ANTIBIOTIC SUSCEPTIBILITY TESTS OF STAPHYLOCOCCUS AUREUS ISOLATED FROM THE NOSTRILS OF PRIMARY SCHOOL PUPILS IN IKWUANO.

ABSTRACT

Staphylococcus aureus has been identified as a risk factor for community acquired and nosocomial infections. It is a common inhabitant of the skin and mucus membrane. Children with nasal carriage of Staphylococcus aureus play an important role in community spread of Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus (MRSA). Staphylococcus aureus is a facultative anaerobic Gram positive bacterium commonly isolated from different parts of human body, particularly the anterior nares, skin and the throat but the nares are the most consistent site of isolation. In this study, 50 nasal swab specimens were collected from asymtomatic school children of Umudike Community Primary School, Umudike, Abia State, Umuariaga Primary School, Umuahia, Abia State, Grace Vine Primary School, Umudike, Abia State, Amaoba Migrant Farmers School, Amaoba, Abia State in Ikwuano Local Government were examined for the presence of Staphylococcus aureus using Mannitol salt agar and it was further identified with Gram staining reaction and Biochemical tests such as catalase and coagulase test. Antibiotic susceptibility tests were performed using Kirby-Bauer’s disc diffusion method according to performance standards of Clinical and Laboratory Standard Institute guidelines. A total of 16 children were found to have Staphylococcus aureus, out of the 50 cases (20 males and 30 females). The results showed that the females had higher percentage of Staphylococcus aureus infection. The carriage rate reduced with increase in age of the children.The antibiotics susceptibility pattern of the isolates revealed that Erythromycin, Ofloxacin, Cefraxone-Sulbactam, Azithromycin and Gentamicin were the most sensitive antibiotics. The study concluded that Nasal colonization by Staphylococcus aureus can provide an indication of a higher risk for subsequent infections, especially when they have developed resistance to wide spectrum of antibiotics.

TABLE OF CONTENTS

Title Page i

Certification Page ii

Dedication iii

Acknowledgements iv

Table of Contents v

List of Tables vii

Abstract viii

CHAPTER ONE

1.1 INTRODUCTION 1

1.2 AIMS AND OBJECTIVES 2

CHAPTER TWO

2.0 LITERATURE REVIEW 3

2.1 ISOLATION OF Staphylococcus aureus 3

2.2 PREVALENCE OF Staphylococcus aureus 6

CHAPTER THREE

3.0 MATERIALS AND METHODS 8

3.1 SPECIMEN COLLECTION AND CULTURING 8

3.2 MEDIA PREPARATION 8

3.3    ISOLATION AND IDENTICATION OF Staphylococcus aureus 8

3.3.1 CULTURE 8

3.4 SMEAR PREPARATION AND GRAM STAINING 9

3.5 BIOCHEMICAL TESTS 9

 3.5.1 CATALASE TEST 9

3.5.2 COAGULASE TEST 10

3.6 ANTIBIOTIC SUSCEPTIBILITY TESTING 10

CHAPTER FOUR

4.0 RESULTS 11

CHAPTER FIVE

5.0 DISCUSSION AND CONCLUSION 16

5.1 DISCUSSION 16

CONCLUSION 17

5.3 RECOMMENDATION 17

REFERENCES

LIST OF TABLES

Table 4.1  Age and Sex distribution of participants 12

Table 4.2  Frequency (%) of Staphylococcus aureus in the nasal specimen 13

Table 4.3    Zone diameters of Antimicrobial Agents when testing 14

Staphylococcus aureus according to CLS1 guidelines, 2011

 (zone diameters are in mm).

Table 4.4: Antimicrobial susceptibility of Staphylococcus aureus strains isolated 15

 from pupil (Expressed in %)

CHAPTER ONE

1.0 INTRODUCTION

Staphylococcus aureus is a facultative anaerobic Gram positive bacterium commonly isolated from different parts of human body, particularly the anterior nares, skin and the throat but the nares are the most consistent site of isolation (Stenehjem and Rimland, 2013).

Staphylococcus aureus can be found on the skin and in the nose of 20-40% of healthy individuals in which it generally causes no harm if it remains on the surface of the skin (Mazzulli, 2017).

Staphylococcus aureus infections range in severity from mild skin infections to severe necrotizing pneumonia. It is simultaneously the leading cause of bacteremia, infective endocarditis (IE) and can also cause skin and soft tissue, pleuropulmonary, and device related infections (Tong et al.,2015).

Staphylococcus aureus nasal colonization has been identified as the most important risk factor for subsequent invasive infections (Akinkunmi and Lamikanra,2012) Staphylococcus aureus by some individuals makes them more prone to skin infections and operational complication caused by the organism (Dinic et al .,2013).

It is one of the most common causes of healthcare and community acquired infections such as localized cutaneous and life threatening systemic infections. Although most community infections are treated in the outpatient setting some invasive infections, including bacteremia, septic arthritis, toxic syndrome, osteomyelitis and endocarditis, have devastating complications and may require hospitalization (Deolo et al., 2011)

However certain strains of bacterium possess virulence factors that make them to cause various types of diseases ranging from mild skin infections such as boils, furuncles, impetigo to more invasive and life threatening systemic infection. (Shen et al., 2013; Ekundayo and Ndubuisi, 2015). Vancomycin is commonly used to treat these infections as this antibiotic is active against both methicillin resistant and methicillin susceptible Staphylococcus aureus (MRSA and MSSA). However, over the years the emergence of resistant strains to antimicrobials has been reported, the main reason why these infections are becoming more difficult to treat (Howden et al, 2010.MCGuiness et al, 2017).

Staphylococcus aureus produces spectrum of exotoxins and other determinant of virulence that contribute to its pathogenicity. Staphylococcus enterotoxins(SEs) are recognized as being the most important virulence factors involved in cases of food poisoning in humans (Durupt et al.,2017 ; Eibach et al., 2017).The prevalence of infection of Staphylococcus aureus in preschool children varies in different Countries; 34% in Belgium(Jourdian et al.,2011),35% in India (Pathak et al.,2010) and in Republic of Korea(Bae et al.,2012).

1.1 AIMS AND OBJECTIVES

The aims and objective of this Study are:

1. To isolate and determine the prevalence of Staphylococcus aureus from the nostrils of school pupils.

2. To carry out the antibiotic susceptibility testing of the Staphylococcus aureus isolates.

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