ABSTRACT
Soil is the major repository of microorganism that clinically useful antibiotics have been isolated for several group of microorganism including bacteria (Streptomyces sp, Bacillus sp). Bacillus and Actinomycetes are the most abundant microorganism present in Soil and inhibit the growth of other microorganisms. Present in soil and inhibit the growth of other microorganism. In this study 5 Soil samples from different ecological sites were obtained and Cultured in microbiological media to isolate microorganisms. Ten fold serial dilution of the Soil samples were prepared and inoculated on nutrient agar and Saboraud Dextrose agar. A total of 15 Isolated were recovered. Ten of the total isolated were sub cultured on nutrient agar and incubated at 37oc for 24hrs. Out of these ten isolates four are Micrococcus, four Bacillus species and two Actinomycetes were obtained. Five different isolates was tested against four test organisms. Namely: Staphylococcus aureus, Pseudomonas aeruginosa, Mycobacterium semgmatis and Escherichia coli for antibacterial activity. Out of the five isolates, only two (NDDCfemale Hostel) exhibitory effect against one of the test organisms (Pseudomonas aeruginosa) with zones of inhibition of 9mm and 13mm respectively. The remaining isolates showed no antimicrobial activity against the test organisms. Therefore, it is confirmed that soil samples for microorganisms collected within the University Premises of Umudike were found to be potential source of OR one of the isolates (NDDC Hostel) that is Actinomyces was able to inhibit growth of Staphylococcus aureus with a clear inhibition range with a clear inhibition range 0-14mm, which indicates an antibacterial activity against S. aureus while others showed no antimicrobial activity against the test organisms. This shows that the isolation of (NDDC Hostel) from this study area may contribute to the discovery of with further extraction and purification of the isolate.
TABLE OF CONTENTS
Title
Page
i
Certification ii
Dedication iii
Acknowledgements iv
Table
of Contents v
List
of Tables vi
Abstract vii
CHAPTER
ONE: INTRODUCTION 1
1.1 Aims And Objective 2
CHAPTER
TWO LITERATURE
2.1 Soil Description and Classification 4
2.2 Microbial Diversity and Sample
Strategy 4
2.3 Types of Microorganisms Found In the
Soil 5
2.3.1 Fungi 5
2.3.2 Algae 6
2.3.3 Protozoa 6
2.4 Sample Collection and Preparation 7
2.5 Fermentation Conditions 8
2.6 Soil Antimicrobial Agents Producing Microbes 9
2.7 Microbial Diversity In Soil 11
2.8 Micro-Organisms As Sources Of Natural
Products 13
2.9 Causes
of improper soil disposal 14
CHAPTER THREE: MATERIALS AND METHODS
3.1 Collection
an Preparation of soil sample 15
3.2 Media
Used 15
3.3
Sterilization of Materials 15
3.4 Samples
Preparation 16
3.4.1 Culturing 16
3.4.2 Colonial Examination 16
3.4.3
Sub-culturing 17
3.5 Gram
Staining 17
3.6 Biochemical
Tests 17
3.6.1.
Starch Hydrolysis 17
3.6.2 Catalase
Tests 18
3.6.3 Citrate
Utilization Test 18
3.6.4 Coagulase
Test 19
3.6.5 Oxidase
Test 19
3.7 Physicochemical
Analysis 19
3.8 Additional
Test 20
3.9 Biochemical
Identification of Organisms Isolates 20
CHAPTER
FOUR: RESULTS 21
CHAPTER FIVE: DISCUSSION,
CONCLUSION AND RECOMMENDATION
5.1
Discussion 28
5.2 Conclusion 29
5.3 Recommendation 29
References 30
LIST
OF TABLES
Table Title Page
1 Collection of Soil Samples from Different
Location in MOUAU 22
2 Physico-Chemical Properties of Soil Sample
of the Different Locations
(Makut, 2011) 23
3 Microbial Count of Soil Samples 24
4 Percentage Occurrence of Microbial Isolated
From Soil Sample in MOUAU 25
5 Biochemical Test Characterization of Isolated
Bacterial from Soil
Sample in MOUAU 26
6 Cultural and Morphological of fungi identification 27
CHAPTER ONE
1.0
INTRODUCTION
The term soil refers to the outer loose
material of the earth crust it maybe regarded as a three phase’s system
composed of solids, liquids and dispersed to form a heterogeneous matrix. On
the whole soil is composed of five major components, these includes; mineral
water, matter, organic matter, air and living organisms. The various component
of the soil environment components constantly changed and the quantity of these
constituents are not the same in all soil body includes. Small animals and
microorganism but is generally considered that it’s in microorganisms that
plays the most important role in the release of Nutrient and Carbondioxide for
plant growth. (Hamlon et al., 2007).
The bacteria are the most abundant group usually more numerous than the four
combined. Soil bacteria can be rod (bacilli)
cocci (Sphereical) spirilla 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 (Bhagabati el al., 2004) which can be found as
single cells or micro colonies development around the soil particles
(Torsuik,2002) they are responsible for many of the key processes in the
biogeochemical cycling on earth such as nitrogen, sulphr and carbon cycle (Lenyeler
et al., 1999) in addition they are a
valuable sources of natural products providing important antibiotic for
pharmaceuticals, important enzymes and bioactive compounds for industries (Strohl,
2000). The number and types 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 et al., 1999). An ecological
niche is composed of many microhabitats; each microhabitat is composed of a
microscopic diversity which includes bacteria, protozoa, fungi, and macroscopic
diversity that that includes plant and insects. Soil is a complex medium in
which one can encounter many kinds of microbial communities. Application of
nucleic acid-based techniques to analyze soil microbial communities has
revealed high prokaryote diversity (Pandey et
al., 2008). The microbial diversity
or communities present in soil principally depend on the composition of the
soil and many physical chemical properties that the medium posses. Also the
flora and decomposing organism matter on the surface of the biowill influence
microbial diversity present. For example, the fallen trees, barks and flowers
provide nutrients both the microbes and plant present, through microbial
degradation of carbohydrates, lipid and proteins to sugars, fatty acids,
glycerol and amino acids and respectively to mineralization. Besides providing
these nutrients plant secondary metabolites that are generally toxic to
microorganisms will need to be degraded or detoxified by certain microbes.
These degraders (microbes) are selectively presented and ultimated evolved to
produce novel secondary metabolites possibly to counteract the toxic plant
secondary metabolites (Pandey et al.,
2008). There are approximately 106 -109 colony forming
unit per gram of soil. Microbes present in medium posses advantages that will
permit or facilitate their survival in that medium. For examples research has
demonstrated that in desert crust or his soil that has low water availability,
gram positive and spore farm microbes are most abundant. The gram positive
bacteria posses a thicker layer of Murein in their cell wall which makes the
cell less vulnerable to the limiting conditions present in these habitat. Also,
spore farming bacteria can resist long period of desiccation and limiting
nutrient conditions smiles they compact and protect their genuine material in
the bacteria spore, until conditions are favorable for sporulation to occur (Sadha
et al.,2000).
1.1 Aims
and Objective
1. To isolate, identify and characterize bacteria
from the soil sample.
2. To isolate, identify and characterize fungi
from the soil sample.
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