THE EFFECT OF METHANOL RHIZOME EXTRACT OF Nymphaea lotus LINN. (NYMPHAEACEAE) IN ANIMAL MODELS OF DIARRHOEA

TABLE OF CONTENTS
Title page
Abstract
Table of Contents
Abbreviations, Definitions, Glossary and Symbols

CHAPTER ONE
1.0       INTRODUCTION
1.1       Statement of Research Problem
1.2       Justification
1.3       Aim and Objectives
1.3.1    Aim
1.3.2    Specific objectives
1.4       Hypothesis

CHAPTER TWO
2.0       LITERATURE REVIEW
2.1       Diarrhoea
2.1.1    Definition
2.1.2    Aetiology
2.1.3    Epidemiology
2.1.4    Pathophysiology/Pathological disorders
2.1.5    Types of diarrhoea
2.1.6    Other types of diarrhoea
2.1.7    Signs and symptoms
2.1.8    Risk factors
2.1.9    Complications of the disease
2.1.10  Diagnosis
2.1.11  Prevention
2.1.12  Management and treatment
2.1.13 Antidiarrhoeal drugs in recent use
2.2       The Plant: Nymphaea lotus
2.2.1    Taxonomy and nomenclature
2.2.2    Synonyms
2.2.3    Common/English names
2.2.4    Vernacular names
2.2.5    Botanical description / morphology
2.2.6    Geographical distribution / habitat
2.2.7    Ethno medical Uses
2.2.8    Pharmacological studies on Nymphaea lotus plant
2.2.9    Other medicinal plants used for management of diarrhoea

CHAPTER THREE
3.0       MATERIALS AND METHODS
3.1       Materials
3.1.1    Plant material
3.1.2    Experimental animals
3.1.3    Equipment and other laboratory materials
3.1.4    Chemicals and drugs
3.2       Methods
3.2.1    Preparation of the plant extract
3.2.2    Preliminary phytochemical screening
3.2.3    Acute Toxicity Studies (LD50)
3.2.4    In vitro Studies
3.2.5    Antidiarrhoeal Studies
3.2.8    Data Analysis

CHAPTER FOUR
4.0       RESULTS
4.1       Percentage yield of crude rhizome extract of Nymphaea lotus
4.2       Phytochemical constituents of methanol rhizome extract of Nymphaea lotus
4.3       Acute toxicity study
4.4       In vitro Studies
4.4.1    Effect of methanol rhizome extract of Nymphaea lotuson isolated rabbit jejunum
4.4.2    Effect on of methanol rhizome extract of Nymphaea lotuson isolated guinea pig ileum
4.5       Castor oil-induceddiarrhoea in mice
4.6       Magnesium sulphate-induceddiarrhoea in mice
4.7       Effect of methanol rhizome extract of Nymphaea lotuson gastric transit time in mice

CHAPTER FIVE
5.0       DISCUSSION

CHAPTER SIX
6.0       SUMMARY, CONCLUSION AND RECOMMENDATIONS
6.1       Summary
6.2       Conclusion
6.3       Recommendation
REFERENCES

Abstract
Nymphaea lotus is one of the foremost aquatic macrophytes that have been identified in Nigerian fresh water bodies. It finds applications in the management of circulatory system disorders, digestive system disorders, infectionsand inflammations. This study aims to evaluate the antidiarrhoeal activity of the methanol rhizome extract of N. lotus Linn plant in laboratory animals. Shade-dried rhizome of N. lotus was extracted with 80% methanol using Soxhlet apparatus. Preliminary phytochemical screening was carried out.This revealed the presence of alkaloids, anthraquinones, carbohydrates, cardiac glycosides, flavonoids, saponins, steroids, tannins and triterpenes. The oral median lethal dose (LD50) was determined andacute toxicity test of the extract gave LD50 value greater than 5,000mg/kg p.o. in mice. The antidiarrhoeal activityof the methanol extract was determined in mice. The extract was screened for activity against castor oil-induced diarrhoea and magnesium sulphate-induced diarrhoea as well as effect on gastric transit time in mice. For castor oil-induced diarrhoea, the extract at doses of 200, 400 and 800mg/kg produced statistical significant reduction in the frequency of diarrhoea (at p<0 .001="" 200="" 200mg="" 3mg="" 400="" 400mg="" 800="" 800mg="" a="" along="" also="" and="" antidiarrhoeal="" antimotility="" at="" atropine="" charcoal="" comparable="" delay="" diarrhoea="" diarrhoeawas="" doses="" drug="" effect="" extract="" frequency="" greater="" groups="" in="" intestinal="" kg="" ll="" loperamide="" magnesium="" meal="" mg="" of="" onset="" p="" produced="" protection="" reduced="" reduction="" respectively.="" respectively="" significant="" significantly="" span="" standard="" statistically="" sulphate-induced="" the="" to="" tract="" transit="" treated="" with=""> compared to the extract. Effect of methanol rhizome extract of N. lotus on isolated rabbit jejunum and guinea pig ileum was determined. The methanol rhizome extract ofNymphaea lotus at low concentration (4×10-4  6.4×10-2mg/ml) had contractile effect on the tone of contraction of the rabbit jejunum while at higher concentrations (8×10-2 -512×10-2 mg/ml) produced significant reduction in the tone and rate of spontaneous contraction of rabbit jejunum. Interaction of the extract,at concentration that causes relaxation, with acetylcholine,attenuated the effect of the latter in a manner similar to atropine.The methanol rhizome extract of N. lotus at lower concentrations (4×10-4 to 2×10-2 mg/ml) has no effect on contraction of the guinea pig ileum while higher concentrations (4×10-2 - 512×10-2 mg/ml) produced significant relaxant activity on guinea pig ileum. Interaction of the extract with histamine attenuated the effect of the latter which may be as a result of physiologic antagonism.This study has shown that the methanol rhizome extract of N. lotus has antidiarrhoeal properties thus justifying its use by the local population for this purpose.

CHAPTER ONE
1.0              INTRODUCTION
Diarrhoea is characterized by increased frequency of bowel movement, wet stool and abdominal pain (Ezekwesili et al., 2004). Diarrhoeal disease account for nearly 1.3 million deaths a year among children under-five years of age, making it the second most common cause of child deaths worldwide, with half of the deaths occuring in just five countries: India, Nigeria, Afghanistan, Pakistan and Ethiopia (UNICEF, 2012).

Diarrhoea occurs most of the time when there is an imbalance between the absorption of and secretion in gastrointestinal tract. When the absorptive capacity of the intestine is exceeded and net secretion is greater than absorption,excess loss of fluid in faeces results (Nigro et al., 2000; Guerrant et al., 2001).

According to the World Health Organization (WHO), diarrhoea is one of the most common causes of morbidity and mortality in many developing countries affecting mainly infants and children (Fernandoet al., 2010). In Nigeria, it is the number one fatal outcome disease among children under five years of age (Ahmed et al., 2007).

The WHO in 2004, introduceda programme for control of diarrhoea which involved and encouraged the use of traditional herbal medicine (Njume and Goduka, 2012). This was introduced keeping in mind centuries‘ old efficacy, experience, accessibility and cost (Yakubuet al., 2012).

Traditional medicine is a widely used and a rapidly growing health system with high economic importance. Up to 80% of the population in Africa uses traditional medicine to help meet their health care needs (WHO, 2002). This is mainly due to the economic viability, accessibility and ancestral experience (Wendell et al., 2008). Traditional Medicine remains an indispensable component of Nigeria‘s health services and practices, patronized by a large percentage of the population. It should therefore be integrated into the nation‘s health delivery system in a manner that protects users of traditional medicine and the general public (APCON, 2014). Local herbalists have depended on medicinal plants as a reliable means of treating diarrhoea. Hence the use of medicinal plants that possess anti-diarrhoeal activities has been explored as a measure that could be of benefit in combating widespread diarrhoea infections especially in third world countries (Adeyemi et al., 2003). Numerous studies have validated the traditional use of antidiarrhoeal medicinal plants by investigating the biological activity of extracts of such plants, which have antispasmodic effects, delay intestinal transit, suppress gut motility, stimulate water absorption or reduce electrolyte secretion (Kambaska et al., 2006)....

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