DETECTION OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS FROM POULTRY, PIG AND FISH FARMS IN MICHAEL OKPARA UNIVERSITY OF AGRICULTURE, UMUDKE

ABSTRACT

The detection of methicillin resistant Staphylococcus aureus (MRSA) in poultry, fish and pig farm droppings were studied. Staphylococcus aureus was isolated from 3 sources including; fish pond, poultry droppings and pig dropping  in 3 weekly tranches which made a total of 15 samples. 3 Staphylococcus species were identified by biochemical methods; which include Staphylococcus aureus, Staphylococcus epidermis and Staphylococcus saprophyticus with gross occurrence of 96.3%, 66.7% and 40.7% respectively. Test for the isolates resistant to methicillin shows that Staphylococcus aureus proved the highest number of resistant strains and had the highest percentage of 42.31%,  Staphylococcus epidermidis had 22.22% while Staphylococcus epidermidis had 22.22% while Staphylococcus saprophyticus had 27.27%. It was observed that the incidence of MRSA was high on the whole and thus has potentials of health risks. The need for improvement of personal and environmental hygiene in the poultry, piggery and fish farms is recommended.

TABLE OF CONTENTS

Title page                                                                                                                                i

Certification                                                                                                                           ii

Dedication                                                                                                                              iii

Acknowledgment                                                                                                                   iv

Table of Contents                                                                                                                   v

List of tables                                                                                                                           vii

Abstract                                                                                                                                   viii

CHAPTER ONE

1.0 Introduction                                                                                                                            1

1.2  Background of the Study                                                                                                        3

1.2 Aim of the study                                                                                                                     3

1.3 Objectives of the Study                                                                                                          3

CHAPTER TWO

2.0 Literature Review                                                                                                                   4

2.1  Historical Background of Methicilin Resistance Staphylococcus aureus (MRSA)                      4

2.2  Epidemiology of MRSA                                                                                                         5

2.3  Livestock Associated MRSA (LA-MRSA)                                                                            6

2.4  History of MRSA infection in animals                                                                                  7

2.5   Prevalence of MRSA in the animal kingdom                                                                        8

2.6  MRSA infection in companion animals                                                                                 10

2.6.1       MRSA infection in horses                                                                                             11

2.6.2   MRSA infection in cattle                                                                                              12

2.6.3       MRSA infection in pigs                                                                                          12

2.6.4      MRSA contamination of meat products                                                                  12

2.7  PVL-producing MRSA strains in animals                                                                       13

2.8  Transmission of MRSA between livestock and owners/attendants                                  14

2.9             MRSA carriage by veterinary and medical personnel                                              15

2.10          MRSA carriage among agricultural personnel                                                    16

2.11 Pitfalls in detection of MRSA in animals                                                                16

2.12  Management of MRSA in animals                                                                          18

2.13Treatment of MRSA infection in animals                                                                      19

2.14 MRSA decolonization of animals and owners                                                              19

2.15 Decolonization of companion animals                                                                     21

2.16 Decolonization of large animals                                                                               22

2.7   Biology and pathogenecity of  S.aureus                                                                          23

CHAPTER THREE

3.0 Materials and Methods                                                                                                     24

3.1 Collection of samples                                                                                                       24       

3.2       Preparation of samples                                                                                               24

3.3       Media preparation                                                                                                      24

3.4       Isolation of Staphylococcus aureus                                                                            24

3.5       Subcuturing of the isolates                                                                                         25

3.6       Identification of S.aureus                                                                                           25

3.7       Gram staining                                                                                                             25

3.8       Biochemical test                                                                                                        26

3.9       Catalase test                                                                                                                26

3.10     Coagulase test                                                                                                             26

3.11     Antibiotic susceptibility testing                                                                                  27

            CHAPTER FOUR

RESULTS                                                                                                                               28       

CHAPTER FIVE

5.1       Discussion                                                                                                                   32

5.2       Conclusion,                                                                                                                 33

5.3       Recommendation                                                                                                       33

References                                                                                                                              35

Appendices                                                                                                                             38

LIST OF TABLES

Table            Title                                                                Page

a:         Prevalence of MRSA infection and carriage in different animals                        22

1:         Characteristics of staphylococcus isolate from different sources for methicilin  resistance test (MRT).                                               29

2:         Occurence of staphylococcus isolate from source                 30

3:         Performance of Staphylococcus isolates to  methicilin                                  31

CHAPTER ONE

1.0 Introduction

Antibiotic resistance is the ability of a microorganism to withstand the effects of an antibiotic. Antibiotic resistance evolves naturally via natural selection through random mutation but it could  also be engineered by applying an evolutionary stress on a population, (Johnson, 2002).

Antibiotic resistance is the product of evolution. Every time a person or animal takes an antibiotic, it kills off only susceptible bacteria while a bacterium with a feature that enables it to resist the drug will survive. Since such a feature is genetically determined, the bacteria’s offspring will inherit it, and will prosper and multiply at the “expense of their unresting cousins and resistant bacteria can transfer their good fortune to others, even others of different species by passing on the “relevant bits” of their DNA. The resistance to antibiotics has been observed since the 1950s (shortly after Flemings discovery of penicillin), but it was ignored as the phenomenon was not widely prevalent. During the 1980s several bacteria have become resistant to the antibiotics which include chloromphenical, tetracycline and streptomycin (Johnson, 2002).

A gene known as MecA is responsible for the protein resistance to methicilin which codes for penicillin-bind protein PBP 2A (Wielder et al., 2002). Lately, a new methicillin resistance mechanism gene, MecC was described in S.aureus (Porrero  et al.,  2014). Garcia-Alvare2  et al (2011), Paterson et al., (2012), Wather  et al; (2012) and  Paterson et al., (2014) reported MRSA isolates carrying MecC gene from humans and animals. Harrison et al., (2013) suggested the public health hazard of MecC-positive MRSA isolates as it has been isolated in human case  and their livestock.

Reports of MRSA isolates in domestic animals seems to be rising in number (Devriese and Hommz, 1975; Hartmana  et al., 1997; Tomlin  et al., 1999; Rich and Roberts, 2004). The epidemiology of MRSA isolates from human and animals sources showed that for certain strains, a cross-infection might have happened (Sqlim  et al., 1999; Stormmenger  et al., 2000; Weese  et al.,  2006). Studies conducted by Feirrara  et a., (2011) and Verkade and Wuytman (2014) suggested that animals can be potential source of MRSA infection to humans. Therefore, knowledge on the epidemiology of MRSA will underpin effective and control strategies including the rational use of antibiotics. Staphylococcus aureus is a Gram-positive cocci bacterium that is a member of firmicues, and is frequently found in the  nose, respiratory tract and on the skin. It is often positive for catalase and nitrate reduction.

 S. aureus is a well known food borne pathogen that produces heat-stable enterotoxins during growth on variety of foods, including meat and poultry product, eggs, cream-filled pasteries, potatoes and some salads (Seo  et al., 2010). Although S. aureus is not always pathogenic.  Although S. aureus is not always pathogenic, it is a common cause  of skin infection such as abscesses, respiratory infections such as sinusitis and food poisoning. It is a bacterium of significant important because of its ability to cause a wide range of disease and capacity to adapt to diverse environmental forms (Lowey, 1998; Waldvogel, 2000). The organism colonies  skin, skin glands and mucous membrane, causing infections both in human and animals such as  rashes, inflammations of bones and the meninges as well as  septicaemia (Aklili  et al., 2010). In addition,  S. aureus  causes inflammation  of the mammary gland in bovine and the lower part of the foot in poultry (Quinn et al 2000).

Pathogenic strain also promote infections by producing potent protein toxins and expressing cell-surface protein that bind and inactive antibodies. The emergence of antibiotics resistant strain of S.aureus  such as MRSA is a worldwide problem in chemical medicine. Penicillin and its derivates, including methicillin have been used for the treatment of infections cause by S. aurues  (Rayner and Munckhof, 2005). However, certain strains  of S. aureus  developed resistance known as methicilin resistant staphylococcus aureus (MRSA). At present, less than 90% of S. aureus  strains  are resistant to most penicillin derivatives (Freeman-Cook, 2006) and Ordinary antimicrobial agents like drugs from the family of aminoglycosides, macrolides, chloramphenicols, tetracycline and fluoroquinolones (Lee, 2003).

1.1       Aim of the study

The aim of this work is to  identify the isolation of methicilin resistance Staphylococcus aureus (MRSA) from poultry, pig and fish farms in MOUAU.

1.2       Objectives of the Study

The main objectives of this work is to:

-       To determine the prevalence of methicilin resistance S .aureus (MRSA) in poultry, pig and fish farm in MOUAU.

-       To detect the presence of  methicilin resistance S .aureus (MRSA) in poultry, pig and fish farms in MOUAU.

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