PREVALENCE AND ANTIMICROBIAL SENSITIVITY PATTERN OF COAGULASE POSITIVE STAPHYLOCOCCI ISOLATED FROM FARM ANIMALS IN MICHAEL OKPARA UNIVERSITY OF AGRICULTURE, UMUDIKE

ABSTRACT

Staphylococcus aureus, an opportunistic pathogen is a Gram positive, spherical bacterium that causes serious diseases in animals. The prevalence and antimicrobial sensitivity pattern of coagulase positive Staphylococci isolated from farm animals in Michael Okpara University of Agriculture, Umudike was investigated. A total of 200 samples were collected from ear and nostrils of five different animals comprising Goat (56), Sheep (16), Rabbit (42), Pig (36) and Cattle (50) were analysed. 108 (54%) of the total number of the samples were coagulase positive Staphylococci while 43 (21.5%) were coagulase negative Staphylococci, 49 (24.5%) showed no growth. Of this 108 positive S. aureus isolates, 37.9% were from goats, 26.8% from cattle, 14% from pigs, 12.1% from rabbits and 9.2% from sheep. All 108 Staphylococcus aureus isolates were subjected to in vitro antibiotic susceptibility test using the Kirby Bauer disc diffusion method. A battery of eight (8) antibiotics containing Ceftazidime (30µg), Cefuroxime, (30µg) Gentamicin (10µg), Ceftriatone (30µg), Erythromycin (5µg), Cloxacillin (5µg), Oflaxacin (5µg) and Augmentin (30µg) was used. Ofloxacin was found to be the most effective chemotherapeutic agent with 60(55.5%) sensitivity, followed by Gentamicin 55(50 ) and Augumentin 40(37  respectively. The highest resistance was recorded against cloxacillin 90 (83.3%), followed by ceftazidime 80(74.1%) and cefuroxime70 (64.8%). Also 86 (79.6%) of the isolates were resistant to two or more of the antibiotics tested. The result of this study identifies the presence of Multiple Drug Resistant (MDR) Staphylococcus aureus in farm animals. This is of a major public health concern due to their zoonotic transmission and also as the farm animals can serve as a reservoir for dissemination of multiple antibiotic resistance Staphylococcus aureus to the community.  

TABLE OF CONTENTS

Title page                                                                                                                    i

Certification                                                                                                               ii

Dedication                                                                                                                  iii

Acknowledgement                                                                                                      iv

Table of Contents                                                                                                       v

List of Tables                                                                                                              viii

List of Figures                                                                                                             ix

List of Plates                                                                                                               x

Abstract                                                                                                                      xi

CHAPTER ONE

INTRODUCTION                                                                                                    1

1.1  Introduction                                                                                                    1

1.2  Objectives of the Study                                                                                  3

CHAPTER TWO

LITERATURE REVIEW                                                                                        4

2.1 Nature of Staphylococcus aureus                                                                         4

2.2 Nature of Staphylococcus aureus                                                                         4

2.3 Growth and Survival Characteristics                                                                   5         

2.4 Virulence factors                                                                                                  5

2.5 Pathogenesis                                                                                                         10

2.6 Epidemiology                                                                                                       11

2.7 Staphylococci Infection in Animals                                                                     11

2.8 Staphylococci Infection in Man                                                                           14

2.9 Antimicrobial Agents                                                                                           15

2.10 Classification of Antimicrobial Agent                                                               15

2.11 Mechanism of Action of Antibiotic                                                                    19

2.12 Antibiotic Resistance                                                                                          21

2.13 Mechanism of Resistance to Antibiotics                                                            22

CHAPTER THREE

MATERIALS AND METHOD                                                                               24

3.1 Research Area                                                                                                       24

3.2 Sample Collection                                                                                                24

3.3 Sterilization of Materials                                                                                      24

3.4 Media used and their preparation                                                                         25

3.5 Microbiological Analysis                                                                                     25

3.6 Isolation of bacteria                                                                                              25

3.7 Identification of Isolates                                                                                       25

3.7.1 Macroscopic Examination                                                                                 26

3.7.2 Microscopic Examination                                                                                  26

3.7.3 Biochemical Tests                                                                                             26

3.7.3.1 Catalase Test                                                                                                   26

3.7.3.2 Coagulase Test                                                                                               27

3.7.3.3 Sugar Fermentation Test                                                                                27

3.8 Antibiotic sensitivity test                                                                                      28

3.9 Statistical analysis                                                                                                28

CHAPTER FOUR   

RESULT                                                                                                                    29

4.1 Results                                                                                                                  29

CHAPTER FIVE

DISCUSSION, CONCLUSION AND RECOMMENDATION                           40

5.1 Discussion                                                                                                             40

5.2 Conclusion                                                                                                            43

5.3 Recommendation                                                                                                 43

REFERENCES        

APPENDIX

LIST OF TABLES

Table                Title                                                                            Page   

4.1       Morphological and biochemical characteristics of Staphylococci isolates                        31

4.2       Percentage Occurrence of Coagulase Positive Staphylococci and Coagulase

Negative Staphylococci in farm animals                                                                    32

4.3       Percentage occurrence of Staphylococcus aureus isolates from the ear and

nostrils of farm animals                                                                                              33

4.4       Antibiotic susceptibility pattern of the Staphylococcus aureus isolates                        34

4.5       Occurrence of Multiple drug resistant Staphylococcus aureus isolated from

 animal samples (%)                                                                                                   35

4.6       Prevalence of Cloxacillin (Methicillin) Resistant Staphylococcus aureus

isolated from animal samples                                                                                     36                                                                                                       

LIST OF FIGURES

Figure                                                 Title                                                                            Page   

1          Percentage distribution of Staphylococcus aureus isolates from farm animals   38

2          Percentage susceptibility pattern of Staphylococcus aureus to antibiotics                   39

LIST OF PLATES

Plate       Title                                                                            Page   

1          Zones of inhibition to antimicrobial drug tested                                           39

CHAPTER ONE

INTRODUCTION

1.3  Introduction

Staphylococcus aureus belong to the family Micrococcaceae and is part of the genus Staphylococcus which contains more than thirty (30) species such as Staphylococcus epidermidis, Staphylococcus saprophyticus and Staphylococcus haemolyticus. Among the Staphylococcus species, Staphylococcus aureus is by far the most virulent and pathogenic for human and animals. Staphylococcus aureus is 1µm in size. The organism is a Gram-positive bacterium which appears in pairs or as grape-like clusters. It is characterized as coagulase and catalase positive, non motile, non spore forming and as facultative anaerobic. It grows in yellow colonies on nutrient rich media and is referred to as the as the yellow Staphylococci (Winn Washington, 2006).

Staphylococcus aureus is a widely distributed bacterium found in the air, soil, water and dust because Staphylococcus aureus has the ability to adapt to different environments and it may colonize the human skin, nails, nares and mucus membrane and  thereby disseminate among recipient host population through physical contact and aerosols (Lowy, 1998). Colonization with Staphylococcus aureus is an important factor for subsequent Staphylococcus aureus infection (Von Eiff et al., 2004; Wertheim et al., 2001). Numerous investigators have reported nasal carriers in normal adults varying from 30% to 80% (Osuide et al., 1996).

Farm animals such as sheep, cattle, goats, pig and rabbits and even man are known to harbor potentially pathogenic Staphylococcus in their body without any apparent clinical signs of illness (Oyekunle and Adetosoye, 1998), but diseases results only when the defensive mechanism of the host animal is broken. As a pathogen of domestic animals, Staphylococcus aureus has been found associated with cases of abortion in cattle as well as documented cases of clinical and subclinical mastitis in dairy farms worldwide (Daniel et al., 1986) and pneumonic lesions in goat ( Ugochukwu,1984).

Staphylococcus aureus causes a wide range of infection in humans, from a variety of skin, wound and deep tissue infection to more life threatening conditions such as pneumonia, endocarditis, septic arthritis and septicemia. The bacterium is also one of the most common species in nosocomial infection, however little is known about the virulence factor behind all these conditions. In addition, Staphylococcus aureus may also cause food poisoning, scalded skin syndrome and toxic shock syndrome through production of different toxins (Winn Washington, 2006). Widespread interchange of Staphylococci between animals and human has been observed possibly due to closeness of animals to human population in their environment (Adekeye, 1981).

Antibiotics are used to control bacterial infection in farm animals. Staphylococcus aureus in general are sensitive to many antibiotics but strains from different patients and carriers differs in the pattern and degree of the sensitivity to different drugs and many strains are now resistant to some of the drugs (Osterlund et al., 2002; O’Neill et al., 2004 ). This has been attributed to multiple use of antibiotics in agriculture as prophylactic agents and as growth promoter has led to the emergence of resistant bacteria in the environment.  As is apparent the use or misuse of antimicrobial agents to control animal infection in farm animals has resulted in development of resistance among microorganisms (Winn Washington, 2006). Moreover, there remains the possibility that resistance may be transmitted from antibiotic resistance bacteria to the susceptible ones (Linton et al., 2000). In line with above, Staphylococcus aureus is one of the bacteria that has remained resistant to antibiotics.

At first, Penicillin was used to treat Staphylococcus aureus infection soon afterwards resistance emerged when strain acquired a genetic element coding for beta-lactamase production and today over 80% of all Staphylococcus aureus strain are resistant to penicillin. The next drug to be introduced for treating infections with Staphylococcus aureus was the semi synthetic, penicillinase resistant penicillin named Oxacillin or Methicillin, but shortly after its introduction, the first isolate with resistance was detected (Winn Washington, 2006). With the emergence of resistance to Penicillinase-resistant Penicillin, the glycopeptides agent Vancomycin became the treatment of choice for infection with methicillin resistant Staphylococcus aureus (MRSA) and in the year 1996, the first isolate with intermediate vancomycin resistance was detected (Brown and Thomas, 2002). Resistance to Methicillin is considered the most important for Staphylococcus aureus. Methicillin resistant Staphylococcus aureus has become endemic in hospitals as well as in health care setting globally (Chamber and Deleo, 2009).

1.4  Objectives of the Study

     a.         To determine the occurrence of Staphylococcus aureus in the ear and nostrils of farm animals in the university farm.

     b.         To determine antibiotic resistance and sensitivity patterns of Staphylococcus aureus.

     c.         To know the prevalence of methicillin resistant Staphylococcus aureus (MRSA) from among the Staphylococcus aureus isolates.

     d.         To provide information on the best choice of antibiotic to be used for empirical treatment of Staphylococcus aureus infections

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