ABSTRACT

This study evaluated various methods of isolation of actinomycetes species from soil of which one soil sample was collected from designated area in Umudike for the purpose of the work. A total of five (5) actinomycetes species were isolated from the soil using the various methods of isolation which included Direct inoculation, Membrane filter and Sprinkle plate methods. The cultural characteristics from this study, using direct inoculation method showed no growth on the media while there was Cottony white appearance on media using both Membrane filter and Sprinkle plate methods. The results from this study showed that a total number of five (5) actinomycetes species were isolated using various isolation methods. Sprinkle plate method had the highest percentage and number of actinomycetes species 4(80%), while the least number of isolates employed in the Membrane filter method 1(20%). No actinomycetes species was isolated using the direct inoculation method. Conclusively, these methods can provide significant impetus towards the isolation and screening of novel actinomycetes which will be ultimately significant for discoveries and other industrially important bioactive compounds like antibiotics. Isolation of rare actinomycetes is difficult using conventional isolation techniques and hence advanced techniques and high screening techniques have been adopted for their isolations. 

TABLE OF CONTENTS

Title Page   i

Certification iii

Dedication iv

Acknowledgements v

Table of Contents vi

List of Tables viii

Abstract ix

CHAPTER ONE

1.0 Introduction 1

1.1 Aim and Objectives 3

CHAPTER TWO

2.1 Literature Review 4

2.2 Brief Description of Actinomycetes 4

2.3 Structure of Actinomycetes 5

2.4 Plant Growth Promoting Activity of Actinomycetes 6

2.5 Production of Plant Growth Hormone by Actinomycetes 8

2.6 Filamentous Actinomycetes 9

2.7 Various Methods of Isolating Rhizospheric Actinomycetes 10

2.7.1 Bacterial Isolation Methods 10

2.7.2 Selective Isolation Methods 10

2.7.3 Nutritional Selection 11

2.7.4 Selective Inhibition 12

2.7.5 Pretreatment of Sample 12

2.7.6 Physical Treatments 13

2.7.7 Chemical Treatments 14

2.7.8 Membrane Filter Method 14

CHAPTER THREE

3.0 Materials and Methods 16

3.1 Sample Collection 16

3.2 Processing of Samples 16

3.3 Microbial Analysis 16

3.3.1 Sterilization Method 16

3.4 Methods of Isolating Actinomycetes from the Soil 16

3.4.1 Serial Dilution Technique 16

3.4.2 Membrane Filter Technique 17

3.4.3 Direct Inoculation Technique 17

3.5 Microbial Characterization and Identification 18

3.5.1 Identification of Rhizospheric Actinomycetes 18

3.5.1.1 Gram Staining 18

3.6.2 Biochemical Tests 18

3.6.2.1 Indole Test 18

3.6.2.2 Carbohydrate Utilization Analysis 18

3.6.2.3 Catalase Test 19

3.6.2.4 Oxidase Test 19

3.6.2.5 Coagulase Test 19

3.6.2.6 Citrate Utilization Test 19

3.6.2.7 Motility Test 20

CHAPTER FOUR

4.0 Results 21

CHAPTER FIVE

5.0 Discussion and Conclusion 26

5.1 Discussion 26

5.2 Conclusion 27

References