ISOLATION OF ANTIBIOTIC PRODUCING STREPTOMYCES FROM THE SOIL AS SOURCE ANTIBIOTICS ACTIVE AGAINST ANTIBIOTICRESISTANT STAPHYLOCOCCUS AUREUS

ABSTRACT

The study involved the isolation of (2) isolates titled RMN1, RMN2,  belonging to the Genus Serratia  from ( 2) soil samples collected from the following two strategic locations in university of Abuja, permanent site: Termite hill besides Girls hostel dump ground and from Faculty of Agricultural Science farm land. The isolates were morphologically distinct on the basis of spore colour, aerial and substrate mycelium formation and production of diffusible pigment, Isolates were tested under a microscope by using  Gram staining technique they were all Gram negative, biochemical test was carried out the two isolate such as catalase test which was catalase positive and there was production of burbles, citrate test it was citrate positive and there was a change of colour to green blue and oxidase test  it was oxidase negative. One of the two isolates were observed to have significant antibiotic producing potential, and the antibacterial activity of the produced antibiotics (Red pigment extracted from cultural supernatants of the isolates grown on Pepton glycerol agar) was assessed using the well-in-agar diffusion method and streaking agar method. The results indicated that the extracted pigment had varying antibacterial activity at different concentration level of 500mg/ml, 250mg/ml 125mg/ml and 62.5mg/ml against 4 strains of the test organism (Staphylococcus aureus) titled S1 to S4, and the extracted pigment was more effective at concentration 500mg/ml against the antibiotic resistant Staphylococcus aureus. The results indicate that the soil of this region is source of prodigiocin having antibacterial activity and thus enable the use of micro-organisms as biological control agents.

 

TABLE OF CONTENT

TITTLE PAGE

DECLERATION..............................................................................i

CERTIFICATION............................................................................ii

DEDICATION.................................................................................iii

ACKNOWLEDGEMENT................................................................iv

ABSTRACT......................................................................................v

TABLE OF CONTENT ....................................................................vi

LIST OF TABLES ............................................................................vii

 

CHAPTER ONE

Introduction…………………………………………………………

Justification…………………………………………………………

Aim…………………………………………………………………..

Objectives…………………………………………………………..

 

CHAPTER TWO

Literature review………………………………………………….

Antibiotics………………………………………………………..

Antibiotics resistances to staphylococcus aureus………………..

Antibiotics production from microorganism…………………….

Historical background…………………………………………….

Why an effort for antibiotics search……………………………….

Concrete objectives of antibiotics research………………………

The Mechanism of action of antibiotics…………………………..

Functions of soil microorganisms…………………………………

Some antibiotic producing microorganisms………………………

Serratia marcesciens……………………………………………..

Prodigiosin……………………………………………………….

 

CHAPTER THREE

Materials and methods…………………………………………….

Material list……………………………………………………….

Sterilisation of glass wears ………………………………………..

Antibiotic sensitivity assay ………………………………………

Soil sample collection ……………………………………………

Drying of soil sample…………………………………………….

Preparation of isolation media …………………………………..

Plating ……………………………………………………………

Identification of growth ………………………………………….

Morphological identification ……………………………………

Physiological identification ……………………………………..

Gram staining …………………………………………………..

Motility test ……………………………………………………

Biochemical identification ……………………………………

Oxidase test …………………………………………………...

Catalase test …………………………………………………

Citrate utilization test ………………………………………..

Media preparation ………………………………………….

Inoculation of medium ……………………………………..

Invitro prodigiosin production ……………………………..

Harvesting of pigment and evaporation to dryness…………..

Determination of minimum inhibitory………………………..

concentration (MIC) of extracted pigment

Antimicrobial activity of extracted red pigment………………

on clinical isolate

Statistical analysis …………………………………………….

 

CHAPTER FOUR

Results …………………………………………………………..

Staphylococcus aureus antibiotic resistance pattern ……………

Physical characteristics of soil samples collected ………………

Morphological characteristics ………………………………….

Biochemical characterisation of RM1 isolate …………………..

Minimum inhibitory concentration of the red…………………..

pigment produced by serratia marcescences

Antimicrobial Assay …………………………………………..

 

CHAPTER FIVE

5.0     Discussion and conclusion................................................................

5.1     Discussion..........................................................................................

5.2     Conclusion..........................................................................................

REFERENCE................................................................................................

APPENDIX 1.................................................................................................xi

APPENDIX 2..................................................................................................xvi

APPENDIX 3..................................................................................................xviii

APPENDIX 4...................................................................................................xxi

STATISTICAL ANALYSIS

Data Analysis....................................................................................................xiii

Introduction ......................................................................................................xiii

Analysis............................................................................................................xiii

Analysis of Variance (ANOVA).....................................................................xiii

To test the effect of extract on S.aureus .............xiii

Descriptive Statistics.......................................................................................xiv

 

GRAPHICAL REPRESENTATION

LIST OF Tables

Antibiotic sensitivity and resistance pattern…………………………….

of four strains of S.aureus against Oxacilin

Characterization of soil samples collect …………………………………

Cultural characteristics of soil isolates ……………………………………

Biochemical characterization of RMN1…………………………………..

Minimum inhibitory concentration (MIC)………………………………..

of Red pigment produced by S.marsceinces  

Antimicrobial activities of red pigment extracted ……………………….

Appendix

Analysis of variance………………………………………………….

Descriptive statistics …………………………………………

Summary ………………………………………………………

  

Appendix

Pictorial Presentations of Plates and Microscopy

Pictorial Presentations of Plates