ANTIMICROBIAL EFFECTS OF ANNONA MURICATA (SOUR SOP) LEAVES, BARKS AND FRUITS ON SOME SELECTED ORGANISMS

ABSTRACT

The antibacterial effect of Annona muricata linn (sour sop) leaf, bark, and fruit on some selected organisms was investigated using Escherichia coll, Salmonella and Shigella. Different concentrations (IQOmg/ml, 50mg/ml and 25mg/ml) of the aqueous and ethanol extracts of the leaf and bark, and the juice extract of the fruit were used. The Kirby-Bauer disc diffusion method was used for the susceptibility test. The results showed that the ethanol bark extract and the aqueous leaf extract at different concentrations showed little inhibition against the organisms. For ethanol bark extract the zone of inhibition observed against the test organisms were 4mm against Salmonella sp and 3mm against Shigella sp at lOOmg/ml concentrations. For the aqueous extract of the leaf, the zones of inhibition were 4mm for E. coli and 3mm for Shigella sp at lOOmg/ml concentration. The fruit juice extract also did not show any inhibition against all the three organisms tested. The antibiotics used as control (gentamicin and chloramphenicol) showed some degree of inhibition against all the organisms. The result of this study shows that this plant cannot be effectively used to treat infections caused by this test organisms, hence it cannot be recommended for use in their treatment.

TABLE OF CONTENTS

Title page i

Certification ii

Dedication iii

Acknowledgement iv

Table of contents v

List of tables viii

Abstract ix
CHAPTER ONE

1.0      Introduction 1

1.1      Aims and objectives 3
CHAPTER TWO

2.0      Literature review 4

2.1       Annona muricata linn 4

2.2.1    Chemical Compositions of Annona Muricata. 4

2.2.2   Chemical components of the leaf. 4

2.2.3    Medicinal properties of sour sop: 5

2.3       Phytochemical constituents of Annona muricato 8

2.3.1    Steroids 8

2.3.2    Cardiac glycosides: 8

2.3.3    Alkaloids 9

2.3.4    Saponins 9

2.3.5    Tannin 10

2.3.6 Flavonoids 11

2.4    Test organisms 12

2.4.1 Shigella 12

2.4.2 Salmonella 13

2.4.3 Escherichia coll 14

2.5 Antibiotics used 15

2.5.1 Chloramphenicol 15

2.5.2 Gentamicin 16

CHAPTER THREE

3.0 Materials and method 17

3.1 Sample collection 17

3.2 Media used. 17

3.3 Collection of test microorganism 17

3.4 Sterilization of equipments 17

3.5      Biochemical identification/confirmation of bacterial isolates       18

3.5.1 Gram Staining: 18

3.5.2 Catalase test. 18

3.5.3 Citrate Test 19

3.5.4 Indoletest .                                                                               19

3.5.5 Methyl red test 19

3.5.6 Voges-Proskauer test 19

3.5.7 Sugar test 20

3.5.8 Motilitytest 20

3.6      Preparation of plant extracts 21

3.6.1    Ethanol Extract Preparation 21

3.6.2    WaterExtract Preparation 21

3.7       Preparation of paper disc 21

3.8       Agar disc diffusion methods 22

CHAPTER FOUR

4.0       Results 23

CHAPTER FIVE

5.0      Discussion, conclusion, and recommendation 28

5.1       Discussion 28

5.2       Conclusion                                         . 29

5.3       Recommendation 29

References 30

LIST OF TABLES
Table Title                                                             Page

1 Characterization and identification of bacterial isolates.                                           24

2         The antibacterial effect of the ethanol leaf and bark extract against the test

organisms recorded as zones of inhibition (mm), 25

3 The antibacterial effect of the aqueous leaf and bark extract against the test

organisms recorded as zones of inhibition (mm). 26

4         The antibacterial effect of the fruit juice extract against the test organisms

recorded as zones of inhibition (mm).                                                               27 

CHAPTER ONE

1.0 INTRODUCTION

For microbial infections either caused by bacteria or fungi, antibiotics are employed. After the discovery of antibiotics, it was thought that infectious diseases will no longer exist. But due to irrational use of antibiotics, a number of bacterial strains with multi-drug resistance have emerged (Khan et al., 2009) and due to infectious diseases millions of people die every year (Dubey et al., 2012). It is a bitter fact that infectious diseases are leading causes of premature deaths which results in approximately 50,000 deaths annually around the globe (Ahmad and Beg., 2001). The unnoticeable use of antimicrobials both in developing countries led to the creation of microbial resistance problems which makes the treatment difficult especially in immune compromised patients (Ahmad and Beg. 2001).

Plants and plants products have been used as medicines since the start of history. Many researchers have conducted research on the plant products to check their antimicrobial effects (Abu-Shanab et al., 2004). The oldest known method for healing is the use of plant. Using higher plants for treatment of diseases had started since man started to live on this planet (Onyeagba et al, 2004). Traditional medicine including the herbal medicines is used at least for primary health care in some domains in almost every country. In the developing countries about 70-95% patients depend on natural medicines. In 2008, the worldwide market of natural medicine was 83 billion U.S dollars and on annual basis there is exponential increase in this bill.

Legal concerns about the herbal/ natural medicines vary widely from state to state and country to country and these medicines are used as self-medicines, health foods, functional foods, homes care remedies, over the counter medicines, prescription medicines etc. The quality of the herbal/traditional medicines is very difficult to control and maintain consistently. World Health Organisation, in cooperation with its local and regional offices has made good agricultural and collection practices (GACP) and good manufacturing practices (GMP) in addition to technical support and assistance for standardization for creation of high quality products. For understanding the approaches of quality, safety and efficacy which are based on research are needed to evaluate the traditional or herbal medicines (Robinson and Zhang. 2011).

Search for the relief from infection from natural resources (plants etc.) is not a new idea. People from all over the world use the plant products for healing. Hippocrates mentioned 300-400 medicinal plants in the late 5^th century B.C (Cowan, 1999). A number of plants contain compounds that have antibacterial property (Khan et al, 2011) compounds such as emetine; berberine and quinine which are derived from plants are very effective for the treatment of infectious microbes (Iwu et al., 1999). On the earth there are more than 300,000 plant species and only about 2% plants have been checked so far, for their antimicrobial properties. Plants extracts from more than 157 plant families have been described which have potential antimicrobial properties (Narayan et al, 2010). In United States of America (USA) about 1/4^th to ½ of the pharmaceuticals dispensed have their origin of higher plants (Cowan, 1999). Medicine which in near past had been derived from natural resources include taxol, camptothecin (anti-cancerous) and artemisinin (antimalarial). These and many other drugs clearly show that plants serve the potential source of medicine today.

Substances derived from plants have become of paramount importance currently because of their numerous functionalities (Baris et al., 2006). Medicinal plants are the greatest source of all kinds of medicines including traditional system of medicines, modern medicines, nutraceuticals and leads for new chemical entities.

Herbal medicines are the main stay of treatment in 75-80% population mainly in the developing countries. Reasons are that these have better acceptability due to the economy and with no or only a few side effects. In the near past the herbal medicines have seen increased use in the developed countries, but due to the emergence of microbial resistance to antimicrobials and side effects associated with synthetic or semi synthetic antimicrobial agents , the main focus is now on the plant derived extracts or crude plant products (Parekh et al., 2005; Khan et al., 2009).

1.1 AIMS AND OBJECTIVES

i To determine the antimicrobial effects of the leaf, bark and fruit of Annona muricata against some organisms (Escherichia coli, Salmonella sp, and Shigella sp)

ii To evaluate the effect of different extracts of the different extracts of the different parts of  Annona muricata (sour sop) on the tested organisms.

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