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PROJECT TOPIC ON ANTIDIARRHOEAL AND ANTIBACTERIAL ACTIVITIES OF METHANOLIC EXTRACTS AND FRACTIONS OF THE PULP AND SEED OF ZIZIPHUS MAURITIANA (CHINEE APPLE)

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ANTIDIARRHOEAL AND ANTIBACTERIAL ACTIVITIES OF METHANOLIC EXTRACTS AND FRACTIONS OF THE PULP AND SEED OF ZIZIPHUS MAURITIANA (CHINEE APPLE)

ABSTRACT

This study evaluated, the antidiarrhoeal and antibacterial activity of the methanolic extracts of seed and pulp of Ziziphus mauritania. The crude extracts were prepared by maceration of seed and pulp powder in methanol. The pulp extract was partitioned into n-hexane, ethylacetate, n-butanol and aqueous fractions. The ethylacetate extract was further fractionated by column chromatography into four fractions A-D. The antibacterial activity of the crude extracts, and fractions were evaluated by Agar well diffusion method .The antidiarrhoeal activity of the crude seed and pulp extracts were evaluated using castor oil-induced diarrheal model, charcoal meal test and anti-fluid accumulation test in rats. The result of phytochemical test indicated that tannins, flavonoids, saponin, cyanogenic glycosides, and terpenoids were present in both seed and pulp extracts. The antibacterial susceptibility test showed that the seed and pulp extracts possess inhibitory activity against Staphyloccus aureus, Eschericial coli and Salmonella spp. The fractions obtained from the pulp extract also showed varied antibacterial activity, the diameter of the zone of inhibition obtained ranged from 29-11 and 27-10 mm for seed and pulp respectively. In the castor oil-induced model, fluid accumulation test, the methanolic extracts of the seed and pulp produced a significant (p<0.05) decline in volume of intestinal contents. Results from the charcoal meal test revealed that all the extract produced a significant (p<0.05) anti-motility effect. Based on the findings of this work, the pulp methanolic extract of this plant possess antibacterial and anti-diarrhoeal properties and validates its use in traditional medicine for the treatment of diarrhoea and a number of ailments caused by microorganisms.

CHAPTER ONE

INTRODUCTION

1.1         Background of the Study

Since the beginning of human civilization, medicinal plants have been used for their therapeutic value. Nature has been a source of medicinal agents for thousands of years and an impressive number of modern drugs have been isolated from natural sources. Many of these isolations were based on the uses of the agents in traditional medicine. The plant-based, traditional medicine system continues to play an essential role in health care, with about 80% of the world‟s inhabitants relying mainly on traditional medicines for their primary health care (Shaikh and Hatcher, 2005).

According to the World Health Organization “a medicinal plant” is any plant, which in one or more of its organs contains substances that can be used for the therapeutic purposes or which, are precursors for the synthesis of useful drugs(WHO, 2001). This definition distinguishes those plants whose therapeutic properties and constituents have been established scientifically and plants that are regarded as medicinal but which have not yet been subjected to thorough investigation. The term “herbal drug” determines the part/parts of a plant (leaves, flowers, seeds, roots, barks, stems, etc.) used for preparing medicines (Anonymous, 2007). Furthermore, WHO defines medicinal plant as herbal preparations produced by subjecting plant materials to extraction, fractionation, purification, concentration or other physical or biological processes which may be produced for immediate consumption or as a basis for herbal products(WHO,2004). Medicinal plants are plants containing inherent active ingredients used to cure disease or relieve pain (Okigboet al., 2005).

The use of traditional medicines and medicinal plants in most developing countries as therapeutic agents for the maintenance of good health has been widely observed (UNESCO, 1998). Modern pharmacopoeia still contains at least 25% of drugs derived from plants and many others, which are synthetic analogues, built on prototype compounds isolated from plants. Interest in medicinal plants as a re-emerging health aid has been fuelled by the rising costs of prescription drugs in the maintenance of personal health and well-being and the bioprospecting of new plant-derived drugs. The growing recognition for medicinal plants use is due to several reasons, including escalating faith in herbal medicine (Kala, 2005). Furthermore, an increasing reliance on the use of medicinal plants in the industrialized societies has been traced to the extraction and development of drugs and chemotherapeutics from these plants as well as from traditionally used herbal remedies (UNESCO, 1998).

The medicinal properties of plants could be based on the antioxidant, antimicrobial, antipyretic effects of the phytochemicals in them (Ayodele, 2005). According to World Health Organization, medicinal plants would be the best source to obtain a variety of drugs. Therefore, such plants should be investigated to better understand their properties, safety and efficacy (Nascimentoet al., 2000).

Medicinal plants produce bioactive compounds used mainly for medicinal purposes. These compounds either act on different systems of animals including man, and/or act through interfering with the metabolism of microbes infecting them. The microbes may be pathogenic or symbiotic. In either way, the bioactive compounds from medicinal plants play a determining role in regulating host-microbe interaction in favour of the host. So the identification of bioactive compound in plants, their isolation, purification and characterization of active ingredients in crude extracts by various analytical methods is important. The instant rising demand of plant-based drugs is unfortunately creating heavy pressure on some selected high-value medicinal plant populations in the wild due to over-harvesting. Several of these medicinal plant species have slow growth rates, low population densities, and narrow geographic ranges (Kala et al., 2006); therefore they are more prone to extinction. Conversely, because information on the use of some plant species for therapeutic purpose has been passed from one generation to the next through oral tradition, this knowledge of therapeutic plants has started to decline and become obsolete through the lack of recognition by younger generations as a result of a shift in attitude and ongoing socioeconomic changes (Kala, 2000).

Phytochemical screening, evaluation of antimicrobial properties and antidiarrhoeal status of medicinal plants are today recognized as the most viable methods of identifying new medicinal plants or refocusing on those earlier reported for bioactive constituents (Adjanahounet al., 2001). Plants which are observed to be efficacious and frequently prescribed may contain compounds that are potential drug candidates and could rightly be recommended for further examination (Tor-Anyiinet al., 2003).

 

The growing resistance of pathogenic organisms against the conventional antibiotics formerly recognized for their efficiency is today a real problem of public health (Nascimentoet al., 2000). In human pathology, microorganisms are responsible for many infections including respiratory tract diseases (pneumonia, bronchitis), skin, wound and mucous infections, sinusitis, endocartidis, osteomyelitis, syphilis, gonorhea, tuberculosis, food poisoning and carbuncles to mention a few. They are also the germs frequently met during surgical wound infections which are often provoked by the use of intravascular catheters or by the spread of bacteria from another source of infection (Landoloet al., 2002). Staphylococcus aureus for instance, one of the most pathogenic among the species of Staphylococcus is responsible for almost 25 % of septicemiasmet in hospitals (Moyenet al., 2003). Generally, the treatment of infections caused by microorganisms is long and expensive. The antimicrobial properties of plants have been investigated by a number of researchers worldwide thorough biological evaluation of plants extracts it is vital to ensure their efficacy and safety. These factors are of importance if plant extracts are to be accepted as valid medical agents for the treatment of infectious diseases especially in the light of the emergence of drug resistant microorganisms (Nascimentoet al., 2000).

Diarrhoea is a common gastrointestinal disorder characterized by an increase in stool frequency and a change in stool consistency (Farthing, 2002). It remains one of the major health threats to populations in the tropical and subtropical poor countries. According to the World Health Organization estimates that 1.7 billion cases occur each year (760 000 in children less than 5 years of age), and that approximately 5 million deaths are due to diarrhoea annually (2.5 million in children less than 5 years of age) (WHO, 2013). Bacteria and some parasitic organismshave long been implicated in the incidence of diarrhoea (WHO, 1999). Diarrhoea is a common symptom of gastrointestinal infections caused by a wide range of pathogens, including bacteria (Escherichia coli, Shigella, Campylobacter, Vibriocholerae), viruses (rotavirus), and protozoa (Cryptosporidium). However, a handful of other organisms are also responsible for most acute cases of childhood diarrhoea. Among these, rotavirus is the leading cause of acute diarrhoea, and responsible for about 40% of all hospital admissions due to diarrhoea among under five children worldwide (CDC, 2008).

Evidence from report of several experimental studies shows that plant material with antimicrobial activity also possesse significant antidiarrhoeal activity particularly in infectious diarrhoea. In both cases, these activities have been attributed to the presence of bioactive agents such as tannins, alkaloids, saponins, flavonoids, steroids, and terpenoids (Longanga Otshudi et al., 2000; Umer et al., 2013; Getnet, 2015).

ZiziphusmauritianaLam belongs to the family Rhamnaceae. It is widely grown in mild-temperate, rather dry areas, of both hemispheres and is adapted to warm climates. Ziziphusmauritianacan grow either as shrublets, shrubs or trees with thornybranches and are used as a hedge to form defensivefences for animals. It is often called merely jujube, Chinese date, Indian plump (Morton, 1987). In Northern Nigeria it is called “Magarya” in Hausa or “Huhue” in bura. The plant finds various uses in traditional medicine for instance; the pulps are applied on cuts and ulcers; are employed in pulmonary ailments and fevers; the dried ripe pulp is a mild laxative. The seeds are sedative and are taken sometimes with butter, to halt nausea, vomiting and abdominal pains in pregnancy. Mixed with oil, they are rubbed on rheumatic areas. The leaves are helpful in liver trouble, asthma and fever. The bitter, astringent bark decoction is taken to halt diarrhoea and dysentery and relieve gingivitis. A root decoction is given as a febrifuge, taenicide and emmenagogue, and the powdered root is dusted on wounds. Juice of the root bark is said to alleviate gout and rheumatism (Morton, 1987). The root is also used in the treatment of epilepsy (Msonthi and Magombo., 1983). The dried root is also used to treat diarrhoea in Northern parts of Nigeria (Personal communication). The leaves are applied as poultices and are helpful in liver troubles, asthma andfever (Michel, 2002).The hepatoprotective activity of ethanol extract of Ziziphus mauritianaleaf against CCl4 – induced liver damage in rats and the antidiarrhoeal activity of the methanol root extract were reported (Dahiruet al., 2005, 2006). The antioxidant activity of the aqueous extract of Ziziphus mauritiana leaf has also been reported(Dahiru and Obida, 2008).

ANTIDIARRHOEAL AND ANTIBACTERIAL ACTIVITIES OF METHANOLIC EXTRACTS AND FRACTIONS OF THE PULP AND SEED OF ZIZIPHUS MAURITIANA (CHINEE APPLE)

1.2         Statement of Research Problem

Diarrhoea is one of the major factors that contribute significantly to high child morbidity and mortality in many developing countries (Onyango and Angienda, 2010). Among children under 5 years old, about 9% of all deaths occur due to diarrhoea and children of under 3 years‟ experience an average of three episodes per year. Annually, at least 1,500 million episodes of diarrhoea occur and at least 1,600 young children die each day or about 580,000 each year (Kumar and Subita, 2012; UNICEF, 2013).An estimated 1.5million children under the age of 5 die from diarrhoea each year more than AIDS, malaria, measles combined according to joint UNICEF-WHO reports (2009).

Although many different antibacterial agents are available in the field of medicine, yet many of these agents are increasingly being incapacitated by microorganisms through the evolution of different mechanisms that amount to resistance to these drugs (Walsh, 2000).In developing countries, synthetic drugs are not only expensive and inadequate for the treatment of diseases but also are often with adulterations and side effects (Shariff, 2001).

1.3         Justification of the Study

Phytomedicine has demonstrated its contribution to the reduction of excessive mortality, morbidity and disability due to diseases such as malaria, tuberculosis, sickle-cell anemia, diabetes, mental disorders and microbial infections (Okigboet al., 2005). It has played a key role in world health care with about 80% of Africans depending on it (WHO, 2004). It hasassisted in reducing poverty by increasing the economic well-being of communities and developed health system by increasing health coverage to the people. Substantial amount of foreign exchange has been earned by exporting medicinal plants to other countries. In this way, indigenous medicinal plants play significant role in the economy of a country (Ghani, 2003).

Research has shown that many herbs possess varying degrees of antimicrobial activities. Kaufman et al., (2009) had reported that more than 25% of the prescribed drugs contained at least one active ingredient of plant origin. Despite widespread use of oral rehydration therapies (ORT) and an increased understanding of the pathogenesis of diarrhoea millions of children still die from this disease condition every year(Thapar and Sanderson,2004).Medicinal plants are now very popular in developing countries with knowledge about the safety, efficacy and quality assurance (Calixto, 2000). Therefore there is a need for a continuous search for new effective and affordable plant derived drugs.

1.4         Aim and Objectives

1.4.1    Aim

The aim of this work was to evaluate the phytochemical constituents, antidiarrhoeal and antibacterial activities ofthe methanolic extracts and fractions of thepulp and seed of Ziziphusmauritiana.

ANTIDIARRHOEAL AND ANTIBACTERIAL ACTIVITIES OF METHANOLIC EXTRACTS AND FRACTIONS OF THE PULP AND SEED OF ZIZIPHUS MAURITIANA (CHINEE APPLE)

1.4.2     Specific Objectives

  1. To determine the phytochemical composition of thepulp and seeds extractsof Z.
  2. To determine the antidiarrhoeal activity of the methanolic extracts of pulp and seed of mauritiana in rats.
  3. To determine the antibacterial activity of the methanolic extracts of thepulp and seed of mauritiana against Staphylococcus aureus, Escherichia coli,and Salmonella spp.
  4. To fractionate the extracts and determine the fraction with the best antibacterial activity.

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