionation, two-dimensional gel electrophoresis, the use of
various mass spectrometers in authentication, the use of lateral flow assay
techniques as point-of-care tools, and detection of additional food frauds
such geographical origin, additives, processing methods, etc. are explored
in this paper.
12.2 TOOLS OF PROTEOMICS
It is difficult to perform proteomics because of the broad variety of proteins.

An animal’s genome, which has about 20,000 genes, has the ability to
generate 1.8 million distinct types of protein (Jensen, 2004). An ideal
proteomics approach can identify as many protein products as possible in
a sensitive, reproducible, and quantitative way. Methods such as protein
extraction, purification, and identification are popular in proteome research.
Proteomics research makes use of a variety of techniques, including 2-DE
gel electrophoresis, mass spectrometry, and bioinformatics. Proteomics is
the study of proteins in a cell under specified conditions (Vaidyanathan
and Goodacre, 2003). Purification of the crude sample is essential for all
proteome investigations before any further analysis and characterization can
be undertaken.
Electrophoresis of proteins using SDS-PAGE (sodium dodecyl sulphate
polyacrylamide gel electrophoresis) has been considered as the best method
for many biochemical studies. Expasy, a publicly accessible collection of
proteomics data, reveals that 2-DE is the most frequently used technique
in large-scale proteomics (http://www.expasy.ch). As of the mid-1970s,
modern 2DE technology uses IEF with highly reproducible immobilized
pH gradient (IPG) strips, a first-dimension isoelctric focusing technology.
The first dimension (IEF) entails the separation of proteins according to
their charge, followed by their molecular weight, which is performed using
SDS-PAGE (2nd dimension). The DIGE (differential gel electrophoresis)
approach, which was very recently created, allows for the direct measure-
ment of abundance changes on a massive level without interference from
gel-to-gel variation. Nanometric detection limits are possible with these dyes.
Lametsch and Bendixen (2001) revealed that for each gel, 1000 individual
protein spots may be separated by 2-DE using optimal extraction, separation,
and staining protocols.
There is a rising need for speedy and accurate protein identification

in light of the rapid advancement of numerous current and accomplished
genome sequencing activities. Proteins in nanogram levels may be identified
using mass spectrometry, which is a very effective method. It is possible for
a basic cell lysate to have hundreds of thousands of peptides after a simple
proteolytic digestion. Since the primary goal of many proteomic approaches
is protein identification using mass spectrometry (MS), it is common practise
to compare the MS results with protein sequence databases, generated by
genomics study (Friedman, 2011).
Mass spectrometry technologies have advanced significantly and are
increasingly being used to authenticate food. The mass spectrometry tech-
nique enables the precise measurement of the molecular mass of charged
molecules. Analytes are ionized in the gas phase and separated by mass-to-
charge ratio using mass analyzers and detectors, and the mass-to-charge ratio
of each analyte is determined by the ionization source and mass analyzer.
These instruments are either one-stage instruments (MSn) or many stages of
instrumentation, depending on how many analyzers are used in tandem or
in hybrid arrangements, such as Q3 or Q-TOF. Because of the simultaneous
advancements in liquid chromatography (LC) and capillary electrophoresis
for in-line separation, mass spectrometry analysis of proteins at the sequence/
peptide level became feasible. It is now possible to identify peptides with
femtomole (I0–15 mole) sensitivity in complicated biological materials using
these recent technical developments in mass-spectrometry methods (Mallick
and Kuster, 2010). Peptide-mass fingerprinting by MALDI-TOF MS is the
most extensively used methodology for protein detection from 2-DE. High-
throughput detection of femtomole amounts of proteins and peptides is partly
owing to this technology’s specificity. The amino acid sequence or peptides,
which are generated after digestion with a protease, can be used to identify
individual proteins in the data obtained by the mass spectrometry (Henzel et
al., 1993). Electrospray-ionization triple quadrupole mass spectrometers are
another frequent type of mass spectrometer. Bioinformatics approaches can
then be used to explore databases which match these mass spectral finger-
prints obtained from the experiment (Eng et al., 1994).
The theory behind spectroscopic meat analysis is that moisture, proteins,
and lipids found in meat produce multiple spectra at different wavelengths
(Wang et al., 2018). In recent years, spectral technologies have been investi-
gated in food quality and safety because of their nondestructive nature. Other
advantages of spectroscopic methods include ease of sample preparation and
the absence of the demand for sample pretreatment (Wang et al., 2018). The
Raman scattering effect is used to analyze the composition and structure of a

material in Raman spectroscopy (Hu et al., 2019). This noninvasive method
does not need sample preparation or a high sample volume, and it may even
be used on opaque materials (Hu et al., 2019; Lee et al., 2017). The type
of the molecules and their concentration may be determined using Raman
spectroscopy’s fingerprints of vibrational frequency and intensity (Lee et al.,
2017).
12.3 MEAT QUALITY AND SAFETY
Meat color is a critical quality characteristic that has a significant impact on

customer purchasing decisions (Nair and Zhai, 2020). The color of raw meat
is influenced by differences in reflection produced by protein denaturation
and myoglobin (Aaslyng, 2002; Miller, 2002). Thus, proteomic techniques
can assist in the comprehension of variations in meat color as well as the
examination of the underlying determinants of muscle-specific meat color.
There were 22 proteins or protein fragments that were found to be differ-
ently expressed in a sarcoplasmic proteins proteomic investigation of pig
Semimembranosus muscle with high and low lightness values (L*) (Sayd
et al., 2006). Overexpression of mitochondrial respiratory chain enzymes,
hemoglobin, and chaperones was found in darker meat with greater levels
of oxidative metabolism, while glycolysis enzymes were found in the
lighter meat. Using pyruvate kinase, which was found to be more abundant
in the Pectoralis major of broilers than in the Supracoracoideus, Zapata et
al. (2012) hypothesized that the fast pH fall associated with “pale poultry
muscle syndrome” might be detected using pyruvate kinase in the Pectoralis
major of broilers.
Researchers have studied the postmortem variance in tenderness among
species, breeds, ages, muscles, and processing settings using various
proteomic methods. Pig meat tenderness may be linked to postmortem
proteome changes, as initially reported by Lametsch et al. (2003). Meat
tenderness is strongly linked to postmortem breakdown of actin and myosin
heavy chain, according to these researchers. Many scientists believe that
heat shock proteins (HSPs) might serve as biomarkers for meat tenderness
(Laville et al., 2009; Polati et al., 2012). To maintain cellular proteins from
stress-induced denaturation and probable loss of function, as well as to stabi-
lize the muscle filament assembly throughout the stressful period, they may
play a role in the tenderization of meat (Feder and Hoffman, 1999; Feasson
et al., 2002; Perng et al., 1999). Several proteins, including uroplakin-1b,

complement C1q subcomponent subunit B, myosin-IIIa, aspartate amino-
transferase, glycogen phosphorylase, cytosolic carboxypeptidase 3, and
phosphatidylinositide transfer protein isoform, were found by Kiran et al.
(2016) that may be suitable biomarkers for the differentiating of young and
spent Indian water buffalo meat.
Water holding capacity (WHC) is significant for a variety of reasons,
including sensory appeal and economics, but it is also important because of
the consequences of this capacity on meat quality. When contractile proteins
are denatured and myofibrils shrink, the result is a loss of water and soluble
components in the form of drip. This occurs as a result of the contraction
of the muscles (Bertram et al., 2004; Offer et al., 1989). In order to better
understand the underlying processes of this characteristic and explore
reasonable protein biomarkers for WHC, a multitude of proteomics studies
have been conducted. 2-DE has revealed a number of protein markers that
may be linked to a decrease in the amount of drip loss in porcine muscle. Yu
et al. (2009) reported that a decrease in heat shock protein expression was
linked to a decrease in the amount of drip loss in muscle, indicating poor
meat quality.
12.4 PROTEOMICS IN FOOD SCIENCE
New and exciting proteomic methods are being used to detect proteins in
food matrixes and to examine protein–protein interactions in foods, along
with protein–protein interactions study among food components. A sensitive
evaluation of changes in protein structure occurring at specific amino acid
locations may be made using this technique to identify components formed
during processing processes.
For understanding the physiological and technical functions of amphi-
philic proteins found in wheat kernels, a proteomic study of these proteins was
carried out (Amiour et al., 2002). Proteomics has resulted in the development
of markers for certain food processing methods as well as indicators for the
quality of processed food products (Van Der Werf et al., 2001). Researchers
were able to detect the distinct proteins generated due to heat stress in tomato
using 2-DE (Iwahashi and Hosoda, 2000). The proteomic techniques can be
used to explore biomarkers that are indicative of certain food spoilage or
pathogenic microorganisms. Food allergies are becoming more and more
of a concern for the food sector as a whole. Genes or allergenic proteins
have been identified and effectively used in the study of allergic illnesses
(Toda and Ono, 2002). The 2-DE and MALDI-MS are useful methods for
the separation and investigation of complex protein mixtures, and they can
also provide crucial information on digestion and bioavailability patterns of
dietary proteins. Biological goods, such as vaccinations, blood products, and
therapies, are made possible by biotechnology-derived therapeutic proteins.
Recently, food quality analysis has seen the application of proteomics, which
shows great promise (Kvasnicka, 2003).
12.5 APPLICATION OF PROTEOMICS IN MEAT QUALITY
An essential step toward a more complete knowledge of the complex

biological systems that govern physiology and disease in animals can be
achieved via the use of proteomics and other postgenomic methods. Animals
are raised on a huge scale for the purpose of producing animal products for
human consumption. Thus, substantial biological investigation is directed
toward elucidating the biological characteristics that influence the quantity
and quality of these products. However, much of the data collected from trials
on farm animals such as swine and cattle is useful not just to farm animal
sciences, but also to our knowledge of complicated molecular systems of
health and illness in humans.
As in many other fields of life science, the application of proteomics
and other postgenomic techniques in animal sciences represents an impor-
tant step toward a more full understanding of the complicated biological
processes that drive physiology and disease in living organisms. Farm
animals are raised on a large scale by large-scale companies with the objec-
tive of generating animal products for human use. As a result, the majority of
biological research is devoted to understanding the biological characteristics
that determine the production and quality of these products. In addition to
advancing our knowledge of human health and disease molecular systems,
much of the data gathered from animal research, such as those on pigs and
cattle, is also important in the field of farm animal science.
Meat quality is a wide phrase used in meat science to refer to a multi-
tude of different factors depending on the objective. The necessity of a safe
product is self-evident to the consumer. In addition, customers are looking
for qualities that are linked to their physiological and hedonic demands in
order to satisfy dietary requirements, but rather to the industry’s competition.
Several proteome reviews published in the 2000s focused on technological
breakthroughs in proteomics, postgenomics technologies, and their limits in

improving product quality (Bendixen, 2005; Mullen et al., 2006; Paredi et
al., 2012). Researchers have employed proteomic research to link one or two
qualitative characteristics to a collection of putative biomarkers in order to
better understand the physiology and biology of animals, muscles, cells, and
molecules. By analyzing the protein content of the Semitendinosus muscle
of Charolais cows using 2DGE, Bouley et al. (2004) uncovered 129 different
proteins. Among the proteins found were those involved in cell structure,
cell defence, muscle metabolism, and the contractile machinery. In addition,
the proteomic method discovered other proteins, including ATP synthase
subunit alpha and alpha actinin 3, that may be involved in the modulation
of muscle calpains (Brule et al., 2010). Postmortem proteomics research
indicated that more than 100 unique protein loci had altered, most likely
owing to proteolysis (Lametsch and Bendixen, 2001). Troponin T, actin,
-crystallin, myokinase, creatine kinase, and mitochondrial ATPase were
shown to be altered in pork aged 72 hours by Morzel et al. (2004). Changes
in cryptic proteins and myozenin were also discovered. It was shown that the
amount of stress-related proteins in the sarcoplasmic fraction reduced during
ageing (Di Luca et al., 2013). It was discovered that Hsp70-1B, a minor
heat shock protein, was a biomarker for decreased tenderness in beef among
the 21 proteins tested in the study (Picard et al., 2014). Morzel et al. (2004)
found that pigs transported the day before slaughter had a lower level of
mitochondrial ATPase activity than pigs transported the day before slaughter.
Protein profiling of chicken hearts was performed by Amid et al. (2012) to
identify troponin I and alpha cardiac muscle actin 1 as putative electrically
stunned markers. A mass spectrometry technique developed by Von Bargen
et al. (2013) can identify even the tiniest amounts of pork and horse meat in
halal beef samples. It was shown that the 2DE protein profiles of raw and
processed meat products differed among pig, chicken, cow, turkey, duck, and
goose species (Montowska and Pospiech, 2013).
Proteins that are critical to meat quality, such as pork color, drip loss,
or shear force, can be identified via proteomic investigations, which look
at a subset of the cellular protein composition at a particular time and often
include several hundred proteins (Laville et al., 2009). Bouley et al. (2004)
used 2-DE and MS to identify 75 gene products in bovine skeletal muscle and
found that considerable cross-species identifications were achievable across
bovine, human, mouse, and rat. Doherty et al. (2004) used 2-DE to separate
soluble extracts from muscle homogenates from broiler and layer chickens,
and then performed MALDI-TOF mass spectrometry on the targeted spots. A
study by Hamelin et al. (2006) employed 2-DE and MALDI-MS to analyze

the molecular basis of muscle fiber type-associated variability and found
the differential expression of proteins involved in oxidative metabolism like
citric acid cycle enzymes, proteins with functions linked to oxidative metabo-
lism, oxidative stress, and higher protein turnover. Houbak et al. (2008) used
2D gel proteome analysis to investigate the breakdown of bovine muscle
proteins by enzymes like proteasome and ubiquitous calpains. Hwang et
al. (2005) used a proteomics technique to examine postmortem proteolysis
in pig longissimus muscle and to investigate the association of proteolytic
degradation to drip loss, WB-shear force, and lightness value during chilling
ageing. It was suggested that developing muscle-specific processing strate-
gies to improve beef color was necessary by Joseph et al. (2010), who studied
the sarcoplasmic proteome to distinguish between muscles that were stable
to liable in color. With DIGE and PMF Hollung et al. (2009) investigated the
effect on adductor muscle size of age and breed.
The welfare of animals, as well as genetics, may have an impact on the
quality of meat. After a time of reduced food intake, compensatory develop-
ment in pigs has been linked to the tenderness of pork (Hollug et al., 2007;
Kristensen et al., 2007). This property was studied by Lametsch et al. (2006)
using proteomics, and they identified a correlation between this trait and the
differential expression of protein. Stress and glycolytic proteins were shown
to be decreased using 2D electrophoresis and MALDI TOF MS, which were
employed in combination with a standard sample preparation. It has been
shown by Liu et al. (2014) that a high-fat diet that alters the expression of
stress proteins in muscle 2-DE was utilized in conjunction with MS to assess
the quality of pig meat. A total of 27 proteins were found to have altered in
response to ageing and changes in meat quality parameters. The majority of
the proteins contained cytoskeletal and metabolic proteins and degradation
products associated with those proteins. Research by Di Luca et al. (2013)
using gel-based techniques (2D-DIGE and WB) indicated that fresh pork
meat had a lower concentration of stress response proteins than dried pork
meat.
12.6 APPLICATION OF PROTEOMICS IN MEAT SAFETY AND

AUTHENTICITY
Authentication of meat has long been a top priority for producers, authorities,
and consumers in the contemporary period. A clear and detailed description
of the meat that people consume, especially in processed forms when visual
distinction of components is only feasible, is required by health-conscious
customers. The issues of food authenticity in meat and meat products are
categorized by Montowska and Pospiech (2012) which are shown in Table
12.1.
TABLE 12.1 Major Categories of Food Authenticity Issues Found in Meat and Meat
Products
Sl. No. Category of food fraud References
1 Substituting cheaper meat species for more Mane et al. (2012); Girish et al. (2015)
expensive ones
2 Substances of animal origin (Offal, mechanically Velioglu et al. (2018); Fengou et al.
recovered meat) that have not been declared (2021); Surowiec et al. (2011)
3 Contradictions in the product’s labeling or Pinto et al. (2015)
specifications
4 Sale of previously frozen fresh beef Alamprese et al. (2016)
5 Inadequate geographic origin labeling of items Zhao et al. (2020)
6 Differentiation between meat from domesticated Amaral et al. (2015)
animals and meat from wild animals
7 Inadvertent inclusion of plant or dairy proteins Salomé et al. (2021)
8 Product sales of nonecological items as ecological Stawitz et al. (2017)
products
Proteomics can significantly contribute in the knowledge of the processes

of infection, antibiotic resistance, and biofilm development of these food-
borne pathogens. An extensive body of literature exists addressing the
discoveries that have been made possible by proteomics methods. A direct
technique for the identification of enterohemorrhagic Escherichia coli
O157:H7 was previously reported by Ochoa and Harrington (2005). In a
recent study, Fagerquist and colleagues employed MALDI-TOF-TOF and
tandem mass spectrometry (MS/MS) to identify distinct subtypes of Shiga
toxin-producing E. coli (STEC) (Fagerquist et al., 2014). Recent studies
employing 2D electrophoresis for the identification of allergens in various
foods include studies on beer (Picariello et al., 2015), milk (Hettinga et al.,
2015), and beef (Apostolovic et al., 2014).
Peptide markers indicating buffalo meat contamination in sheep meat
combinations as low as 0.5% were identified using a proteomic technique,
which was successful in both raw and cooked meat mixtures (Naveena et
al., 2017; 2018). The researchers Fornal and Montowska (2019) used liquid
chromatography and high-resolution Q-TOF mass analyzers to effectively
detect unique peptide markers in highly processed composite food products

from duck, goose, and chicken. Montowska and Pospiec (2013) discovered
interspecies differences in 2-DE protein patterns in raw and processed meat
from cow, pig, chicken, turkey, duck, and goose. They also revealed changes
in 2-DE protein patterns in raw and processed meat from turkey, duck,
and geese (2013). As potential markers, scientists suggested employing
proteins that have been shown to be thermally stable and stable over long
periods of time in the beef resource. Chicken contamination was detected
by Sentandreu et al. (2010) utilizing OFFGEL isoelectric focusing, followed
by MALDI–TOF MS and LC–ESI–MS/MS/MS analysis of species-specific
peptide biomarkers in a combination of pig and chicken meat. A metabolo-
mics and lipidomics technique was used by Trivedi et al. (2016) to identify
pork adulteration in beef. Volatile molecules can be used as a biomarker
for the detection of meat adulteration since various meat species have
distinct flavors. Using an electronic nose and GC-MS, the research teams
of Nurjulianaet al. (2011) and Haddie et al. (2015) were able to detect meat
adulteration. Hu et al. (2017) used Fourier transform infrared spectroscopy
to identify 10% of animal offal in ground beef with 99% accuracy (FT-IRS).
Lee et al. (2018) recently used the RS approach to effectively identify beef
tallow from pork lard, chicken fat from duck oil, and other animal fats as
well. It was shown that 46 proteins were differently expressed in the muscle
proteome of sheep exposed to stunning compared to non-stunning, showing
alterations in the quantity of proteins involved in structural, catalytic, and
stress-related processes (Kiran et al., 2019; 2021).
12.7 FUTURE TRENDS
Protein/peptide biomarkers can be used to verify the authenticity of food,

although they are still a relatively new approach. However, they have the
potential to be used in the future because of their high sensitivity, resilience,
multiplexing capacity, and high-throughput and predictive power. A key
limitation of DNA approaches is that DNA degradation occurs in highly
processed samples (Buckey et al., 2009), peptide-based methods avoid this
problem. Proteomic tool advancements have also led to their application
in addressing safety and authenticity concerns such as the identification
of meat species, detection of animal byproducts, non-meat components
and tissues, traceability, identification of genetically modified ingredients,
chemical residues, and other harmful substances. Understanding the impact
on transportation, stunning, slaughter stress, halal verification, and difficul-

ties linked to animal welfare are some of the possible areas where proteomic
methods are being applied (Banerjee et al., 2022). Researchers should make
an attempt in future research to include essential aspects that have been
overlooked in the majority of previous studies. The tests will become more
sensitive and specific when high-resolution MS instruments improve their
resolution. Between initial discovery investigations and final validation
trials, targeted MS/MS acquisition methods can help to fill the gap between
the two. Innovative food proteome/matrix bioinformatic tools, as well as the
existing ones, will be critical in advancing the fields of integrative biology
and systems biology in the food industry. The use of high-throughput tech-
nologies, such as mass spectrometry and NMR-based metabolomics, mass
spectrometry and NMR-based metabolomics, quantitative proteomics, and
lipidomics, is a recent trend in farm animal research (Wang et al., 2012;
Zhao et al., 2012). The scientific community should work together to share
publicly available databases and data sets and to produce certified reference
materials and standard operating procedures (Ortea et al., 2016).
12.8 CONCLUSION
The use of proteomic techniques is speeding up the understanding of the

biological pathways and mechanisms that support meat quality. The meat
industry has a long-term need and a significant technological difficulty
in analyzing several targets on a single-test platform. As a potent tool for
studying protein alterations generated by varied situations, proteomics is
indispensable. Global protein analyses are supposed to provide a more direct
knowledge of protein function and regulation than gene analyses. Genes and
proteins that govern crop design and/or stress resistance in a wide variety
of settings must be discovered in order to promote biological crop improve-
ment in the midst of present food insecurity issues. The use of a wide array
of procedures to protect consumers by screening for adulteration in the
meat value supply chain is underway. In order to improve the precision and
reproducibility of proteomic and metabolomic approaches, efforts should
be made to standardize the extraction of analytes. Integrating genomes,
transcriptomics, proteomics, and metabolomics will improve the accuracy
and precision of authentication results, as would a more comprehensive
approach. Proteomics investigations in meat science have been limited by a
lack of access to high-throughput proteomic approaches that need expensive
apparatus and highly qualified technical staff that are typically unaffordable
among most meat science research centers. It is also necessary to expand
collaboration between scientists, proteomics platforms, and the meat industry
itself in order to solve these obstacles.
KEYWORDS
• food production
• omics technology
• bioinformatics
• proteomics methods
• meat quality
• meat safety
REFERENCES
Aaslyng, M. D. Quality Indicators for Raw Meat. Meat Processing: Improving Quality,
157–174.
Alamprese, C.; Amigo, J. M.; Casiraghi, E.; Engelsen, S. B. Identification and Quantification
of Turkey Meat Adulteration in Fresh, Frozen-Thawed and Cooked Minced Beef by FT-NIR
Spectroscopy and Chemometrics. Meat Sci. 2016, 121, 175–181.
Amaral, J. S.; Santos, C. G.; Melo, V. S.; Costa, J.; Oliveira, M. B. P.; Mafra, I. Identification
of Duck, Partridge, Pheasant, Quail, Chicken and Turkey Meats by Species-Specific PCR
Assays to Assess the Authenticity of Traditional Game Meat Alheira Sausages. Food
Control 2015, 47, 190–195.
Amid, A.; Samah, N. A.; Yusof, F. Identification of Troponin I and Actin, Alpha Cardiac
Muscle 1 as Potential Biomarkers for Hearts of Electrically Stimulated Chickens. Proteome
Sci. 2012, 10 (1), 1–8.
Amiour, N.; Merlino, M.; Leroy, P.; Branlard, G. Proteomic Analysis of Amphiphilic Proteins
of Hexaploid Wheat Kernels. Proteomics 2002, 2(6), 632–641.
Apostolovic, D.; Tran, T. A. T.; Hamsten, C.; Starkhammar, M.; Cirkovic Velickovic, T.; van
Hage, M. Immunoproteomics of Processed Beef Proteins Reveal Novel Galactose‐α‐1, 3‐
Galactose‐Containing Allergens. Allergy 2014, 69 (10), 1308–1315.
Banerjee, R.; Maheswarappa, N. B.; Mohan, K.; Biswas, S.; Batabyal, S. Proteomic
Technologies and Their Application for Ensuring Meat Quality, Safety and Authenticity.
Curr. Proteomics 2022, 19 (2), 128–141.
Bendixen, E. The Use of Proteomics in Meat Science. Meat Sci. 2005, 71 (1), 138–149.
Bertram, H. C.; Whittaker, A. K.; Shorthose, W. R.; Andersen, H. J.; Karlsson, A. H. Water
Characteristics in Cooked Beef as Influenced by Ageing and High-Pressure Treatment—An
NMR micro imaging study. Meat Sci. 2004, 66 (2), 301–306.
Bouley, J.; Chambon, C.; Picard, B. Mapping of Bovine Skeletal Muscle Proteins Using
Two‐Dimensional Gel Electrophoresis and Mass Spectrometry. Proteomics 2004, 4 (6),
1811–1824.
Brulé, C.; Dargelos, E.; Diallo, R.; Listrat, A.; Bechet, D.; Cottin, P.; Poussard, S. Proteomic
Study of Calpain Interacting Proteins During Skeletal Muscle Aging. Biochimie 2010, 92
(12), 1923–1933.
Buckley, M.; Collins, M.; Thomas‐Oates, J.; Wilson, J. C. Species Identification by Analysis
of Bone Collagen Using Matrix‐Assisted Laser Desorption/Ionisation Time‐of‐Flight Mass
Spectrometry. Rapid Commun. Mass Spectrometry 2009, 23 (23), 3843–3854.
Campbell, P. A Cast of Thousands. Nat Biotechnol. 2003, 21, 213.
Di Luca, A.; Elia, G.; Mullen, A. M.; Hamill, R. M. Monitoring Post Mortem Changes in
Porcine Muscle Through 2-D DIGE Proteome Analysis of Longissimus Muscle Exudate.
Proteome Sci. 2013, 11 (1), 1–14.
Di Pinto, A.; Bottaro, M.; Bonerba, E.; Bozzo, G.; Ceci, E.; Marchetti, P. et al. Occurrence
of Mislabeling in Meat Products Using DNA-Based Assay. J. Food Sci. Technol. 2015, 52
(4), 2479–2484.
Doherty, M. K.; McLean, L.; Beynon, R. J. Avian Proteomics: Advances, Challenges and New
Technologies. Cytogenetic Genome Res. 2007, 117 (1–4), 358–369.
Eng, J. K.; McCormack, A. L.; Yates, J. R. An Approach to Correlate Tandem Mass Spectral
Data of Peptides with Amino Acid Sequences in a Protein Database. J. Am. Soc. Mass
Spectrometry 1994, 5 (11), 976–989.
Fagerquist, C. K.; Zaragoza, W. J.; Sultan, O.; Woo, N.; Quiñones, B.; Cooley, M. B.;
Mandrell, R. E. Top-Down Proteomic Identification of Shiga Toxin 2 Subtypes from Shiga
Toxin-Producing Escherichia coli by Matrix-Assisted Laser Desorption Ionization–Tandem
Time of Flight Mass Spectrometry. Appl. Environ. Microbiol. 2014, 80 (9), 2928–2940.
Feasson, L.; Stockholm, D.; Freyssenet, D.; Richard, I.; Duguez, S.; Beckmann, J. S.; Denis,
C. Molecular Adaptations of Neuromuscular Disease‐Associated Proteins in Response to
Eccentric Exercise in Human Skeletal Muscle. J. Physiol. 2002, 543 (1), 297–306.
Feder, M. E.; Hofmann, G. E. Heat-Shock Proteins, Molecular Chaperones, and the Stress
Response: Evolutionary and Ecological Physiology. Annu. Rev. Physiol. 1999, 61 (1),
243–282.
Fengou, L. C.; Lianou, A.; Tsakanikas, P.; Mohareb, F.; Nychas, G. J. E. Detection of Meat
Adulteration Using Spectroscopy-Based Sensors. Foods 2021, 10 (4), 861.
Fornal, E.; Montowska, M. Species-Specific Peptide-Based Liquid Chromatography–Mass
Spectrometry Monitoring of Three Poultry Species in Processed Meat Products. Food
Chem. 2019, 283, 489–498.
Friedman, D. B. An Introduction to Proteomics Technologies for the Genomics Scientist.
Genomics: Essential Methods 2010, 307–323.
Girish, P. S.; Anjaneyulu, A. S. R.; Viswas, K. N.; Shivakumar, B. M.; Anand, M.; Patel,
M.; Sharma, B. Meat Species Identification by Polymerase Chain Reaction-Restriction
Fragment Length Polymorphism (PCR-RFLP) of Mitochondrial 12S rRNA Gene. Meat
Sci. 2005, 70 (1), 107–112.
Haddi, Z.; El Barbri, N.; Tahri, K.; Bougrini, M.; El Bari, N.; Llobet, E.; Bouchikhi, B.
Instrumental Assessment of Red Meat Origins and Their Storage Time Using Electronic
Sensing Systems. Anal. Methods 2015, 7(12), 5193–5203.
Hamelin, M.; Sayd, T.; Chambon, C.; Bouix, J.; Bibé, B.; Milenkovic, D. et al. Proteomic
Analysis of Ovine Muscle Hypertrophy. J. Anim. Sci. 2006, 84 (12), 3266–3276.
Henzel, W. J.; Billeci, T. M.; Stults, J. T.; Wong, S. C.; Grimley, C.; Watanabe, C. Identifying
Proteins from Two-Dimensional Gels by Molecular Mass Searching of Peptide Fragments
in Protein Sequence Databases. Proc. Natl. Acad. Sci. 1993, 90 (11), 5011–5015.
Hettinga, K. A.; Reina, F. M.; Boeren, S.; Zhang, L.; Koppelman, G. H.; Postma, D. Set al.
Difference in the Breast Milk Proteome Between Allergic and Non-Allergic Mothers. PloS
One 2015, 10 (3), e0122234.
Hollung, K.; Grove, H.; Færgestad, E. M.; Sidhu, M. S.; Berg, P. Comparison of Muscle
Proteome Profiles in Pure Breeds of Norwegian Landrace and Duroc at Three Different
Ages. Meat Sci. 2009, 81 (3), 487–492.
Houbak, M. B.; Ertbjerg, P.; Therkildsen, M. In Vitro Study to Evaluate the Degradation of
Bovine Muscle Proteins Post-Mortem by Proteasome and μ-Calpain. Meat Sci. 2008, 79
(1), 77–85.
Hu, R.; He, T.; Zhang, Z.; Yang, Y.; Liu, M. Safety Analysis of Edible Oil Products via Raman
Spectroscopy. Talanta 2019, 191, 324–332.
Hu, Y.; Zou, L.; Huang, X.; Lu, X. Detection and Quantification of Offal Content in Ground
Beef Meat Using Vibrational Spectroscopic-Based Chemometric Analysis. Sci. Rep. 2017,
7 (1), 1–9.
Hwang, I. H.; Park, B. Y.; Kim, J. H.; Cho, S. H.; Lee, J. M. Assessment of Postmortem
Proteolysis by Gel-Based Proteome Analysis and Its Relationship to Meat Quality Traits in
Pig Longissimus. Meat Sci. 2005, 69 (1), 79–91.
Iwahashi, Y.; Hosoda, H. Effect of Heat Stress on Tomato Fruit Protein Expression.
Electrophoresis 2000, 21 (9), 1766–1771.
Jensen, O. N. Modification-Specific Proteomics: Characterization of Post-Translational
Modifications by Mass Spectrometry. Curr. Opin. Chem. Biol. 2004, 8 (1), 33–41.
Joseph, P.; Suman, S. P.; Rentfrow, G.; Li, S.; Beach, C. M. Proteomics of Muscle-Specific
Beef Color Stability. J. Agric. Food Chem. 2012, 60 (12), 3196–3203.
Kiran, M.; Maheswarappa, N. B.; Banerjee, R.; Ch, V.; Rapole, S. Impact of Stunning Before
Slaughter on Expression of Skeletal Muscles Proteome in Sheep. Anim. Biotechnol. 2021,
1–8.
Kiran, M.; Naveena, B. M.; Reddy, K. S.; Shahikumar, M.; Reddy, V. R.; Kulkarni, V. V. et al.
Understanding Tenderness Variability and Ageing Changes in Buffalo Meat: Biochemical,
Ultrastructural and Proteome Characterization. Animal 2016, 10 (6), 1007–1015.
Kiran, M.; Naveena, B. M.; Reddy, K. S.; Shashikumar, M.; Reddy, V. R.; Kulkarni, V. V. et
al. Muscle‐Specific Variation in Buffalo (B ubalus bubalis) Meat Texture: Biochemical,
Ultrastructural and Proteome Characterization. J. Text. Stud. 2015, 46 (4), 254–261.
Kiran, M.; Naveena, B. M.; Smrutirekha, M.; Reddy, P. B.; Rituparna, B.; Kumar, Y. P. et al.
Traditional Halal Slaughter Without Stunning Versus Slaughter with Electrical Stunning of
Sheep (Ovis aries). Meat Sci. 2019, 148, 127–136.
Kristensen, L.; Therkildsen, M.; Aaslyng, M. D.; Oksbjerg, N.; Ertbjerg, P. Compensatory
Growth Improves Meat Tenderness in Gilts But Not in Barrows. J. Anim. Sci. 2004, 82
(12), 3617–3624.
Kvasnička, F. Proteomics: General Strategies and Application to Nutritionally Relevant
Proteins. J. Chromatogr. B 2003, 787 (1), 77–89.
Lametsch, R.; Bendixen, E. Proteome Analysis Applied to Meat Science: Characterizing Post
Mortem Changes in Porcine Muscle. J. Agric. Food Chem. 2001, 49 (10), 4531–4537.
Lametsch, R.; Karlsson, A.; Rosenvold, K.; Andersen, H. J.; Roepstorff, P.; Bendixen, E.
Postmortem Proteome Changes of Porcine Muscle Related to Tenderness. J. Agric. Food
Chem. 2003, 51 (24), 6992–6997.
Lametsch, R.; Kristensen, L.; Larsen, M. R.; Therkildsen, M.; Oksbjerg, N.; Ertbjerg, P.
Changes in the Muscle Proteome After Compensatory Growth in Pigs. J. Anim. Sci. 2006,
84 (4), 918–924.
Laville, E.; Sayd, T.; Morzel, M.; Blinet, S.; Chambon, C.; Lepetit, J. et al. Proteome Changes
During Meat Aging in Tough and Tender Beef Suggest the Importance of Apoptosis and
Protein Solubility for Beef Aging and Tenderization. J. Agric. Food Chem. 2009, 57 (22),
10755–10764.
Lee, J. Y.; Park, J. H.; Mun, H.; Shim, W. B.; Lim, S. H.; Kim, M. G. Quantitative Analysis
of Lard in Animal Fat Mixture Using Visible Raman Spectroscopy. Food Chem. 2018, 254,
109–114.
Lee, T. H.; Wani, W. A.; Koay, Y. S.; Kavita, S.; Tan, E. T. T.; Shreaz, S. Recent Advances in
the Identification and Authentication Methods of Edible Bird’s Nest. Food Res. Int. 2017,
100, 14–27.
Leheska, J. M.; Thompson, L. D.; Howe, J. C.; Hentges, E.; Boyce, J.; Brooks, J. C. et al.
Effects of Conventional and Grass-Feeding Systems on the Nutrient Composition of Beef.
J. Anim. Sci. 2008, 86 (12), 3575–3585.
Liu, J.; He, J.; Yu, J.; Mao, X.; Zheng, P.; Huang, Z. et al. Birth Weight Alters the Response
to Postnatal High-Fat Diet-Induced Changes in Meat Quality Traits and Skeletal Muscle
Proteome of Pigs. Br. J. Nutr. 2014, 111 (10), 1738–1747.
Mallick, P.; Kuster, B. Proteomics: A Pragmatic Perspective. Nat. Biotechnol. 2010, 28 (7),
695–709.
Mane, B. G.; Mendiratta, S. K.; Tiwari, A. K.; Bhilegaokar, K. N. Detection of Adulteration of
Meat and Meat Products with Buffalo Meat Employing Polymerase Chain Reaction Assay.
Food Anal. Methods 2012, 5 (2), 296–300.
Miller, R. K. Factors Affecting the Quality of Raw Meat. Meat Processing 2002, 27–63.
Montowska, M.; Pospiech, E. Is Authentication of Regional and Traditional Food Made of
Meat Possible? Crit. Rev. Food Sci. Nutr. 2012, 52 (6), 475–487.
Montowska, M.; Pospiech, E. Species-Specific Expression of Various Proteins in Meat
Tissue: Proteomic Analysis of Raw and Cooked Meat and Meat Products Made from Beef,
Pork and Selected Poultry Species. Food Chem. 2013, 136 (3–4), 1461–1469.
Morzel, M.; Chambon, C.; Hamelin, M.; Santé-Lhoutellier, V.; Sayd, T.; Monin, G. Proteome
Changes During Pork Meat Ageing Following Use of Two Different Pre-Slaughter Handling
Procedures. Meat Sci. 2004, 67 (4), 689–696.
Mullen, A. M.; Stapleton, P. C.; Corcoran, D.; Hamill, R. M.; White, A. Understanding Meat
Quality Through the Application of Genomic and Proteomic Approaches. Meat Sci. 2006,
74 (1), 3–16.
Nair, M. N.; Zhai, C. Application of Proteomic Tools in Meat Quality Evaluation. In Meat
Quality Analysis; Academic Press, 2020; pp 353–368.
Naveena, B. M.; Jagadeesh, D. S.; Babu, A. J.; Rao, T. M.; Kamuni, V.; Vaithiyanathan,
S. et al. OFFGEL Electrophoresis and Tandem Mass Spectrometry Approach Compared
with DNA-Based PCR Method for Authentication of Meat Species from Raw and Cooked
Ground Meat Mixtures Containing Cattle Meat, Water Buffalo Meat and Sheep Meat. Food
Chem. 2017, 233, 311–320.
Naveena, B. M.; Jagadeesh, D. S.; Kamuni, V.; Muthukumar, M.; Kulkarni, V. V.; Kiran, M.;
Rapole, S. In‐Gel and OFFGEL‐Based Proteomic Approach for Authentication of Meat
Species from Minced Meat and Meat Products. J. Sci. Food Agric. 2018, 98 (3), 1188–1196.
Nurjuliana, M.; Man, Y. C.; Hashim, D. M.; Mohamed, A. K. S. Rapid Identification of
Pork for Halal Authentication Using the Electronic Nose and Gas Chromatography Mass
Spectrometer with Headspace Analyzer. Meat Sci. 2011, 88 (4), 638–644.
Ochoa, M. L.; Harrington, P. B. Immunomagnetic Isolation of Enterohemorrhagic Escherichia
coli O157: H7 from Ground Beef and Identification by Matrix-Assisted Laser Desorption/
Ionization Time-of-Flight Mass Spectrometry and Database Searches. Anal. Chem. 2005,
77 (16), 5258–5267.
Offer, G.; Knight, P.; Jeacocke, R.; Almond, R.; Cousins, T.; Elsey, J. et al. The Structural
Basis of the Water-Holding, Appearance and Toughness of Meat and Meat Products. Food
Struct. 1989, 8 (1), 17.
Ortea, I.; Cañas, B.; Gallardo, J. M. Selected Tandem Mass Spectrometry Ion Monitoring for
the Fast Identification of Seafood Species. J. Chromatogr. A 2011, 1218 (28), 4445–4451.
Paredi, G.; Sentandreu, M. A.; Mozzarelli, A.; Fadda, S.; Hollung, K.; de Almeida, A. M.
Muscle and Meat: New Horizons and Applications for Proteomics on a Farm to Fork
Perspective. J. Proteomics 2013, 88, 58–82.
Perng, M. D.; Cairns, L.; Van Den IJssel, P.; Prescott, A.; Hutcheson, A. M.; Quinlan, R. A.
Intermediate Filament Interactions Can be Altered by HSP27 and alphaB-Crystallin. J. Cell
Sci. 1999, 112 (13), 2099–2112.
Picard, B.; Gagaoua, M.; Micol, D.; Cassar-Malek, I.; Hocquette, J. F.; Terlouw, C. E. Inverse
Relationships Between Biomarkers and Beef Tenderness According to Contractile and
Metabolic Properties of the Muscle. J. Agric. Food Chem. 2014, 62 (40), 9808–9818.
Picariello, G.; Mamone, G.; Nitride, C.; Ferranti, P. Proteomic Analysis of Beer. In Proteomics
in Food Science; Academic Press, 2017; pp 383–403.
Polati, R.; Menini, M.; Robotti, E.; Millioni, R.; Marengo, E.; Novelli, E. et al. Proteomic
Changes Involved in Tenderization of Bovine Longissimus Dorsi Muscle During Prolonged
Ageing. Food Chem. 2012, 135 (3), 2052–2069.
Salomé, M.; Huneau, J. F.; Le Baron, C.; Kesse-Guyot, E.; Fouillet, H.; Mariotti, F. Substituting
Meat or Dairy Products with Plant-Based Substitutes Has Small and Heterogeneous Effects
on Diet Quality and Nutrient Security: A Simulation Study in French Adults (INCA3). J.
Nutr. 2021, 151 (8), 2435–2445.
Sayd, T.; Morzel, M.; Chambon, C.; Franck, M.; Figwer, P.; Larzul, C. et al. Proteome
Analysis of the Sarcoplasmic Fraction of Pig Semimembranosus Muscle: Implications on
Meat Color Development. J. Agric. Food Chem. 2006, 54 (7), 2732–2737.
Sentandreu, M. A.; Fraser, P. D.; Halket, J.; Patel, R.; Bramley, P. M. A proteomic-Based
Approach for Detection of Chicken in Meat Mixes. J. Proteome Res. 2010, 9 (7), 3374–3383.
Stawitz, C. C.; Siple, M. C.; Munsch, S. H.; Lee, Q.; Derby, S. R. Financial and Ecological
Implications of Global Seafood Mislabeling. Conserv. Lett. 2017, 10 (6), 681–689.
Surowiec, I.; Fraser, P. D.; Patel, R.; Halket, J.; Bramley, P. M. Metabolomic Approach for
the Detection of Mechanically Recovered Meat in Food Products. Food Chem. 2011, 125
(4), 1468–1475.
Toda, M.; Ono, S. J. Genomics and Proteomics of Allergic Disease. Immunology 2002, 106
(1), 1–10.
Trivedi, D. K.; Hollywood, K. A.; Rattray, N. J.; Ward, H.; Trivedi, D. K.; Greenwood, J. et
al. Meat, the Metabolites: An Integrated Metabolite Profiling and Lipidomics Approach for
the Detection of the Adulteration of Beef with Pork. Analyst 2016, 141 (7), 2155–2164.
Vaidyanathan, S.; Goodacre, R. Proteome and Metabolome Analyses for Food Authentication.
In Food Authenticity and Traceability; Michele L. Wood Head Publishing, 2003.
Vanderwerf, S. M.; Cooper, M. J.; Stetsenko, I. V.; Lutsenko, S. Copper Specifically
Regulates Intracellular Phosphorylation of the Wilson’s Disease Protein, a Human Copper-
Transporting ATPase. J. Biol. Chem. 2001, 276 (39), 36289–36294.
Velioglu, H. M.; Sezer, B.; Bilge, G.; Baytur, S. E.; Boyaci, I. H. Identification of Offal
Adulteration in Beef by Laser Induced Breakdown Spectroscopy (LIBS). Meat Sci. 2018,
138, 28–33.
Verbeke, W.; Pérez-Cueto, F. J.; de Barcellos, M. D.; Krystallis, A.; Grunert, K. G. European
Citizen and Consumer Attitudes and Preferences Regarding Beef and Pork. Meat Sci. 2010,
84 (2), 284–292.
von Bargen, C.; Dojahn, J.; Waidelich, D.; Humpf, H. U.; Brockmeyer, J. New Sensitive
High-Performance Liquid Chromatography–Tandem Mass Spectrometry Method for
the Detection of Horse and Pork in Halal Beef. J. Agric. Food Chem. 2013, 61 (49),
11986–11994.
Wang, G. J.; Zhou, G. Y.; Ren, H. W.; Xu, Y.; Yang, Y.; Guo, L. H.; Liu, N. Peptide Biomarkers
Identified by LC–MS in Processed Meats of Five Animal Species. J. Food Compos. Anal.,
73, 47–54.
Yu, J.; Tang, S.; Bao, E.; Zhang, M.; Hao, Q.; Yue, Z. The Effect of Transportation on the
Expression of Heat Shock Proteins and Meat Quality of M. Longissimus dorsi in Pigs. Meat
Sci. 2009, 83 (3), 474–478.
Zapata, I.; Reddish, J. M.; Miller, M. A.; Lilburn, M. S.; Wick, M. Comparative Proteomic
Characterization of the Sarcoplasmic Proteins in the Pectoralis Major and Supracoracoideus
Breast Muscles in 2 Chicken Genotypes. Poultry Sci. 2012, 91 (7), 1654–1659.
Zhao, J.; Li, A.; Jin, X.; Pan, L. Technologies in Individual Animal Identification and Meat
Products Traceability. Biotechnol. Biotechnol. Equip. 2020, 34 (1), 48–57.
Zhao, Y.; Zhang, B.; Chen, G.; Chen, A.; Yang, S.; Ye, Z. Tracing the Geographic Origin
of Beef in China on the Basis of the Combination of Stable Isotopes and Multielement
Analysis. J. Agric. Food Chem. 2013, 61 (29), 7055–7060.
CHAPTER 13
Food Waste Revalorization as a

Functional Food or Ingredient
ESTRADA-GIL L. E.1, N. D. CERDA-CEJUDO1, CHÁVEZ-GONZÁLEZ M. L.1,
A. C. FLORES-GALLEGOS A. C.1, M. GOVEA-SALAS2,
CONTRERAS-ESQUIVEL J. C.1, C. N. AGUILAR-GONZÁLEZ1, and J. A.
ASCACIO-VALDÉS1
1
Bioprocesses & Bioproducts Group, Food Research Department, School of
Chemistry, Universidad Autónoma de Coahuila, Saltillo, Coahuila, México
2
Laboratory of Nanobiociences, School of Chemistry,
Universidad Autónoma de Coahuila, Ing. J. Cárdenas Valdéz S/N,
República, Saltillo, México
ABSTRACT
One of the greatest problems humanity faces nowadays is the loss of food and
the discarding of many different food sources that are not appealing to the
consumer, with little knowledge that these so-called “wastes” are what holds
one of the greatest sources of high-value bioactive compounds. Peels, leaves,
stems, and seeds of many fruits and vegetables have already been studied
and they possess phenolic compounds, and dietetic fibers among many other
biologically active compounds that may prove to benefit the consumer. In
this research the different methods to apply agricultural by-products from
different fruits and vegetables are shown, varying from their use in bakery
products to yogurt ingredients to fortify and enhance the already healthy
attributes some foods have.
13.1 INTRODUCTION
One of the world’s greatest problems in modern times is not the production
of food, as we can produce enough food to nourish every single person on
the planet, but instead, the biggest problem humanity faces at this time is
the untimely loss of food before it can be consumed as it is estimated by the
World Food Programme that at least one-third of the food produced globally
is lost in retail and the fridges of many people around the world (WFP, 2020).
Data from the Food and Agriculture Organization (FAO) estimates that
around 14% of the food of the world is lost in the process of the shipping
of the products from harvest to retail which represents a loss of $400 billion
annually and an estimated amount of 17% of all the food purchased and kept
in households and retail goes to waste (FAO, 2021).
COVID-19 also changed the amount of waste that is produced world-
wide, as people stayed locked down in their households many food products
like fruits, vegetables, poultry, and beef were lost as the government lock-
down caused huge food losses in food market retail as well as in agricultural
production.
Not only food waste is an economical problem, but it is also an ethical,
environmental, and social problem that will just keep on increasing as time
goes by and food prices keep going up. International organizations, NGOs,
governments, and many branches of the United Nations have declared that
this is an issue of utmost importance as it affects all levels of society. As an
ethical and social problem, it must be addressed that many people around
the globe do not possess access to a source of food or are unable to buy food
products for the constant uprising in prices (Edjabou et al., 2016; Ponis et
al., 2017; Roodhuyzen et al., 2017). As an environmental problem, the high
amounts of food wastes such as peels, seeds, pulps, and rotten or spoiled
foods produce high amounts of greenhouse gases and not only that, as an
example, many residues from the coffee industry are left behind in roads just
to decompose, this waste does not only contaminate the air with gases but
also the soil due to high amounts of antinutrients present in the pulp and peel
of the coffee, just to state an example, contributing to climate change and
future problems in the use of soil (Arya et al., 2022).
The valorization approach for food waste can differ from each different
kind of waste as not all of them are suitable for human consumption as they
are produced, some may contain carbohydrates, proteins, fats, and other
nutraceutical compounds that can be reused to obtain energy, biofuels,
enzymes, antioxidant extracts, novel biodegradable materials, and other
commercial products (Duenas and Garciá-Estévez, 2020).
Food Waste Revalorization as a Functional Food or Ingredient 323
Food waste is by no means, inevitable as it is a vicious circle, where the

production and consumption of food will always produce waste, but how we
approach this waste is part of the answer. Nowadays, companies, producers,
and retailers possess new policies for zero waste, for example, the wine
industry donates the bagasse of the grapes that were used in the production
of wine to extract and obtain high-value compounds which can be used to
elaborate different products like fungicides (da Silva et al., 2022), feed for
shrimps (Rosas et al., 2022), and even cookies with antioxidant activity
(Fontana et al., 2022).
Functional ingredients represent one of the best alternatives in the treat-
ment of food waste, as many of the wastes such as peels, seeds, bagasse,
and leaves of fruits, vegetables, cereals, and legumes have many bioactive
compound molecules which may confer the consumer great properties to
maintain health and prevent chronic diseases such as diabetes type 2, cardio-
vascular disease, mental diseases, and some kinds of cancer. The usage of
these compounds has an extensive background as there are studies that show
the possibility of the usage of certain agricultural food wastes (AFW) to
enrich pasta with phenolic compounds (Melini et al., 2020). Another usage
for agricultural waste is the elaboration of kombucha as stated in a study
by Liu et al., 2022. Another option for the usage of agricultural waste is the
elaboration of a functional bakery, where bananas, mango, pineapple, pome-
granate, and other peels were used in the elaboration of bakery products
(Ajila et al., 2008; Eshak, 2016; Ismail et al., 2014; Noor Aziah et al., 2011;
Wu and Shiau, 2015).
In this work, the usage of food waste will be explored in novel ways
to reuse and reduce the waste produced by the agricultural industry as
new functional foods, ingredients, and extracts to obtain high-added value
biological compounds will be reviewed.
13.2 FRUIT AND VEGETABLE WASTE REVALORIZATION
Fruits and vegetables are highly perishable food that has a high index of
being thrown to waste or considered as a loss in certain cases, but it must
be considered that loss and waste are not the same as loss can be considered
when the food is no longer edible, and waste is when the food does not get
eaten once in a household or retail market. Food loss is a problem that is
bigger in developing countries as they do not possess the technology, infra-
structure, or production practices that ensure the safety of food production.
Food waste is most likely to happen in developed countries as the consumer

has higher aesthetical standards where a fruit or vegetable that is not eye-
appealing to the customer will not get purchased and sooner than later it will
rot. In East and Southeast Asia as well as in sub-Saharan Africa some studies
show around 50% of the fruit and vegetables in those regions are lost due to
poor storage and production conditions; this calls for measures not only in
the valorization of this waste but also on the improvement of conditions on
which food is produced and stored (FAO, 2017, 2020).
The use of fruit and vegetable wastes as a functional ingredient has been
studied for a long time; this makes it possible to find different studies that take
advantage of these wastes, and here are some of the most recent examples of
how these wastes have been used.
13.3 FRUIT WASTE AS FUNCTIONAL INGREDIENTS
13.3.1 PROSOPIS NIGRA

Prosopis nigra also known as “Algarrobo negro” is a legume tree from South
America that can produce a pod that is very sweet, yellow and with purple
spots and it is used to produce flours and a fermented alcoholic beverage
(María J. Pérez et al., 2018).
This tree is very much appreciated in the regions of Argentina where it is
grown because practically all of the trees can be very well exploited, the bark
of this tree is very useful as it can be used as a great astringent and dye, its
wood is very useful as it has many uses for example construction, charcoal
and firewood, and the leaves are great fertilizers (María J. Pérez et al., 2014).
A study by María J. Pérez et al. (2020) utilized the waste obtained from
the flour manufacturing process, which was processed to obtain powders
with a fine particle size where F1, F2, and F3 were obtained, each with a
different size. All powders had similar properties as they all possessed great
fiber and fructose content, which are some desirable characteristics a powder
destined to be consumed can have. As macronutrients correspond the three
samples contained a similar amount of glucose around 65–67 g/100 g of
powder, 11.5–12.84 g/ 100 g of powder of proteins, and a low-fat content
with barely 0.87–1.44 g g/100 g of powder, presence of Mg, K, Na, Cu, Fe,
and Zn can be detected in this potential functional ingredient which makes it
a great option for revalorization and usage in food production.
HPLC/ESI/MS analysis of the powder detected the presence of various
polyphenolic compounds such as quercetin, kaempferol, coumaric acid,
ferulic acid, and vicenin II among others, the same compounds which
have been studied and proven to possess antioxidant, anti-inflammatory,
neuroprotective, vascular-protective, anticarcinogenic, antiplatelet, antihy-
pertensive and hypoglycemic activities just to mention some of them that can
grant a great positive effect on the consumer if this functional ingredient is
consumed (Bento-Silva et al., 2018; Cattaneo et al., 2019; Xiao et al., 2016).
13.3.2 APPLE WASTE
One of the most common kinds of waste in the food industry is the peel of the
fruit, as it is often considered to not be useful anymore, although some are
a great font of fiber and edible. In some fruits the peels constitute as much
as 40 to 50% of the total weight of the product and in others it is just around
20%, and this varies from fruit to fruit. Taking this information into account
fruit processing industries that only use the pulp of the fruit to elaborate their
products, generate a great deal of peel waste that just goes into the trash
to generate more pollution and create a bigger environmental problem for
future generations. Apple is by far one of the world’s most popular fruits as it
is the fourth most widely produced crop fruit in the world with a production
of 87 million tons (Wani et al., 2022).
A study by Lazari et al. (2019) demonstrated that it was possible to
incorporate peel and pulp from apples into a low-calorie ice cream product,
the peel was added in two different ways, cooked and natural, and three
formulations were made, each different in the amount and kind of ingredi-
ents used but they all contained cream, apple peel and pulp, skimmed milk,
condensed milk, and emulsifier. Among the three formulations, the best one
obtained an amount of 80.8 kcal, 11.3 g of carbohydrates, 1.36 g of protein,
and 2.92 g of lipids for every 100 g of ice cream. In this same study also,
sensory evaluations were performed obtaining great results for acceptations
with high values of 92% for sweetness, 92.2% of acceptation on flavor, and
84.7 acceptations for texture.
Apple pomace is by far one of the most popular functional ingredients
obtained from agricultural waste, as it is composed of the core, peel, seed,
pulp, and kernel of the fruit and it represents almost 20–35% of the fruit’s
weight when it is fresh (Rabetafika et al., 2014). It is estimated that around 4
million tons of apple pomace are produced worldwide by the juice industry,
making it a great problem to dispose of this waste effectively (J. M. Costa et
al., 2022).
A study by Curutchet et al. (2021) demonstrates how useful apple pomace

is, in the elaboration of cakes as a fiber source, as 40% of it is constituted
of dietary fiber which in long term can help in the treatment of obesity,
atherosclerosis, coronary heart diseases, intestine cancer, diabetes and can be
used as a potential prebiotic by stimulating the growth of probiotic bacteria
(Kruczek et al., 2017). For every 100 g, the fiber-enriched cake had 275 kcal,
43 g of carbohydrate, 4.8 g of protein, and 9.5 g of lipids of which 1.1 g are
from saturated fat and 6.3 g of fiber, compared to the regular cake which
had 332 kcal, 52 g of carbohydrate, 11 g of protein, 8.7 g of lipids out of
which 3.2 are from saturated fat and 1 g of fiber. The results benefit each
individually as a regular cake possesses more protein than its counterpart,
but its saturated fat content is higher than the fiber-enriched cake, also it is
important to remark the fact that the fiber content cannot be compared as this
cake made with apple pomace possesses greater fiber content with 6.3 g. A
sensorial evaluation was also performed in this test and the results show that
both cakes are very well accepted by consumers.
13.3.3 MANGO WASTE
Mango (Mangifera indica) is one of the most popular fruits harvested in

tropical countries around the world, with a production of 2.3 million tons per
year in 2021 (FAO, 2022b), mango is most commonly used in the elabora-
tion of liquors, juices, and for household consumption, but only the pulp of
the fruit is used in these types of consumption, leaving behind both the peel
and the seed which represent a great deal of the weight of the fruits with
around 50–60% of the total weight (Tirado-Kulieva et al., 2022).
The usage of mango peel flour has been reported as an ingredient in the
elaboration of yogurt, as it possesses a great amount of fiber which can be
considered prebiotic and polyphenols which may confer the product with
antioxidant activity among others that may be of interest to the consumer
(Pérez-Chabela et al., 2022). Yogurt is a fermented food that is very much
consumed worldwide as it is a great source of probiotics; it is a fermented
food with the presence of lactic acid bacteria such as the Lactobacillus
family. The introduction of mango peel as a fiber source is capable of making
yogurt formulations a type of symbiotic food, meaning that it is a combina-
tion of probiotics and prebiotics that give the consumer/host great health
benefits by improving the survival and colonization of selected bacteria in
the gastrointestinal tract (Fazilah et al., 2018).
It is well established that mango peel is a great source of dietary fiber,

but it is also a great antioxidant, as extracts with high polyphenolic content
can be obtained from it. Mango peel nanoparticles have been used in many
different studies regarding quality enhancement and shelf-life improvement,
for example in the elaboration of beef sausages where mango peel nanopar-
ticles proved to be a great way to maintain microbial quality for this product
when used in a concentration of 2% for 9 days (Boff et al., 2022).
Another study by Aamir et al. (2022) determined that mango peel could
be additioned to multigrain cookies and fortify them with their nutrients.
Cookies made with mango peel powder were analyzed and it was determined
that they contained 6.66% moisture, 2.17% ashes, protein in 8.17% of, and
fat at 1.67%. Sensory evaluation was performed on the cookies they proved
to possess high sensory scores and showed better results for physicochemical
and antioxidant analysis.
13.3.4 GRAPE WASTE
Grape (Vitis vinifera sativa) is one of the world’s most cultivated fruits all
around the world and it is of high relevance, not only for its consumption in
wine but it is also consumed fresh and dried and the fruit’s pomace possesses
great nutritional value as well as biological compounds with pharmaceu-
tical activities among which we can find, anti-inflammatory, antioxidant,
antihypertensive, among many others. The pomace is constituted of skin,
stems, seeds, and pulp which represents around 20% of the total weight of
the fruit (M. Costa et al., 2022), all of this is obtained in the natural process
of winemaking, as it is obtained by the crushing and pressing of the fruits,
considering that the world's production of wine in 2021 was 260 million
hectoliters, this represents a huge amount of waste produced every year.
With pomace there is a potential to fabricate flours, which can potentially
be used in the elaboration of cookies, bread, cereal bars, pasta, vitamins, and
juices (Oliveira et al., 2016) when pomace is used in the elaboration of these
products they are conferred with high polyphenolic content as anthocyanins,
flavonols, catechins, and proanthocyanins are present in this agricultural
by-product. These compounds are the ones that confer the pomace of their
important biological activities.
In a study by Marchiani et al. (2016) grape pomace is used as a functional
ingredient in the elaboration of yogurt to improve and add dietary fiber and
some antioxidant compounds, same that can help the probiotic bacteria in
the yogurt to colonize the gastrointestinal tract. This yogurt was prepared
using UHT whole milk with 36 g/kg of fat, 31 g/kg of protein, and 48 g/kg
of carbohydrates, milk powder at 3% was added and the milk was inoculated
with a culture of YO-MIX 401 (Santamaria, Burago di Molgora, Italy)
which contained a mixture of Streptococcus thermophilus and Lactobacillus
dellbrueckii. Milk was fermented at 42°C until a pH of 4.8 was obtained and
then the grape pomace was added to the mix; there were 3 types of pomace
used from different varieties such as Chardonnay, Moscato, and Pinot noir.
The addition of the pomace to the yogurt resulted in the finding of various
phenolic compounds such as gallic acid, protocatechuic acid, PB1, THA,
catechin, vanillic acid, epicatechin, rutin, and quercetin, same which are not
present in normal yogurt. Although the presence of grape pomace changed
its organoleptic characteristics, they were not very appealing to the sensory
test subjects, as pinot noir additioned yogurt was not suitable for tests due to
its strong smell and flavor, Chardonnay and Moscato varieties were able to
be evaluated in sensory tests, but the results were not all that favorable. The
addition of pomace to yogurt can be done but organoleptic characteristics
were not of the liking of the evaluators, properties like sweetness, acidity,
smell, and taste were not very favorable, a new strategy for the elaboration
of yogurt additioned with grape pomace should be considered.
Pasta is one of the most consumed products worldwide as it is present in
many places, from restaurants to households and retail markets and it can
be acquired by almost everyone, so it has a great reach in the food industry,
so the addition of enriched flours that may help in the enhancement of the
nutritional traits of the product is of utmost importance. Grape pomace as it
can be made into fine flour is ideal to do as the addition of this ingredient
modifies the cooking properties, starch digestibility, and glycemic index of
the pasta. Tolve et al. (2020) analyzed the elaboration of pasta by replacing
durum wheat semolina with grape pomace; the pomace used was from the
Corvina variety grape. The pasta obtained was of reddish color which only
increased with the concentration of grape pomace used, this was due to the
natural pigments of the fruit, the addition of pomace also added phenolic
compounds to the pasta with the best treatment obtaining 2.57 mg of gallic
acid equivalents (GAE)/g while uncooked and 1.81 mg GAE/g after cooking
the pasta, this loss is normal as the boiling of water can degrade biological
compounds such as phenolic compounds, the pasta also presented antioxidant
activities in FRAP and ABTS assays. Fiber content changed in comparison
to normal pasta, as grape pomace additioned pasta had an amount of 8.2 g of
fiber/100 g of pasta compared to 3 g of fiber for the normal semolina pasta.
The results found in this study show the importance of grape pomace as the
addition of it to pasta can increase its nutritional properties.
13.4 VEGETABLE WASTE AS FUNCTIONAL INGREDIENTS
13.4.1 CARROT WASTE

Carrots are a very popular vegetable as it is produced in many regions of the
world; they originated in the region now known as Afghanistan and some
parts of northern Iran and Pakistan (Simon, 2020). The global production
of this vegetable was around 40 million tons per year in 2020 and China
is the main producer with almost 50% of the total production, followed by
Uzbekistan, Russia, the United States (USA), Ukraine, and Poland (FAO,
2022a). There are many kinds of carrot as there are many varieties around
the world, the most dominating one is the orange kind as it can be seen
everywhere but, white types of carrots still are being consumed in western
and Eastern Europe, mainly as feed for livestock, red carrots can be found in
Japan and yellow and purple in the Middle East.
Carrot pomace is an industrial by-product obtained by the elaboration
of juices, as only 60–70% of juice yield is obtained from a carrot, although
carrot pomace is a high nutritional value by-product as it has large amounts
of carotenoids, vitamins, dietary fiber, and minerals (Umair et al., 2021). The
utilization of carrot pomace can be done as a functional ingredient, one of the
ways it can be used is to enhance the nutritional properties of baked products
such as cakes. Kamiloglu et al. (2017) used the pomace of black carrot to
improve the nutritional characteristics of a cake and analysis showed that
the cake had the presence of the five major anthocyanins, neochlorogenic
acid, chlorogenic acid, and caffeic acid were detected in the enriched cakes
conferring this product great antioxidant activities.
Smoothies are a popular way to elaborate beverages that contain many
kinds of fruits and vegetables, some are made with just water but others
are made with a great variety of ingredients such as milk, ice cream, sugar,
yogurt, and other milk derivates, the composition of some smoothies makes
it viable to add some ingredients that may negatively alter the taste as they
get covered up by the ingredients previously mentioned, this is the case for
black carrot a study by Kidoń and Uwineza (2022) aimed to use black carrot
with a mix of banana and pumpkin in 3 concentrations of sugar, high, low,
and no sugar. The smoothies produced with these ingredients proved to be
a great source of polyphenols as the total polyphenolic content ranged from
39.2 to 55.8 mg/100 g with a strong correlation with antioxidant activity. The
mix of pumpkin and carrot had better carotenoid content while the mixture
of banana and carrot showed better antioxidant activity, while the sensory
evaluation was a success as the judges gave excellent scores of the smoothies
in categories such as smell, taste, consistency, and overall acceptance.
13.4.2 CASSAVA WASTE
Cassava is a very starchy root and its leaves can be eaten as a source of
protein, vitamins, and minerals; it originated in South America and has had
a great journey all around the world as it is present also in Asia and Africa,
ever since the production of cassava has been increasing mainly because
it is a relatively easy crop to work with and it is one of the main foods for
more than 800 million people worldwide. Cassava is now considered to have
the potential to fight food insecurity, poverty, and hunger in many African
countries (Otun et al., 2022).
Cassava peel is a material that has been studied for its high content of
fibers, minerals, and sugars and has been used in animal feed, in a study
by Rodrigues et al. (2021) cassava peel was used in the elaboration of flour
which was packed with high polyphenolic content, some of the main one’s
found were gallic acid at 270.62 mg/100 g, catechin 14.87 mg/100 g, chloro-
genic acid 0.79 mg/100 g, caffeic acid 0.19 mg/100 g, and rutin 0.96 mg/100
g. Also, antioxidant activity was evaluated in ABTS and DPPH assays with
good results with 26.31 μM Trolox/g and 24.13 μM Trolox/g respectively.
13.4.3 BROCCOLI WASTE
Brassica oleracea is a biennial plant that belongs to the Brassicaceae family,

it is eaten as a vegetable all over the world, and it is a popular vegetable
that has been consumed ever since the time of the Greeks 2500 years ago.
China is the largest producer of broccoli followed by India, the USA, Spain,
Mexico, and Italy, not so long ago, broccoli consumption spiked due to
the now popular awareness of healthy lifestyle and diet, in which broccoli
apports an ingredient with high content of polyphenols, vitamins, mineral,
fibers, and very low caloric content (Nagraj et al., 2020).
Recently, the scientific community has taken into consideration that
not only the head and inflorescence of the vegetable contain the nutritional
content mentioned, but also the leaves and stalks which represent a 70%
of the broccoli plant. This causes a great environmental problem that can
be fixed by taking advantage of this waste; a study by Costa-Pérez et al.
(2022) set out to identify which biological compounds were present in the
stalk of the vegetable. To mention some of the polyphenolic content found
the stalk had the presence of 5-caffeoylquinic acid, feruloyl-caffeoyl deriva-
tive, di-sinapoyl-gentiobioside II, 1,2,3’-Tri-sinapoyl-gentiobioside among
others, which make the stalk an option to make a functional ingredient.
Núñez-Gómez et al. (2022) conducted a study to obtain different dietary
fiber fractions from broccoli stalks to revalorize the vegetable waste and
potentially obtain a new functional ingredient. The fiber fractions obtained
from broccoli yielded 67% total fiber fraction and 70% for insoluble fiber
fraction which makes this a great material to obtain fiber from; these frac-
tions also contained a great amount of nutritional value as protein had 3.8–4
g/100 g, 19.3–24.4 g/100 g of total carbohydrates, 60.8–68.9 g/100 g of
fiber, 54.0–54.3 g/100 g of total insoluble fiber, and 7.5–10.8 g/100 g of total
ash. The implementation of a functional ingredient of these results found
can be very beneficial to consumers as the number of nutrients found in
this by-product of the food industry is astounding, and the amount of fibers
present makes it an ideal candidate to elaborate bread with it.
The elaboration of bread with broccoli by-products has been studied by
Lafarga et al. (2019) which demonstrated that the addition of powdered broc-
coli stalks and leaves into the formulation of bread at a concentration of 2%
does not affect the acceptance of the product by the consumers, but there are
notable changes such as color and texture. The addition of these compounds
to the bread confers the product of increased nutritional characteristics and
against all odds the antioxidant and phenolic compounds found in the stalks,
resist the baking process making this a great option for consumption.
13.5 FUTURE PERSPECTIVES
New functional ingredients are found every day in the least of the expected
places, the revalorization of the wastes that the food industry generates is
now more important than ever as new options to lower the carbon footprint
and lower the emission of greenhouse gases arise. As we have seen in this
study it is possible to find gold, where you least expect it as new functional
ingredients, which can be obtained from agricultural by-products which
are often considered as low-value waste, but studies indicate that most
of the biological importance factors reside in this so-called waste. Many
researchers have found that in the peels of fruits we can find polyphenolic
content which depending on the compound is the range of biological activi-
ties which we may find and may vary from antioxidant, antiproliferative,
anti-inflammatory, antibiotic, prebiotic, etc.
Much research ends up just finding what biological properties are present
in the studied subject and they never move up to the next stage, finding a
practical use for the newfound information, more research is needed in func-
tional food development as there is so much potential in the new ingredients
that every day are being discovered out of food wastes. Environmentally this
could create a long-term solution to one of humanity’s greatest problems,
food loss, and food discarding, as the valorization of every single aspect of
an ingredient, from the peel, seed, pulp, stem, stalk, leaves, everything is an
important aspect of a whole matrix. Ethically this could be a new beginning
for new ways to open new food resources to entire populations who live
in conditions where food is more like a luxury instead of a human right.
Economically food industries would benefit so much from the usage of all
the food source as waste management and food loss represents a big invest-
ment every year.
13.6 CONCLUSIONS
The usage of waste as a functional ingredient or to obtain new functional

foods is one of the best ways technology can be applied to solve a problem,
millions of tons of food are wasted every year and every year it turns worse,
the valorization of the industry’s by-products is one of the best strategies
that can be performed to reduce the amount of waste that is generated, as
we have seen it is possible to turn peels of fruit into new ingredients such as
flour to prepare additioned and fortified products such as bakery products,
yogurts, ice cream. Research is needed to find new possible ingredients out
of materials usually considered waste to fight back food loss and while doing
so, also fight contamination and global pollution.
ACKNOWLEDGMENTS
Autor Estrada-Gil L.E. thanks the Mexican Council for Science and Tech-
nology (CONACYT) for the financial support for the development of the
project (Ph.D.) in Food Science and Technology offered by the Autonomous
University of Coahuila.
KEYWORDS
• food loss
• waste
• bioactive compounds
• phenolic compounds
• agricultural by-products
• yogurt ingredients
• healthy foods
REFERENCES
Aamir, M.; Saleem, A.; Rakha, A.; Nadeem, M.; Ateeq, H.; Saeed, F.; Asghar, A.; Shah, Y.
A.; Afzaal, M.; Javed, A.; Tufail, T.; Qamar, A. Functional Utilization of Mango Waste for
Improving the Nutritional and Sensorial Properties of Multigrain Cookies. J. Food Process.
Preserv. 2022. https://doi.org/10.1111/jfpp.17173
Ajila, C. M.; Leelavathi, K.; Prasada Rao, U. J. S. Improvement of Dietary Fiber Content
and Antioxidant Properties in Soft Dough Biscuits with the Incorporation of Mango Peel
Powder. J. Cereal Sci. 2008, 48 (2), 319–326. https://doi.org/10.1016/j.jcs.2007.10.001
Arya, S. S.; Venkatram, R.; More, P. R.; Vijayan, P. The Wastes of Coffee Bean Processing
for Utilization in Food: A Review. J. Food Sci. Technol. 2022, 59 (2), 429–444. https://doi.
org/10.1007/s13197-021-05032-5
Bento-Silva, A.; Vaz Patto, M. C.; do Rosário Bronze, M. Relevance, Structure and Analysis
of Ferulic Acid in Maize Cell Walls. Food Chem. 2018, 246, 360–378. https://doi.
org/10.1016/j.foodchem.2017.11.012
Boff, J. M.; Strasburg, V. J.; Ferrari, G. T.; de Oliveira Schmidt, H.; Manfroi, V.; de Oliveira, V.
R. Chemical, Technological, and Sensory Quality of Pasta and Bakery Products Made with
the Addition of Grape Pomace Flour. Foods 2022, 11 (23), 3812. https://doi.org/10.3390/
foods11233812
Cattaneo, F.; Roco, J.; Alarcón, G.; Isla, M. I.; Jeréz, S. Prosopis Alba Seed Flour Improves
Vascular Function in a Rabbit Model of High Fat Diet-Induced Metabolic Syndrome.
Heliyon 2019, 5 (8), e01967. https://doi.org/10.1016/j.heliyon.2019.e01967
Costa-Pérez, A.; Moreno, D. A.; Periago, P. M.; García-Viguera, C.; Domínguez-Perles, R. A
New Food Ingredient Rich in Bioaccessible (poly)Phenols (and Glucosinolates) Obtained
from Stabilized Broccoli Stalks. Foods 2022, 11 (12), 1–17. https://doi.org/10.3390/
foods11121734
Costa, J. M.; Ampese, L. C.; Ziero, H. D. D.; Sganzerla, W. G.; Forster-Carneiro, T. Apple
Pomace Biorefinery: Integrated Approaches for the Production of Bioenergy, Biochemicals,
and Value-Added Products—An Updated Review. J. Environ. Chem. Eng. 2022, 10 (5),
108358. https://doi.org/10.1016/j.jece.2022.108358
Costa, M.; Alfaia, C. M.; Lopes, P. A.; Pestana, J. M.; Prates, J. A. M. Grape By-Products
as Feedstuff for Pig and Poultry Production. Animals 2022, 12 (17), 1–18. https://doi.
org/10.3390/ani12172239
Curutchet, A.; Trias, J.; Tárrega, A.; Arcia, P. Consumer Response to Cake with Apple
Pomace as a Sustainable Source of Fibre. Foods 2021, 10 (3), 1–12. https://doi.org/10.3390/
foods10030499
da Silva, C. M. S.; da Boit Martinello, K.; Lütke, S. F.; Godinho, M.; Perondi, D.; Silva, L.
F. O.; Dotto, G. L. Pyrolysis of Grape Bagasse to Produce Char for Cu (II) Adsorption:
A Circular Economy Perspective. Biomass Convers. Biorefinery 2022, Ii. https://doi.
org/10.1007/s13399-022-02792-8
Duenas, M.; Garciá-Estévez, I. Agricultural and Food Waste: Analysis, Characterization and
Extraction of Bioactive Compounds and Their Possible Utilization. Foods 2020, 9 (6), 4–6.
https://doi.org/10.3390/foods9060817
Edjabou, M. E.; Petersen, C.; Scheutz, C.; Astrup, T. F. Food Waste from Danish Households:
Generation and Composition. Waste Manage. 2016, 52, 256–268. https://doi.org/10.1016/j.
wasman.2016.03.032
Eshak, N. S. Sensory Evaluation and Nutritional Value of Balady Flat Bread Supplemented
with Banana Peels as a Natural Source of Dietary Fiber. Ann. Agric. Sci. 2016, 61 (2),
229–235. https://doi.org/10.1016/j.aoas.2016.07.002
FAO. Policy Measures for Managing Quality and Reducing Post-Harvest Losses in Fresh
Produce Supply Chains in South Asian Countries; 2017.
FAO. Fruit and Vegetables—Your Dietary Essentials. In Fruit and Vegetables—Your Dietary
Essentials; FAO, 2020. https://doi.org/10.4060/cb2395en
FAO. Food Loss and Waste; Food and Agriculture Organization, 2021. https://www.fao.org/
nutrition/capacity-development/food-loss-and-waste/en/
FAO. FAOSTAT; 2022a. https://www.fao.org/faostat/en/#data/QCL
FAO. Major Tropical Fruits: Market Review 2021. Statistical Compendium Rome FAO, 01,
18; 2022b.
Fazilah, N. F.; Ariff, A. B.; Khayat, M. E.; Rios-Solis, L.; Halim, M. Influence of Probiotics,
Prebiotics, Synbiotics and Bioactive Phytochemicals on the Formulation of Functional
Yogurt. J. Funct. Foods 2018, 48, 387–399. https://doi.org/10.1016/j.jff.2018.07.039
Fontana, M.; Murowaniecki Otero, D.; Pereira, A. M.; Santos, R. B.; Gularte, M. A. Grape
Pomace Flour for Incorporation into Cookies: Evaluation of Nutritional, Sensory and
Technological Characteristics. J. Culinary Sci. Technol. 2022, 1–20. https://doi.org/10.108
0/15428052.2022.2086956
Ismail, T.; Akhtar, S.; Riaz, M.; Ismail, A. Effect of Pomegranate Peel Supplementation on
Nutritional, Organoleptic and Stability Properties of Cookies. Int. J. Food Sci. Nutr. 2014,
65 (6), 661–666. https://doi.org/10.3109/09637486.2014.908170
Kamiloglu, S.; Ozkan, G.; Isik, H.; Horoz, O.; Van Camp, J.; Capanoglu, E. Black Carrot
Pomace as a Source of Polyphenols for Enhancing the Nutritional Value of Cake: An In
Vitro Digestion Study with a Standardized Static Model. LWT 2017, 77, 475–481. https://
doi.org/10.1016/j.lwt.2016.12.002
Kidoń, M.; Uwineza, P. A. New Smoothie Products Based on Pumpkin, Banana, and
Purple Carrot as a Source of Bioactive Compounds. Molecules 2022, 27 (10). https://doi.
org/10.3390/molecules27103049
Kruczek, M.; Gumul, D.; Kačániová, M.; Ivanišhová, E.; Mareček, J.; Gambuś, H. Industrial
Apple Pomace By-Products as a Potential Source of Pro-Health Compounds in Functional
Food. J. Microbiol. Biotechnol. Food Sci. 2017, 7 (1), 22–26. https://doi.org/10.15414/

jmbfs.2017.7.1.22-26
Lafarga, T.; Gallagher, E.; Bademunt, A.; Viñas, I.; Bobo, G.; Villaró, S.; Aguiló-Aguayo,
I. Bioaccessibility, Physicochemical, Sensorial, and Nutritional Characteristics of Bread
Containing Broccoli Co-Products. J. Food Process. Preserv. 2019, 43 (2), 1–11. https://doi.
org/10.1111/jfpp.13861
Lazari, M.; Aguiar-Oliveira, E.; de Oliveira, D. S.; Kamimura, E. S.; Maldonado, R. R.
Production of Low-Calorie Ice Cream Utilizing Apple Peel and Pulp. J. Culinary Sci.
Technol. 2019, 17 (6), 481–490. https://doi.org/10.1080/15428052.2018.1489323
Liu, Y.; Zheng, Y.; Yang, T.; Mac Regenstein, J.; Zhou, P. Functional Properties and Sensory
Characteristics of Kombucha Analogs Prepared with Alternative Materials. Trends Food
Sci. Technol. 2022, 129, 608–616. https://doi.org/10.1016/j.tifs.2022.11.001
Marchiani, R.; Bertolino, M.; Belviso, S.; Giordano, M.; Ghirardello, D.; Torri, L.; Piochi,
M.; Zeppa, G. Yogurt Enrichment with Grape Pomace: Effect of Grape Cultivar on
Physicochemical, Microbiological and Sensory Properties. J. Food Qual. 2016, 39 (2),
77–89. https://doi.org/10.1111/jfq.12181
Melini, V.; Melini, F.; Acquistucci, R. Phenolic Compounds and Bioaccessibility Thereof in
Functional Pasta. Antioxidants 2020, 9 (4), 1–30. https://doi.org/10.3390/antiox9040343
Nagraj, G. S.; Chouksey, A.; Jaiswal, S.; Jaiswal, A. K. Broccoli. In: Nutritional Composition
and Antioxidant Properties of Fruits and Vegetables; 2020; pp 5–17. https://doi.org/10.1016/
B978-0-12-812780-3.00001-5
Noor Aziah, A. A.; Lee Min, W.; Bhat, R. Nutritional and Sensory Quality Evaluation of
Sponge Cake Prepared by Incorporation of High Dietary Fiber Containing Mango
(Mangifera indica var. Chokanan) Pulp and Peel Flours. Int. J. Food Sci. Nutr. 2011, 62 (6),
559–567. https://doi.org/10.3109/09637486.2011.562883
Núñez-Gómez, V.; González-Barrio, R.; Baenas, N.; Moreno, D. A.; Periago, M. J. Dietary-
Fibre-Rich Fractions Isolated from Broccoli Stalks as a Potential Functional Ingredient with
Phenolic Compounds and Glucosinolates. Int. J. Mol. Sci. 2022, 23 (21), 13309. https://doi.
org/10.3390/ijms232113309
Oliveira, R. M.; Oliveira, F. M.; Hernandes, J. V.; Jacques, A. C. Composição Centesimal
De Farinha De Uva Elaborada Com Bagaço Da Indústria Vitivinícola. In: Mio Cid e D.
Sebastião, Vol. 1; Peter Lang, 2016; pp 2–7. https://doi.org/10.3726/978-3-653-06456-8/4
Otun, S.; Escrich, A.; Achilonu, I.; Rauwane, M.; Alexis, J.; Lerma-Escalera, Morones-
Ramírez, J. R.; Rios-Solis, L. The Future of Cassava in the Era of Biotechnology in
Southern Africa. Crit. Rev. Biotechnol. 2022, 0 (0), 1–19. https://doi.org/10.1080/0738855
1.2022.2048791
Pérez-Chabela, M. de L.; Cebollón-Juárez, A.; Bosquez-Molina, E.; Totosaus, A. Mango
Peel Flour and Potato Peel Flour as Bioactive Ingredients in the Formulation of Functional
Yogurt. Food Sci. Technol. (Brazil) 2022, 42. https://doi.org/10.1590/fst.38220
Pérez, M. J.; Zampini, I. C.; Alberto, M. R.; Isla, M. I. Prosopis nigra Mesocarp Fine Flour, A
Source of Phytochemicals with Potential Effect on Enzymes Linked to Metabolic Syndrome,
Oxidative Stress, and Inflammatory Process. J. Food Sci. 2018, 83 (5), 1454–1462. https://
doi.org/10.1111/1750-3841.14113
Pérez, M. J.; Cuello, A. S.; Zampini, I. C.; Ordoñez, R. M.; Alberto, M. R.; Quispe, C.;
Schmeda-Hirschmann, G.; Isla, M. I. Polyphenolic Compounds and Anthocyanin Content of
Prosopis nigra and Prosopis alba Pods Flour and Their Antioxidant and Anti-Inflammatory
Capacities. Food Res. Int. 2014, 64, 762–771. https://doi.org/10.1016/j.foodres.2014.08.013
Pérez, M. J.; Rodriguez, I. F.; Zampini, I. C.; Cattaneo, F.; Mercado, M. I.; Ponessa, G.;
Isla, M. I. Prosopis nigra Fruits Waste Characterization, a Potential Source of Functional
Ingredients for Food Formulations. LWT 2020, 132 (June). https://doi.org/10.1016/j.
lwt.2020.109828
Ponis, S. T.; Papanikolaou, P.-A.; Katimertzoglou, P.; Ntalla, A. C.; Xenos, K. I. Household
Food Waste in Greece: A Questionnaire Survey. J. Clean. Product. 2017, 149, 1268–1277.
https://doi.org/10.1016/j.jclepro.2017.02.165
Rabetafika, H. N.; Bchir, B.; Blecker, C.; Richel, A. Fractionation of Apple By-Products as
Source of New Ingredients: Current Situation and Perspectives. Trends Food Sci. Technol.
2014, 40 (1), 99–114. https://doi.org/10.1016/j.tifs.2014.08.004
Rodrigues, C.; Silva, V.; Loyola, A.; Silva, M.; Augusti, R.; Melo, J.; Carlos, L.; Fante, C.
Characterization and Identification of Bioactive Compounds in Agro-Food Waste Flours.
Química Nova 2021, 45 (4), 403–409. https://doi.org/10.21577/0100-4042.20170853
Roodhuyzen, D. M. A.; Luning, P. A.; Fogliano, V.; Steenbekkers, L. P. A. Putting Together
the Puzzle of Consumer Food Waste: Towards an Integral Perspective. Trends Food Sci.
Technol. 2017, 68, 37–50. https://doi.org/10.1016/j.tifs.2017.07.009
Rosas, V. T.; Mureb, R. A.; Monserrat, J. M.; Wasielesky, W.; Tesser, M. B. Inclusion of Grape
Bagasse (Vitis sp.) in the Diet of White Shrimp (Litopenaeus vannamei) and Its Effects
on Growth and Antioxidant System. Aquac. Res. 2022, 53 (13), 4805–4813. https://doi.
org/10.1111/are.15972
Simon, P. W. Carrot Facts; USDA, 2020. https://www.ars.usda.gov/midwest-area/
madison-wi/vegetable-crops-research/docs/simon-carrot-facts/
Tirado-Kulieva, V. A.; Gutiérrez-Valverde, K. S.; Villegas-Yarlequé, M.; Camacho-Orbegoso,
E. W.; Villegas-Aguilar, G. F. Research Trends on Mango By-Products: A Literature Review
with Bibliometric Analysis. J. Food Measure. Character. 2022, 16 (4), 2760–2771. https://
doi.org/10.1007/s11694-022-01400-7
Tolve, R.; Pasini, G.; Vignale, F.; Favati, F.; Simonato, B. Effect of Grape Pomace Addition
on the Technological, Sensory, and Nutritional Properties of Durum Wheat Pasta. Foods
2020, 9 (3), 354. https://doi.org/10.3390/foods9030354
Umair, M.; Jabbar, S.; Nasiru, M. M.; Lu, Z.; Zhang, J.; Abid, M.; Murtaza, M. A.; Kieliszek,
M.; Zhao, L. Ultrasound-Assisted Extraction of Carotenoids from Carrot Pomace and Their
Optimization through Response Surface Methodology. Molecules 2021, 26 (22), 6763.
https://doi.org/10.3390/molecules26226763
Wani, S. A.; Naqash, F.; Wani, F. J. Current World Trade and Market Trends. In: Apples;
Wani, S. A., Naqash, F., Wani, F. J., Eds., 1st ed.; Taylor & Francis, 2022; p 346. https://doi.
org/https://doi.org/10.1201/9781003239925
WFP. 5 Facts About Food Waste and Hunger, 2020. https://www.wfp.org/
stories/5-facts-about-food-waste-and-hunger
Wu, M.-Y.; Shiau, S.-Y. Effect of the Amount and Particle Size of Pineapple Peel Fiber on
Dough Rheology and Steamed Bread Quality. J. Food Process. Preserv. 2015, 39 (6),
549–558. https://doi.org/10.1111/jfpp.12260
Xiao, J.; Capanoglu, E.; Jassbi, A. R.; Miron, A. Advance on the Flavonoid C -glycosides and
Health Benefits. Crit. Rev. Food Sci. Nutr. 2016, 56 (sup 1), S29–S45. https://doi.org/10.1
080/10408398.2015.1067595
Index
A C
Aeromonas, 273–274 C. difficile, 228–229
Agriculture C. perfringens, 228–229
antiprotozoa activity, 186 Campylobacter, 226–227
Agro-industrial wastes, 251–252 Centers for Disease Control and Prevention
bioactive compounds, 253 (CDC), 222
β-glucosidase enzyme, 255 Clostridium botulinum, 277–278
bioprocess, 255 Clostridium perfringens (C. perfringens),
emerging technologies, 255 277
extraction, 254, 255 Cnidoscolus quercifolius, 206
polyurethane foam (PUF), 256 Controlled atmosphere (CA) technology,
solid state fermentation, 255 136
Algae polysaccharides, 12 application, 142–143
Algarrobo negro, 324–325. See also bacterial rot/soft rot, 150–151
Prosopis nigra conventional CA (CCA), 140
Alginate, 12–13 disadvantages
Anaerobiosis, 144 anaerobiosis, 144
Anthelmintic, 181 bacteriological issues, 144
Antifungal, 185–186 bad flavors and odors, 144
Antimicrobial activity, 178–181 high initial cost, 145
Antiprotozoa activity, 181–182 inadequate gas levels, 144
Antiviral activity, 182–183 irregular ripening, 145
physiological disorders, 144
B dynamic control system CA (DCS), 142
fast conventional CA (FCCA), 140
Bacillus cereus (B. cereus), 276–277 gases employed
Bacterial Rot/Soft Rot, 150–151 carbon dioxide (CO2), 139
Bacteriological issues, 144 nitrogen, 139
Bad flavors and odors, 144 oxygen, 139
β-glucosidase enzyme, 255 high carbon dioxide, 141
Bioactive compounds, 253 insecticidal, 141–142
β-glucosidase enzyme, 255 internal changes, 138
bioprocess, 255 low-ethylene CA (LECA), 141
carbon dioxide (CO2), 139 onion preservation, 137
emerging technologies, 255 conservation, 151–153
extraction, 254, 255 fungal damage, 150
nitrogen, 139 postharvest quality, 145
oxygen, 139 quality criteria, 146–149
polyurethane foam (PUF), 256 storage, 153–157
solid state fermentation, 255 quality losses, 137
Brucellosis, 274–275 ultra-low oxygen CA (ULOCA), 140–141
338 Index
Conventional CA (CCA), 140 M. bovis (bovine TB), 278–279

Croton betaceus, 206 norovirus, 281–282
Croton dichogamous, 207 Paratuberculosis (MAP), 279–280
rotavirus, 284
D Staphylococcus aureus (S. aureus),
DIGE (differential gel electrophoresis), 305 275–276
Dynamic control system CA (DCS), 142 Vibrio Spp., 272–273
virology research, 281
E Yersinia enterocolitica, 280
Flax cellulose nanocrystals (FCNs), 44
Ebracteolata hayata, 205 Food packaging
Environmental impact, 8 active and intelligent packagings, 54–56
animal-derived proteins, 9 coatings, 52–53
pea proteins, 10–11 flexible packaging, 54
plant derived proteins, 10 rigid packaging, 53–54
proteins, 9 Food safety, 304
soybean proteins, 10 Food spoilage, 198
Essential oils, 198 Euphorbiaceae plants
Essential oils of garlic (GEO), 23 Cnidoscolus quercifolius, 206
Euphorbia heterophylla, 199, 204–205 Croton betaceus, 206
Euphorbia neriifolia L., 205 Croton dichogamous, 207
Euphorbiaceae plants Ebracteolata hayata, 205
Cnidoscolus quercifolius, 206 Euphorbia heterophylla, 204–205
Croton betaceus, 206 Euphorbia neriifolia L., 205
Croton dichogamous, 207 industrial applications, 208, 211–212
Ebracteolata hayata, 205 Jatropha curcas, 207
Euphorbia heterophylla, 204–205 Jatropha dioica, 208
Euphorbia neriifolia L., 205 Jatropha neopauciflora, 207
industrial applications, 208, 211–212 Mallotus oppositifolius, 206
Jatropha curcas, 207 Manihot multifida (L.) crantz, 205–206
Jatropha dioica, 208 preservation methods, 200–201
Jatropha neopauciflora, 207 natural alternatives, 201–203
Mallotus oppositifolius, 206 Food waste, 322
Manihot multifida (L.) crantz, 205–206 fruits functional ingredients, 323–324
apple waste, 325–326
F grape waste, 327–329
Fast conventional CA (FCCA), 140 mango waste, 326–327
FBDS functional ingredients, 323
Aeromonas, 273–274 Prosopis nigra, 324–325
Bacillus cereus (B. cereus), 276–277 valorization approach, 322
brucellosis, 274–275 vegetable functional ingredients, 323–324
Clostridium botulinum, 277–278 broccoli waste, 330–331
Clostridium perfringens (C. perfringens), carrot waste, 329–330
277 cassava waste, 330
foodborne parasites, 285–287 Fruits functional ingredients, 323–324
global status, 287–288 apple waste, 325–326
hepatitis E virus (HEV), 282–284 grape waste, 327–329
Indian status, 288–289 mango waste, 326–327
Index 339
H cellulose, 11–12
microorganism-derived, 14
Hazard analysis and critical control points
plant-derived, 11
(HACCP), 221
starch, 12
Heat shock proteins (HSPs), 307
synergies, 14
Hepatitis E virus (HEV), 234, 282–284
smart hydrogels, 15
Hexahydroxydiphenic acid (HHDP), 253
antimicrobial properties, 20
High carbon dioxide, 141
color properties, 21–22
High initial cost, 145
design, 16
Hydrogels, 4–5
electrosensitive hydrogels, 18
applications, 7–8
flavor properties, 21
bacterial metabolites, 26
food characteristics, 19
essential oils, 22–23
functional properties, 21
extracts, 25
gas barrier properties, 20
magnetic response, 27
nutritional properties, 20–21
natural extracts, 23–24
photosensitive hydrogels, 18–19
packaging labels, 26
pH-sensitive hydrogels, 16–17
superabsorbent polymer (SAP), 6–7
salt-sensitive hydrogels, 18
superporous hydrogels (SPH), 6
thermosensitive hydrogels, 16–17
tea polyphenols (TP), 24
Irregular ripening, 145
I
J
Inadequate gas levels, 144
Intelligent food packaging Jatropha curcas, 207
chemical and physical crosslinking, 5–6 Jatropha dioica, 208
environmental impact, 8 Jatropha neopauciflora, 207
animal-derived proteins, 9
pea proteins, 10–11
L
plant derived proteins, 10 Larrea tridentata (LT)
proteins, 9 agriculture, 184
soybean proteins, 10 antifungal, 185–186
hydrogels, 4–5 antiprotozoa activity, 186
applications, 7–8 medicinal application
bacterial metabolites, 26 anthelmintic, 181
essential oils, 22–23 antimicrobial activity, 178–181
extracts, 25 antiprotozoa activity, 181–182
magnetic response, 27 antiviral activity, 182–183
natural extracts, 23–24 cytotoxicity study, 176–178
packaging labels, 26 toxicity, 183–184
superabsorbent polymer (SAP), 6–7 NMR-identified compounds, 171–173
superporous hydrogels (SPH), 6 plant composition and antioxidant
tea polyphenols (TP), 24 capacity, 168
polysaccharides antioxidant activity, 174–176
algae polysaccharides, 12 bioactive compounds, 170
alginate, 12–13 lignans, 174
animal-derived, 13–14 nordihydroguaiaretic acid, 174
biopolymers, 15 Lavender essential oil (LEO), 23
carrageenans, 13 Liquid chromatography (LC), 306
340 Index
Listeria, 227–228 O
Low-ethylene CA (LECA), 141
OMICs technologies, 304
Onion preservation, 137
M
conservation, 151–153
M. bovis (bovine TB), 278–279 fungal damage, 150
Mallotus oppositifolius, 206 postharvest quality, 145
Manihot esculenta, 199 quality criteria, 146–149
Manihot multifida (L.) crantz, 205–206 storage, 153–157
Mass spectrometry (MS), 306 Oxidative stabilizer
Meat authentication, 304 in oils, 124–125
Meat quality, 309
ATPase activity, 310 P
authentication, 311–313
Paratuberculosis (MAP), 279–280
employed 2-DE and MALDI-MS, 311
Pea proteins, 10–11
heat shock proteins (HSPs), 307
Physiological disorders, 144
MALDI TOF MS, 311
Polylactic acid (PLA)
proteins, 310
biopolymers, 68
safety, 307–308
3D printing, 82
water holding capacity (WHC), 308
food derivatives
Moringa oleifera, 112
sugar bagasse pulp (SCB), 69
antimicrobial effects, 125–126
materials, 68
bioactive compounds, 116–118 natural fibers, 75–77
extraction methods, 118–119 natural oils, 72
oxidative stabilizer organic material, 73–74
in oils, 124–125 Polysaccharides
uses, 113–116 algae polysaccharides, 12
dairy foods, 120–121 alginate, 12–13
meat foods, 121–123 animal-derived, 13–14
sea foods, 123–124 biopolymers, 15
carrageenans, 13
N cellulose, 11–12
Nanocomposites microorganism-derived, 14
techniques plant-derived, 11
bio-nanocomposite films, 44 starch, 12
casting, 43–45 synergies, 14
extensive utilization, 45 Polyurethane foam (PUF), 256
extrusion compounding, 45–46 Pomegranate (Punica granatum L.), 249
flax cellulose nanocrystals (FCNs), 44 bioactive compounds, 251
nanocrystalline cellulose (NCC), 44 nutraceuticals and functional foods, 259
Nanocrystalline cellulose (NCC), 44 biological activities, 256
Nanocrystals anti-inflammatory activity, 258–259
characterization, 42 antimicrobial activity/prebiotic effect,
synthesis, 39–40 258
Nanoparticles antioxidant activity, 257
characterization, 42 chemical composition, 252
synthesis, 40–41 pomegranate peel (PP), 252
Nanotechnology, 38 hexahydroxydiphenic acid (HHDP), 253
Index 341
purify ellagitannins, 256 Q

Pomegranate peel (PP), 252
Quality losses, 137
hexahydroxydiphenic acid (HHDP), 253
Pork, 221 R
diverse farm and husbandry systems, 223
hepatitis E virus (HEV), 234 Raman scattering effect, 307
safety, pathogens, 234–235
zoonotic pathogens, 223
S
C. difficile, 228–229 S. aureus, 229
C. perfringens, 228–229 S. suis, 230–231
Campylobacter, 226–227 Salmonella, 224–225
Listeria, 227–228 SDS-PAGE (sodium dodecyl sulphate
S. aureus, 229 polyacrylamide gel electrophoresis), 305
Shigatoxigenic Escherichia Coli (STEC),
S. suis, 230–231
229–230
Salmonella, 224–225
Smart hydrogels, 15
Shigatoxigenic Escherichia Coli antimicrobial properties, 20
(STEC), 229–230 color properties, 21–22
T. gondii, 232–233 design, 16
T. solium, 231–232 electrosensitive hydrogels, 18
T. spiralis, 233–234 flavor properties, 21
Y. enterocolitica, 225–226 food characteristics, 19
Prosopis nigra, 324–325 functional properties, 21
Proteomics, 304 gas barrier properties, 20
in food science nutritional properties, 20–21
2-DE and MALDI-MS, 309 photosensitive hydrogels, 18–19
food allergies, 308 pH-sensitive hydrogels, 16–17
meat quality, 309 salt-sensitive hydrogels, 18
ATPase activity, 310 thermosensitive hydrogels, 16–17
authentication, 311–313 Solid state fermentation, 255
Soybean proteins, 10
employed 2-DE and MALDI-MS, 311
Staphylococcus aureus (S. aureus), 275–276
heat shock proteins (HSPs), 307
Starch nanocomposites, 38, 46
MALDI TOF MS, 311
barrier properties, 49–50
proteins, 310 food packaging
safety, 307–308 active and intelligent packagings, 54–56
water holding capacity (WHC), 308 coatings, 52–53
tools flexible packaging, 54
DIGE (differential gel electrophoresis), rigid packaging, 53–54
305 mechanical properties, 48–49
liquid chromatography (LC), 306 rheological and pasting properties, 47–48
MALDI-TOF MS, 306 thermal properties, 50–52
mass spectrometry (MS), 306 Sugar bagasse pulp (SCB), 69
nanometric detection limits, 305 Superabsorbent polymer (SAP), 6–7
Raman scattering effect, 307 Superporous hydrogels (SPH), 6
SDS-PAGE (sodium dodecyl sulphate
polyacrylamide gel electrophoresis), T
305 T. gondii, 232–233
Purify ellagitannins, 256 T. solium, 231–232
342 Index
T. spiralis, 233–234 brucellosis, 274–275

Tea polyphenols (TP), 24 Clostridium botulinum, 277–278
Thermal properties, 88 Clostridium perfringens (C.
PLA perfringens), 277
3D printing, 99–104 foodborne parasites, 285–287
fibers incorporating, 94–96 global status, 287–288
foods and natural oils, 89–91 hepatitis E virus (HEV), 282–284
nanocomposites in, 97–98 Indian status, 288–289
organic materials, 91, 93 M. bovis (bovine TB), 278–279
Thermoplastic starch (TPS), 38 norovirus, 281–282
Paratuberculosis (MAP), 279–280
U rotavirus, 284
Staphylococcus aureus (S. aureus),
Ultra-low oxygen CA (ULOCA), 140–141
275–276
Vibrio Spp., 272–273
V virology research, 281
Valorization approach, 322 Yersinia enterocolitica, 280
Vegetable functional ingredients, 323–324 strategies and preventive approaches, 289
broccoli waste, 330–331 applied research, 291
carrot waste, 329–330 awareness campaigns, 291
cassava waste, 330 coordinated approach, 291
microbiological risk assessment, 290
W recommendations/actionable points, 292
risk management, 290
Water holding capacity (WHC), 308
surveillance and response, 290
Y Zoonotic pathogens, 223
C. difficile, 228–229
Y. enterocolitica, 225–226 C. perfringens, 228–229
Yersinia enterocolitica, 280 Campylobacter, 226–227
Listeria, 227–228
Z S. aureus, 229
Zoonoses S. suis, 230–231
bacterial foodborne pathogens Salmonella, 224–225
Campylobacter, 268–269 Shigatoxigenic Escherichia Coli (STEC),
E.Coli infection, 270–271 229–230
Listeria monocytogenes, 271–272 T. gondii, 232–233
Salmonella, 269–270 T. solium, 231–232
FBDS T. spiralis, 233–234
Aeromonas, 273–274 Y. enterocolitica, 225–226
Bacillus cereus (B. cereus), 276–277

Engineering Principles For Food Processing Technology and Product Realization - Sanet.st

Uploaded by

Copyright:

Available Formats

Engineering Principles For Food Processing Technology and Product Realization - Sanet.st

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Engineering Principles For Food Processing Technology and Product Realization - Sanet.st

Uploaded by

Copyright:

Available Formats

ENGINEERING PRINCIPLES FOR

FOOD PROCESSING TECHNOLOGY

© 2025 by Apple Academic Press, Inc.

Library and Archives Canada Cataloguing in Publication

CIP data on file with Canada Library and Archives

Library of Congress Cataloging-in-Publication Data

CIP data on file with US Library of Congress

ISBN: 978-1-77491-575-2 (hbk)

Leonardo Sepúlveda Torre, PhD

Leonardo Sepúlveda Torre, PhD, is a full Professor at the School of Chemistry

Porteen Kannan, PhD

Porteen Kannan, PhD, is an Assistant Professor in the Department of Veteri-

A. K. Haghi, PhD, is a retired professor and has written, co-written, edited

SECTION 1: Food Packaging and Storage Engineering ...................................1

2. Bionanocomposites Based on Thermoplastic Starch for the

3. Release Studies to Improve the Mechanical Properties of the

4. Thermal Properties by Adding Natural Oils, Foods,

5. Antioxidant and Antimicrobial Properties of Moringa

6. Storage of Fruits and Vegetables in Controlled Atmospheres and

SECTION 2: Food Biotechnology .....................................................................165

8. Perspectives for Innovation of New Food Preservatives from

9. Biotechnology and Food Safety: Zoonotic Perspectives ..........................221

SECTION 3: Case Studies..................................................................................247

11. Foodborne Zoonoses: Current Status and Control Strategies................265

12. Omics Technologies: An Overview of Application of Proteomics

13. Food Waste Revalorization as a Functional Food or Ingredient ............321

Sonia Yesenia Silva Belmares

Teresa Darlen Carrillo-Castillo

Rebeca Villanueva Chávez

Luis Enrique Cobos-Puc

Cristian Torres León

Erika Flores Loyola

Diana B. Muñiz Márquez

A. Arun Prince Milton

Elda Patricia Segura-Ceniceros

José Eduardo Urquizo-Contreras

ABF alkali-treated bamboo fibers

FDM fused deposition modeling

OHF olive husk flour

UAE ultrasound-assisted extraction

This title is a comprehensive research-oriented book designed to cover essen-

It is an indispensable reference book for food industry professionals and

Advances in the Development of

Hydrogels are networks formed by polymeric chains, cross-linked chemically

1.1.1 HYDROGELS CHARACTERISTICS AND THEIR APPLICATION

1.1.1.1 CHEMICAL AND PHYSICAL CROSSLINKING

1.1.1.2 SUPERPOROUS AND SUPERABSORBENT HYDROGELS

1.1.1.3 APPLICATIONS AREAS

FIGURE 1.1 Application of hydrogels in the food field.

The method of making films for the application of hydrogels in food

1.2 HYDROGELS BASED ON BIOPOLYMERS TO REDUCE

polymers of biological origin instead of those produced synthetically, since

1.2.1.1 ANIMAL-DERIVED PROTEINS

1.2.1.2 PLANT-DERIVED PROTEINS

TABLE 1.1 Biopolymers Derived from Animal Proteins to Form Hydrogels.

1.2.1.2.1 Soybean Proteins

The main component of soybeans is protein (35–40%). This fraction is

1.2.1.2.2 Pea Proteins

mainly because of its sustainable production, non-allergenic status, and