ANTIFUNGAL EFFECTS OF MICROORGANISMS ISOLATED FROM SNAIL GUT

ABSTRACT

The quest for search of a new microbial active agent has led to the exploitation of the snail .Achatina fulica, this species is known to carry parasites and harbor a dense metabolically active community. The microorganisms isolated from snail gut samples gotten from Ahiaeke Market, Abia State, Nigeria was studied. The bacterial isolated from the snail gut were Micrococcus sp, Bacillius sp, Pseudomonas sp, and Enterobacter sp. And fungi isolated were Aspergillis sp, Penicillium sp, and Fusarium sp.  Aspergillius sp and Penicillium sp were used as test organisms against the isolated bacterial. To know which inhibits the growth and shows antifungal effect against test organisms. Aspegillus sp and Penicillum sp with D1 showing the highest zone of inhibition (16mm) and E1 showing the least zone of inhibition (9mm)

TABLE OF CONTENTS

Title page                                                                                                                                i

Declaration                                                                                                                             ii

Certification                                                                                                                           iii

Dedication                                                                                                                               iv

Acknowledgements                                                                                                                v

Table of Contents                                                                                                                   vi

List of Tables                                                                                                                          viii

Abstract                                                                                                                                  ix

CHAPTER ONE

1.1           INTRODUCTION                                                                           1

1.1        Aim                                                                                             3

1.2        Objective                                                                   3                                     

CHAPTER TWO

LITERATURE REVIEW

2.1       Snail Description                                                                                                        4

2.1.1    Diet                                                                                                                             4

2.1.2    Damage                                                                                                                       5

2.1.3   Prevention and Avoidance                                                                                         7

2.1.4    Physical Barriers                                                                                                         7

2.1.5    Cultural Practices                                                                                                       8

2.1.6    Biological Control                                                                                                      9

2.2       Overview of Fungi                                                                                                      9

2.2.1    Characteristic/functions of fungi life                                                                          10

2.2.2    Life water associated fungi (aquatic fungi)                                                                10

2.3       Overview of bacteria                                                                                                  12

2.3.1    Growth                                                                                                                        12

2.3.2    Factors that affect the generation time of bacteria                                                     13

2.3.3    Bacterial metabolism                                                                                                  14

2.3.4    Human bacterial diseases                                                                                           14

2.3.5    Importance of bacteria                                                                                                15

2.3.6    Secondary metabolites of fungi                                                                                  15

2.4       Aspergillus                                                                                                                 17

2.5       Penicillium                                                                                                                18

CHAPTER THREE

MATERIALS AND METHODS

3.1       Collection of samples                                                                                                 20

3.2       Isolation of Microorganism from the Gut of Snail                                                     20

3.2.1    Screening of isolate for antagonism (antimicrobial effect)                                        20

3.2.2    Identification and Characterization of isolate                                                           21

3.3       BIOCHEMICAL TESTS                                                                                           21

3..3.1   Gram Staining                                                                                                             22

3.3.2    Catalase Test                                                                                                               22

3.3.3    Indole Test                                                                                                                  22

3.3.4    Citrate Utilization Test                                                                                               22

3.3.5    Hydrogen Sulphide (H₂S) Production Test                                                               22

3.3.6    Oxidase Test                                                                                                               23

3.3.7    Starch Hydrolysis                                                                                                       23

3.4.      Antifungal Screening                                                                                                  24

3.5       Agar well diffusion method                                                                                        24

3.6       Measuring zone of inhibition                                                                                     24

CHAPTER FOUR

4.1       RESULTS                                                                                                                   26

CHAPTER FIVE

DISCUSSION, CONCLUSION AND RECOMMENDATIONS

 5.1      Discussion                                                                                                                   30       

5.2       Conclusion                                                                                                                  32

REFERENCES

APPENDIX

List of Tables

Table 1:  Diameter zone of inhibition (mm) of isolates against test fungi                27

Table 2: Description of culture Morphology and Biochemical features of the isolates.                                    28

Table 3: Identification of fungi isolated                                      29

CHAPTER ONE

            1.1           INTRODUCTION

Microorganism in nature competes for survival in their habitats.  The biologics took of antagonistic nature is achieved by the development of competitive mechanisms such as the production of antimicrobial agents like antibiotics, sideophores, dioxin and enzymes, etc. antagonistic microorganism, by their interactive with various pathogens play a major role in microbial equilibrium and serves as powerful agent for biological disease control.

The interaction between antimicrobial agents produced by bacteria and plant pathogen has been studied extensively, and the application of antimicrobial agents in the protection of crops or promising, besides their pharmaceutical application.

In the post-industrial information age, biotechnological era is replaced largely the mechanical and chemical eras of the industrial age.  In the back drop of globalization, there is a reawakening to our ultimate reliance on biological processes.  These has been a rise in studies of relationship between invertebrates, microorganism and plants/soil development (Edward et al.,1988).  Investigation of the interaction of microorganism with soil invertebrates is one of the ways to study the development in biogeocenoses (Szabo, 2001, Ehu et al., 2000). As snails are bred in bioconversion of organic works their gut flora must be explored as they are ecofriendly and farmers best friends. So the present study was focused on the gut bacteria, fungal and their activities.

Since the time of the “green Revolution” promoted in early part of 20^th century, the food production was boosted but without forisecing some of its oil effects.  With the recent realization to maintain ecological balance for the substances of agriculture production, farmers and scientist alots are aiming of finding an alternative to chemical agriculture.  The potential use of microbes based biocontrol agents as replacement or supplement for agrochemical has been addressed in many recent reports (Shimizu et al., 2000, Aghighi et al., 2004, Shahidi Bonjor et al., 2006).

The green snail, Turbo marmorates (Gastropoda Turbinidae), has been widely used as food items in tropical and subtropical western pacific regions, as well as a material for shell craft in Korea, China, Japan and Europe for hundreds of years (Avakawa 1985).  Over the past twenty years, the population of the green snail in the Ryukyu Island (Southern Japan) and Southern Ashier have markedly diseased are become endangered because of heavy over fishing (Chantrapornsyl 1995).  Pactectard sea farming co-ogeratives in both Okinawa and Kageshina initiated seed production and release project of the green snail to help sustainable propagation of the natural resources of this species (Komatsu et al., 1995, Manu et al., 1996).  In order to select the most suitable site for releasing green snail seed, it is necessary to detail the distribution, habitat and food habit of the animal (Yoshiga et al., 1987).  Although the habitat o the green snail in Tokunoshima Island has been preliminarily reported by Homma (1987), Yamagachi (1988), and yamagachi and Kikutani (1989), its vertical distribution was not well known.  However, this information is necessary to select the optimal site for release of seeds, further, a survey on the gut center of the green snail, as well as the distribution of the marine algae award their natural habitat, was necessary to clarify the nutrient sources for the culture of green snail.

Microorganisms lives in biosphere, including soil, hot springs, several miles deep in the ocean, 40 miles high in the atmosphere. The total amount of soil and sub-surface bacterial carbon is estimated as 5x10¹⁷g.

Microorganisms are crucial to nutrient recycling in ecosystem as they act as decomposers. Fix nitrogen, they are as vital parts of the nitrogen cycle, microorganism are also exploited in biotechnology, even in traditional food and beverages preparation. A small proportion of microorganism are pathogenic and causes disease and even death in plants and animals. Microorganisms can also be called Microbes.

1.2   Aim

i.               To evaluate the Anti-fungal effect of microorganisms isolated from snail gut.

1.3   Objective            

·       Isolation of microorganisms from snail gut.

·       Characterization of isolated organisms.

·       Evaluation of antifungal activities  of isolates

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