ANTIBIOTICS RESISTANCE PROFILE OF E. COLI ISOLATED FROM FARM ANIMAL IN UMUAHIA METROPOLIS

ABSTRACT

This study was to determine the antibiotics resistance profile of E. coli isolated from farm animal in Umuahia metropolis. A total of Forty (40) samples were analyzed in the study of which ten (10) swab samples each were collected from Goats, Sheep, Pigs and Rabbits. These samples were cultured on Blood and MacConkey agar plates using streak plate technique and incubated at 37oC for 24hrs. Escherichia coli strains were identified by standard techniques on the basis of their colonial morphology, Gram staining reaction, motility and biochemical characteristics. Among the forty (40) samples analysed, pig had the highest number positive for E. coli with high percentage occurrence 8(42.1%). The antibiotic susceptibility testing, using disc diffusion techniques reveals patterns with high resistant rates of the isolates to Nalidixic acid, Amoxicillin, Septrin and Ampicilin. The highest sensitivity rates on E. coli were recorded with Ciprofloxacin (94.7%), Streptomycin (78.9%), Oflaxacin (89.5%), and gentamicin (84.2%). The results from this study demonstrated the high antimicrobial resistance amongst E. coli to the commonly prescribed antimicrobial drugs which substantiates the alarming occurrence and ongoing spread of various multi resistant Enterobacteriaceae strains in the human and animal population. There is need for continuous surveillance of antimicrobial resistance trends particularly among organisms resident in the gastrointestinal tract of farm-animals which are implicated in infectious diseases in human.

TABLE OF CONTENTS

Title Page                                                                                                                                  i

Certification                                                                                                                           iii

Dedication                                                                                                                              iv

Acknowledgement                                                                                                                  v

Table of Contents                                                                                                                   vi

List of Tables                                                                                                                          vii

Abstract                                                                                                                                  ix

CHAPTER ONE

1.0       Introduction                                                                                                                1

1.1       Aim and Objectives                                                                                                    3

CHAPTER TWO

2.0       Literature Review                                                                                                       4

2.1       Modes of Action of Antibacterial Agents                                                                  4

2.1.1    Inhibition of Cell-Wall Synthesis                                                                               5

2.1.2    Inhibition of Protein Synthesis                                                                                   6

2.1.3    Injury to the Plasma Membrane                                                                                 7

2.1.4    Inhibition of Nucleic Acid Synthesis                                                                         7

2.1.5    Inhibition of Essential Metabolites                                                                            8

2.2       Antibiotics Resistance Mechanisms                                                                           8

2.2.1    Resistance Based on Altered Receptors for a Drug                                                    9

2.2.2    Decreased Entry of Antibiotics                                                                                  10

2.2.3    Synthesis of Resistance or Alternative Pathway                                                        10

2.3       Resistance to B-Lactam Antibiotics                                                                           11

2.3.1    Tetracycline Resistance                                                                                              11

2.3.2    Chloramphenicol Resistance                                                                                      13

2.4       Factors Contributing to the Emergence of Resistance                                               13

2.5       Containment of Antimicrobial Resistance                                                                 14

2.5.1    Infection Control                                                                                                        14

2.5.2    Rational Use of Antibiotics                                                                                        15

2.6       Biology and Pathogenecity of E. coli                                                                         16

2.6.1    Escherichia coli                                                                                                          16

2.6.1.1 Diseases Caused by Escherichia coli O157: H7                                                        17

CHAPTER THREE

3.0       Materials and Methods                                                                                               18

3.1       Study Location                                                                                                           18

3.2       Sample Collection                                                                                                      18

3.3       Sample Processing                                                                                                      18

3.4       Isolation and Identification of E. coli Isolates                                                            18

3.4.1    Colonial Morphology                                                                                                 18

3.4.2    Gram Staining                                                                                                            19

3.4.3    Motility Test                                                                                                               19

3.5       Biochemical Tests                                                                                                      19

3.5.1    Catalase Test                                                                                                               19

3.5.2    Methyl Red Test                                                                                                         20

3.5.3    Voges-Proskauer Test                                                                                                 20

3.5.4    Indole Test                                                                                                                  20

3.5.5    Citrate Utilization Test                                                                                               20

3.5.6    Urease Test                                                                                                                 21

3.5.7    Triple Sugar Iron Agar Test                                                                                       21

3.6       Antibiotic Sensitivity Testing                                                                                     21

3.7       Data Analysis                                                                                                              22

CHAPTER FOUR

4.0       Results                                                                                                                        23

CHAPTER FIVE

5.0       Discussion, Conclusion and Recommendation                                                          27

5.1       Discussion                                                                                                                   27

5.2       Conclusion                                                                                                                  28

5.3       Recommendation                                                                                                        28

              References

LIST OF TABLES

CHAPTER ONE

1.0       INTRODUCTION

Antibiotic usage is possibly the most important factor that promotes the emergence, selection and dissemination of antibiotic-resistant microorganisms in both veterinary and human medicine (Daniels et al., 2009). This acquired resistance occurs not only in pathogenic bacteria but also in the endogenous flora of exposed individuals (animals and humans). In intensively reared food animals, antibiotics may be administered to whole flocks rather than individual animals, and antimicrobial agents may be continuously fed to food animals such as poultry, goats, and cattle as growth promoters. Therefore, the antibiotic selection pressure for bacterial drug resistance in the animal is high and invariably their faecal flora contains a relatively high proportion of resistant bacteria (Whitworth et al., 2008).

Antibiotic resistance among microorganisms is a major problem, both in human and livestock industry. And the problem is usually attributed to unregulated and inappropriate use of antibiotics (Lawson, 2008). It has been severally reported that commensal Escherichia coli isolates from healthy animals like cattle, swine as well as from humans play significant roles in the perpetration of drug resistant pathogens and subsequent infections (Tian et al., 2012).

The mechanism for spreading antibiotic resistance from animals to humans and vice versa remains controversial. Colonization of the intestinal tract with resistant Escherichia coli from chicken has been shown in human volunteers and there is historical evidence that animals are a reservoir for E. coli found in humans (Akwar et al., 2008). Furthermore, spread of antibiotic resistance plasmids in E. coli from chickens to human handlers or of antibiotics - resistant microorganisms from animal to humans in various countries has been reported (Fang et al., 2008). Resistance has been found in organisms common to both humans and animals, such as E. coli, Salmonella spp., Campylobacter spp. and Enterococcus among others (Davis et al., 2009). Due to the intricate balance of microflora of different habitats within the ecosystem, the transfer of resistance genes among bacteria occupying different habitats has the potential to occur frequently. Resistance genes may be transferred vertically among bacteria of different genera and families or horizontally among different bacterial species within the same genus or family (Call et al., 2008).

Widespread reliance on antimicrobials in food animal production has resulted in a considerable rise of antimicrobial-resistant strains of bacteria, complicating the treatment of infectious diseases in livestock, companion animals, and humans. This has led to important changes in the perceptions and priorities of regulatory agencies with regard to antimicrobial usage, particularly the use of antimicrobials as growth promoters and prophylactic agents. The selective pressure from the use of antimicrobial agents at sub therapeutic levels in dairy cattle could result in the selection of those strains that contain genes for antimicrobial resistance (Call et al., 2008).

Molecular tools have been used to correlate animal associated pathogens with similar pathogens affecting humans and to clearly demonstrate transferable resistant genes carried by plasmid\s common to both animals and humans (Pitout et al., 2009). The possibility of antibiotic resistance genes circulating among humans, animals and the environment constitutes a direct threat to public health. This threat prompts research into emerging resistance mechanisms, novel approaches to antimicrobial efficacy and stringent control measures in the prudent use of antimicrobials in animal medicine.

In Nigeria livestock industry, the problem of occurrence of multidrug resistant E. coli is becoming very rampart, because they are often encountered in routine diagnoses of disease conditions from livestock brought for confirmatory diagnosis in microbiology diagnostic units of some Tertiary Veterinary Teaching Hospitals in Nigeria (unpublished data). Earlier in Nigeria, Ogunleye et al. (2008), reported nineteen different multidrug resistant patterns to commonly available antibiotic in E. coli isolated from some diseased poultry samples from eleven poultry farms in Abeokuta, Ogun State, Nigeria. The 39 E. coli were isolated from various organs like liver, lungs, kidneys, ovary, intestine and colorectum of birds that died of septicaemic conditions. Each of the E. coli isolates studied were resistant to between 5 and 12 com-monly used antibiotics such as nitrofurantoin, cefuroxime, norfloxacin, cotrimoxazole, nalidixic acid, chloramphe-nicol, ampicilin, ofloxacin, penincilin G, amoxylin, cloxa-cilin and ciprofloxacin (Ogunleye et al., 2008).

1.1       AIM AND OBJECTIVES

To determine the antibiotics resistance profile of E. coli isolated from farm animal in Umuahia metropolis, while the specific objectives are;

·       To isolate E-coli strains from selected farm animals

·       To determine the antibiotic sensitivity pattern of E. coli strains isolated from the farm animals

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