Evaluation Of Different Methods Of Isolation Of Actinomycetes

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