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CHAPTER ONE

1.0     INTRODUCTION

Medicinal plants are useful for healing as well as for curing of human diseases because of the presence of phytochemical constituents. Phytochemicals are naturally occurring chemical in the medicinal plants, leaves, vegetables and roots that have defense mechanism and protective against various diseases. Chlorophyll, proteins andcommon sugars are included in primary constituents and secondary compounds have terpenoid, alkaloids and phenolic compounds. Terpenoids exhibit variousimportant pharmacological activities i.e., anti-inflammatory, anti-cancer, anti-malarial, inhibition of cholesterol synthesis, anti-viral and anti-bacterial activities. Terpenoids are very important inattracting useful mites and fight against herbivorous insects.  Alkaloids are used as anaesthetic agents and are found in medicinal plants.[1]

Phytochemicals are defined as bioactive non-nutrient plant compounds in fruits, vegetables, grains, and other plant foods that have been linked to reducing the risk of major chronic diseases. The word ‘phyto-’ is derived from the Greek phytowhichmeans – plant. The presence of these bioactive components are said to confer the plant with resistance against bacterial, fungal and pesticidal pathogens. These bioactive components are said to be responsible for the antimicrobial effects of plant extracts.The interest in plants with antimicrobial properties has been revived due to current problems associated with the use of antibiotics with the increased prevalence of multidrug resistant (MDR) strains of a number of pathogenic bacteria. [2]

1.1     Statement of Problem

There is an ever continuous and urgent need to discover new antimicrobial compounds with diverse chemical structures and novel mechanisms of action due to the alarming increase that has been witnessed in the incidence of both new and reemerging infectious diseases. A further concern is the development of resistance to the antibiotics in current clinical use, and the problem of availability and affordability of antibiotics.

1.2 Aim and Objectives

The research tends to investigate the pharmaceutical active component of ethanol leaf extract of Corchorusolitorius so as to identify potential bioactive compounds for drug synthesis.

Objectives

The objectives of the study will be

  • To determine phytochemical composition of the leaf using some standard methods.
  • To extract the active plant constituents using ethanol
  • To investigate the antimicrobial properties of the extract using disc diffusion methods on some isolated micro-organisms
  • To compare the antimicrobial activities of the pharmaceuticals active compounds presents in the leaf with some known commercial antibiotic.

1.3     SIGNIFICANCE OF STUDY

Corchorusolitorius (Ahihara) plant has been widely acclaimed as a potent medicine traditionally therefore is a necessity to prove it scientifically, if it is found to be highly potentit may be incorporated into the health care systems after the necessary toxicity studies have been carried out. This study can afford the possibility of discovering new organic substances that are useful against disease for which suitable cure are not yet known.

1.4     SCOPE OF STUDY

This work is expected to cover

       Collection and identification of the leaf Corchorusolitorius(Ahihara) plant

       Extraction of active compounds in the leaf using ethano

1.5.    Taxonomy of Corchorusolitorius

KINGDOM: Plantae

PHYLUM: Tracheophyta

CLASS: Magnoliopsida

ORDER:Malvales

FAMILY: Malvaceae

GENUS: Corchorus L.

SPECIES: Olitorius[3]

Plate 1.1 Picture of Corchorusolitorius leaf

 

1.5.1  Description of Corchorusolitorius

Is an annual herb with slender stem with height of 6-10 cm. Corchorusolitorius is an important green leafy vegetable in many tropical areas including Egypt, Sudan, India, Bangladesh, in tropical Asia in such countries as the Philippines and Malaysia, as well as in tropical Africa, Japan, South America, the Caribbean and Cyprus. In West African countries particularly Ghana, Nigeria and Sierra Leone, where staple diets consist of starchy food-stuffs such as rice, cassava, maize and yams, leafy vegetables are used to complement such staple foods. It is cultivated to provide bark for the production of fibres (Jute) and its mucilaginous leaves are used in food as a vegetable.The crop is an excellent source of vitamins A and C, fiber, minerals including calcium and iron and other micronutrients. Corchorus. olitorius L. is extensively consumed as a “healthy vegetable” in Japan, because it contains abundant carotenoids, vitamin B1,B2, C and E, and minerals.Jute contains high levels of all essential amino acids except methionine which is at marginal concentrations.It has highprotein levels and is, along with other leafy species, the main source of dietary protein in many tropical countries. The seeds are used as a purgative and leaves as demulscent, diuretic, febrifuge (infusion) and in chronic cystitis and dysuria. On preliminary analysis, seeds have been found to contain cardenolide glycosides.Based onthe fact that there is no scientific research reporting on the antifungal activity of this plant, we decided to take this opportunity to screen for both its potential both antibacterial and antifungal activity.[4]

1.5.2  Medicinal uses of Corchorusolitorius

Corchorusolitorius leaf is used in folklore for managing diabetes and hypertension[5]CorchorusOlitorius leaf prevent internal bleeding, protects eye health, reduces restless leg syndrome, supports skin health and cell growth, fights off colds and flu, reduces cholesterol, prevent Cancer, maintains healthy Teeth and Gums, Prevents Asthma, Protects Healthy Hair and Skin. [6]

Other traditional uses and benefits of Corchorusolitorius includes

       Corchorusolitorius is a traditional remedy for aches and pains, fever, dysentery, enteritis, pectoral pains, and tumors.

       Leaves are used for ascites, pain, piles, and tumors.

       The cold infusion is said to restore the appetite and strength.

       An infusion is used in the treatment of dysentery, fevers, liver disorders and dyspepsia.

       A decoction of the roots and unripe fruits is used in the treatment of dysentery.

       The Leaf is also used as ready to cystitis, dysuria, fever, and gonorrhea.[6]

 

1.6     PHYTOCHEMICALS

Phytochemicals (from the Greek word phyto, meaning plant) are biologically active, naturally occurring chemical compounds found in plants, which provide health benefits for humans further than those attributed to macronutrients and micronutrients. They protect plants from disease and damage and contribute to the plant’s color, aroma and flavor. In general, the plant chemicals that protect plant cells from environmental hazards such as pollution, stress, drought, UV exposure and pathogenic attack are called phytochemicals. Recently, it is clearly known that they have roles in the protection of human health, when their dietary intake is significant. More than 4,000 phytochemicals have been cataloged and are classified by protective function, physical characteristics and chemical characteristics and about 150 phytochemicals have been studied in detail. [7] In wide ranging dietary phytochemicals are found in fruits, vegetables, legumes, whole grains, nuts, seeds, fungi, herbs and spices. Broccoli, cabbage, carrots, onions, garlic, whole wheat bread, tomatoes, grapes, cherries, strawberries, raspberries, beans, legumes, and soy foods are common sources. Phytochemicals  accumulate in  different  parts  of  the  plants,  such  as  in  the roots, stems, leaves, flowers, fruits or seeds. Many phytochemicals, particularly the pigment molecules, are often concentrated in the outer layers of the various plant tissues. Levels vary from plant to plant depending upon the variety, processing, cooking and growing conditions. Phytochemicals are also available in supplementary forms, but evidence is lacking that they provide the same health benefits as dietary phytochemicals. [7]

These compounds are known as secondary plant metabolites and have biological properties such as antioxidant activity, antimicrobial effect, modulation of detoxification enzymes, stimulation of the immune system, decrease of platelet aggregation and modulation of hormone metabolism and anticancer property.  There are more than thousand known and many unknown phytochemicals. It is well-known that plants produce these chemicals to protect themselves, but recent   researches demonstrate that many phytochemicals can also protect human against diseases. [7]

Phytochemicals are not essential nutrients and are not required by the human body for sustaining life, but have important properties to prevent or to fight some   common diseases. Many of these benefits suggest a possible role for phytochemicals in the prevention and treatment of disease, Because of this  property many researches have been performed to reveal the beneficial health effects of phytochemicals. [7]

1.6.1  Biological Activities of Phytochemicals

Phytochemicals present in plants which are responsible for preventing disease and promoting health have been studied extensively to establish their efficacy and to understand the underlying mechanism of their action. Such studies have included    identification and isolation of the chemical components, establishment of their biological potency both by in-vitro and in-vivo studies in experimental animals   and through epidemiological and clinical case control studies in man. Study findings suggest that phytochemicals may reduce the risk of coronary heart disease by preventing the oxidation of low-density lipoprotein (LDL) cholesterol, reducing the synthesis or absorption of cholesterol,normalizing blood pressure and clotting, and improving arterial elasticity. Phytochemicals may detoxify substances that cause cancer. They neutralize free radicals, inhibit enzymes that activate carcinogens, and activate enzymes that detoxify carcinogens. Phytochemicals have also been promoted for the prevention and treatment of diabetes, high blood pressure, and macular degeneration. Phytochemicals are classified by function an individual compound may have more than one biological function serving as both an antioxidant and antibacterial agent. [7]

1.6.2  Classification of Phytochemicals

The exact classification of phytochemicals could have not been performed so far, because of the wide variety of them. In recent year Phytochemicals are classified as primary or secondary constituents, depending on their role in plant metabolism. Primary constituents include the common sugars, amino acids, proteins, purines and pyrimidines of nucleic acids, chlorophyll’s etc. Secondary constituents are the remaining plant chemicals such as alkaloids, terpenes,lignans, plant steroids, curcumines, saponins, phenolics, flavonoids and glucosides. [7] Literature survey indicate that phenolics are the most numerous and structurally diverse plant phytoconstituents. [7]

1.6.2.1        Phenolics

Phenolics are the largest category of phytochemicals and the most widely distributed in the plant kingdom. The three most important groups of dietary phenolics are flavonoids, phenolic acids, and polyphenols. Phenolic are hydroxyl group (-OH) containing class of chemical compounds where the (-OH) bonded directly to an aromatic hydrocarbon group. Phenol (C6H5OH) is considered the simplest class of this group of natural compounds. Phenolic compounds are a large and complex group of chemical constituents found in plants. They are plant secondary metabolites, and they have an important role as defence compounds.Phenolics exhibit several properties beneficial to humans and its antioxidant properties are important in determining their role as protecting agents against free radical-mediated disease processes. Flavonoids are the largest group of plant phenols and the most studied. Phenolic acids form a diverse group that includes the widely distributed hydroxybenzoic and hydroxycinnamic acids. Phenolic polymers, commonly known as tannins, are compounds of high molecular weight. [7]

1.6.2.2        Phenolic acids

The term “phenolic acids”, in general, designates phenols that possess one carboxylic acid functional group. Naturally occurring phenolic acids contain two distinctive carbon frameworks: the hydroxycinnamic and hydroxybenzoic structures Hydroxycinnamic acid compounds are produced as simple esters with glucose or hydroxyl carboxylic acids. Plant phenolic compounds are different in molecular structure, and are characterized by hydroxylated aromatic rings. These compounds have been studied mainly fortheir properties against oxidative damage leading to various degenerative diseases, such as cardiovascular diseases, inflammation and cancer. Indeed, tumour cells, including leukaemiacells, typically have higher levels of reactive oxygen species (ROS) than normal cells so that they are particularly sensitive to oxidative stress. Many papers and reviews describe studies on bioavailability of phenolic acids, emphasizing both the direct intake through food consumption and the indirect bioavailability deriving by gastric, intestinal and hepatic metabolism.In recent years, the importance of antioxidant activities of phenolic compounds and their potential usage in processed foods as a natural antioxidant compounds has reached a new level and some evidence suggests that the biological actions of these compounds are related to their antioxidant activity. [7]

1.6.2.3        Activity of Phenolic Acids

Phenolic acid compounds and functions have been the   subject of a great number of agricultural, biological, chemical and medical studies.  Phenolic compounds in many plants are polymerized into larger molecules such as the proanthocyanidins (PA; condensed tannins) and lignins. Moreover, phenolic acids may arise in food plants as glycosides or esters with other natural compounds such as sterols, alcohols, glucosides and hydroxyfatty acids. Increases bile secretion, reduces blood cholesterol and lipid levels and antimicrobial activity against some strains of bacteria such as staphylococcus aureus are some of biological activities of phenolic acids. Phenolics acid possesses diverse biological activities, like antiulcer, anti-inflammatory, antioxidant,cytotoxic and antitumor, antispasmodic, and antidepressant activities. [7]

1.6.3 Flavonoid

Flavonoids are polyphenolic compounds that are ubiquitous in nature. More than 4,000 flavonoids have been recognised, many of which occur in vegetables, fruits and beverages like tea, coffee and fruit drinks. The flavonoids appear to have played a major role in successful medical treatments of ancient times, and their use has persisted up to now. Flavonoids are ubiquitous among vascular plants and occur as aglycones, glucosides and methylated derivatives. More than 4000   flavonoids have been described so far within the parts of plants normally consumed by humans and approximately 650 flavones and 1030 flavanols are known Small amount of aglycones (i.e., flavonoids without attached sugar) are frequently present and occasionally represent a considerably important proportion of the total flavonoid compounds in the plant. [7]

 1.6.3.1        Activity of Flavonoids 

Flavonoids have gained recent attention because of their broad biological and pharmacological activities, Flavonoids have been reported to exert multiple biological property including antimicrobial, cytotoxicity, anti-inflammatory as well as antitumor activities but the best described property of almost every group of flavonoids is their capacity to act as powerful antioxidants which can protect the human body from free radicals and reactive oxygen species. The position of hydroxyl groups and other features in the chemical structure of flavonoids are important for their antioxidant and free radical scavenging activities. On the other hand flavonoids such as luteolin and cathechins, are better antioxidants than the nutrients antioxidants such as vitamin C, vitamin E and β-carotene. Flavonoids have been stated to possess many useful properties, containing anti-inflammatory activity, enzyme inhibition, antimicrobial activity, oestrogenic activity, anti-allergic activity, antioxidant activity, vascular activity and cytotoxic antitumor activity. Flavonoids constitute a wide range of substances  that  play important role  in  protecting  biological systems against the harmful effects of oxidative processes on macromolecules, such as carbohydrates, proteins, lipids and DNA. [7]

 1.6.4  Tannin

From a chemical point of view it is difficult to define tannins since the term encompasses some very diverse oligomers and polymers. It might be said that the tannins are a heterogeneous group of high molecular weight polyphenolic compounds with the capacity to form reversible and irreversible complexes with proteins (mainly), polysaccharides (cellulose, hemicellulose, pectin, etc.),   alkaloids, nucleic acids and minerals, etc. On the basis of their structural   characteristics it is therefore possible to divide the tannins into four major groups: Gallotannins,ellagitannins, complex tannins, and condensed tannins. Tannins are  found  commonly  in  fruits  such  as grapes, persimmon, blueberry, tea, chocolate, legume forages, legume trees like Acacia spp., Sesbania spp., in grasses i.e; sorghum, corn, etc. Several health benefits have been recognized for the intake of tannins and some epidemiological associations with the decreased frequency of chronic diseases have been established. [7]

1.6.4.1        Activity of Tannins

In medicine, especially in Asian (Japanese and Chinese) natural healing, the tannin-containing plant extracts are used as astringents, against diarrhoea, as diuretics, against stomach and duodenal tumours, and as anti-inflammatory, antiseptic, antioxidant and haemostatic pharmaceuticals. Tannins are used in the dyestuff industry as caustics for cationic dyes (tannin dyes), and also in the production of inks (iron gallate ink). In the food industry tannins are used to clarify wine, beer, and fruit juices. Other industrial uses of tannins include textile dyes, as antioxidants in the fruit juice, beer, and wine industries, and as coagulants in rubber Production. Recently the tannins have attracted scientific interest, especially due to the increased incidence of deadly illnesses such as AIDS and various   cancers. The search for new lead compounds for the development of novel pharmaceuticals has become increasingly important, especially as the biological action of tannin-containing plant extracts has been well documented. [7]

1.6.5  Alkaloids

Alkaloids are natural product that contains heterocyclic nitrogen atoms, are basic in character. The name of alkaloids derives from the “alkaline” and it was used to   describe any nitrogen-containing base. Alkaloids are naturally synthesis by a large numbers of organisms, including animals, plants, bacteria and fungi. Some of  the fires natural products to beisolated   from   medicinal  plants  were   alkaloids when they first obtained from the plants materials in  the  early years of 19th century,  it  was  found that they  were  nitrogen  containing  bases  which formed  salts with  acid.  Hence  they  were  known as  the  vegetable  alkalis  or  alkaloids  and  these alkaloids  are  used  as  the  local  anesthetic  and stimulant as cocaine.  Almost all the alkaloids have a bitter taste. The alkaloid quinine for example is one of the bitterest tasting substances known and is significantly bitter (1×10-5) at a molar concentration. Alkaloids are so numerous and involve such a variety of molecular structure that their rational classification is difficult.   However, the best approach to the problem is to group them into families, depending on the type of heterocyclic ring system present in the molecule. [7] For historical reasons as also because of their structural complexities, the nomenclature of alkaloids has not been systematized. The names of individual members are, therefore, generally derived from the name of the plant in which they occur, or from their characteristic physiological activity.[7]

1.6.5.1        Activity of Alkaloids

Alkaloids are significant for the protecting and survival of plant because they ensure their survival against micro-organisms (antibacterial and antifungal activities), insects and herbivores (feedingdeterrens) and also against other plants by means of allelopathically active chemicals. The use of alkaloids containing plants as dyes, spices, drugs or poisons can be traced back almost to the beginning of civilization. Alkaloids have many pharmacological activities including antihypertensive effects (many indole alkaloids), antiarrhythmic effect (quinidine, spareien), antimalarial activity (quinine),and anticancer actions (dimeric indoles, vincristine, vinblastine). These are just a few examples illustrating the great economic Importance of this group of plant constituents. Some alkaloids have stimulant property as caffeine and nicotine, morphine are used as the analgesic and quinine as the antimalarial drug [20]

1.6.6  Terpenoids

The terpenoids are aclasses of natural products which have been derived from five-carbon isoprene units. Most  of  the  terpenoids  have multi   cyclic   structures   that   differ   from   one another  by  their  functional  groups  and  basic carbon  skeletons.  These  types  of  natural  lipids can be found in every class of living things, and therefore  considered  as  the  largest  group  of natural  products.  Many  of  the  terpenoids  are commercially interesting because of their use as flavours  and  fragrances  in  foods  and  cosmetics examples  menthol  and  sclareol  or  because  they are  important  for  the  quality  of  agricultural products,  such  as  the flavour  of  fruits  and  the fragrance  of  flowers  like  linalool.  Terpenesare  widespread  in  nature,  mainly  in  plants  as constituents  of  essential  oils.  Their building block is the hydrocarbon isoprene, CH2=C(CH3)-CH=CH2. Terpene hydrocarbons therefore  have  molecular  formula  (C5H8)n  and they  are  classified  according  to  the  number  of isoprene units. [7]

1.6.6.1        Activity of Terpenes

Among  plant  secondary  metabolites  terpenoidsare   a   structurally   most   diverse   group;   they function  as  phytoalexins  in  plant  direct  defense, or  as  signals  in  indirect  defense  responses  which involves  herbivores and  their  natural  enemies.Many plants produce volatile terpenes in order to attract specific insects for pollination or otherwise to expel certain animals using these plants as food. Less volatile but strongly bitter-tasting or toxic terpenes also protect some plants from being eaten by animals (antifeedants) last, but not least, terpenes play an important role as signal compounds and growth regulators (phytohormones) of plants, as shown by preliminary investigations. In addition, terpenoids can have medicinal properties such as anti-carcinogenic (e.g. perilla alcohol), antimalarial (e.g. artemisinin), anti-ulcer, hepaticidal, antimicrobial or diuretic (e.g. glycyrrhizin) activity and the sesquiterpenoid antimalarial drug artimisinin and the diterpenoid anticancer drug taxol.

1.6.7. Saponin

Saponins are a group of secondary metabolites found widely distributed in the plant kingdom. They form a stable foam in aqueous solutions such as soap, hence    the name “saponin”. Chemically, saponins as a group include compounds that are    glycosylated steroids, triterpenoids, and steroid alkaloids. Two main types of steroid aglycones are known, spirostan and furostan derivative. The main triterpene aglycone is a derivative of oleanane. The carbohydrate part consists of oneor   more sugar moieties containing glucose, galactose, xylose, arabinose, rhamnose, or glucuronic acid glycosidically linked to a sapogenin(aglycone).  Saponinshave one sugar molecule attached at  the  C-3  position  are called  monodesmosidesaponins, and  those that have a minimum of two sugars, one attached to the C-3  and one at C-22, are calledbidesmosidesaponins[7]

1.6.7.1        Activity of Saponins

The physiological role of saponins in plants is not yet fully understood. While there is a number of a publication describing their identification inplants, and their multiple effects in animal cells and on fungi and bacteria, only a few have addressed their function in plant cells. Many saponins are known to be antimicrobial, to inhibitmould, and to protect plants from insect attack. Saponins may be considered a part of plants’ defencesystems, and as such have been included in a large group of protective molecules found in plants named phytoanticipins or phytoprotectants. Saponin mixtures present in plants and plant products possess diverse biological effects when present in the animal body. Extensive research has been carried out into the membrane-permeabilising, immunostimulant, hypocholesterolaemic and anticarcinogenicproperties of saponins and they have also been found to   significantly affect growth, feed intake and reproduction in animals. These  structurally  diverse  compounds  have  also been observed to kill protozoans and molluscs, to be antioxidants, to impair the digestion of protein and  the  uptake  of  vitamins  and  minerals  in  the gut, to causehypoglycaemia, and to act as antifungal and antiviral.

1.7 Microbes

A microorganism or microbe is a microscopic organism, which may be single celled[8]. or multicellular. The study of microorganisms is called microbiology, a subject that began with the discovery of microorganisms in the 1670s by Antonie van Leeuwenhoek[9].Robert Hooke coined the term “cell” after viewing plant cells under his microscope. Later, in the 19th century, Louis Pasteur found that microorganisms caused food spoilage, debunking the theory of spontaneous generation. In 1876 Robert Koch discovered that microorganisms cause diseases.

Microorganisms are very diverse and include all bacteria, fungi and most protozoa. This group also contains some fungi, algae, and some micro-animals such as rotifers. Many macroscopic animals and plants have microscopic juvenile stages. Some microbiologists classify viruses and viroids as microorganisms, but others consider these as non-living[10, 11]. In July 2016, scientists identified a set of 355 genes from the last universal common ancestor of all life, including microorganisms, living on earth[12].

1.7.1  Bacteria

Bacteria (common noubacteria, singular bacterium) constitute a large domain of prokaryotic microorganisms. Typically a few micrometers in length, bacteria have a number of shapes, ranging from spheres to rods and spirals. Bacteria were among the first life forms to appear on Earth, and are present in most of its habitats. Bacteria inhabit soil, water, acidic hot springs, radioactive waste. [13] And the deep portions of earth’s crust. Bacteria also live in symbiotic and parasitic relationships with plants and animals. Most bacteria have not been characterized, and only about half of the bacterial phyla have species that can be grown in the laboratory [14].The study of bacteria is known as bacteriology, a branch of microbiology.

Types of bacteria

Gram Positive and Gram negative bacteria

Gram positive bacteria are those bacteria with thick cell wall. These wall contain many layers of peptidoglycan and teichoic acids.

Gram negative bacteria are those bacteria with thinner cell wall. These wall consist of a few layers of peptidoglycan surrounded by a second lipid membrane containing lipopolysaccharides and lipoproteins

These names originate from the reaction of cells to the Gram stain, a long-standing test for the classification of bacterial species[15].

. Lipopolysaccharides, also called endotoxins, are composed of polysaccharides and lipid A that is responsible for much of the toxicity of gram-negative bacteria. Most bacteria have the gram-negative cell wall, and only the Firmicutes and Actinobacteria have the alternative gram-positive arrangement[16]. These differences in structure can produce differences in antibiotic susceptibility; for instance, vancomycin can kill only gram-positive bacteria and is ineffective against gram-negative pathogens, such as Haemophilusinfluenzae or Pseudomonas aeruginosa[17].

1.7.1.1Eschericia coli

It is naturally found in the intestine tract, in soil and water, it is the commonest pathogen associated with patient with cystitis, wound, meningitis, diarrhea disease like dysentery[17].

1.7.1.2 Streptococcus species

They are parasites of humans and animals, they can cause streptococcal sore throat, scarlet fever, erysipelas, acute glomerulonephritis, rheumatic fever, endocarchitis and nearly 70% of lobal pneumonia in human [18].

1.7.1.3Shigella

It is a genus of Gram-negative, facultative anaerobic, nonspore-forming, nonmotile, rod-shaped bacteria genetically closely related to E. coli. The genus is named after Kiyoshi Shiga, who first discovered it in 1897.[19].

The causative agent of human shigellosis, Shigella causes disease in primates, but not in other mammals.It is only naturally found in humans and gorillas.During infection, it typically causes dysentery[20].Shigella is one of the leading bacterial causes of diarrhea worldwide, causing an estimated 80–165 million cases.The number of deaths it causes each year is estimated at between 74,000 and 600,000 deaths.It is in the top four pathogens that cause moderate-to-severe diarrhea in African and South Asian children [21].

1.7.1.4Staphylococcus aureus

It is a gram-positive, round-shaped bacterium that is a member of the Firmicutes, and is frequently found in the nose, respiratory tract, and on the skin. It is often positive for catalase and nitrate reduction and is a facultative anaerobe that can grow without the need for oxygen.Although S. aureus is not always pathogenic, it is a common cause of skin infections including abscesses, respiratory infections such as sinusitis, and food poisoning. Pathogenic strains often promote infections by producing virulence factors such as potent protein toxins, and the expression of a cell-surface protein that binds and inactivates antibodies. The emergence of antibiotic-resistant strains of S. aureus such as methicillin-resistant S. aureus (MRSA) is a worldwide problem in clinical medicine. Despite much research and development there is no approved vaccine for S. aureus.

  1. aureus can cause a range of illnesses, from minor skin infections, such as pimples,impetigo, boils, cellulitis, folliculitis, carbuncles, scalded skin syndrome, and abscesses, to life-threatening diseases such as pneumonia, meningitis, osteomyelitis, endocarditis, toxic shock syndrome, bacteremia, and sepsis. It is still one of the five most common causes of hospital-acquired infections and is often the cause of wound infections following surgery. Each year, around 500,000 patients in hospitals of the United States contract a staphylococcal infection, chiefly by S. aureus[22].

1.7.1.5Candida alblicans:

This is often isolated from warm blooded animals which include human. It exist as part of the normal microbiota of mucus membranes. Sometimes this fungus may become pathogenic causing candidiasis, a disease of the mucus membranes of the mouth, vagina and alimentary tract. More serious infections can involve the heart, the blood and the brain. Candida alblican can be obtained from the urine sample of an infected patient [18].

1.7.1.6AspergillusFlavus and Niger

Aspergillus was first catalogued in 1929 by Italian priest and biologist Antonio Micheli. Group of diseases caused by Aspergillus are called Aspergillosis and their symptom include fever, cough, chest pain, breathlessness. They are highly aerobic and found in almost all oxygen rich environments. They are important medically and commercially and can cause infections in humans and other animals[23].

 1.8     Literature Review

Ilhanet at (2007) investigated the antimicrobial activity of three extracts of Corchorusolitorius usingpetroleum ether, methanol and ethyl acetate+water extracts of C. olitoriususing agar-well diffusion method.The result revealed that all the extracts displayed varied levels of zones of inhibition against bacteria and fungiused [4].

Ganiyuet al(2012) assessed the Inhibitory effect of polyphenol-rich extracts of Corchorusolitorius on key enzyme linked to type 2 diabetes (α-amylase and α-glucosidase) and hypertension the result revealed that the extracts inhibited α-amylase and α-glucosidase[5].

Rehab, M.O. (2016) studied the phytochemical, Antimicrobial and Antioxidant Activity of Corchorusolitorius leave extractHis result revealed thatthe 96% ethanolic, Ethyl acetate and chloroform extracts exhibit significant antimicrobial activity and it show the presence of sterols and triterpenes,Carotenoids, coumarins, alkaloids, saponins, tannins,flavonoid and carbohydrates[24].

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