ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

REVIEW OF LITERATURE
The current research on antimicrobial activity of selected Indian spices
against multi drug resistant pathogens has been substantiated by many
researchers and a few literature quoted below have helped in a successful
completion of the research.

Staphylococcus aureus is considered to be a major pathogen of both

humans and animals and is causative agent of contagious bovine mastitis, a
serious disease of livestock resulting in both reduced milk production and
quality. Staphylococcus aureus is found naturally on the skin, mucous
membranes and in the nasopharynx of the human body. Some members are
normal micro-flora of skin and mucous membranes; while other causes skin
pustules, impetigo, suppuration, abscesses, septicemia, pneumonia,
osteomyelitis, renal abscess, endocarditis, meningitis, gastroenteritis and toxic
shock syndrome. Pathogenic Staphylococcus aureus produces variety of
extracellular enzymes and heat stable enterotoxins which cause food
poisoning.

In fact, according to the Center for Disease Control and Prevention

(CDC) about 25-30% of the population carries Staphylococcus aureus as
normal flora in their nasal cavity. These people are referred to as
Staphylococcus aureus carriers. Among the healthy carriers, this bacterium is
usually harmless and causes no disease; however, upon entrance into a sterile
body environment, Staphylococcus aureus may cause minor subcutaneous
infections such as folliculitis, pustules and boils. These conditions can
typically be treated with incision and drainage of the wound followed by
antibiotic treatment.

According to the CDC, Staphylococcus species are the most common

cause of skin infections in the United States. When left untreated, these

Review of Literature 12
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

Staphylococcal infections may result in serious illnesses by causing

septicemia and pneumonia, which ultimately may be fatal. Even though most
Staphylococcal infections have historically been treated with antibiotics, in
recent years, an increasing number of Staphylococcus aureus strains have
become resistant to a wide array of antibiotics, including Methicillin. While
Methicillin-Resistant Staphylococcus aureus (MRSA) infection is not any
more infectious than other strains of Staphylococcus aureus infections, the
MRSA infection are much more difficult to treat due to their inherent
resistance to many antibiotics.

Fang et al, (2008) reported that in 1980’s and 1990’s, MRSA

infections were commonly associated with hospitals and long-term healthcare
facilities, but more recently the worldwide emergence and spread of MRSA
strains that are independent of healthcare facilities has been widely reported
and is commonly referred to as Community Acquired MRSA. Molecular
epidemiology studies have shown that CA-MRSA differ both phenotypically
and genotypically from HA-MRSA.

Brantner and Grein, (1994); Somchit et al, (2003) discussed the

antimicrobial activity of different plant extracts. Some plants exhibited a good
treatment for the urinary tract infections, gastrointestinal disorders, respiratory
diseases and cutaneous infections. Using plants for medicinal purposes is an
important part of the culture and the tradition in India.

Thomas et al, (2012) mentioned that many spices have antimicrobial

properties. Spices have many other uses like medicinal, religious ritual,
cosmetics or perfume production, or as a vegetable. For example, turmeric is
used for cooking recipe and garlic as an antibiotic. A variety of spices grown
across the Indian subcontinent (a sub-region of South Asia) are used in the
diet. With different climates in different parts of country, India produces a
variety of spices, many of which are native of the subcontinent, while others

Review of Literature 13
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

were imported from similar climates and have since been established and
cultivated locally for centuries.

Mohamed Helal El-Sayed et al, (2014) reported that the antimicrobial

resistance is a subject of great concern in public health and also in the
designing of strategies for current therapeutic protocols all over the world.
Plants used for traditional medicine contain a wide range of substances which
can be used to treat various infectious diseases. Hence, antibacterial activities
of aqueous extracts of 10 plant species were studied against 25 multidrug
resistant (MDR) clinical isolates using the agar well diffusion method. The
most resistant organisms were Acinetobacter baumanii (resistant to 16
different antibiotics), Enterococcus faecium (resistant to 15 different
antibiotics), Pseudomonas aeruginosa (resistant to 15 antibiotics), Gemella
morbillorum (resistant to 14 different antibiotics), Enterobacter cloacae
(resistant to 13 different antibiotics) respectively. Among the tested plant
extracts Allium cepa, Allium sativum, Foeniculum vulgare, Matricaria
chmomilla, Salvia offcinalis and Thymus vulgaris showed strong antibacterial
activity against MDR isolates with inhibition zones ranging from 8.33 to 26
mm. The minimum inhibitory concentration (MIC) and minimum bactericidal
concentration (MBC) of the most active plant extracts; Allium cepa and
Foeniculum vulgare were ranged from 0.062 to 0.25 mg/ml and 0.031 to
0.125 mg/ml, respectively. Foeniculum vulgare extract was bactericidal for all
bacteria while Allium cepa extract was bacteriostatic. Hence, the discovered
compounds from these plants can use as templates for the development of
new antibacterial agents.

Hemali Padalia et al, (2015) reported that there is a global increase in

the occurrence of fungal and bacterial infections and development of
microbial resistance to antibiotics is a global concern. Apart from infectious
diseases, food spoilage also occurs because of contamination caused by
microorganisms. The quality, taste, colour, flavour and shelf life of food is
Review of Literature 14
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

decreased. The best way to solve this problem is the use of natural compounds
especially essential oils. One tactic employed to overcome the resistance
mechanisms is the use of plant extracts or essential oil singly or in
combination i.e., synergistic approach. Synergistic effects may be due to
certain complex formations that become more effective in the inhibition of
microorganisms. It has been reported that several medicine plants, spices and
especially herbs containing essential oils significantly inhibited a broad range
of microorganisms. Essential oils are the volatile oily liquids of the secondary
metabolism of plants which are obtained from almost all plant parts like
flowers, buds, leaves, stems, seeds, bark, fruits and roots. Essential oils can be
individually effective or they may be combined with antibiotics or plant
extracts. Traditional healers often use combinations of plants to treat or cure
diseases and found that synergy was most often quite promising. This review
describes some promising medicinal plant essential oils and their synergistic
effects. This may be a novel approach with promising results to deal with
multi drug resistant microorganisms and new ways of treating resistant
bacteria and fungi.

Sundar et al, (2014) investigated the antimicrobial activity of Nutmeg

(Myristica fragrans Houtt) seed extract against the lower respiratory pathogen
Acinetobacter baumannii. The extract was subjected to phytochemical
detection which confirms the presence of alkaloids, terpenoids, flavonoids,
phenols and quinones, with the exception of Phlobatanins. The seed extract
was tested against sixteen isolates of Acinetobacter baumannii. The
susceptibility of these isolates towards the seed extracts was compared with
each other and the antibiotic (Meropenem) which was used a as a positive
control. Results in the former showed that toulene, tetrahydrofuran and
methanol extracts of the seed exhibited antibacterial activity against the
bacterial isolates. The bioactive components present in the methanol extract
were also evaluated by GC-MS analysis and the analysis revealed the

Review of Literature 15
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

presence of thirteen compounds. The study concludes that the secondary

metabolites present in the plant can be an indispensible source of
anitimicrobial compounds.

Surbhi Kaushik et al, (2012) described the in vitro antibacterial

activity, minimum inhibitory concentration (MIC) and phytochemical analysis
of the extracts of different solvent extracts of nutmeg (Myristica fragrans)
evaluated against different strains of Gram positive Staphylococcus aureus
and Gram negative Escherichia coli through agar well diffusion method. All
the extracts displayed antibacterial properties with the effective zone of
inhibition range from 21.0 mm to 8.0 mm. The active extracts exhibited MIC
ranges against various strains of Staphylococcus aureus and E. coli from ˃512
μg/ml-128 μg/ml.

Wright and Philipson, (1990) mentioned that the spices are classified
as major and minor spice. The major spices: Include black pepper, capsicum,
ginger, turmeric, cardamom whereas the minor ones are ajowan, aniseed,
caraway, celery seed, cumin, coriander, and dill seed. Since ancient times the
spices and condiments are been used not only for flavoring food but also for
preservatives and medicinal purpose. A large number medicinal plant has
been used for many centuries for treating various diseases.

Hema et al, (2009) explained the antimicrobial activity of 25, 50, 75

and 100% alcohol extract of Piper nigrum (Black Pepper), Psidium guajava
(Guava), Amomum cardamom (Cardamom) and Cynogen dactylon (Bermuda
grass) has been evaluated against Pseudomonas lundensis, Bacillus cereus,
Aspergillus niger and Aspergillus flavus. Psidium guajava extract showed
excellent antimicrobial activity against all the test organisms. In 25%
concentration, Psidium guajava showed the highest 11mm antimicrobial zone
against Pseudomonas lundensis, Aspergillus niger and Aspergillus flavus. In
50% concentration, Amomum cardamom showed the highest 16mm

Review of Literature 16
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

antimicrobial zone against Aspergillus niger. In 75 and 100% concentration,

Psidium guajava showed the highest 14mm, 19mm antimicrobial zone
respectively against Pseudomonas lundensis and Aspergillus niger. Among all
the results obtained, the maximum antimicrobial zone formation with minimal
concentration was recorded in Psidium guajava with 11mm of antimicrobial
zone against Pseudomonas lundensis, Aspergillus niger and Aspergillus
flavus.

Melvin Joe et al, (2009) studied the antibacterial activity of Allium

sativum (garlic), Zingiber officinale (ginger) and Piper nigrum (pepper)
extracts against Klebsiella pneumoniae, Staphylococcus aureus, Morganella
morgani, Candida albicans, Escherichia coli and Proteus vulgaris. Among
ten extracts evaluated for antimicrobial activity garlic extract showed good
antimicrobial activity against all the test organisms but excellent antibacterial
activity against Proteus vulgaris and Morganella morgani. The ginger extract,
however, showed only a moderate antimicrobial activity against
Staphylococcus aureus, whereas the pepper extract showed the least activity
against the test organisms.

Chitanand et al, (2014) designed to evaluate the antimicrobial activity

of eight Indian spices against Gram positive and Gram negative pathogenic
bacteria viz., Staphylococcus aureus, Bacillus subtilis, Bacillus cereus,
Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa using aqueous,
ethanolic and methanolic extracts. Among all extracts tested alcoholic extract
of Star anise (Ilicium verum), and black pepper (Piper nigrum) showed
maximum antimicrobial activity against gram negative bacteria while
alcoholic extract of clove (Syzygium arromaticum) and black pepper showed
maximum activity against gram positive bacteria. By and large, all spices
used during the studies proved as antibacterial compounds with maximum
activity

Review of Literature 17
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

Vahidi et al, (2002) studied the antimicrobial activity of different parts

of Crocus sativus L. (saffron) including stigma, stamen, leaves and colora
extracted by various solvents, were tested against different bacteria
(Micrococcus luteus, Staphylococcus epidermis, Staphylococcus aureus and
E. coli) and fungal ( Candida albicans, Aspergillus niger and Cladospourium
sp) by cup plate diffusion method. Minimal inhibitory concentration (MIC)
values of each active extracts were determined. The results obtained show
strong activity of the ethyl acetate extract of various plant parts against
bacteria and fungi used as test organisms.

Abdul Wadood et al, (2013) explained that Medicinal plants have bioactive
compounds which are used for curing of various human diseases and also play
an important role in healing. Phytochemicals have two categories i.e., primary
and secondary constituents. Primary constituents have chlorophyll, proteins
sugar and amino acids. Secondary constituents contain terpenoids and
alkaloids. Medicinal plants have antifungal, antibacterial and anti-
inflammation activities. The present study involves ten different medicinal
plants Acacia nilotica, Psidium gujauva, Luffa cylindrical, Morus alba,
Morus nigra, Momordica charantia, Fagonia cretica, Punica granatum,
Ficus palmate and Prunus persica locally available in Mardan region of
Pakistan. The leaves of the selected medicinal plants were washed, air dried
and then powdered. The aqueous extract of leaf samples were used for the
phytochemical analysis to find out the phytochemical constituents in the
plants.

The phytochemical analysis of the plants is very important

commercially and has great interest in pharmaceutical companies for the
production of the new drugs for curing of various diseases.

Sivapriya et al, (2016) explained that there has been an increasing

interest in the development of neutracueticals to combat the drug resistant

Review of Literature 18
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

pathogens. Although spices have been well accepted as traditional medicine,

antimicrobial effect and isolation of its active constituents are still in infancy.
The antimicrobial activity of the aqueous extracts of commonly used spices
against Escherichia coli were determined and compared with antibiotic
amikacin. The spices exhibited antibiotic efficacy in various magnitudes.
Among spices, Tamarindus indica and Cinnmomum zeylanicum were
extremely sensitive against Escherichia coli followed by Curcuma longa,
Zingiber officinale and Allium sativum which were very sensitive. Syzium
aromaticum and Capsicum chinese also exhibited antibiotic activity.

Ranganathan Kapilan et al, (2015) studied the antimicrobial activity of

the natural spices was tested against gram positive bacteria Bacillus pumilus,
Bacillus cereus and Staphylococcus aureus and gram negative bacteria
Escherichia coli, Salmonella typhi and Pseudomonas aeruginosa using
aqueous, ethanolic, methanolic and liquid nutrient extracts. Among all the
extracts tested alcoholic extracts of Cardamom (Elettaria cardamom), Clove
(Eugenia caryophyllus) and Lemongrass (Cymbopogon citratus) showed
maximum antimicrobial activity against gram negative bacteria while
alcoholic extract of cardamom and lemongrass showed maximum activity
against gram positive bacteria. All the spices tested in this study proved that
they have antibacterial activity.

Anna Witkowska et al, (2013) evaluated and compare the

antimicrobial properties of extracts of thirty types of commercial herbs and
spices commonly used in the production of ready meals. Various extracts of
spices were evaluated for their antimicrobial activities against Escherichia
coli, Listeria innocua, Staphylococcus aureus and Pseudomonas fluorescens
using a microdilution broth method. Ethanol and hexane extracts of oregano,
clove, sage, rosemary and celery showed relatively strong antimicrobial
activities against all bacteria tested. In contrast, water extracts displayed little
or no antimicrobial activity. Flow cytometry revealed that cell membrane
Review of Literature 19
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

structures were damaged by spice and herb active extracts, while analysis of
intra- and extracellular ATP contents of bacteria indicated that an increase in
extracellular ATP was partially due to intracellular leakage. Extract
combinations assessed using the checkerboard method did not display
synergistic effects, however, some additive effects were observed when
oregano was combined with sage or rosemary against Listeria innocua or
Staphylococcus aureus. This study has demonstrated that some commercial
spice extracts have antimicrobial activity against food-borne bacterial species
and should be considered as potential antibacterial agents for addition to
ready meals.

Rehnuma Sharmeen et al, (2012) evaluated the antibacterial activity of

35 aqueous herbal extracts against a total of 20 clinical Klebsiella sp. isolates.
The maximum antibacterial activity was found as 90% in crude extracts of
Syzygium aromaticum (leaf) and Citrus limon L. (fruit) followed by 85% in
Spondias pinnata (leaf). Sensitivity of these isolates was also evaluated for
eight commercial antibiotic discs following disc diffusion assay where most
of the isolates found to develop resistance against multiple commercial
antibiotics. 85% of isolates exhibited resistant to chloramphenicol and
erythromycin and 80% were resiatant to sulfamethoxazole and cephradine.
The isolates showed their resistance between 55-60 % to the other four
antibiotic discs, viz; gentamycin, streptomycin, ciprofloxacin and
azithromycin. Among 35 herbal extracts tested, 19 herbal extrats were found
to possess antimicrobial activity in all multi-drug resistant isolates. Therefore
these herbal extracts could be used in future direction as alternative
therapeutic agents for the treatment of human diseases caused by Klebsiella
sp.

Rahat Ejaz et al, (2014) determine the resistance patterns of

Staphylococcus aureus isolated from Pakistan and to identify antimicrobial
agents against multi-drug resistant Staphylococcus aureus strains. Plant
Review of Literature 20
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

extracts of 5 medicinal plants (Psidium guajava, Nigella sativa, Piper nigrum,

Valeriana jatamansi, and Cucurbita pepo) exhibited antibacterial activity
against locally isolated multi drug resistant strains of Staphylococcus aureus.
The minimum inhibitory concentration of these extracts was ranged from
0.328 to 5,000 mg/ml. Plant extracts of Psidium guajava, Piper nigrum seed,
Valeriana jatamansi, Cucurbita pepo and Nigella sativa showed significant in
vitro antibacterial activity and thus, such findings may serve as valuable
contribution in the treatment of infection and may contribute to the
development of potential antimicrobial agents against multi drug resistant
strains of Staphylococcus aureus.

Bakkali et al, (2008) explained many methods for the extraction of

essential oils. They can be obtained by steam distillation, mechanical
expression, hydro distillation, fermentation or extraction but the method of
steam distillation is most commonly used for commercial production. During
distillation, water condensate and is separated by gravity leaving a very small
amount of volatile liquid that is the essential oil, these may be contain a
variety of volatile molecules such as terpenoids, terpenes, aromatic
compounds and aliphatic components.

Sun et al, (2015) reported antibacterial activity of root essential oil of

Dictamnus angustifolius against the Staphylococcus aureus, Escherichia coli
and Pseudomonas aeruginosa. The antimicrobial activity showed that the
essential oil possess significant inhibition in Escherichia coli and
Staphylococcus aureus.

Burt and Reinders (2003) analyzed the antibacterial properties of five

essential oils bay (Pimenta racemosa), clove bud (Eugenia caryophyllata,
Syzygium aromaticum), oregano (Origanum vulgare), red and light thyme oils
(Thymus vulgaris) against Escherichia coli O157:H7strain. Oregano and light

Review of Literature 21
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

thyme essential oils had the potential of preventing the growth of Escherichia
coli O157:H7 in foods.

Ghalem and Mohamed (2014) found that Eucalyptus camaldulensis

leaves essential oil showed good antibacterial activity against Escherichia coli
and Staphylococcus aureus.

Sousa et al, (2015) reported antimicrobial activity of essential oil and

its four fractions from Eugenia calycina Cambess leaves against oral bacteria.
The results showed that essential oil exhibited strong antibacterial activity
against anaerobic Gram-negative bacteria Prevotella nigrescens and
Porphyromonas gingivalis and Fraction 3 and Fraction 4, composed of
oxygenated sequiterpenes showed higher activity against all the bacteria.

Matan et al, (2006) reported that mixture of cinnamon and clove

essential oils showed inhibitory activity against food spoilage
microorganisms. Clostridium rubum and Escherichia coli were the most
susceptible bacterial pathogens.

Trivedy et al, (2011) investigated the antimicrobial activity of two

Piper species namely Piper nigrum and Piper longum. Aqueous and
methanolic extract were analyzed for antimicrobial activity by agar cup plate
method. All the extracts of Piper species exhibited a variable spectrum of
antimicrobial activity. Piper species extracts were active against gram
positive, gram negative bacteria and fungi. The antibacterial activity of both
plants extracts was maximum against Staphylococcus aureus (gram+ve) and
Escherichia coli (gram - ve) respectively.

Dahiya and Purkayastha (2011) elucidated the antimicrobial activities

of various solvent extracts of Piper longum against clinically proved multi-
drug resistant bacteria (Methicillin-resistant Staphylococcus aureus,
Enterococcus sp., Klebsiella sp., Escherichia coli, Pseudomonas aeruginosa

Review of Literature 22
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

and Acinetobacter sp. and reference strains of bacteria (Staphylococcus

aureus ATCC 25923 and Escherichia coli ATCC 25922) by using agar well
diffusion assay. The highest antimicrobial potentials were observed for the
hexane and acetone extracts of Alangium salviifolium and Piper longum,
displaying maximum inhibitory zone of 18 mm against Enterococcus sp.

According to Ibrahim et al, (2011) Myristica fragrans showed good

antibacterial activity against MDR strains viz: Staphylococcus aureus, Proteus
vulgaris, and Klebsiella pneumoniae of clinical origin. Its MIC was found to
be 25mg/ml for Staphylococcus aureus.

Gulten Okmen et al, (2016) investigate the antibacterial effects of

Crocus sativus extracts against mastitis pathogens, and its other biological
activities. The aqueous extract showed maximum inhibition zone against one
bacteria (Coagulase- Negative Staphylococci- 32; CNS 32), and the zone was
11 mm. Two bacteria (CNS – 33 and 36) showed the lowest sensitivity to
3250 μg/mL /mL concentration for ethanol. In addition, the extracts were
tested against the stable DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) free-
radical for antioxidant activity. As a result, the ethanol extract observed a
strong antioxidant activity (trolox equivalent: 2,4 mM). The extracts of C.
sativus have antibacterial and antioxidant activities.

Asad Khan et al, (2009) determined the antimicrobial activities of the

crude ethanolic extracts of five plants against multidrug resistant (MDR)
strains of Escherichia coli, Klebsiella pneumoniae and Candida albicans.
ATCC strains of Streptococcus mutans, Staphylococcus aureus, Enterococcus
faecalis, Streptococcus bovis, Pseudomonas aeruginosa, Salmonella
typhimurium, Escherichia coli, Klebsiella pneumoniae and Candida albicans
were also tested. The strains that showed resistance against the maximum
number of antibiotics tested were selected for an antibacterial assay. The
MDR strains were sensitive to the antimicrobial activity of Acacia nilotica,

Review of Literature 23
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

Syzygium aromaticum and Cinnamum zeylanicum, whereas they exhibited

strong resistance to the extracts of Terminalia arjuna and Eucalyptus
globulus. Community-acquired infections showed higher sensitivity than the
nosocomial infections against these extracts. The most potent antimicrobial
plant was Acacia nilotica (MIC range 9.75-313μg/ml), whereas other crude
plant extracts studied in this report were found to exhibit higher MIC values
than Acacia nilotica against community acquired as well as nosocomial
infection. This study concludes that Acacia nilotica, Cinnamum zeylanicum
and Syzygium aromaticum can be used against multidrug resistant microbes
causing nosocomial and community acquired infections.

Mahmoud Nassar et al, (2007) isolated sixteen volatile compounds

from the n-hexane extract of the buds of Syzygium aromaticum by using gas
chromatography-mass spectroscopy (GC-MS). The major components were
eugenol (71.56 %) and eugenol acetate (8.99 %). The dichloromethane extract
of the buds yielded limonin and ferulic aldehyde, along with eugenol. The
ethanol extract of the clove buds showed remarkable scavenging activity, as
compared with synthetic antioxidants such as butylated hydroxyl toluene
(BHT).

Chaieb et al, (2007) studied the antimicrobial activity of clove

essentials oil against a large number of multi-resistant Staphylococcus
epidermidis as well as the composition of the oil by GC/MS analysis.

Arora Pooja et al, (2013) studied the chemical composition of

cinnamon oil and karpur tulsi oil was subjected to Gas chromatography- Mass
spectroscopy (GC-MS) analysis. The major component of cinnamon oil was
found to be cinnamaldehyde (91.82%). Minor compounds were found to be 1-
8 cineole, α-muurolene, selinene, geraniol, E-cinnamic acid, vinyl trans-
cinnamate. The major component of kapur tulsi oil was found to be camphor

Review of Literature 24
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

(46.14%). Other components were found to be eugenol, 1-8 cineole,

limonene, α- pinene, camphene, β- myrecene and α-terpineol.

Bizuneh Adinew, (2013) isolated the essential oil of cinnamon bark

growing in Tepi (south-west, Ethiopia) by hydrodistillation and gas
chromatography-mass spectrometry was used to identify the component. Six
components were identified and 2-propenal, 3-phenyl was the major
constituent (87.013%) while Eugenol (9.317%), O-Methoxy cinnamic
aldehyde (0.236%), α-muurolene (0.133%), Naphthalene, 1,2,3,4-tetrahydro-
1,6-dime (0.195%), Tricyclo [3.3.1.0 (2,8)] nona-3,6-dien-9-on (0.173%),
were minor constituents. 2-propenal, 3-phenyl comprises 87.013% of the
essential oil extracted from cinnamon bark, and is the compound most
responsible for the cinnamon barks aroma and therapeutic effect.

Javid Parray et al, (2014) investigated the antibacterial and antioxidant

activities of Crocus sativus L. Kashmirianus extract (callus and stigmas).
Profuse callus was obtained on Murashige and Skoog medium enriched with
6-benzyl aminopurine (BAP) (20μM) + α-naphthalene acetic acid (NAA)
(15μM) under in vitro conditions from corm slices. Four pathogenic bacterial
strains (Staphylococcus aureus CD0001, Escherichia coli CD0006,
Pseudomonas aeruginosa CD0023 and Shigella flexneri CD0033) were used
for determining the antibacterial activity of extracts. The antioxidant activity
was determined by DPPH (2,2- diphenyl-1-picrylhydrazyl) assay, DNA
(Deoxyribonucleic acid) protection assay, FTC (Ferricthiocyanate) method,
TBA (Thiobarbituric acid) assay and lipid peroxidation assay. The methanol
stigma extract of saffron was found to be more effective in inhibiting all the
pathogenic strains. The stigma extract also showed significant radical
scavenging or chelation capacities in four of the methods; however, callus
extract exhibited maximum inhibition of peroxy- radicals in lipid peroxidation
assay. The protocol for callus production is described. It was concluded that
as well as the specific parts of plants displaying diverse pharmacological
Review of Literature 25
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

activities, callus produced under in vitro conditions will assist in enhancing

the production of secondary metabolites, which will reduce the pressure on
natural saffron.

Nisa et al, (2013) studied the extraction of possible bioactive

compounds from the saffron stigma and callus. The extraction of saffron was
carried out in methanol as well as on other plant materials.

Anand et al, (2011) studied the bioactive chemical compounds present

in Zehneria scabra identified by Nuclear Magnetic Resonance (NMR)
spectral analysis. The proton nuclear magnetic resonance spectra of the root
sample were recorded and the chemical shift values of the various signals are
identified by the presence of Gypenoside. This bioactive compound is
confirmed as the valuable medicinal compound likely to cure vast array of
disease.

Chidambaram et al, (2015) analyze the major phytochemical

compounds of Indian spices, cumin (Cuminum cyminum L.), fenugreek
(Trigonella foenum-graecum L.) and fennel (Foeniculum vulgare Mill.).
Hydro-alcoholic (30% ethanol in water, v/v) extracts were prepared from
selected spices and analyzed for extract yield, loss on drying (LOD), pH and
phytochemical compounds such as total alkaloids, phenols, flavonoids and
tannins were quantified by spectrometric methods. High performance thin
layer chromatography (HPTLC) and gas chromatography coupled with mass
spectrometer (GC-MS) techniques were employed to reveal the
phytochemicals of selected spice extracts. Hydro-alcoholic extracts from
selected spices revealed the pH to be 5.36-5.62, loss on drying (12.54–
15.41%) and extract yield (8.62–29.19%). Among the investigated samples,
higher levels of alkaloids (2.032%) and tannins (0.809%) were found in
fenugreek while fennel exhibited highest content of total phenolics (3.39%)
and flavanoids (2.621%). The hydro-alcoholic extracts were subjected to

Review of Literature 26
ANTIMICROBIAL ACTIVITY OF SELECTED INDIAN SPICES AGAINST MULTI-DRUG RESISTANT PATHOGENS

HPTLC analysis and the results suggested the presence of three different
major phytochemical compounds in cumin and fenugreek, whereas fennel
extract displayed only one major peak. Cumin, fenugreek and fennel extracts
showed relatively similar spots with Rf values of 0.51, 0.62, 0.90 and 0.97,
which indicates the presence of four similar type of flavonoids in each extract.
Presence of some volatile compounds in extracts was identified by GC-MS
analysis.

Review of Literature 27