SCREENING, ISOLATION, AND CHARACTERIZATION OF ANTIBIOTIC PRODUCING MICROORGANISMS FROM SOIL SAMPLES IN MICHAEL OKPARA UNIVERSITY OF AGRICULTURE, UMUDIKE ABIA STATE.

ABSTRACT

This study was carried out to find the antimicrobial effects of soil microorganisms that have the potential to produce antibiotics. Ten soil samples were collected from four different (Rhizosphere, Waterlog, Back of COLNAS, Forest soil) sites of Umudike. The isolated strains were obtained by ten-fold serial dilution method plated on Sabrouraud Dextrose agar and Nutrient agar by spread plate method. These were then assessed for antibacterial activity against four pathogenic bacteria. Twenty six isolates were recovered and four showed evidence of antibiotic. However, result indicated that three out of the four that showed evidence of antibiotic were active against at least one of the test organisms, Escherichia coli (ATCC 25922),Staphylococcus aureus (ATCC25923), Pseudomonas aeruginosa (ATCC 27853),Mycobacterium smegmatis (ATCC 28735).S1A2 and S3A19 exhibited high antibacterial activity against Staphylococcus aureus, S4A24 showed slight zone of inhibition against Pseudomonas aeruginosa, while S2A15 shown no clear zone of inhibition against test organisms. Therefore, it is confirmed that soil samples screened for microorganisms producing antibiotic collected within the University premises of Umudike were found to be potential source of novel antibiotics.

TABLE OF CONTENTS

Title page ii

Certification iii

Dedication iv

Acknowledgements v

Table of contents vi

List of Tables viii

Abstract ix

CHAPTER ONE

1.0 INTRODUCTION 1

1.1 Antibiotics 2

1.2 Aims and Objectives 4

CHAPTER TWO

2.0 LITERATURE REVIEW 5

2.1  Occurences of Antibiotic producing Bacteria from Soils 6

2.2 Factors Considering in Selection of Antibiotics 9

2.3  Antibiotics and its Classification 9

2.4  Physiology of Antibiotic Production 11

2.5 Factors affecting Antibiotic Production  11

2.6 Fermentation Conditions 13

2.7  Biology and Pathogenicity of some Test Organisms 15

CHAPTER THREE

MATERIALS AND METHODS

3.1  Collection of Soil Samples 18

3.2 Media Used 18

3.3 Media Preparation 18

3.4 Collection of Test Organisms 18

3,5 Sample Preparation 19

3.6 Sub culturing 19

3.7 Gram Staining 19

3.8 Biochemical Identification of Bacterial Isolates ` 20

3.9 Biochemical Tests 20

3.10 Confirmation of Antibacterial Activity

CHAPTER FOUR

RESULTS 23

CHAPTER FIVE

 DISCUSSION 28

Conclusion 30

References 31

LIST OF TABLES

TABLES TITLE PAGES   

 1;           Description of Samples collected from different sites in

Michael Okpara University of Agriculture,Umudike. 24

 2;          Description of Samples collected from different sites in Umudike, and isolate actinomycetes. 25

 3;            Morphology, and Biochemical Characterization of the

Bacterial Isolates 26

 4;                   Inhibition of test organisms by  bacterial isolates 27

CHAPTER ONE

INTRODUCTION

1.0  SOIL:  

The term soil refers to the outer loose material of the earth crust. It may be regarded as a three phase’s system composes of solids, liquids and gases, dispersed to form a heterogeneous matrix. On the whole the soil is composed of five major components, these include; Mineral matter, Water, Organic matter, Air and living Organisms. The various component of the soil environment constantly changed and the quantity of these constituents are not the same in all soil, but vary with locality. Living portion of the soil body includes small animals and microorganisms but it is generally considered that its microorganisms that plays the most important role in the release of nutrient and carbondioxide for plant growth. The bacteria are the most abundant group usually more numerous than the four combined. Soil bacteria can be rod, (bacilli),cocci (spherical), spirilla (spirals), of these, Bacillus are more numerous than the others. They are one of the major groups of soil bacteria population and are very widely distributed. The number and type of bacteria present in a particular soil would be greatly influenced by geographical location such as soil temperature, soil type, soil pH, organic matters contents, cultivation, aeration and moisture content (Davies and Williams, 1999). An important factor influencing the productivity of our planets various ecosystem is the mixture of their soils.

Therefore, soils are vital for the existence of many forms of life that have evolved on our planets. For example, soil provides vascular plants with a medium for growth and supply these organisms with rest of their nutritional requirement. Soil itself is very complex, it would be very wrong to think of soils as just a collection of the mineral particles. The formation of   soil is influenced by organisms, climate, topography, parent material and time. Soil can be considered essentially as a non-renewable resource. Also soil has a role as a Habitat and gene pool, serves as platform for human activities, landscape and heritage and acts as a provider of raw materials. It contains about twice the amount of carbon in the atmosphere and three times  the amount to be found in vegetation. These functions are worthy of   protection because of their socio-economic as well as environmental importance.  

1.1 ANTIBIOTICS:

The term antibiotic means against life. In our every day usage however, we use the word to describe a set of chemical that inhibit or kill bacteria. Antibiotics are one of the most important commercially exploited secondary metabolites produced by the bacteria and employed in a wide range. The British scientist Alexander Fleming is credited with being first to notice that another organism could inhibit by bacteria growth in 1928. He noticed that growth of bacterium Staphylococcus aureus was inhibited a mold (fungus) that contaminated his plate. The mold was later identified as Penicillin notatum and the antibiotic isolated a short time later was named penicillin (Nester et al., 2009). There are numbers of bacteria having potential to produce antibiotic example of which is Bacillus species which produce antibiotic like bacitracin, pumulin and gramicidin which are active against Gram positive bacteria such as Staphylococci, Streptococci, Corynebacter, Streptomyces species which produce antibiotic like tetracycline, chloramphenicol, vancomycin, gentamycin which are active against Gram negative bacteria and Lactobacillus species which antibiotic like nisin which is produce by Lactobacillus lactis (Waites et al., 2008).

Antibiotics are the best known products of actinomycete. Over 5,000 antibiotics have been identified from the cultures of Gram-positive and Gram-negative organisms, and filamentous fungi, but only about 100 antibiotics have been commercially used to treat human, animal and plant diseases. The genus, Streptomycete, is responsible for the formation of more than 60 % of known antibiotics while a further 15 % are made by a number of related Actinomycetes, Micromonospora, Actinomadura, Streptoverticillium and Thermoactinomycetes.

Antibiotics, because of their industrial importance, are the best known products of actinomycetes. The actinomycetes produce an enormous variety of bioactive molecules, e.g., antimicrobial compounds. One of the first antibiotics used is streptomycin produced by Streptomycin griseus.(Waksman,1954). On the whole, the last 55 years have seen the discovery of more than 12,000 antibiotics.

Actinomycetes are Grams-positive bacteria frequently filamentous and sporulating with DNA rich in G+C from 55-75% (Ho et al., 2002). The name Actinomycetes derived from Greek aktis (a ray) and mykes (fungus) was given to these organisms from initial observation of their morphology. Streptomycetes is the dominant among Actinomycetes. The non-streptomycetes are called rare Actinomycetes, comparing approximately 100 genera numbers of the Actinomycetes, which live in marine environment, are poorly understood and only few reports are available pertaining to Actinomycetes from mangroves (Kumar, 2001).

Antibiotics are one of the most important commercially exploited secondary metabolites produced by bacteria and employed in a wide range. Most of the antibiotics used today are from the microbes. Bacteria are easy to isolate, culture, maintain and to improve their strain. Bacillus species being the predominant soil bacteria because of their resistant endospore formation and production of vital antibiotic like bacitracin etc. are always found inhibiting the growth of the other organisms. In the present research study, soil bacteria with the antibiotic activity was screened and isolated

1.2 AIMS AND OBJECTIVES

1. To isolate the various micro organisms capable of producing antibiotics.

2. To characterize and identify the micro organisms

3. To identify possible micro organisms that produced antibiotics from soil samples.

Buyers has the right to create dispute within seven (7) days of purchase for 100% refund request when you experience issue with the file received. 

Dispute can only be created when you receive a corrupt file, a wrong file or irregularities in the table of contents and content of the file you received. 

ProjectShelve.com shall either provide the appropriate file within 48hrs or send refund excluding your bank transaction charges. Term and Conditions are applied.

Buyers are expected to confirm that the material you are paying for is available on our website ProjectShelve.com and you have selected the right material, you have also gone through the preliminary pages and it interests you before payment. DO NOT MAKE BANK PAYMENT IF YOUR TOPIC IS NOT ON THE WEBSITE.

In case of payment for a material not available on ProjectShelve.com, the management of ProjectShelve.com has the right to keep your money until you send a topic that is available on our website within 48 hours.

You cannot change topic after receiving material of the topic you ordered and paid for.