PREVALENCE AND ANTIBIOGRAM OF STAPHYLOCOCCUS AUREUS ISOLATED FROM PIGS AND PIG FARMERS IN MICHAEL OKPARA UNIVERSITY OF AGRICULTURE, UMUDIKE

ABSTRACT

Staphylococcus aureus is an important human pathogen that causes serious infections both in hospitals and communities globally due to its multi-drug resistance tendency. This study was undertaken to determine the prevalence of Staphylococcus aureus  and antibiotic sensitivity pattern among pigs and pig farmers in Michael Okpara University of Agriculture, Umudike (MOUAU). Sixty (60) swab samples were collected from various sites of the pigs of 2 species (Duroc and Large-white) and from the nostrils of the pig farmers. The samples were subjected to standard microbiological techniques to identify Staphylococcus aureus. Resistance to Methicillin was determined by using Cefoxitin (30µg). A total of 48 isolates of Staphylococcus species were recovered representing 80% (48/60) of total isolates and MRSA carriage of 0% (0/48) was obtained showing no prevalence of MRSA. Large white specie of the pigs had a higher prevalence of 70.83% (34/48) compared with the isolates from Duroc pigs which had a lower prevalence of 29.16% (14/48). The antibiotic susceptibility profiles of the isolates to the commonly used drugs showed high resistance to Ceftazidime (95.83%) followed by Erythromycin (47.90%). All isolates were susceptible to Cefuroxime (100%), Ceftriaxone (100%) and Cefoxitin (100%). The prevalence of Staphylococcus aureus was more in the nasal discharge of the pigs (61.54%) than in the pig farmers (38.46%). This should therefore call for urgent intervention because pigs can serve as reservoir through which infections related to Staphylococcus aureus  can spread to other animals, humans and community at large. Therefore, proper hygiene practices, control of indiscriminate use of antibiotics, and frequent screening of this population for  Staphylococcus aureus  related infections, are hereby recommended both for prevention and control of livestock acquired Staphylococcus aureus infections.

TABLE OF CONTENTS

Title page                                                                                                                              i

Certification                                                                                                                      ii 

Dedication                                                                                                                          iii

Acknowledgment                                                                                                                iv

Table of contents                                                                                                                  v

Abstract                                                                                                                                ix

CHAPTER ONE

1.0           INTRODUCTION                                                                                                      1

1.1    Aims and objectives of the study                                                                                  3

CHAPTER TWO

2.0           LITERATURE REVIEW                                                                                           4

2.1       Staphylococcaceae                                                                                                     4

v

2.2       Staphylococcus aureus                                                                                               5

2.3       Morphology and identification of Staphylococcus aureus                                            5

2.4       Biology and pathogenesis of Staphylococcus aureus                                                 6

2.5       Methicillin Resistant Staphylococcus aureus (MRSA)                                              7

2.6       Mechanism of resistance of MRSA                                                                            9

2.7       Historical Background of Methicillin Resistant Staphylococcus aureus (MRSA)          10

2.8       Epidemiology of MRSA                                                                                             11

2.9       MRSA and its association with Pigs                                                                          12

2.10       Antibiotics                                                                                                                12

2.10.1   Classes of Antibiotics                                                                                               13

2.10.2  β-Lactam Antibiotics                                                                                                 13

2.11     Penicillin                                                                                                                    14

2.12     Methicillin                                                                                                                  15

2.13     Mode of action of β-Lactam Antibiotics                                                                    16

2.14     Mechanism of resistance of β-Lactam antibiotics                                                      17

vi

CHAPTER THREE

3.0           MATERIALS AND METHODS                                                                               18

3.1.1    Culture media                                                                                                             18

3.1.2     Reagents used                                                                                                                        18       

3.1.3    Equipment and materials used                                                                                    18

3.1.4    Antibiotic discs used                                                                                                   18

3.2       Collection of samples                                                                                                 18

3.3       Media preparation                                                                                                      19

3.4       Characterization and identification of the isolates                                                     19

3.4.1    Growth on Mannitol Salt Agar (MSA)                                                                       19

3.4.2    Growth on blood agar (Sub-culturing of the isolates)                                                20

3.4.3    Identification of Staphylococcus aureus                                                                    20

3.5       Gram staining                                                                                                             20

3.6       Biochemical characterization of isolates                                                                    21

3.6.1    Catalase Test                                                                                                               21

3.6.2    Coagulase Test                                                                                                           21

vii

3.7       Antibiotic Sensitivity Testing                                                                                     22

3.8       Detection of MRSA (using disc diffusion method)                                                    22

CHAPTER FOUR

RESULTS                                                                                                                              23

CHAPTER FIVE

5.0 DISCUSSION AND CONCLUSION                                                                               32       

5.1 Discussion                                                                                                                                                 32

5.2 Conclusion                                                                                                                                                33

References                                                                                                                             35

CHAPTER ONE

1.0       INRODUCTION

Staphylococcus aureus is a well-known food-borne pathogen that produces heat-stable enterotoxin during growth on varieties of food, including meat (especially pork) and poultry product, eggs, cream filled pasteries, potatoes and some salad (Cefai et al; 2010). It is a bacterium of significant importance because of its ability to cause a wide range of disease and capacity to adapt to diverse environmental forms (Aarestrup et al., 2010). 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 (Adjitev et al; 2013). In addition, Staphylococcus aureus causes inflammation of the mammary gland in bovine and the lower part of the foot in poultry (Burke et al; 2000). Penicillin and its derivatives, including methicillin have been used for the treatment of infections caused by Staphylococcus aureus (Cavaco LM et al., 2014). However, certain strains of Staphylococcus aureus are resistant to most penicillin derivatives (Cosgrove et al., 2003) and ordinary antimicrobial agents like drugs from the family of aminoglycosides, macrolides, chloramphenicol, tetracycline and fluroquinolone (Cuny et al., 2010).The development of antibiotic resistance has become a global public health challenge which is causing ineffectiveness of antibacterial agents leading to increase in diseases and death rate (Dam-Deisz WD et al., 2009).

  Over the last 50 years Methicillin resistant Staphylococcus aureus (MRSA) has emerged as a major public health issue in both community and nosocomial infections. It has become one of the leading nosocomial pathogen and recently has been responsible for life threatening infections. In the past few decades, resistance towards methicillin by MRSA has been accompanied by resistance towards wide range of other antibiotics such as gentamycin, erythromycin, and tetracycline (Dand MZ et al., 2010).

    The wide use of antibiotics in the treatment of bacterial infections has led to the emergence and spread of resistant strains. Methicillin resistant Staphylococcus aureus (MRSA) is a major cause of infections in farm animals especially pigs. MRSA infections are very difficult to cure because MRSA strains are resistance against almost all clinically available antibiotics. For most MRSA strains, glycopeptide-type drugs such as Vancomycin are the only effective antimicrobial agents. However, Vancomycin resistant Staphylococcus aureus (VRSA) has been reported (Deresnski, 2005). According to Ekklenkamp et al. (2006), it is extremely important to find new antimicrobial agents or new ways that are effective for the treatment of infectious diseases caused by drug-resistant bacteria including MRSA.

     A gene known as MecA is responsible for the protein resistance to methicilin which codes for penicillin-binding protein PBA2A (Etebu et al; 2016). Lately, a new methicillin resistant gene, MecC was described in Staphylococcus aureus (Gibs ST et al., 2006). Graham, D et al (2009), Gravel H et al; (2011) reported MRSA isolates carrying MecC gene from humans and animals Haddadin et al (2012) suggested the public health hazard of MecC positive MRSA isolates as it has been isolated in human case and livestock (especially pigs). Reports of MRSA isolated in pigs seems to be rising in number (Hartmeyer et al., 1997; Tomlin et al; 1999; Rich and Robert, 2004).  The epidemiology of MRSA isolates from human and animals sources showed that for certain strains; a cross-infection might have happen (Heedeick D et al; 2011 Ike et al; 2016; Kar et al; 2016). Studies conducted by Khana T et al; (2008) and animals can be potential source of MRSA infection to human. Therefore, knowledge on the epidemiology of MRSA will underpin effective and control strategies including the rational use of antibiotics.

        Resistance to penicillins and other β-lactams is due to one of four general mechanisms like; inactivation of antibiotic by β-lactamase, modification of target PBPs, impaired penetration of drug to target PBPs and efflux. β-lactamase production is the most common mechanism of resistance. β-lactamases produced by Staphylococcus aureus, Haemophilus species. and Escherichia coli are relatively narrow in substrate specificity preferring penicillins to cephalosporins. Altered target PBPs are the basis of methicillin resistance in Staphylococcci and of penicillin resistance in pneumococci and Enterococci. These resistant organisms produce PBPs that have low affinity for binding β-lactam antibiotics; consequently they are not inhibited except at relatively high drug concentrations (Van Cleef et al., 2010).

1.3       Aims and Objectives of the Study

This aim and objective of this study is:

1.     To isolate and identify Staphylococcus aureus and determine its prevalence as well as its antibiogram in pigs and pig farmers of Michael Okpara University of Agriculture, Umudike (MOUAU).

2.     To determine the level of Methicillin resistance in MOUAU pig farm.

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