ANTIBIOTIC PRODUCING MICRO-ORGANISMS FROM SOILS OF FARMLANDS IN AMAKAMA, UMUAHIA SOUTH LGA OF ABIA STATE

ABSTRACT

The quest to harness the microbial world's potential for antibiotic discovery has led to extensive research endeavors, particularly in regions abundant with microbial diversity such as the soils of farmlands. This study aimed to isolate and screen soil microorganisms from farmlands in Amakama, Umuahia South Local Government Area of Abia State, Nigeria, for their antibiotic-producing capabilities.

Soil samples were meticulously collected from various farmlands in the designated area, focusing on the rhizosphere, where microbial activity is enriched. Isolates were purified through sub-culturing on nutrient agar plates, and the resulting colonies were stored for further analysis. Test organisms including Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, and Salmonella typhi were prepared for sensitivity testing.

The antibacterial potential of the isolates was assessed using the paper disc diffusion method. Clear zones of inhibition around the impregnated discs indicated the presence of antimicrobial compounds. Furthermore, the isolated bacteria were characterized and identified through gram staining and a series of biochemical tests including the indole test, citrate utilization test, starch hydrolysis test, catalase test, and hydrogen sulfide test.

Results revealed the isolation of thirty-five bacterial isolates from twenty soil samples, highlighting the rich microbial diversity within the sampled farmlands. The colonial morphology and biochemical characteristics of the isolates with antimicrobial activity were meticulously documented. Sensitivity testing provided insights into the efficacy of the isolated compounds against a panel of pathogenic bacteria.

The findings of this study underscore the immense potential of soil microorganisms from farmlands in Amakama for antibiotic production. While the discovery of antibiotics remains one of the most significant achievements in microbiology, the journey from discovery to application in chemotherapy demands rigorous exploration and refinement. The study's results emphasize the necessity of further exploration into the antimicrobial capabilities of these bacteria, highlighting avenues for future research and drug development.

In conclusion, the search for novel antibiotics in microbial ecosystems such as farmland soils holds promise for addressing the global challenge of antimicrobial resistance. The findings of this study provide a foundation for continued investigation into the diverse microbial communities residing in agricultural soils, with the potential to unlock new avenues for combating infectious diseases and improving public health outcomes.

TABLE OF CONTENTS

CHAPTER ONE

1.0       INTRODUCTION

1.1       AIMS AND OBJECTIVES

CHAPTER TWO

2.0       LITERATURE REVIEW

2.1       ANTIBIOTIC RESISTANCE

2.2       MICROBIAL DIVERSITY IN SOIL

2.3       FACTORS AFFECTING THE ISOLATION OF SOIL MICROORGANISMS

CHAPTER THREE

3.0       MATERIALS AND METHODS

3.1       SAMPLE COLLECTION

3.2       ISOLATION OF SOIL MICROORGANISMS

3.3       PURIFICATION OF ISOLATES

3.4       PREPARATION OF INOCULUM OF TEST ORGANISMS

3.5       PREPARATION OF INOCULUM OF EACH ISOLATE

3.6       CHARACTERIZATION AND IDENTIFICATION OF ISOLATES

3.6.1    GRAM STAINING

3.6.2    BIOCHEMICAL TESTS

3.6.2.1 INDOLE TEST

3.6.2.2 CITRATE UTILIZATION TEST

3.6.2.3 STARCH HYDROLYSIS TEST

3.6.2.4 CATALASE TEST

3.6.2.5 HYDROGEN SULPHIDE TEST 

CHAPTER 4

4.0       RESULTS

CHAPTER 5

5.0       DISCUSSION

5.1       CONCLUSION

REFERENCES

LIST OF TABLES

Table 1:           Some clinically important antibiotics produced by microorganisms

Table 2:           Antibiotics produced by Bacillus species

Table 3:           Description of soil samples and number of organisms isolated from each.

Table 4:           Cultural morphology and biochemical characterization of isolated organisms

with antimicrobial activity.

Table 5:           Zones of inhibition (mm) produced by isolates against test organisms.

LIST OF FIGURES

Fig.1 Antagonistic effect of bacterial suspension on different pathogenic bacteria:

CHAPTER ONE

1.0       INTRODUCTION

The soil being the outermost layer of the earth crust is a natural reservoir for microorganisms and their antimicrobial products. The numbers and species of microbes in the soil is dependent on environmental conditions like nutrient availability, soil texture, presence of moisture in soil and type of vegetation cover, and their number varies according to the type of environmental condition. The soil is a favourable habitat for the growth of microorganisms including bacteria, algae, fungi, viruses and protozoa. Microorganisms are found in large numbers in soil, usually several millions are present per gram of soil with bacteria and fungi being the most prevalent.

An antibiotic was in previous times defined as chemical substances produced by various species of microorganisms that is capable, in low concentrations, of inhibiting the growth of or killing other microorganisms. Today, this definition has been replaced by any substance produced by a microorganism or a similar product produced wholly (synthetic) or partially (semi-synthetic) by chemical synthesis and in low concentrations inhibits the growth of or kills microorganisms. Antibiotics are one of the most important commercially exploited secondary metabolites produced by bacteria and employed in a wide range use in Medicine and Agriculture (Musliu and Salawudeen, 2012).

However, soil microorganisms with known antibiotic properties are mainly bacteria and fungi. A group of Gram-posititve bacteria distinguished from other bacteria by their morphology called actinomycetes have been identified as the major producers of antibiotics (Abebe et al., 2013). More than 70% of naturally occurring antibiotics have been isolated from different genera of actinomycetes especially Streptomyces spp (Khanna et al., 2011). Other bacterial species that have the ability to produce antibiotics are Bacillus species and Lactobacillus species (Musliu and Salawudeen, 2012) while Penicillium and Cephalosporium, terrestrial and marine molds respectively are antibiotic-producing fungi.

The worldwide use of antibiotics has rapidly increased since the accidental discovery of Penicillin by Alexander Fleming in 1928. Since then, thousands of useful secondary metabolites of microbial origin have been discovered (Nordenfjäll, 2014). While many antibiotics are known to exist, efforts to discover new antibiotics still continue. Therefore many species such a Streptomyces, Bacillus and Penicillium have been studied continuously for their ability to produce antibiotics (Sandeepta et al., 2015). Some antibiotics like Penicillin, Erythromycin and Methicillin which used to be one-time effective treatment against infectious diseases are now less effective because bacteria have become more resistant to such antibiotics (Raja et al., 2010).

Pathogenic bacteria are acquiring resistance to existing antibiotics, most of which are expensive and have been associated with side effects like nephrotoxicity. Bacteria have evolved numerous strategies for resisting the action of antibiotics and antibacterial agents (Sandeepta et al., 2015). Antibiotic resistant pathogens such as Methicillin and Vancomycin resistant strains of Staphylococcus aureus and others are enormous threat to the treatment of serious infections and to avoid this happening, immediate replacement of the existing antibiotic is necessary.

The problem of resistance against the present antibiotics in bacteria increases day by day. So there is an urgent need to search new antibiotics or the sources of new antibiotics. A lot of work has been done during last few decades that has witnessed the production of novel antibiotics from different microorganisms (Mashoria et al., 2014). Considerable research is being done in order to find new antimicrobial producing bacteria isolated from soil. One of the possible ways to increase the chance of finding novel antibiotics is to find new approaches for isolating interesting bacteria and fungi or at least make the existing methods more efficient (Nordenfjäll, 2014).

1.1       AIMS AND OBJECTIVES

1.      To isolate soil microorganisms from soil samples collected from farmlands in Amakama.

2.      To screen isolates for production of antibiotics against several bacterial pathogens.

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