Evaluation of antimicrobial effects of watermelon rind

Folorunso Toyin Evelyn

2015070503391

Department of Chemistry
Faculty of Science
The Polytechnic Ibadan

December 2023
 Title page
Certification
Dedication
Acknowledgment
Table of contents
List of Tables
Abstract

CHAPTER 1
Introduction
1.1 Background on watermelon
1.2 watermelon rind
1.3 Medicinal values of watermelon rind
1.4 Antimicrobial
1.5 Aim and Objectives
1.6 Scope of study
1.7 Justification

CHAPTER 2
Literature review
2.1 Health benefits of watermelon rind
2.2 Antimicrobial effects of watermelon rind

CHAPTER 3
Methodology
3.1 Sample collection
3.2 Sample preparation
3.2.1 Preparation of watermelon rind
3.2.2 preparation of watermelon rind extracts
3.3 Culturing of microbes(salmonella typhi)
3.4 Antimicrobial Activities

Reference

Abstract

Watermelon (Citrullus lanatus) is a popular fruit worldwide due to its refreshing taste and its high water content
(92% of its weight). According to the phytochemistry of the plant, carbohydrates, saponins, glycosides, steroids,
alkaloids, polyphenols, flavonoids, and tannins can be detected in watermelons. C. lanatus generates by-products
and, as such, can be characterized by its seeds and rinds. These by-products’ phytochemicals are nutritious and
functional. Watermelon seeds contain many phytochemical compounds with beneficial biological activities,
according to numerous scientific studies. This fact promotes watermelon seed consumption and encourages food and
pharmaceutical companies to use this innovative ingredient. The watermelon rind has all the nutrients of the pulp
and also more antioxidants, minerals, vitamins, and bioactive substances. Numerous studies show that watermelon
peel is high in fiber and nutrients and can be used in a variety of culinary products. These residues need to be
utilized for creating innovative functional food products with added value for the food chain’s sustainability. More
studies of watermelon by-products are required to promote functional food, nutraceutical, and pharmaceutical
applications. This review aims to shed light on the underutilized portion of watermelon and its chemical properties
in order to pave the way for future research.

Keywords: Cucurbitaceae; watermelon; peel; rind; extract; antibacterial activities

CHAPTER 1

Introduction

1.1 Background of study

Watermelon (Citrullus lanatus), family of Cucurbitacea is known as a popular fruit crop that is originally from
tropical and subtropical countries [1]. With a global production of almost 103 million tons in 2018, watermelon
(Citrullus lanatus) is considered the second-largest fruit in the world by production [2]. The cultivated area of
watermelon in Iran is around 132,786 ha, and its average yield is 26.8 tons per hectare, which ranks second in the
world with a production of 3,568,134 tons in 2017 [3}

The Cucurbitaceae family is considered to be the most extensive family, encompassing a total of 120 genera and an
estimated 825 species [1]. These plants are mainly found in tropical areas, with limited representation in temperate
regions. The vegetable crops belonging to the Cucurbitaceae family hold significant importance as horticulture
crops, mostly cultivated for their succulent and delicious fruits in various warm regions across the globe [2].
Citrullus lanatus, commonly known as watermelon, is classified as a distinct type of berry, characterized by its
spherical or oblong shape [3,4]. Its diameter typically ranges from 30 to 60 cm, while the thickness of its peel varies
between 10 and 40 mm. The fruit in question exhibits a dark green hue, frequently adorned with white marbling,
while its inner flesh displays a vibrant range of red and yellow tones. The presence of carotenoids, specifically β-
carotene and lycopene, is responsible for the coloring of the watermelon [5]. Watermelon, commonly consumed
throughout the summer, is widely valued by individuals for its sweet flavor and invigorating properties. This is due
to its high water content, which is approximately 92% of its total weight [6]. The fruit is utilized in the preparation
of jam and juice, and the rind is also subjected to pickling [7]. As depicted in Figure 1, the fruit consists of four
basic parts. These are the skin or peel, which corresponds to the outermost layer of the fruit; the rind or exocarp,
which is the green and white outer part of the fruit; the flesh, which consists of the endocarp and mesocarp and is the
edible and colored part of the fruit that contains the juice primarily; and the seeds, which are typically discarded
when consuming the fruit [6].

Figure 1. Watermelon anatomy.

Carbohydrates account for 7.6% of the fruit’s composition. Within the carbohydrate content, sugars make up 6.2%,
and dietary fiber constitutes 0.4% of it. Watermelon is devoid of lipids and cholesterol. Hence, it may be inferred
that this particular fruit possesses a relatively low caloric content while exhibiting a high sugar content.
Additionally, this food item serves as a valuable source of β-carotene, an antioxidant compound that acts as a
precursor to vitamin A [8]. The fruit is rich in amino acids, including citrulline and arginine, which are present in
high proportions. It also contains carotenoids, with the red-fleshed form having an extraordinarily high content of
lycopene and the yellow-fleshed kind containing β-carotene. Additionally, watermelon contains phenolic
compounds, more specifically 2.6 ± 0.3 mg gallic acid equivalents (GAE)/100 g in rind, as determined by Neglo et
al. [9], and 89.5 ± 0.06 mg GAE/100 g in seed oil, as stated by Ouassor et al. [10]. Watermelon is a fruit that
contains numerous minerals, such as calcium, potassium, magnesium, and iron. Furthermore, it is well
acknowledged as a valuable source of vitamins A, E, C, and B complex. The by-products of the fruit also contain
valuable components. Watermelon seeds include substantial quantities of proteins, lipids, carbohydrates, ash, and
minerals. Hence, seeds are considered to be a valuable source of nutrition and may also possess significance in
relation to their fiber, mineral, and phenolic compositions, thereby contributing to both health advantages and
economic value [7,8]. The composition of watermelon rind mostly consists of celluloses, hemicelluloses, pectins,
and lignins, along with entrapped sugars, lycopene, carotenoids, citrulline, and phenolics [11]. The watermelon rind
extract demonstrates antioxidant and antibacterial properties because of the presence of polyphenols.

The plant itself has been utilized for ages in the traditional treatment of diverse health conditions. In particular, it
holds significant medical value within the Ayurvedic and Indian traditional systems of medicine. Watermelon is a
fruit that possesses nutritional properties, including minerals and bioactive compounds that have ethnomedicinal
advantages for human health [12]. Among them, the antibacterial activity of watermelon has gained popularity lately
[13]. Due to their high value and potential as viable additions to the human diet, watermelon by-products with
substantial nutritional value have attracted significant attention. Utilizing watermelon rind and watermelon seeds as
a cost-effective raw material holds promise due to their abundance and potential to contain valuable components that
can be utilized in various industries [14,15]. The objective of this review is to examine the current state of research
pertaining to the antibacterial properties of watermelon by-products.

1.2 Watermelon rind

Watermelon consists of three major parts: the pulp (about 68% of the total weight), rind (approximately 30% of the
total weight), and seed (about 2% of the total weight) [14,15]. The outer rind has a dark green color or pale green
stripes, which can change to a yellowish-green color after ripening [16]. Watermelon rinds are typically consumed
as pickles in the southern United States [17]. It also can be used in the formulation of jellies, fruit preserves, and
conserves [18,19]. Although the watermelon rind is edible, many consumers avoid eating it because of its
undesirable flavor. The inner rind consists of a high amount of citrulline, a non-essential amino acid, which has been
conveyed to arginine (utilized in the urea cycle for the elimination of ammonia from the body) [20]. Some people
suffer from an arginine deficiency, so the production of extracts or nutritional supplements containing watermelon
rind has been considered to treat this specific type of deficiency [21]. This amino acid is capable of dilation of the
arteries and improving blood circulation [22]. Environmental issues are raised because watermelon rind, which
makes up around 30% of the entire fruit, is frequently thrown irresponsibly into the environment [23]. Due to the
lack of understanding regarding potential conversion strategies of highly valuable components, the valorisation of
this waste is limited [24,25].

According to Athmaselvi et al. [26], the watermelon rind contains approximately 95% water, which makes it
vulnerable to deterioration. Therefore, it is vital to minimize the moisture content for the production of longer shelf-
life by-products from watermelon rind [81]. Also, considering that the rind covers the about one-third total weight of
this fruit and contains a significant amount of nutrients (protein, carbohydrate, fiber, and phenolic compounds
[27,28], it is required to utilize watermelon rinds in the formulation of various food products. By using the drying
process, the rind can become value-added and the more sustainable food product and can be employed in order to
prepare bakery products [27].

1.3 Medicinal value of watermelon rind

When eating fresh watermelon, most people stick to the red or pink flesh. But you can eat the rind and seeds, too, as
they offer their own holistic health benefits.Rinds are lower in sugar and higher in fiber than the flesh of a
watermelon, Meyer-Jax says, “When eaten with the rest of the melon, it helps slow down sugar absorption in the gut
and mellows the rise in blood sugar.” Watermelon rinds also contain L-citrulline, which may reduce blood pressure
and boost athletic performance.

1.4 Antimicrobial

An antimicrobial is an agent that kills microorganisms (microbicide) or stops their growth (bacteriostatic agent).[1]
Antimicrobial medicines can be grouped according to the microorganisms they act primarily against. For example,
antibiotics are used against bacteria, and antifungals are used against fungi. They can also be classified according to
their function. The use of antimicrobial medicines to treat infection is known as antimicrobial chemotherapy, while
the use of antimicrobial medicines to prevent infection is known as antimicrobial prophylaxis.[2]

The main classes of antimicrobial agents are disinfectants (non-selective agents, such as bleach), which kill a wide
range of microbes on non-living surfaces to prevent the spread of illness, antiseptics (which are applied to living
tissue and help reduce infection during surgery), and antibiotics (which destroy microorganisms within the body).
The term antibiotic originally described only those formulations derived from living microorganisms but is now also
applied to synthetic agents, such as sulfonamides or fluoroquinolones. Though the term used to be restricted to
antibacterials (and is often used as a synonym for them by medical professionals and in medical literature), its
context has broadened to include all antimicrobials. Antibacterial agents can be further subdivided into bactericidal
agents, which kill bacteria, and bacteriostatic agents, which slow down or stall bacterial growth. In response, further
advancements in antimicrobial technologies have resulted in solutions that can go beyond simply inhibiting
microbial growth. Instead, certain types of porous media have been developed to kill microbes on contact.[3]
Overuse or misuse of antimicrobials can lead to the development of antimicrobial resistance. Use

1.5 Aim of study

The aim of the study is to evaluate the antimicrobial effects of watermelon rind on salmonella typhoid

1.6 Objectives of study

The objective of this study includes

● To investigate the antimicrobial effects of watermelon rind on a selected bacterial

● To create awareness on the use of watermelon rind in the treatment of bacteria infection .

1.7 Scope of study

This research is restricted to the investigation of the antimicrobial property of watermelon rind using salmonella
typhi bacteria.

1.8 Justification of study

The increasing prevalence of infection and antibiotics induced side effects calls for the search for plant based and
side effects free alternative to orthodox antibiotic. Watermelon rind is considered as waste by majority ,this research
will show the possibility of using watermelon rind as a cure for typhoid fever.

CHAPTER 2

LITERATURE REVIEW

2.1 Health benefits of watermelon rind

Watermelon rind contains Citrulline which is an essential amino acid that regulates heart functions, circulatory

system, and immune system. The other nutrients found in the watermelon skin are:

● Vitamin A

● Vitamin C

● Potassium

● Magnesium

● Zinc

● Carbohydrates

Some health benefits of eating watermelon rind ars:

1.Improves heart health

Watermelon rinds can help keep your heart healthy. It encourages blood circulation, which is beneficial for the

heart. In addition, the citrulline present in it is beneficial in improving blood flow and heart failure, coronary artery

disease diseases.

2. Good for kidney health

Watermelon rinds contain potassium, which is beneficial for healthy kidneys. Watermelon peels have diuretic and

hydrating properties, which are also beneficial in UTIs. In case of a UTI(urinary tract infection), drink a glass of

fresh watermelon juice regularly.

3. Reduce inflammation

Watermelon rinds contain lycopene, which helps reduce inflammation. Eating peel of watermelon helps in reducing

skin inflammation to arthritis pain. It is also effective in treating acne by reducing your inflammation, for this, you

make a mask by mixing it with avocado or banana with melon peel and apply it on the face

4. Weight loss and blood pressure control

Watermelon rinds from watermelon can help reduce your high BP. Watermelon and its peel contain L-citrulline,

which helps to thin the blood vessels and in turn helps to normalize blood pressure by decreasing it. The citrulline

content of melon peel also helps in weight loss. There is fiber in its peel, which helps you to stay full for longer.

Besides, watermelon and its peel also help you in better workout performance

5. Improve sleep

Watermelon rind contain magnesium. It is a mineral that can help you get better results. Watermelon also helps in

regulating your metabolism, which helps to overcome the problem of sleep disturbances and insomnia.

6.It can reduce your blood pressure

Some research has shown that watermelon extract supplements are able to help obese adults control their blood
pressure.

However, citrulline supplements are likely more effective. Most studies suggest citrulline supplements reduce blood
pressure in people with hypertension.

Watermelon is also a potential diuretic, which often is prescribed for people with high blood pressure.

7. It’s rich in fiber

Another benefit of watermelon rind is that it’s a rich source of fiber. A diet high in fiber has a whole host of health
benefits, including the following:

● Fiber helps maintain regular bowel movements and may help reduce the risk of developing diseases of the
colon.

● Fiber can help lower cholesterol and blood sugar levels.

● Foods with fiber fill you up faster, helping achieve and maintain a healthy weight.

Summarily, Watermelon rind is high in vitamin A, vitamin C, potassium and magnesium. They are also high in
fibre, which aids in digestion and subsequently helps control blood sugar levels and cholesterol levels. Eating
watermelon rinds has also been shown to lower blood pressure due to their high levels of citrulline. Citrulline
supplements have been linked to heart health and improved blood circulation overall.It's a good source of fiber and.
Helps to control blood pressure,it Contains amino acid (which is good for health), Aids in digestion,Low cholesterol
levels,Aid skin health,and improves the immune system . It is rich in antioxidants, minerals, vitamins and some
active ingredients. These peels are low in calories, but they are rich in vitamin C, vitamin A, vitamin B6, potassium
and zinc. Apart from this, nutrients like chlorophyll, citrulline, lycopene, amino acids, flavonoids and phenolic
compounds are also found in watermelon peels.

2.2 Antimicrobial effects of watermelon rinds .

Polyphenols are abundant in plant extracts and are frequently acknowledged as antioxidants, but they can also be
classified as antimicrobials. These compounds demonstrate inhibitory properties against both bacteria responsible
for food spoilage and pathogens that might cause food-borne illnesses [16]. In recent years, there has been a shift
among the scientific community towards the utilization of this kind of antibacterial compound. The excessive use of
antibiotics has led to the emergence of bacteria that are resistant to multiple drugs, necessitating the development of
more potent or intricate antibiotic compositions to effectively combat them [17,18]. The notion of distinguishing
between bactericidal and bacteriostatic medicines has proven to be effective in differentiating antibiotics that have
the ability to kill bacteria, known as bactericidal, from antibiotics that just suppress bacterial growth, referred to as
bacteriostatic. The principle that distinguishes the two types of antibiotics is that bactericidal medications possess a
greater capacity for antibacterial activity, enabling them to effectively eradicate bacteria. Another way to fight
bacteria is by damaging their biofilm. Biofilm is a term used to describe clusters of microbial cells that are
enveloped by bacterial extracellular polymers. The process of biofilm creation encompasses three key stages:
bacterial adhesion, biofilm development, and maturity. Additionally, the process of biofilm formation has the
potential to enhance the resistance of bacteria against drugs and mechanical damage [19,20]. Biofilms consist of a
diverse array of polymers, including alginate, exopolysaccharides, and proteins. In contrast to planktonic growth,
biofilms have been found to enhance the resilience of the host immune system against antibiotics and provide a
significant challenge to clinical therapy [21].

There are several antibacterial mechanisms, such as inhibition of cell proliferation, inhibition of nucleic acid
synthesis, inhibition of energy metabolism, attenuation of pathogenicity, reduction in biofilm formation and cell
adhesion, and damage to membranes possibly by producing hydrogen peroxide [22]. The process of suppressing cell
proliferation involves the inhibition or destruction of a critical cellular organ, resulting in the suppression of cell
proliferation. This inhibition can be achieved through many mechanisms. Several methods of action can be
employed to achieve desired effects, including the inhibition or destruction of enzymes and proteins [23,24,25],
tissue-specific inhibition [26], the cell autophagy signaling pathway [27], the utilization of long non-coding RNA
(lncRNA) [28], as well as the application of extracts or drugs that impede certain cellular functions [29,30,31]. More
specifically, Buranrat et al. [24] proved in their study that the Oroxylum indicum extracts inhibit the growth of MCF-
7 cells. In another study, Mo et al. [28] discovered that the liver-specific lcRNA FAM99B was downregulated in
hepatocellular carcinoma tissues relative to adjacent normal tissues. There are additional antibacterial assays that
function by blocking the production of nucleic acids, specifically DNA and RNA [32,33,34,35,36,37,38].
Consequently, these agents disrupt regular cellular processes as well as their proliferation, ultimately resulting in cell
death. Another effective way to fight bacteria is by inhibiting energy metabolism [39,40,41,42,43,44,45,46]. A
Metabolic Inhibitor diminishes enzymatic activity by typically forming a relatively weak interaction with the
enzyme. This pertains to competitive inhibitors. Enzymatic productivity can be diminished by impeding the entry of
substrates into the active sites.

The detection of bacterial pathogens from clinical samples in clinical microbiology is predominantly dependent
on culture-based methods. Historically, culture has been conducted with agar-based media of a generic nature (e.g.,
blood agar) that could facilitate the proliferation of a broad spectrum of pathogens. The virulence of a
microorganism is determined by various factors, including the presence of specific species, strains, or genes, as well
as their relative abundance. Hence, the differential proliferation of a single microorganism can lead to the transition
of additional microorganisms into or out of a harmful state. This is called attenuation of pathogenicity, and it is
widely utilized nowadays [47,48,49,50,51,52,53,54]. Another effective antibacterial method is the inhibition of
biofilm development and cell adhesion [55,56,57,58,59,60,61,62,63,64]. The subsequent phase of biofilm formation
is distinguished by the gathering of cellular aggregates on the surface [65]. The current stage is seemingly facilitated
by a polysaccharide antigen that enhances intercellular adhesion. The implementation of anti-adhesive surfaces has
been shown to effectively mitigate the adhesion of bacteria to solid surfaces, thereby facilitating the elimination of
germs prior to their adherence and subsequent multiplication. Finally, comes the damage to membranes by
producing hydrogen peroxide [66,67,68,69,70,71,72,73,74,75]. Hydrogen peroxide (H2O2) is an oxidizing agent that,
when present in excessive quantities, leads to cellular damage. This oxidative stress disrupts normal cell cycle
progression, resulting in cell death. Emerging research suggests that hydrogen peroxide has the capacity to promote
the progression of the cell cycle by oxidizing specific thiol proteins.

The peels, rind and seeds are usually discarded or used as a feed supplement for animals. These often-neglected
parts are claimed to possess health benefits. The rind is known to possess a vasorelaxant effect [10]; the seed
regulates blood sugar by triggering insulin release from the β-cell of the islet of Langerhans [11] whereas the peels
are known to have analgesic activities [12]. However, research on their biological effects is scarce. It is therefore
important to carry out studies on the health benefits of these neglected parts to increase awareness of consumers on
the beneficial effects of all the various parts of watermelon [13]
 CHAPTER 3

METHODOLOGY

3.1 Samples collection

Fresh fully grown watermelons were obtained from Oje market located in mapo , Ibadan.

3.2 Samples Preparation

The obtained fruits will first be washed thoroughly with distilled water and cut into sections.The outerskin will be
removed and the remaining rind ( the thick whitish inner portion of the exocarp) and the pulp( the red and juicy part)
will be carefully separated from each other.The rind will then be cut into bits and juiced by blending and after that,it
will be sieved and air dried to get the rind powder.

3.2.1 Extraction of Watermelon rinds

The watermelon extraction can be done using either Cold maceration process or the Soxhlet Extraction process.

Cold maceration process will be used.In this process,The watermelon rind powdered will be soaked in 100ml each
of methanol and aqueous solution for 24hrs on a mechanical shaker at room temperature and then filtered with the
whatmanNo.1 filter paper.A rotary evaporator will be used to concentrate the filtrate.The Aqueous extract will be
concentrated in a water-bath at 90 c for 30 mins.The extract will then be stored in a Airtight container until it is
ready to be used .

3.2.2 Culturing of Microbes ( salmonella typhi)

Strains of salmonella typhi will be obtained and sub-cultured on Mueller Hinton Nutrient agar .The organism will be
grown overnight at 37 c in the Mueller Hinton broth and stored in the refrigerator until further use.

3.3 Antimicrobial Activities

The antimicrobial activity of the extract will be carried out by Agar well diffusion technique against Salmonella
typhi bacteria. Approximately 150 μL of extract solution will be poured into the well (of 8 mm in diameter) on Petri
dishes with nutrient agar. The inoculated plates will be put in an incubator (37 °C for 24 h), then the inhibition area
surrounding the wells will be assessed and recorded.

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