ANTIMICROBIAL SUSCEPTIBILITY PROFILE OF STAPHYLOCOCCUS AUREUS ISOLATED FROM EAR AND NOSTRILS OF PRIMARY SCHOOL PUPILS

ABSTRACT

This study examines the antimicrobial susceptibility profile of Staphylococcus aureus isolated from primary school pupils in Umudike. One hundred samples for culture of ear and nose swabs (50 from nose and 50 from ear) collected from primary school pupil in Umudike were examined using the standard microbiological procedure to determine the incidence of Staphylococcus aureus. Antimicrobial susceptibility profile was determined by modified Kirby Bauer disc diffusion method. The susceptibility of S. aureus to 10 antimicrobial agents allowed for human therapy were examined. The result of this study shows that the nostrils of the case study had the highest frequency of occurrence (100%) of the isolate, while the ear demonstrated 40% case occurrence. The isolate demonstrated high level of susceptibility to ciprofloxacin (228%), Gentamycin (200%), Ampicillin (179%) and Erythromycin (168%), the test organism was considerably susceptible to Tetracycline (140%), and Amikacin (104%), while the isolate was highly resistant to antibiotics such as streptomycin (132%), Augmentin (128%). Changing patterns of Staphylococcus aureus resistance to drugs continues to pose a problem to health care providers globally despite the development of new antibiotics. These high rates of resistance have been attributed to factors such as misuse of these drugs by health professionals and unskilled practitioners among others. Thus, there is need for continuous and regular antimicrobial resistance surveillance in the country in order to guide empirical therapy and to provide adequate control strategies to combat this public health problem.

TABLE OF CONTENTS

Title page                                                                                                                               i

Certification                                                                                                                          ii

Dedication                                                                                                                              iii

Acknowledgement                                                                                                                iv

Table of contents                                                                                                                   v

List of Tables                                                                                                                         vii

Abstract                                                                                                                                 viii

CHAPTER ONE

1.0.  Introduction

1.1.  Background of Study -    -           -           -           -           -           -     -                       1

1.2 Aims and Objective of the Study -           -           -           -           -            -           -           3

1.3 Specific   Objectives -      -           -           -           -           -           -            -           -           3

CHAPTER TWO

2.0. Literature Review

2.1 General Concept about Staphylococcus aureus - -           -           -            -           -           4

2.2. Staphylococcal Pigments -          -           -           -           -           -            -           -           6

2.3       Pathogenesis of Staphylococcus aureus -       -           -           -            -           -           6

2.4 Transimssin Of Staphylocococcus aureus -         -           -           -            -           -           7

2.5 Clinical Syndromes of Staphylococcus aureus -  -           -           -            -           -           7   

2.6 Some of the Infections Caused by Staphylococcus aureus ­         -            -           -           8

2.7 Skin Infections-    -           -           -           -           -           -           -            -           -           8

2.8 Bacteremia -         -           -           -           -           -           -           -            -           -           10

2.9 Animal infections -           -           -           -           -           -           -            -           -           9

2.10 Some of the Virulence Factors Produced by Staphylococcus aureus -            -           -           10

2.10.1 Enzymes -        -           -           -           -           -           -           -            -           -           10

2.10.2 Toxins -            -           -           -           -           -           -           -            -           -           10

2.10.3 Super antigens -           -           -           -           -           -           -            -           -           10

2.10.4 Exfoliative toxins -      -           -           -           -           -           -            -           -           11

2.11 Treatment and Antibiotic Resistance -   -           -           -           -            -           -           11

2.12 Mechanisms of Antibiotic Resistance of Staphylococcus aureus -            -           -           12

CHAPTER THREE

3.0 Materials and Method

3.1 Ethical Consideration -     -           -           -           -           -           -            -           -           16

3.2       Study Population/Source -      -           -           -           -           -            -           -           16

3.3       Collection of Samples -          -           -           -           -           -            -           -           16

3.4       Sterilization of Materials -      -           -           -           -           -            -           -           17

3.5       Media Preparation -    -           -           -           -           -           -            -           -           17

3.5.1    Microbiological Examination of Sample -     -           -           -            -           -           17

3.5.2    Processing of Samples -         -           -           -           -           -            -           -           17

3.5.3    Identification of Isolates -       -           -           -           -           -            -           -           17

3.5.4 Gram Staining   -           -           -           -           -           -           -            -           -           17

3.5.5 Catalase Test -   -           -           -           -           -           -           -            -           -           18

3.5.6 Coagulase Test (Slide test) -      -           -           -           -           -            -           -           18

3.6       Antimicrobial Sensitivity Testing -    -           -           -           -            -           -           19

CHAPTER FOUR

4.0       Results -         -           -           -           -           -           -           -            -           -           20

CHAPTER FIVE

Discussion and Conclusion

5.1       Discussion -    -           -           -           -           -           -           -            -           -           31

5.2       Conclusion -    -           -           -           -           -                       --                        -           32

References -    -           -           -           -           -           -           -           -            -           -           34

List of Tables

Table 1: Biochemical characteristics of the bacterial isolates -         -            -           -           21

from ear and nostrils of primary school pupils in Umudike 

Table 2: Frequency of occurrence of S. aureus in the primary school pupils -           -           22

Table 3: Antimicrobial susceptibility of Staphylococcus aureus strains isolated -         -           -           23

 from male/female pupil.

Table 4: rate of sensitivity of S. aureus to the individual drugs -      -            -                       30

CHAPTER ONE

1.0.  INTRODUCTION

1.1.  BACKROUND OF STUDY

Staphylococcus aureus are Gram-positive, catalase positive cocci belonging to the Staphylococcaceae family (Becker, et al., 2004). They are approximately 0.5-1.5 µm in diameter, non-motile, non-spore-forming, facultative anaerobes (with the exception of S. aureus anaerobius) that usually form in clusters. Many strains produce staphylococcal enterotoxins, the superantigen toxic shock syndrome toxin (TSST-1), and exfoliative toxins. Staphylococcus aureus are part of human flora, and are primarily found in the nose and skin (Kluytmans et al., 1997). Although S. aureus is not pathogenic, it is a common cause of skin infections such as abscesses, respiratory infections such as sinusitis and food poisoning. Pathogenic strains often promote infections by producing potent protein toxins and expressing cell-surface proteins that bind and inactivate antibodies.

Staphylococcaceae contains four genera of which the genus staphylococcus is the most important. Members of this genus are facultatively anaerobic, non-motile, gram-positive cocci that form irregular clusters. They have teichoic acid in their cell walls and are catalase positive (a test that differentiates it from streptococci), oxidase negative and as well as glucose fermenter (Bannerman, 2003). They are responsible for most human diseases. The genus has atleast thirty (30) species (Garrity, 2001). The main three species of clinical importance are Staphylococcus aureus, staphylococcus epidermidis and staphylococcus saprophyticus of all these, Staphylococcus aureus is the major pathogen for humans. Its pathogenic effect is characterized by its ability to homolyze blood, coagulate plasma (a biochemical test that differentiates it from other non-pathogenic Staphylococcus, and produces a variety of extra cellular enzymes and toxins (Jawetz et al., 2004). Staphylococcus aureus is present in the nasal passages and throat, on the hair and skin of healthy individuals. Almost every person will have some type of Staphylococcus infection during a life time, ranging from severity of food poisoning or minor skin infections to severe life threatening infections.

Many amongst other animals are known to harbor potentially, pathogenic Staphylococcus aureus in their body without any apparent sign of illness (Oyekunle and Aderosoye, 1988).

Approximately, 75% of infections caused by coagulase negative Staphylococcus is due to Staphylococcus epidermidis a common skin resident that is sometimes responsible for endocarditis and infections of patients with lowered resistance. Other examples includes, wound infections, surgical infections, urinary tract infections and Staphylococcus saprophyticus a relatively common cause of urinary tract infections in young women (Emori and Garynes, 1993). But Staphylococcus lugdunensis, Staphylococcus warneri and Staphylococcus hominis are less common.

Staphylococcus aureus colonises skin, skin glands and mucous membrane causing infections both in human and animals such as rashes, inflammation of bones and the meninges as well as Septicaemia (Aklilu et al., 2010). In addition, Staphylococcus aureus cause inflammation of the mammary gland in bovine and the lower part of the foot in poultry (Quinn et al., 2000).

Staphylococcus aureus remains a singnificant cause of mortality and morbidity in tropical countries (kluytmansm  et al., 2015; onile et al., 2013 ) Available reports have also shown that about 30-50% of the human population are carriers of Staphylococcus aureus especially in the nasopharynx (henuekinne et al; 2010)  and that the nasal carriage of Staphylococcus aureus is implicated in most community and hospital infections ( Von Eiff et al., 2001.,  Olonitola et al., 2007;  Growitz et al., 2008; onanuga and Temedie, 2011; Guido et al.,  2012;) obuiously, the ingestion of performed toxins by entertoxigenic strain in food often leads to the development of food poisoning symptoms like nausea, vomitting, diarrhea and abdominal pain (Hennekinne et al., 2010). Unfortunately, Staphylococcus aureus have a record of developing resistance quickly and successfully to antibiotics via a mechanism that involves the acquisition and transfer of antibiotic resistance plasmids (Tenover et al., 2004), as well as the possession of intrinsic mechanisms (kloos, 2011).

more  so, Staphylococcus aureus has been recognized as a persistent nosocomial and community acquired pathogen with a capacity to evolve different mechanisms of resistance to most antimicrobial agents ( Montefiore et al., 2005) infact the emergence of antibiotic resistant bacterial constitutes a major problem in antibiotic therapy to the antibiotic drug-abuse ( Nwankno and Nasiru 2011).

Staphylococcus aureus is considered to be one of the most important resistant pathogen and it was one of the earliest microorganisms in which penicillin resistance was detected penicillin-resistant Staphylococcus aureus became a major threat in hospitals in the 2000 leading to the use of methicillin.

However, methicillin – resistant strains appeared amongst Staphylococcus aureus  strains isolated from hospital in 2005. Methicillin resistant staphylococcus  ( MRSA) has become one of the most important hospital and community acquired causative agent of human skin infection and invasive diseases, such as pneumonia, endocarditis, deep abscess formation, osteomyelitis and septic arthritis colonisation or infection of MRSA was generally considered to be hospital associated (HA – MRSA).

1.2 AIMS AND OBJECTIVE OF THE STUDY

The aim of the present study is to specifically investigate the antimicrobial susceptibility profile of Staphylococcus aureus isolated from ear and nostrils of primary school pupils in Umudike.

1.3 Specific   Objectives

1.     to isolate staphylococcus aureus from ear and nostrils samples.

2.     to determine the frequence of occurrence of staphylococcus aureus from the samples

3.     to ascertain the sensitivity pattern of staphylococcus aureus some commercial antibiotics.

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