ISOLATION AND IDENTIFICATION OF MICROORGANISMS FROM PALM WINE SOLD IN UMUDIKE, ABIA STATE

ABSTRACT

Microbiological analysis was conducted on fresh and fermented palm wine in Umudike, Abia state. Fresh and fermented palm wine samples were collected from a palm wine tapper immediately after tapping and from different palm wine bars respectively, using tightly capped sterile containers and was taken to a microbiology laboratory for analysis. Spread plate and streak plate method was used for the isolation of microorganisms. The microorganisms isolated were identified based on their cultural, morphological and bio-chemical characteristics. The microorganisms include Bacillus spp, Streptococcus spp, Lactobacillus spp, Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Acetobacter spp, Candida spp, and Saccharomyces spp. The mean average microbial counts of the sample were 2.7 × 104cfu/ml, 3.1 × 104cfu/ml, 0.3 × 104cfu/ml, and 3.2× 104cfu/ml for heterotrophic count, fungal isolates, coliform bacteria count and isolates on De Man, Regosa and Sharpe agar respectively. Saccharomyces spp occurred in all the samples (100.0% occurrence) while Bacillus spp, Klebsiella spp and Escherichia coli had the least occurrence (20.0%). In the assessment of the survival of isolates in relation to time (0-96hrs) Klebsiella pneumoniae, Staphylococcus aureus were eliminated from the sample after 24hours while Streptococcus spp, Lactobacillus spp, Escherichia coli and Candida spp were eliminated after 48hours. Bacillus spp, Acetobacter spp and Saccharomyces spp survived beyond 72 hours of fermentation. The presence of Bacillus spp, Staphylococcus aureus, Klebsiella pneumoniae, and Candida spp indicated unhygienic handling of the beverage. It is recommended that appropriate agencies should ensure that proper personal and environmental hygiene is enforced to prevent outbreak of disease.

TABLE OF CONTENTS

Title page ﾿ i ﾿

Certification ﾿ ii

Dedication ﾿ iii

Acknowledgements ﾿ iv

Table of Contents ﾿ v

List of Tables ﾿ vii

Abstract ﾿ viii

CHAPTER ONE ﾿ 1

INTRODUCTION ﾿ 1

Background of the study ﾿ 1

General objective ﾿ 2

Specific objectives ﾿ 3

CHAPTER TWO ﾿ 4

LITERATURE REVIEW ﾿ 4

2.1 Production of palm wine ﾿ 4

2.2 Biochemical constituents of palm wine ﾿ 5

2.2.1 Sugar identified and their concentration in palm wine. ﾿ 6

2.2.2 pH and organic acid concentration in palm wine ﾿ 7

2.2.3 Ethanol concentration in palm wine ﾿ 8

2.2.4 Minerals and trace elements present in palm wine ﾿ 9

2.2.5 Odorant of palm wine ﾿ 9

2.3 Microbial communities in palm wine ﾿ 10

2.4 Fermentation of palm wine ﾿ 12 ﾿

2.5 Uses of palm wine ﾿ 13

CHAPTER THREE ﾿ 14

MATERIALS AND METHODS ﾿ 14

3.1 Study Area ﾿ 14

3.2 Sample collection ﾿ 14

3.3 Preparation of media ﾿ 14

3.4 Serial dilution ﾿ 15

3.5 Isolation of Microorganisms ﾿ 15

3.6 Characterization and identification of isolates ﾿ 15

3.6.1 Fungi ﾿ 15

3.6.2 Bacteria ﾿ 16

3.6.2.1 Gram staining ﾿ 16

3.6.2.2 Spore staining ﾿ 16 ﾿

3.6.2.3 Catalase test ﾿ 17

3.6.2.4 Coagulase test ﾿ 17

3.6.2.5 Oxidase test ﾿ 17

3.6.2.6 Citrate test ﾿ 17

3.6.2.7 Indole test ﾿ 18

3.6.2.8 Carbohydrate fermentation test ﾿ 18 ﾿

CHAPTER FOUR ﾿ 19

Results ﾿ 19 ﾿

CHAPTER FIVE ﾿ 25

DISCUSSION, CONCLUSION AND SUMMARY ﾿ 25

5.1 Discussion ﾿ 25

5.2 Conclusion ﾿ 26

5.3 Recommendation ﾿ 26

REFERENCES ﾿ 27