CHAPTER ONE

INTRODUCTION

BACKGROUND OF THE STUDY

In a hungry world with an ever-increasing population, it becomes imperative

that efforts are made at reducing crop loss due to pathogen infestation. Much of

the yield of watermelon fruits is lost to Bacterial Fruit Blotch (BFB), a yield-

reducing disease. Watermelon (Citrullus lanatus) belongs to the family

Cucurbitaceae. According to Dutta (1979), plants in this family are mostly used as

vege- tables, a few yield delicious summer fruits and a few are medicinal. Other

members of this group include sweet gourd or muskmelon (Cucurbita moschata),

pumpkin (C. pepo), Cucumis spp. and Momordica species. Water- melon

originated in Africa and has been in cultivation for more than 4, 000 years in the

drier parts of the continent and throughout India and parts of Asia (Cobley and

Steele, 1976). It is used as a dessert fruit and a thirst quencher and in the very dry

parts of Africa, it is relished by both man and his animals as a source of water.

Tiger nut (Cyperus esculentus) belongs to the Division–Magnoliophyta,

Class–Liliopsida, Order– cyperales and Family–Cyperaceae, Species- Cyperus

esculentus and was found to be a cosmopolitan, perennial crop of the same genus

as the papyrus plant (Hankus and Sarret, 1967). The plant was introduced by the

Arabs, first in the Valencia region. It is native to most of the Western Hemisphere

as well as Southern Europe, Africa, Madagascar, the Middle East and the Indian
1
Subcontinent (Abaejoh et al., 2006). The tuber is known by various names in

Nigeria, as “Aya” in Hausa, “Imumu” in Yoruba and “Aki Hausa” in Igbo. Tiger

nuts can be eaten raw, roasted, dried, or baked (Belewu and Abodunrin, 2006;

Oladele and Aina, 2007; Rita, 2009). It can also be used for preparation of “kunu

aya” (a local beverage in Nigeria) (Belewu and Abodunrin, 2008). Tiger nut milk

or “kunu aya” is mostly consumed in the afternoon to cool the body from the hot

weather, it is cheap and popular; available, affordable, drink of both the poor and

the rich (Bamishaiye and Bamishaiye, 2011). Tiger nut milk is a nutritive and

energetic drink both for old and young people (Abaejoh et al., 2006; David, 2010),

is rich in energy content (starch, fat, sugar and protein), minerals (phosphorus,

potassium) and vitamins E and C (Belewu and Belewu, 2007; Oladele and Aina,

2007).

The milk is underutilized due to its short shelf life and lack of information

on its nutritional potentials (Omode et al., 1995; Cortes et al., 2005). Apart from

use as a beverage, tiger nut milk is thought to be beneficial to diabetic patients

(if sugar-free) (Anderson et al., 2009) and those seeking to reduce cholesterol or

lose weight (Martinez, 2003; Oladele and Aina, 2007; Borges et al., 2008), due

to its content of carbohydrates with a base of sucrose and starch (without

glucose), and its high content of arginine, which liberates the hormone that

produces insulin (Chevallier, 1996; Alegría-Torán and Farré- Rovira, 2003). It

was reported that tiger nut is high in dietary fiber content (Alegría-Torán and
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Farré-Rovira, 2003), which could be effective in the treatment and prevention of

many diseases including colon cancer (Adejuyitan et al., 2009), coronary heart

diseases (Chukwuma et al., 2010), obesity, diabetics and gastro-intestinal

disorders (Anderson et al., 2009) and treatment of flatulence, indigestion,

diarrhoea and dysentery (Bixquert-Jiménez, 2003) because it provides digestive

enzymes like the catalase, lipase and amylase (Bixquert-Jiménez, 2003;

Adejuyitan, 2011).

Tiger nut milk has a very short shelf life of often less than 24 hours

depending on the condition of storage (Akoma et al., 2006).

Watermelon plant is a trailing hairy annual with rough, angular stems and dark

green alternate leaves carried on fairly long petioles. Watermelons are

monoecious and have pale yellow flowers which are smaller than those of

pumpkins and squashes. The fruit is large and rounded or oblong with a hard,

smooth rind often mottled light and dark green. The fruit contains a red, pink or

yellowish- white flesh with numerous flattened, black, brown or white seeds.

There is little food value in the flesh, with very little flavour and about 95% or

more being made up of water (Cobley and Steele, 1976). The seeds, however,

contain a considerable amount of oil.

STATEMENT OF THE PROBLEM

Tiger nuts are super high in resistant starch fiber, which has been getting a lot of

buzz for its weight loss benefits, says Gina Consalvo, RD. Resistant starch passes
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through the stomach and small intestine without being digested, and may even help

you lose weight by reducing blood sugar spikes and keeping you fuller longer than

other foods with the same number of calories, she says. It also benefits your gut by

acting as a prebiotic, stimulating the growth of good bacteria in the digestive

tract."Since the gut acts as our immune system's first line of defense, eating foods

like tiger nuts that are loaded with resistant starch help our natural protective

bacteria," says Jaclyn London, MS, RD, CDN. "Since tiger nuts are so loaded with

the stuff, you’re essentially set for the day just by consuming one serving."

One ounce, or about 50 chickpea-sized pieces of plain, raw tiger nuts, contains 120

calories, 10 grams of fiber, 9 grams of naturally occurring sugars, tons of vitamins

and minerals such as magnesium and iron, and 7 grams of fat, most of which is

oleic acid, a monounsaturated fat that research shows can help reduce appetite and

promote weight loss. For some context, a serving of almonds — about 23 nuts —

has 163 calories and about a third of the filling fiber of tiger nuts.

People with digestive issues such as IBS or sensitivities to high-fiber foods might

experience some gas, bloating, cramping, or diarrhea if you eat too many tiger

nuts, says Consalvo. Raw tiger nuts are also higher in anti-nutrients like phytates,

oxalates, saponins, and tannins that may reduce the amount of vitamins and

minerals your gut can absorb. You can soak them, boil them, or toast them to

increase their nutritional value if you'd like. Other than that, there aren’t many

drawbacks.
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However, high levels of consumption could cause some discomfort. For instance,

the consumption of more than 30 mg of lycopene daily could potentially cause

nausea, diarrhea, indigestion and bloating, according to the American Cancer

Society.

Loading up on water-dense foods like watermelon can be tempting for those

looking to lose weight because they help you feel full, but Lemond cautions

against going to extremes. "Eating more fruits and vegetables of any kind naturally

helps decrease overall calories (energy) of the diet," she said. "We know that

people that eat higher quantities of fruits and vegetables typically have healthier

body weights. However, I do not recommend eating only watermelon … You will

lose weight, but that weight will be mostly muscle."

Jarzabkowski also warns watermelon lovers to be mindful of their sugar intake.

"Though watermelon's sugar is naturally occurring, [watermelon] is still relatively

high in sugar."

OBJECTIVE OF THE STUDY

The main objective of this study is to isolate and identify the bacterials on tigernut

and watermelon.

SIGNIFICANCE OF THE STUDY

Like many plants, tiger nuts are rich in antioxidants, including vitamins C and E.

“Antioxidants help prevent cell damage and may offer protective benefits against

the effects of aging, as well as diseases like cancer and heart disease,” notes
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Culbertson. Roasted tiger nuts tend to have higher amounts of antioxidants than

raw or boiled ones.

"Watermelons help with overall hydration, and that is a great thing," said Lemond.

"They say we can get 20-30 percent of our fluid needs through our diet alone, and

foods like these certainly help." Additionally, their juice is full of good

electrolytes. This can even help prevent heat stroke.  

The watermelon also contains fiber, which encourages a healthy digestive tract and

helps keep you regular.  

A cup of watermelon contains nearly one-quarter of your recommended daily

intake of Vitamin A. 

Like other fruits and vegetables, watermelons may be helpful in reducing the risk

of cancer through their antioxidant properties. In lab dish and animal studies,

lycopene reduced prostate cancer cell proliferation, according to the National

Cancer Institute. 

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 CHAPTER TWO

LITERATURE REVIEW

Tigernut (Cyperus esculentus) is a rhizome spherical crop that can be eaten raw,

dry or processed (Bazine and Arslanoğlu, 2020). It is actually not a nut and it calls

with other names such as nutgrass, Chupa, nutsedge, earth almond, etc. (Samuel,

2016). It is native to most tropical and temperate regions of the world (Rubert et

al., 2017; Bazine and Arslanoğlu, 2020). It is majorly produced in Africa,

Madagascar, Middle-East, Southern Europe and Indian subcontinent, the leading

producing nations are Nigeria, Niger, Togo, Benin, United States, Iran, Iraq and

Morocco. The plant can re-sprout severally, the leaves are very tough and fibrous,

single plant can produce up to 2420 seeds, the tuber size ranges from 0.3 to 1.9 cm

(“Cyperus esculentus”, 2021). It grows in large quantities in many West-African

countries and Spain (Rosell, 2020). Thermal stress can cause changes to the tuber

membrane properties, this accounts for thermal adaptation of tigernut and makes it

possible for the tuber to thrive under various environmental conditions (Rubert et

al., 2018).

There are three cultivars of tigernut; yellow, brown and black cultivars

(Figure 1). The cultivars possessed different physicochemical properties (Ayo et

al., 2016; Nina et al., 2019; Ayaşan et al., 2020) and functional properties (Nina et

al., 2019; Ismaila et al., 2020). The major factors that account for the chemical

7
variation in tigernut are genetic makeup, production location (Ihenetu et al., 2021),

environment and growing conditions (Duman, 2019).

The tuber can be oval, ovoid or oblong (Asare et al., 2020). Moisture

content is a key determinant factor on the rheological, functional (Gasparre and

Rosell, 2019) and thermos-physical properties of tigernut. (Usman et al., 2019)

The geometry, porosity, density and mechanical strength of the tuber are affected

by freshness and moisture content (Emurigho et al., 2020). Physical properties of

tigernut such as thickness, geometric mean diameter, length, width, bulk density,

tuber density and surface area are also affected by its moisture content (İnce et al.,

2017). Thermal conductivity, diffusivity, specific heat, bulk density and surface

area increase with an increase in moisture content, while tuber porosity decrease

with an increase in moisture content (Usman et al., 2019). The black variety

possessed higher loose and bulk densities, swelling capacity and water absorption

capacity (Ayo et al., 2016).

Raw tigernut produced sweet-nutty flavor while chewing (Maduka and Ire,

2018). The sweet taste resulted from the activities of various endogenous enzymes

that act internally on the starch (Owuama and Owuama, 2020). Minimal

processing such as soaking and mild roasting are employed to facilitate edibility

(Umaru et al., 2018). It is preserved by drying and the tuber is available year-round

in many places (Maduka and Ire, 2018). Brown tigernut is smaller in size, hence,

dry faster and possesses low moisture content (Evbuomwan and Alabi, 2020).
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 Tigernut is an underutilized tuber rich in many essential nutrients including

proteins, carbohydrates, vitamins, minerals (Mohdaly, 2019), phytochemicals, oil

and fiber, (Ihenetu et al., 2021). The tuber was reported to have numerous

nutritional and health benefits (Bazine and Arslanoğlu, 2020). The lipid and

dietary fiber of tigernut resemble that of nut and starch content resembles that of

tuber (Roselló-Soto et al., 2019). Tigernut oil is rich in phytosterols, tocopherols

and essential unsaturated fatty acids (Roselló- Soto et al., 2018; Duman, 2019;

Mohdaly, 2019). Muhammad et al. (2019) reported that total color value, total

flavonoid, pH, and Ferric Reducing Antioxidant power (FRAP) of tigernut tuber

are affected by blanching and plasma-activated water treatment.

NUTRITION AND HEALTH BENEFITS OF TIGERNUT

The tuber is grown for its nutritional and health benefits (Achoribo and Ong, 2017;

Asare et al., 2020). It contains significant amounts of fiber, unsaturated fat and

moderate amounts of protein (Rosell, 2020). The tuber contains 45.73%

carbohydrate, 30.01 % oil, 5.08 % protein, 2.23 % ash and 14.80 % crude fiber

(Sabah et al., 2019). Ismaila et al. (2020) reported 7.90 and 10.25 % protein

contents in yellow and black cultivar respectively. Tigernut contains 77.49 - 80.01

% essential fatty acids and 31.32 - 34.03 mg/100 g essential amino acids (Ijarotimi

et al., 2018). The tuber is rich in disaccharide; D-saccharose, which yielded D-

glucose, D- galactose, D-xylose and D-arabinose upon hydrolysis (Marchyshyn et

al., 2021). Total sugar content between 10.09 - 12.64 % was reported in the yellow
9
variety (Obinna-echem et al., 2019). The tuber can contains up to 13.49 %

fructans (Marchyshyn et al., 2021). The brown variety is richer in fat and energy

(Ayo et al., 2016) while the black variety is richer in minerals (Nina et al., 2019),

protein, carbohydrates (Ayo et al., 2016) and fiber (Evbuomwan and Alabi, 2020).

It is also rich in P and Ca (Roselló-Soto et al., 2019). Ismaila et al. (2020) reported

Ca, Na, P, K, Fe, Zn and Cu in both yellow and black cultivars.

The tuber contains active ingredients such as sterols, alkaloids, tannins, saponins,

resins and vitamins E and C (Marchyshyn et al., 2021). The phytochemicals in

tigernut are exceptional and can be used in the production of drugs and therapeutic

diets (Ihenetu et al., 2021). Tigernut contains 62% flavonoids compounds, 23 %

phenolic acids and their derivatives and 15 % phenylethanoid glycosides (Mayer,

2019). The black variety contains more tannin, phytate, oxalate and saponin and

the brown variety contains more flavonoids, polyphenols and alkaloids (Ayo et al.,

2016). Processing generally reduces the phytochemicals content of tigernut

(Uchechi, et al., 2020a). The concentration of saponin, tannin, phytate, oxalate,

hydrogen cyanide and hemagglutinin reduces after fermentation (Ji and Gi, 2018).

Tannin, oxalate and saponin can be reduced by soaking and roasting (Umaru et al.,

2018).

The presence of numerous phytochemicals with antioxidant potentials

account for the health benefits of tigernut (Roselló- Soto et al., 2019; Willis et al.,

2019). Tigernut drinks can be used as functional food based on their chemical
10
composition (Oluwadunsin et al., 2021). Olagunju and Oyewumi (2019)

recommended the use of a beverage containing tigernut in the prevention of

cardiovascular diseases. Gugsa and Yaya (2018) reported several compounds with

antioxidant, anti- inflammatory, anticancer, antimicrobial and antiseptic properties

in smoke from burned tigernut. Consumption of tigernut improves antioxidant

mechanisms and can also lower the risk of obesity and diabetes due to its α-

amylase and lipase inhibition capacity (Willis et al., 2019). The phytochemicals in

tigernut milk were reported to prevent drug-induced liver damage in rats by either

inducing glutathione synthesis or by functioning as antioxidants (Onuoha et al.,

2017). The quercetin and beta-sitosterol present in tigernut are known for their

anti-cancer properties (Achoribo and Ong, 2019). The antioxidant activities of 24.5

- 54.9 % and 10.8 - 12.1% were reported for DPPH radical scavenging and iron

chelation respectively (Ijarotimi et al., 2018). Higher DPPH radical scavenging

was reported in germinated tuber (Adebayo and Arinola, 2017). Tigernut aqueous

extracts showed an anti- proliferative effect on cancer cells 48 h post-treatment

(Achoribo and Ong, 2019). Consumption of tigernut and its products prevent colon

cancer, thrombosis, heart attack (Sethi et al., 2016) and can cure diarrhea and

inflammation (Bazine and Arslanoğlu, 2020). Arogundade et al. (2018) associated

an increase in memory function in rats fed with tigernut extract with antioxidant

neurotherapeutic properties of tigernut. More researches are needed to fully

understand the anti-cancer mechanisms of tigernut (Achoribo and Ong, 2017).

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TIGERNUT PRODUCTS

Over the years, tigernut was consumed raw, little attention was given to its

processing and content extraction (Maduka and Ire, 2018). The three most

important products obtained from tigernut processing which can further be

processed into wide varieties of foods are tigernut milk, tigernut oil and tigernut

flour (Maduka and Ire, 2018).

TIGERNUT MILK

Tigernut milk (Kunun-aya) is a traditional non-alcoholic beverage

commonly consumed in West-African countries, it is an aqueous extract of

tigernut, coconut, date and spices blend (Ibrahim et al., 2016a; Kayode et al.,

2017). The milk is also used as an ingredient in the production of Kunu, a well-

known traditional beverage in West Africa (Ezekiel et al., 2019). Its low acid

vegetable milk rich in protein and starch (Elbrhami, 2016), also contains less

energy when compared with dairy milk (Amponsah et al., 2017). Tigernut milk is a

good substitute for vegetarians and persons with lactose intolerance (Amponsah et

al., 2017). Beverages with acceptable sensory properties were developed by

combining tigernut with other crops, these include tigernut and coconut (Echem

and Torporo, 2018), tigernut and pineapple (Elizabeth and Tijesuni, 2020), tigernut

and sweet potato (Idris et al., 2019), tigernut and cocoa (Oluwadunsin et al., 2021)

and tigernut and baobab (Badejo et al., 2020).

A probiotic beverage produced from tigernut possessed sensory qualities

12
similar to that of dairy probiotic beverage (Amponsah et al., 2017). The findings of

Wongnaa et al. (2019) showed that yogurt consumers in Kumasi, Ghana are

positive about the nutritional and sensory qualities of tigernut yogurt and are

willing to pay more for it. Optimum conditions for the production of tigernut

yogurt are incubation for 3.12 h at 35 °C and a starter culture concentration of 2.74

% (Odejobi et al., 2018).

WATERMELON

Watermelon (Citrullus lanatus) is a member of the family Cucurbitaceae native to

tropical Africa near the Kalahari Desert. Botanists refer to it as a "pepo," a fruit

with a thick rind and a fleshy centre. It is commonly consumed as a refreshing

summer fruit, much appreciated by consumers for its refreshing ability, attractive

colour, delicate taste and high water content to quench the summer thirst.

Watermelon's sweetness is due primarily to a mixture of sucrose, glucose, and

fructose. Sucrose and glucose account in a ripe watermelon for 20–40 per cent and

fructose for 30–50 per cent of total sugars Athens et al (2006).

HEALTH BENEFITS

The latest dietary recommendation to increase diet-rich intake of natural

antioxidants has created interest in replacing energy-dense snack per day with

antioxidant-possessing fruit. Watermelon juice and pulp contain considerable

amounts of fibre and carbohydrates. Fibre plays a significant role in blood

cholesterol, which helps in the prevention of large bowel diseases, while

13
carbohydrates are the source of energy for the cells. Watermelon is a

quintessential exotic fruit containing nutrients and phytochemicals which have

been reported to support human health. It is a good source of B, C, and E

vitamins, as well as minerals like phosphorus, magnesium, calcium, and iron.

Watermelon has been identified as a good source of vitamin C (3.72 mg/100 g).

Vitamin C also has the potential to cut off the blood supply to growing cancers

and therefore inhibits cancer cell growth. Epidemiological studies have shown

that it has antioxidants with anti-inflammatory, antihypertensive properties, and a

defensive effect against toxicity caused by carbon tetrachloride. Bailey et al.

found that supplementation with watermelon juice improves aspects of vascular

health in an individual with hypertension.

All the biochemical processes that occur in the cells and organelles of our bodies

maintain a stable and normal human body. Ijah et al. stated that an oxidation

reaction in a human body contributes to the formation of free radicals. Free

radicals are highly unstable atoms, ions, or molecules that interfere aggressively

with other molecules, affecting the normal functioning of deoxyribonucleic acid

(DNA) and cell membranes. The cellular redox cycle of free radicals and reactive

molecules contributes to the development of reactive oxygen species (ROS),

which as toxic as well as beneficial compounds, play a double role. In addition,

chemical reactions, free radicals and certain redox reactions are a cause of

oxidative stress of macromolecules in living cells. This results in damage to

14
cellular components like lipids, DNA, and proteins. Disorders such as CVD,

cancer, and neurodegenerative diseases arise from this degradation. CVD is

estimated to be the largest cause of death in European countries; the number of

deaths due to cardiovascular diseases is further expected to be around 23.6

million by 2030 Choudhury et al (2000).

ECONOMIC IMPORTANCE

Marketing efficient distribution of watermelon [Citrullus lanatus] is required to

optimize economic returns for marketers and consumers. Around January and

February 2018, an economic study of watermelon marketing was carried out in

Lagos State, Nigeria. Data were collected using a questionnaire to obtain

information from 56 study area marketers. Half of the advertisers used urban

markets as points of sale and were supplied regularly with watermelon.

Marketing for watermelons has been successful. The main restriction marketers

have faced their inability to access credit. Regardless of educational level, ample

consumer knowledge improved the economic returns of watermelon marketers.

15
 CHAPTER THREE

MATERIALS AND METHODS

3.1 COLLECTIONS OF SAMPLE

Two types of various unwashed fruits were collected in leather bag from Ugbokolo

Market, watermelon and tigernut were brought in the laboratory for further

analysis.

The bacteria were isolated from the two fruits by using serial dilution method the

fruits were crushed into presterilized mortal and pestle separately with distilled

water to form 10-1 to 10-5 2.8g of nutrients agar was measured using weighing

Balance and was transferred into conical flask using measuring cylinder 1000ml of

water was measured ad was transferred into the conical flask containing the

nutrient agar and was stirred until the agar was completely dissolved and was

allowed to cool to 47oc and dispensed into sterile petri dishes and allowed to

solidify the nutrient agar was supplemented with amphotericin B (10Ng/ml) before

pouring to prevent fungi growth then inoculation take place to inoculated

petriplates were incubated at 37oc for 24hours for bacteria growth after incubation

the morphologically different colonies of bacteria were isolated and different agars

were prepared for subculturing MSA 6.7gm, SSA 3-8gm, cled 2.3gm after

preparation it was allowed to cool at 47oc and then each bacteria colony was

picked using a sterilized inoculating wireloop and streaked on the agars an

incubated at 37oc for 24hours.

16
ANTIMICROBIAL TEST

Pure culture of clinical isolate of salmonella and shiegella were subculture medium

dry using streak plate method.

0.2ml of tigernut, 0.2ml of watermelon was pipette using syringe and dispensed

into different plate and incubated for 24hours after which the plate were observed

for zone inhibition

GRAM STAINING

- Crystal violet stain was added over the fixed culture

- After 10-60seconds, the stain was poured off, and the excess stain is rinsed

with water. The goal is to wash off the stain without losing the fixed culture.

- Iodine solution was used to cover the smear for 10 to 60 seconds. This step

is known as “Fixing the dye’s”

MICROSCOPIC EXAMINATION

- The slide undergo examination under a microscope under oil immersion.

The initial slide examination used was x40 objective to evaluate the smear

distribution, and then examined using x100 oil immersion objective.

17
CHAPTER FOUR

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