CHAPTER ONE

1.0 INTRODUCTION

The increasing prevalence of multidrug-resistant strains of bacteria and the recent appearance

of strains with reduced susceptibility to antibiotics raised the specter of ‘untreatable’

bacterial infections and adds urgency to the search for new infection-fighting strategies (El

Astal et al., 2020; Eleyinmi, 2022). Plants are known to produce a variety of compounds to

protect themselves against a variety of pathogens. It is expected that plant extracts showing

target sites other than those used by antibiotics will be active against drug-resistant pathogens

(Guevaraa et al., 2019). For a long time, plants have been an important source of natural

products for human health. The antimicrobial properties of plants have been investigated by a

number of studies worldwide and many of them have been used as therapeutic alternatives

because of their antimicrobial properties (Bugno et al., 2020). There are several reports on the

presence of antimicrobial compounds in various plant parts like leaves, bark, fruit, root, and

flowers (Tsaknis et al., 2019). Moringa species are well-documented plant herbs due to their

extraordinary nutritional and medicinal properties. Moringa oleifera Lam. and Moringa

stenopetala are the most widely cultivated species of the monogenic family, the Moringaceae

(Walter et al., 2021).

Moringa oleifera is one of the species of family Moringaceae, native to, Africa, Arabia, South

Asia, South America, Himalaya region, India, Pakistan, the Pacific, and Caribbean Islands.

Moringa oleifera has been naturalized in many tropic and subtropic regions worldwide, the

plant is referred to number of names such as horseradish tree, drumstick tree, Ben oil tree,

miracle tree, and “Mothers best friend” (Pinal et al., 2014). They have long been known in folk

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medicine as having value in treating a wide variety of ailments. They are known to be

antihelminthic, antibiotic, detoxifiers, immune builders and have been used to treat malaria

(Thilza et al., 2022). The Moringa plant (Moringa oleifera) has been the object of much

research due to its multiple uses and well-known bactericidal potential (Viera et al., 2020).

1.2 Justification

The primary benefits of using local/herbal medicines are that they are safer than synthetic

alternatives, offering profound therapeutic benefits and more affordable. They are effective with

minimal or no side effects in the treatment of infectious diseases, while simultaneously

mitigating many of the side effects that are often associated with synthetic drugs. Many

local/herbal medicines have tropism to specific organs or systems in the body.

1.3 Statement of Problem

Pathogenic bacteria have become an increasing problem in the health care units and communities

around the world. This has a serious threat to the public health sector. The non- availability and

high cost of new generation antibiotics have resulted in increase in morbidity and mortality rate.

Therefore, testing the antibacterial activities of some selected pathogenic bacteria on honey may

serve as a suitable alternative measures in future.

1.4 Aim of the study

The research work was aimed at reviewing of the antimicrobial activities of Morning oleifera

leaf extract against some pathogenic bacterial.

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 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 MORINGA

Moringa oleifera (MO) is an aboriginal of Indian subcontinent and has become naturalized

in the tropical and subtropical areas around the world (Guevaraa et al., 2019). Nearly thirteen

species of Moringa are included in the family Moringaceae (Nadkarni, 2020). Indians have

been using it as a regular component of conventional eatables for nearly 5000 years (Anwar et

al., 2019; Anwar and Bhanger, 2017). Moringa tree can grow well in the humid tropic or hot dry

land with average height that ranges from 5 to 10 m. It can survive in harsh climatic condition

including destitute soil without being much affected by drought (Morton, 2021). It can tolerate

wide range of rainfall requirements estimated at 250 mm and maximum at over 3000 mm and a

pH of 5.0 to 9.0 (Muangnoi et al., 2018).

Its trunk is soft, white corky and branches bearing a gummy bark. Each tripinnately

compound leaves bear several small leaf legs (Guevaraa et al., 2019). The flowers are white and

the three wings seeds are scattered by the winds. The flowers, tenders leaves and pods are eaten

as vegetables. The leaves are rich in iron and therefore highly recommended for expected

mothers. In some part of the world, MO is referred to as the ‘drum stick tree’ or the ‘horse radish

tree’, whereas in others, it is known as the kelor, marango, mlonge, moonga, mulangay,

nebeday, saijhan, sajna or Ben oil tree (Anwar and Bhanger, 2017; Prabhu et al., 2022). In India

and Pakistan, MO is locally known as Sohanjna and is grown and cultivated all over the country

(Anwar et al., 2019).

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It has been reported by Bureau of plant industry that Maringa is an outstanding source nutritional

components. Its leaves (weight per weight) have the calcium equivalent of four times that of

milk, the vitamin C content is seven times that of oranges, while its potassium is three times that

of bananas, three times the iron of spinach, four times the amount of vitamin A in carrots, and

two times the protein in milk (Kamal, 2020). Besides, Moringa is also suggested as a viable

supplement of dietary minerals. The pods and leaves of Moringa contains high amount of Ca,

Mg, K, Mn, P, Zn, Na, Cu, and Fe (Bhattacharya et al., 2019). Although, minerals content of

Moringa shows variation in composition with changes in location (Anjorin et al., 2021).

Ancient medicinal system relies on several plant products used by traditionally human

communities in many parts of the world for different diseases. Among these plants, MO has its

great contribution from ancient time (Guevaraa et al., 2019). It is a plant with exceptional

medicinal properties which can resolves the health care needs in several situations. Easy

cultivation of Moringa within adverse environmental condition and wide availability attract

attention for economic and health related potential in resource limited developing

countries. This study discusses medicinal potential of this exceptional plant and its

potential as a commercial medicinal and nutritional supplement (Guevaraa et al., 2019).

2.2 PLANT DESCRIPTION

M. oleifera is a short, slender, deciduous, perennial tree that grows to about 10 m tall, slender

with drooping branches; branches and stem are brittle, with corky bark. Leaves are feathery, pale

green, compound, tripinnate, (30–60 cm long), with many small leaflets, 1.3–2 cm long, 0.6–0.3

cm wide, lateral ones slightly elliptic, terminal ones obovate, and slightly larger (Donli and

Dauda, 2020). Flowers are fragrant, white, or creamy-white, 2.5 cm in diameter, and borne in

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sprays. The stamens are yellow, and the pods are pendulous, brown, triangular, splitting

lengthwise into three parts when dry, and containing about 20 seeds embedded in the pith. The

pod has nine ribs on both ends and the seeds are dark brown with three papery wings (Donli and

Dauda, 2020).

M. oleifera tree, leaves, seeds, and flowers (Donli and Dauda, 2020).

2.3 SCIENTIFIC CLASSIFICATION

Kingdom: Plantae
Clade: Tracheophytes
Clade: Eudicots
Clade: Rosides
Order: Brassicales
Family: Moringaceae

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 Genus: Moringa
Species: Moringa oleifera

2.4 BENEFITS OF MORINGA OLEIFERA

2.4.1 Nutritional properties

Moringa oleifera is called a ‘Miracle Tree’ due to its multipurpose nutritional uses and capacity

to cure many diseases. Every part of plant has reserved nutrients. The Leaves of Moringa

oleifera are rich source of minerals like calcium, potassium, zinc, magnesium, iron and copper.

The Leaves has low calorific value so can be used in the diet of obese. The leaves also contain all

essential amino acids and are rich in protein and minerals (Hukkeri et al., 2022). Pods contains

around 46.78% fiber, 20.66% protein and are highly valued for curing digestive problem and

colon cancer (Kamal, 2020). Vitamins like vitamin A, Vitamin B, Beta-carotene, pyridoxine,

nicotinic acid, vitamin C, Vitamin D and E also present in abundant amount in Moringa oleifera.

Moringa oleifera also contains major phytochemicals such as: tannin, sterols, terpenoids,

flavonoids, saponins, anthraquinones, alkaloids and reducing sugar is also present along with

anti-cancerous agents like glucosinolates, isothio cyanates, glycoside compounds and glycerol-1-

9-octadecanoate. Oligosaccharides and oxalate were reported as anti-nutrient factor in Moringa

leaves (Mahajan et al., 2021). The Dried leaves (M. oleifera) retained 87.5% and 50% of β‐

carotene after 4 and 3 month storage, respectively, and could therefore be processed for

convenient use (Moharram et al., 2019).

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2.4.3 Nutrient Compositions Of Leaves, Leaf Powder, Seeds And Pods

(Prashith et al., 2022)

The use of different parts of Moringa oleifera is described as below:

Leaves: The leaves of MO are considered to be a rich source of vitamins and minerals and

exhibits strong antioxidant activity, often attributed to the plants’ vitamins and phenolic

compounds such asquercetin and kaempferol (Prashith et al., 2022). The leaves can be also taken

as vegetable and processed into tea, powder and other pharmaceuticals purpose. Furthermore,

juice can be extracted from fresh leaves and act like growth harm one and increase crop yield by

25-35% (Ndong et al., 2019). According to Japanese study in 2009, leaves act as a good sink for

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carbon dioxide absorption and utilisation. The rate of assimilation of carbon dioxide by Moringa

tree is 20 times higher then general vegetation (Nepolean et al., 2019). Moringa is said to

provide 7 times more vitamin C than oranges, 10 times more vitamin A than carrots, 17 times

more calcium than milk, 9 times more protein than yoghurt, 15 times more potassium than

bananas and 25 times more iron than spinach (Pari and Kumar, 2021).

Root: The root bark of Moringa has potential to cure gastric ulcers and gastric mucosal lesions.

It also decreases the acidity and increases the pH of gastric juice (Prashith et al., 2022). Hence,

MO possesses antiulcer and anti-secretary activity and hence, can be used as a source for

antiulcer drugs in future.

POD husks: MO pod husks contain alkaloids, flavonoids, tannins, tritepenoids, diterpenoids and

cardiac glycosides. MO pod husks extract show potential antimicrobial action against some gram

positive staphylococcus epidermidis, Enterococcus faecalis and Gram negative Bacteria

Klebsiella pneumonie and Salmonella typhimurium (Prashith et al., 2022).

Seed: Moringa oleifera seed contains a range of phytochemicals, including antioxidants such as

vitamin C, β-carotene, α and γ-tocopherol, β-sitosterol, vitamin A, the phenolic compounds

quercetin and kaempferol, flavonoids, and anthocyanins, along with a few rare classes of

compounds, including alkaloids, glucosinolates, and isothiocyanates. The mature seed of MO is

rich in oil, containing between 22 and 40% crude fat (Rastogi et al., 2019). Examination of the

oil composition indicates that the oil has a high proportion of monounsaturated fatty acids,

particularly oleic acid. It has been demonstrated that a higher dietary intake of monounsaturated

fatty acid (mainly oleic acid) is associated with decreased risk in coronary heart disease

(Siddhuraju and Becker, 2022). The seed extract could be used to prevent some vectors such as

for malaria Anopheles stephensi mosquitoes. The phytochemicals derived from MO seeds

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extracts are effective mosquito vector control agents and the plant extracts may be used for

further integrated pest management programs (Rao et al., 2019).

2.5 MEDICINAL PROPERTIES OF MORINGA

Moringa oleifera has enormous medicinal potential, which has long been recognized in the

Ayurveda and Unani system (Chuang et al., 2021). Nearly every part of this plant, including

root, bark, gum, leaf, fruit (pods), flowers, seed, and seed oil have been used for various ailments

in the indigenous medicine (Chumark et al., 2020), but recent research is also indicating about

several active constituents for accepting its applicability in modern medicine .

2.5.1 Antimicrobial and antihelmintic effects

Antimicrobial components of MO have been validated after the discovery of inhibitory activity

against several microorganisms. In a recent study, aqueous extracts of MO was found to be

inhibitory against many pathogenic bacteria, including Staphylococcus aureus, Bacillus

subtilis, Escherichia coli, and Pseudomonas aeruginosa in dose dependent manner (Saadabi

and Abu Zaid, 2019). MO extracts was also found to be inhibitory against Mycobacterium

phlei and B. subtilis (Eilert et al., 2018). Leaf extract of MO was found to be effective in

checking growth of fungi Basidiobolus haptosporus and Basidiobolus ranarums (Fahey, 2020).

Another study involving aqueous methanolic extract and fixed oil against microorganisms was

performed using Scenedesmus obliquus (green algae), E. coli ATCC 13706, P. aeruginosa

ATCC10145, S. aureus NAMRU 3 25923, Bacillus stearothermophilus (bacterial strains) and

Herpes Simplex virus type 1 (HSV 1) and Polio virus type 1 (sabin vaccine). Varying degree of

antimicrobial activity was observed ranging from sensitive for B. stearothermophilus to

resistant for P. aeruginosa (Al-Awwadi et al., 2022).

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Beside antibacterial activity of MO oils, it also posses anti-fungal activity (Chuang et al., 2021).

Study comparing relative antimicrobial activity of seed extracts against bacteria (Pasturella

multocida, E. coli, B. subtilis and S. aureus) and fungi (Fusarium solani and Rhizopus solani)

revealed that P. multocida and B. subtilis were the most sensitive strains, and their activity was

influenced by cations (Na+, K+, Mg2+ and Ca2+) (Jabeen et al., 2020). Another relative

comparison of antibacterial and antifungal efficacy of MO steam distillate observed more

inhibition for E. coli followed by S. aureus, Klebsiella pneumoniae, P. aeruginosa and B.

subtilis (Prashith et al., 2022). In case of fungi, Aspergillus niger was strongly inhibited followed

by Aspergillus oryzae, Aspergillus terreus and Aspergillus nidulans (Prashith Kekuda et al.,

2022). Contrary to resistance against P. aeruginosa and Candida albicans for MO in other

studies, one study using ethanolic extract of leaves, seeds and flowers showed the antimicrobial

activity against E. coli, K. pneumoniae, Enterobacter species, Proteus mirabilis, P.

aeruginosa, Salmonella typhi A, S. aureus, Streptococcus and Candida albicans (Nepolean et

al., 2021). Moringa contains pterygospermin (originally found in Moringa pterygosperma) which

has powerful antibacterial and fungicidal effects (Rao et al., 2019).

Several other specific components of Moringa have been reported with antibacterial activity,

including 4-(4'-O-acetyl-a-L-rhamnopyranosyloxy)benzyl isothiocyanate, 4-(a-L-

rhamnopyranosyloxy) benzyl isothiocyanate, niazimicin, benzyl isothiocyanate, and 4- (a-L-

rhamnopyranosyloxy) benzyl glucosinolate (Fahey, 2020). Other bioactive compounds, such

as Spirochin and Anthonine are found in root and are active against several bacteria. Anthonine

has potent inhibitory activity against Vibrio cholerae (Ghasi et al, 2020). MO flower and leaves

are also capable of controlling parasitic worms; their antihelmintic activity has been

demonstrated during several studies (Bhattacharya et al., 2019). Moreover, it has also been

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reported to inhibit Indian earthworm Pheritima posthuma with MO leaves ethanolic extracts

(Rastogi et al., 2019).

2.5.2 Anti-inflammatory activity

Moringa plant parts have substantial anti-inflammatory activity. For instance, the root extract

exhibits significant anti-inflammatory activity in carrageenan induced rat paw oedema (Singh et

al., 2020). The crude methanol extract of the root inhibits carrageenan- induced rat paw oedema

in a dose dependent manner after oral administration (Valarmathy et al., 2020). Moreover, n-

butanol extract of the seeds of MO shows anti- inflammatory activity against ovalbumin-

induced airway inflammation in guinea pigs (Mahajan et al., 2021). Amelioration of

inflammation associated chronic diseases can be possible with the potent anti-inflammatory

activity of MO bioactive compounds (Muangnoi et al., 2022).

Considering potent anti-inflammatory activity of Moringa plant, it can be surmised that this plant

shows profound influence on inflammation associated diseases and resultant symptoms. As a

consequence, this plant shows beneficial effects on asthma, pain, and other resultant symptoms

(Prashith et al., 2022).

2.5.3 Anti-asthmatic activity

It has been reported a long time ago that Moringa plant alkaloid closely resembles ephedrine in

action and can be used for the treatment of asthma. Alkaloid moringine relaxes bronchioles

(Ndong et al., 2019). The seed kernels of MO also showed promising effect in the treatment

of bronchial asthma, during a study to analyze efficacy and safety of seed kernels for the

management of asthmatic patients. The study showed significant decrease in the severity of

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asthma symptoms and also concurrent respiratory functions improvement (Agrawal and Mehta,

2020).

2.5.4 Analgesic activity

The analgesic activity of Moringa has been reported in several Moringa species. In a study

using ethanolic extracts of Moringa concanensis tender pod-like fruits in experimental animals,

a significant analgesic activity was observed (Ndong et al., 2019). Furthermore, alcoholic

extract of the leaves and seeds of MO also possess marked analgesic activity as evidenced

through hot plate and tail immersion method (Ndong et al., 2019).

2.5.5 Antipyretic activity

As a result of anti-inflammatory action of Moringa bioactive constituents, the antipyretic

activity can be hypothesized. A study was designed to assess antipyretic effect of ethanol,

petroleum ether, solvent ether and ethyl acetate extracts of MO seeds using yeast induced

hyperpyrexia method. Paracetamol was used as control during the study. Not surprisingly,

ethanol and ethyl acetate extracts of seeds showed significant antipyretic activity in rats (Hukkeri

et al., 2022).

2.5.6 Antihypertensive, diuretic and cholesterol lowering activities

Moringa leaves contain several bio active compounds, they exert direct effect on blood pressure,

and thus these can be used for stabilizing blood pressure. MO com- pounds leading to blood

pressure lowering effect includes nitrile, mustard oil glycosides and thiocarbamate glycosides

present in Moringa leaves (Anwar et al., 2021). In addition, diuretic activity of Moringa exists

in its roots, leaves, flowers, gum and the aqueous infusion of seeds (Morton, 2021). Moreover,

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Maringa leaves also contain bioactive phytoconstituent, (that is, b-sitosterol) with cholesterol

lowering effect. This compound is capable to reduce cholesterol level from the serum of high fat

diet fed rats (Ghasi et al., 2020).

2.5.7 Antidiabetic activity

Several medicinal plants have been evaluated for their potential as therapeutic agent for diabetes.

MO is also an important component in this category. MO leaves significantly decrease blood

glucose concentration in Wistar rats and Goto-Kakizaki (GK) rats, modeled type 2 diabetes

(Ndong et al., 2019). Another study indicated that the extract from Moringa leaf is effective in

lowering blood sugar levels within 3 h after ingestion (Mittal and Agarwal, 2023). As a

mechanistic model for antidiabetic activity of MO, it has been indicated that dark chocolate

polyphenols (Grassi et al., 2020) and other polyphenols (Al-Awwadi et al., 2022; Moharram

et al., 2019) are responsible for hypoglycemic activity. Moringa leaves are potent source of

polyphenols, including quercetin-3- glycoside, rutin, kaempferol glycosides, and other

polyphenols (Ndong et al., 2019). Thus, potential anti- diabetic activity of MO can be

commercialized through the development of suitable technology with achieving anti-diabetic

activity up to conventional drugs.

2.5.8 Antioxidant activity

MO is a rich source of antioxidant (Chumark et al., 2018). It has been reported that aqueous

extracts of leaf, fruit and seed of MO act as an antioxidant (Singh et al., 2020). During a

study reporting antioxidant property of freeze dried Moringa leaves from different extraction

procedures, it was found that methanol and ethanol extracts of Indian origin MO have the

highest antioxidant activity with 65.1 and 66.8%, respectively (Lalas and Tsaknis, 2019;

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Siddhuraju and Becker, 2022). It was also reported that the major bioactive compounds of

phenolics, such as quercetin and kaempferol are responsible for antioxidant activity (Bajpai

et al., 2015; Siddhuraju and Becker, 2013). During another study, quercetin and kaempferol

have shown good antioxidant activity on hepatocyte growth factor (HGF) induced Met

phosphorylation with IC50 value for 12 and ~6 µM/L, respectively (Labbe et al., 2019).

Another recent study comparing palm oil with MO seeds for their antioxidant potential found out

that MO seed are superiors for radical scavenging (Selvakumar and Natarajan, 2018).

2.5.9 Hepatoprotective activity

MO has shown significant hepatoprotective activity in several studies (Selvakumar and

Natarajan, 2018). MO leaves ethanolic extracts showed significant protection against liver

damage induced by antitubercular drugs [isoniazid (INH), rifampicin (RMP), and pyrazinamide

(PZA)] in rats. It was found that hepatoprotective activity of MO is medicated by its effect on

the levels of glutamic oxaloacetic transaminase (aspartate aminotransferase), glutamic

pyruvic transaminase (alanine aminotransferase), alkaline phosphatase, and bilirubin in the

serum; lipids, and lipid peroxidation levels in liver (Pari and Kumar, 2021). Moreover,

methanolic and chloroform extracts of MO leaves also showed significant protection against

CCl4 induced liver damage in albino rats (Hamza, 2020). Besides hepatoprotective activity

of MO leaves, its root and flowers also possess strong hepatoprotective activity. Moringa

flowers contain a well-recognized flavonoid (Quercetin), which may be responsible for its

potent hepatoprotective activity (Selvakumar and Natarajan, 2018). In a recent study

evaluating the effect of MO seed extract on liver fibrosis, it was found that MO seed extract

has the ability to subside liver fibrosis. This study involved CCl4 induced liver fibrosis and

concurrent administration of MO seed extract. MO seed extract control the elevation of serum

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aminotransferase activities and globulin level induced by CCl4. Moreover,

immunohistochemical studies also showed that MO reduces liver fibrosis (Hamza, 2020).

2.5.10 Antitumor activity

MO has been found as a potent anticancer plant and several bioactive compounds with

significant antitumor activity have been discovered from MO. Among bioactive compounds

from MO, niazimicin, a MO leaves thiocarbamate was found to have potent anticancer

activity (Guevaraa et al., 2019). Furthermore, niazimicin also shows the inhibition of tumor

promoter teleocidin B- 4-induced Epstein-Barr virus (EBV) activation (Jabeen et al., 2020).

Another study involving 11 plants used in Bangladeshi folk medicine, MO was considered as

potential source of anticancer compounds. During this study, the plant extract were

analyzed for cytotoxicity through brine shrimp lethality assay, sea urchin eggs assay,

hemolysis assay and MTT assay using tumor cell lines. The study also indicated the potential

cytotoxic effects of MO leaf extract on human multiple myeloma cell lines (Costa-Lotufo et

al., 2015; Parvathy and Umamaheshwari, 2017). Beside leaves, MO seed extracts also have

anticancer activity through its effects on hepatic carcinogen metabolizing enzymes, and

antioxidant property (Bharali et al., 2019).

2.5.11 Antifertility activity

MO plant also has pertinent antifertility activity. The aqueous extract obtained from root and

bark of MO showed post-coital antifertility effect in rat and also induced foetal resorption

at late pregnancy (Prakash et al., 2022). Moreover, aqueous extract of MO roots was also

evaluated for estrogenic, anti-estrogenic, pro- gestational and antiprogestational activities. This

extract induces several consequences for affecting its antifertility property (Pullakhandam and

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Failla 2017). During another study analyzing anti reproductive potential of folk medicine

plants, MO leaf extracts were found to be 100% abortive with doses equivalent to 175 mg/kg

of starting dry material (Nath et al., 2019).

2.5.12 Antispasmodic and antiulcer effects

Moringa root and leaves contain several compounds with spasmolytic activity. These compounds

include 4- (alpha- L-rhamnosyloxybenzyl)-o-methyl thiocarbamate which is possibly affected

through calcium channel blockade, niazinin A, niazinin B, niazimicin, etc., with hypotensive

and bradycardiac effect. The spasmolytic activity of different constituents support for

traditional uses of this plant in gastrointestinal motility disorder (Gilani et al., 2016). MO

methanolic extract is also capable in protecting experimental rats from gastric lesions induced

by acetylsalicylic acid, serotonin and indomethacin. In addition, it also enhances healing

process of chronic gastric lesions induced by acetic acid in experimental animals

(Eleyinmi, 2022). Another study has reported the antiulcer effect of MO leaves aqueous extract

on adult Holtzman albino rats (Debnath and Guha, 2017).

2.5.13 Cardiac and Circulatory Stimulant

In addition to earlier mentioned bradycardiac effect of MO leaves, all parts of MO are

reported with somewhat cardiac and circulatory stimulant activity. Root bark of Moringa

contains alkaloid moringinine which acts as cardiac stimulant through its effect on

sympathetic nervous system (Nepolean et al., 2019). The aforementioned effects can also

result due to the prevention of hyperlipidemia. It has been demonstrated that MO prevent

hyperlipidemia in male Wister rat due to iron deficiency (Ndong et al., 2019). During a

study performing comparison of MO leaf extract with antenolol (a selective β1 receptor

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antagonist drug, used for cardiovascular diseases) on serum cholesterol level, serum

triglyceride level, blood glucose level, heart weight and body weight of adrenaline induced rats,

it was found that MO leaf extract cause significant changes in cardiovascular parameters.

This study reported MO leaf extract as hypolipidimic, lowering body weight, heart weight,

serum triglyceride level and serum cholesterol level in experimental animals (Labbe et al.,

2019).

In addition to the aforementioned studies, ant atherosclerotic and hypolipidaemic effect of MO

leaves were alsoanalyzed in another study using simvastatin as control (Chumark et al., 2020).

MO also causes cardio protective effects in isoproterenol (ISP)-induced myocardial infarction in

male Wistar albino rats. It was reported that MO treatment plays favorable modulation on

biochemical enzymatic parameters including, superoxide dismutase, catalase, glutathione

peroxidase, lactate dehydrogenase, and creatine kinase-MB. Moreover, it also prevents

histopathological damage and ultra-structure perturbation caused due to ISP induced

myocardial infarction (Nandave et al., 2019).

In ocular diseases Vitamin A deficiency is a major cause of blindness, which ranges from

impaired dark adaptation to night blindness. Consumption of MO leaves, and pods and leaf

powder which contain high proportion of vitamin A can help to prevent night blindness and eye

problems in children. Ingesting drumstick leaves with oils can improve vitamin A nutrition and

can delay the development of cataract (Pullakhandam and Failla, 2017). In fact the use of MO as

a supplementary food was highly accepted for integrated child development scheme

supplementary food (ICDS-SFP) for its potential as vitamin A source (Nambiar et al., 2020).

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2.6 Other Medicinal Properties

Moringa oleifera has several medicinal properties and has potentiality to cure many diseases. It is

used to treat diseases such as diabetes, heart disease, anaemia, arthritis, respiratory problems,

skin, liver problems, paralysis, sterility, rheumatism, digestive disorders and many more. In

India, it was named the ‘plant of the year’ in 2008 by the National Institute of Health and Family

Care (Thilza et al., 2022). Several other countries like Africa, it is also used for the treatment of

ascites, pneumonia and venomous bites. According to various research, the leaves are said to be

anti-fungal, anti-viral, anti-abortifacient, and act as flocculating agent and stimulants

(Pullakhandam and Failla, 2017). Moringa powder can be used as a substitute for iron tablet,

hence a treatment for anaemia. The health benefits of this wonderful tree appear to be boundless.

Apart from all the goodness discussed above, the regular intake of Moringa is said to help further

(Rastogi et al., 2019).

2.6.1 It fights Against free radicals

The antioxidants are popular due to the fact as they fight against free radicals that cause

oxidative stress, cell damage, and inflammation (Pullakhandam and Failla, 2017). In addition to

this, Moringa contains antioxidants called flavonoids, polyphenols, and ascorbic acid in the

leaves, flowers, and seeds which are beneficial in many ways (Thilza et al., 2022). A study found

that leaf extracts has higher antioxidant activity, free-radical-scavenging capacity, and higher

inhibition of lipid, protein, and DNA oxidation than flowers and seeds. This means it prevents

the damage and degradation that free radicals cause in the cells of different organs in the body,

keeping them healthy and functioning at their best (Tsaknis et al., 2019).

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2.6.2 It protects the cardiovascular system

Moringa leaf powder has heart-healthy benefits, particularly to control blood lipid, the

prevention of plaque formation in the arteries, and reduced cholesterol levels (Tsaknis et al.,

2019). The great combination of diuretic along with lipid and blood pressure lowering

components make this plant incredibly beneficial in cardiovascular disorders. Moringa leaf juice

has very important role on stabilizing blood pressure. Mustard oil glycosides and thiocarbamate

glycosides have been isolated from Moringa leaves, which had been observed to be responsible

for lowering of blood pressure (Al-Awwadi et al., 2022).

2.6.3 Supports brain health

Moringa strongly support for brain health and boost cognitive power due to its antioxidant and

neuro-enhancer activities. It has shown several preliminary result as a treatment of Alzheimer’s

disease (Tsaknis et al., 2019). The high amount of vitamin C and E helps to improve brain

function and also normalize the neurotransmitters serotonin, dopamine and noradrenalin in the

brain which play a key role in memory, mood, organ function, responses to stimulus such as

stress and pleasure, and mental health like depression and psychosis (Agrawal and Mehta, 2020).

2.6.4 Enhances wound healing

Moringa has blood-clotting properties in its leaves, roots, and seeds that benefit wound healing

and can reduce clotting time, which means it reduces the time it takes for scratches, cuts, or

wounds to stop bleeding. Antipyretic and wound healing properties from the ethyl acetate and

ethanolic extracts of Moringa oleifera leaves were described by Hukkeri et al., (2022). The

ethanolic and ethyl acetate extracts of seeds defines significant antipyretic activity in rats, where

ethyl acetate extract of dried leaves presents wound healing activity (10% extracts in the form of

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ointment) on excision, incision and dead space (granuloma) wound models in rats (Viera et al.,

2020).

2.7 TRADITIONAL USES Morning oleifera Leaf

Moringa and its healing potential were documented for the first time around 5000 years ago in

the Vedic litrature in India (Guevaraa et al., 2019). In folk medicine, M. peregrina leaf extract is

rubbed over skin to treat paralysis and skin rashes (Hamza, 2020). The pod oil is used to treat the

convulsions or infantile paralysis in the northern region of Oman (Miller and Morris, 1988). Its

seeds are most commonly used to control diabetes in Sultanate of Oman (Jabeen et al., 2020). It

is also effectively used for the diabetes related symptoms such as hyperlipidemia and

hyperglycemia in the Indian sub-continent. The young leaves of M. peregrina are used

traditionally in folk medicine as antioxidant and wound healing in Arab countries. The bark juice

is also used as disinfectant (Labbe et al., 2019) and also to treat fever, headache, constipation,

back and muscle pains, slimness, burns and labor pain (Muangnoi et al., 2022). The leaves are

used for wound healing (Moharram et al., 2019) and seeds are used for abdominal pain. The

roots and leaves of M. peregrina are mixed together with water and used to treat hypertension,

malaria, asthma, stomach disorders, diabetes, and to expel a retained placenta (Nambiar et

al., 2020). Traditionally, the oil of this plant is used to treat skin problems such as freckles,

itches, and scabies (Jabeen et al., 2020).

In addition to their medicinal importance, M. peregrina has significant nutritional importance.

The young leaves of M. peregrina can be used as a vegetable (El Astal et al., 2020). The

immature seeds are eaten in India and mature seeds are consumed either roasted or fried in

Malawi (Eleyinmi, 2022). In traditional herbal medication, the seeds of the plant are mixed with

other herbs and used as food for anti-malnutrition (Nambiar et al., 2020). In addition, M.

20
peregrina is one of the important native trees in the UAE as it has cultural, spiritual, and

religious connections. Locally, the leaves of the plant are used to flavor the meat during smoked

meat (tanour) preparation. This traditional practice is still followed by the native people of the

UAE (Hamza, 2020).

2.7.1 Side Effects

When taken by mouth: Moringa is likely safe when the leaves, fruit, and seeds are eaten as food.

Moringa leaf and seeds are possibly safe when used as medicine, short term. Products containing

moringa leaf have been used for up to 6 months. Products containing moringa seed have been

used for up to 3 weeks. Moringa root and root bark are possibly unsafe. The roots and root bark

contain toxic substances (Prabhu et al., 2022).

When applied to the skin: Moringa leaf is possibly safe when used on the skin, short term. A

product containing moringa leaf extract has been used for up to 3 months.

2.7.2 Special Precautions and Warnings

When taken by mouth: Moringa is likely safe when the leaves, fruit, and seeds are eaten as food.

Moringa leaf and seeds are possibly safe when used as medicine, short term. Products containing

moringa leaf have been used for up to 6 months. Products containing moringa seed have been

used for up to 3 weeks. Moringa root and root bark are possibly unsafe. The roots and root bark

contain toxic substances (Parvathy et al. 2017).

When applied to the skin: Moringa leaf is possibly safe when used on the skin, short term. A

product containing moringa leaf extract has been used for up to 3 months. Pregnancy: It is

possibly safe to use moringa leaves in pregnancy during the second or third trimester. But it's

possibly unsafe to use the root, bark, or flowers of moringa when pregnant (Nath et al., 2019).

Chemicals in the root, bark, and flowers might make the uterus contract. In traditional medicine,

21
the root and bark were used to cause miscarriages. There isn't enough reliable information to

know if other parts of moringa are safe to use when pregnant. Stay on the safe side and avoid use

(Nath et al., 2019).

Breast-feeding: Moringa leaf is possibly safe to use while breastfeeding for up to 4 months.

There isn't enough reliable information to know if other parts of moringa are safe to use when

breast-feeding. Stay on the safe side and avoid use (Chumark et al., 2020).

Children: Moringa leaf is possibly safe when taken by mouth, short-term. Moringa leaf has been

used with apparent safety in children for up to 2 months (Chuang et al., 2021).

2.7.3 Dosing

Moringa is an important food source in some parts of the world. The immature green pods

(drumsticks) are prepared similarly to green beans, while the seeds are removed from more

mature pods and cooked like peas or roasted like nuts. The leaves are cooked and used like

spinach, and they are also dried and powdered for use as a condiment (Bugno et al., 2020).

Moringa is also available in supplements. Moringa leaves and seeds have most often been used

by adults in powders or extracts, in doses of 6-10 grams by mouth daily. Speak with a healthcare

provider to find out what dose might be best for a specific condition (Bharali et al., 2019).

22
 CHAPTER THREE

3.0 METHODOLOGY

3.1 Plant collection

Fresh leaves of M. oleifera were harvested at California area, Ogbomoso, Oyo State, Nigeria

(8.1333°N, 4.2356°E). Plants were collected monthly throughout year 2020 and mixed together

to avoid the influence of seasonal variation. The plant was identified and authenticated by Dr.

Mrs. Ogundola of the Botany Unit, Department of Pure and Applied Biology, Ladoke Akintola

University of Technology, Ogbomoso. The name of the botanical was confirmed

on [Link] A voucher specimen, LHO 616, was obtained and kept in the

Herbarium at the Department of Botany, Ladoke Akintola University of Technology, Ogbomoso,

Nigeria.

3.2 Preparation of plant extract

Methanolic M. oleifera leaf extract (MMOLE) was made as earlier reported (Walter et al., 2021).

Fresh leaves of Moringa oleifera were air-dried for 2 weeks and pulverized using electric

blender. About 500 g of the obtained sample was soaked in 70% methanol for 72 h (3 days).

After 3 days, the samples were filtered with muslin paper to separate the residue from the filtrate.

The filtrate was then poured inside the round bottom flask of the Soxhlet apparatus and heated

for 1 hour at 60°C, then poured inside a beaker and placed in a water bath for concentration at

100°C for 24 h. The obtained yield was 11.72%.

23
 CHAPTER FOUR

4.0 RESULT AND DISCUSSION

4.1 Discussion

Concern has been expressed about the rising prevalence of pathogenic microorganisms,

which are resistant to the newer or modern antibiotics that have been produced in the last three

decades (Valarmathy et al., 2010). Plant-based antimicrobials have enormous therapeutic

potential as they can serve the purpose with lesser side effects that are often associated with

synthetic antimicrobials. M. oleifera is highly valued plant, with impressive range of medicinal

uses and high nutritional value (Tamanna, 2010).

In this study, we evaluated the antimicrobial activity of Moringa oleifera leaf extracts

against pathogenic bacteria isolated from urinary tract infections and five standard strains of

American type culture collection. The three concentration of water extract had inhibitory effects

only on P. vulgaris standard strain. The petroleum ether extracts showed no inhibitory activity on

any organism. These results were compared with standard antibiotics Amikacin, Ciprofloxacin,

and Norfloxacin which showed moderate sensitivity against S. aureus and Amikacin was

completely resistant to K. pneumoniae isolated from UTI. These varying antimicrobial

activity of Moringa oleifera leaf extracts showed on different microorganisms was also

reported by other workers. Devendra et al., (2011) mentioned that chloroform extract of

Moringa oleifera Lam. plant leaves showed antibiotic property against wide range of pathogens

like Escherichie coli (MTCC 443), Pseudomonas aeruginosa (MTCC 424), Staphylococcus

aureus (MTCC3160) and Streptococcus pyogenes (MTCC 442). Eleyinmi (2007) and Bukar et

al., (2010) reported antimicrobial activities of Moringa oleifera leaves plant extracts on

foodborne pathogens. Thus, M. oleifera Lam. could become promising natural antimicrobial

24
agent with potential applications in pharmaceutical industry for controlling the pathogenic

bacteria. However, if plant extracts are to be used for medicinal purposes, issues of safety and

toxicity will always need to be considered.

25
 CHAPTER FIVE

5.0 CONCLUSION AND RECOMMENDATIONS

5.1 CONCLUSION

As Moringa oleifera could be grown and propagated with easy to do cultivation practices,

initiatives should be taken for its large-scale extension activities with distribution of either seed

or stem cuttings among the large population, both rural and urban setup. Owing to its richness

with varieties of essential nutrients and bioactive molecules, tree parts (leaves, drumstick, flower,

root) could be the used for solving health problems such as malnutrition, stunted growth, vitamin

deficiency, mineral deficiency and lifestyle diseases. Stress is one that is faced by all classes of

people across the world irrespective age group. As Moringa seed and leaves are rich in

antioxidant, several value-added products could be generated to augment the stress. Taking into

account the knowledge of traditional medicines coupled with growing demands of plant sourced

bioactive molecules, the Moringa oleifera perfectly emerges as an ideal tree for extensive

research and investigation for variety development, agronomic practices, harvesting protocol,

value added product generation and testing, real time data on benefits of Moringa consumption,

commercialization etc.

5.2 Recommendations

Based on the findings of this study further research will be encourage to achieve that Moringa

oleifera can be used to discover antibacterial agent for developing new pharmaceuticals using

another solvent for the extracts as water and petroleum extract only showed little or no activity

against the test organisms. Also, the extract could be against other sets of pathogenic bacteria for

26
better understanding on the extent to which Moringa oleifera can be used in curing different

disease.

27
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