ABSTRACT

Actinomyces sp. And Streptomyces sp. was isolated from the soil of Okwuta in Abia State. Physiological and biochemical analysis strongly suggested that the isolate belonged to the strains ofActinomycete .Nitrogen source is a critical component of cultivation medium and almost the useful tool for stimulation of the antifungal metabolites production. The aim of this study was to select the best nitrogen sources in medium for the production of antifungal compounds effective against Collectotrichum and Alternaria sp. Activity of the cultivation liquids on Alternairia and Collectotrichum isolates was tested in vitro using Agar well diffusion method. The results  indicate that maximum inhibition zone was reached in medium with soybean meal as nitrogen source with inhibition diameter 25.3mm for Alternaria sp.,15mm for Collectotrichum sp. in isolate 1( Actinomyces sp) and 17.5mm for Collectotrichum sp., 27.7mm for Collectotrichum sp in Isolate 2 (Streptomyces spp). Malt and urea showed the  least minimum  inhibition  zone for both tested fungi and isolates.

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 

1.3 Aim and objectives 4

1.4 Objectives      4

CHAPTER TWO

2.0    Literature Review 4 

2.1    Taxanomy             8 

2.2    Bioactive Agents In Actinomycetes    10

2.2.1 Antibiotics       10

2.2.2 Antifungals       10 

2.3    Plant Growth Bioactive Molecules 11

2.3.1 Indole Acetic Acid   11 

2.3.2 Hydrogen Cyanide 12

2.4   Enzyme Production from Actinomycetes 13 

2.5   Actinomycetes as Source of Agroactive Compounds 15 

2.6   Actinomycetes As Biocontrol Tools 17

2.6.1 Plant colonization and biocontrol 18 

2.6.2 Actinomycetes as Biopesticide Agents 19 

2.7   Optimization Of Actinomycetes 21 

2.8   Colletotrichum 22 

2.9   Alternaria 23

CHAPTER THREE

3.0      Materials and Methods 25

3.1 Sample Collection 25

3.2 Sterilization of Materials  25

3.3       Pre-treatment of Soil Samples 25

3.4 Media Used 25

3.4.1 Media Preparation                    25

3.4.2    Test Organisms 26

3.5 Serial Dilution 26

3.5.1 Culturing 26

3.5.2 Identification 27

3.5.3 Sub-culturing 27

3.5.4 Gram Staining 27

3.6 Biochemical Test 28

3.6.1 Starch hydrolysis test 28

3.6.2 Hydrogen sulfide test. 28

3.6.3    Gelatin hydrolysis 29

3.6.4 Catalase test 29

3.6.5 Oxidase test 29

3.6.6 Carbohydrate fermentation test 30

3.7       Optimization of Different Nitrogen Source 30

3.8      Crude Extraction 31

3.9      In-Vitro Antimicrobial Test 31

CHAPTER FOUR

4.0      Results 32

CHAPTER FIVE

5.0   Discussion, Conclusion and Recommendation 37

5.1   Discussion 37

5.2   Conclusion 40

5.3   Recommendation 40

References 41 

Appendix 50 

LIST OF TABLES

  Table                                        Title  Page

 1:    Shows the Morphology of isolated Organisms    33

2:    Shows the Biochemical Characterization of isolated organisms  34

3:  Shows the effects of different nitrogen sources on Actinomyces spp 35

(isolate 1) and   their impact on Antimicrobial agent production  

measured by inhibition zones against Alternaria sp. And  Collectotrichum sp. 

By agar well diffusion method.

 4   Shows the Effects of Different Nitrogen Sources on Streptomyces spp.  36

(isolate 2) and Their Impacts on Antimicrobial Agent Production Measured By Inhibition Zones       Against  Alternaria sp. and Collectotrichum sp. by Agar well diffusion method.