PREVALENCE AND ANTIBIOTIC SUSCEPTIBILITY PATTERN OF SALMONELLA AND SHIGELLA ISOLATED FROM STOOL OF STUDENTS

ABSTRACT

 Salmonellosis and shigellosis are still global health problem, especially, in developing countries where poor sanitation, lack of clean water supply and proper sewage disposal system exist. The emergence of increased antimicrobial susceptibility to Salmonella and Shigella spp are global challenges particularly in developing countries like Nigeria where misused antibiotics by human beings occur. This study aims to investigate the antibiotics susceptibility pattern of these microbes. A total of 60 stool samples were isolated from healthy students of Michael Okpara University of Agriculture Umudike and cultured on Salmonella-Shigella agar for screening of microbes and the antibiotic susceptibility pattern of the isolates were determined following standard bacteriological method.  Antibiotic were selected for the test (Amoxicilin, Ampicilin, Nalixidic acid, Ciprofloxacin, Gentamicin, Chloramphenicol and Tetracycline). 9 (15%) Samonella and 7(11.67%) Shigella was isolated from stool samples of both male and female students. Sensitivity of Salmonella was 100% to Chloramphenicol, 60% to Nalixidic acid, Ampicillin and Gentamicin, 40% to Ciprofloxacin and Tetracycline, and 0% Amoxicillin in female. And 75% to Amoxicillin, Nalixidic acid and Gentamicin, 100% to Ciprofloxacin and Chloramphenicol, 25% to Ampicillin and 0% to Tetracycline in male. Shigella had sensitivities of 75% Amoxicillin, Nalixidic acid and Gentamicin and Tetracycline in female and 100% to Nalixidic acids, Chloramphenicol and Ciprofloxacin, 33.3% to Amoxicillin and Ampicillin, 0.0% to Gentamycin and Tetracycline. However, isolates of Samonella and Shigella were 100% susceptible to Ciprofloxacin, Chloramphenicol and Nalixidic acid. This study revealed that isolates of Shigella and Salmonella showed high rate of drug resistance  to commonly used   antibiotics and all isolates were susceptible to Ciprofloxacin and Chloramphenicol. Therefore Ciprofloxacin and Chloramphenicol can be used as drugs of choice for the treatment of Shigellosis and Salmonellosis.  

TABLE OF CONTENTS

Title page                                                                                                                               

Certification                                                                                                                           i

Dedication                                                                                                                              ii

Acknowledgement                                                                                                                  iii

Table of content                                                                                                                      iv

List of tables                                                                                                                           viii

Abstract                                                                                                                                  ix

CHAPTER ONE

1.0 Introduction                                                                                                                      1

1.1 Aims and objectives                                                                                                         3

CHAPTER TWO

2.0 Literature review                                                                                                              4

2.1 Morphology and culture                                                                                                   4

2.2 Characteristics of the genius Salmonella and Shigella                                                    5

2.3 Geographical distribution                                                                                                 7

2.4 Pathogenicity of the genus Salmonella                                                                            7

2.4.1 Enteric fever                                                                                                                  8

2.4.2 Gastroenteritis                                                                                                               8

2.5  Pathogenicity of genus Shigella                                                                                      9

2.6 Transmission of salmonellosis and shigellosis                                                                 10

2.7 Clinical signs and symptoms of Salmonella infection                                                     10

2.8 Clinical signs of Shigella infection                                                                                  11

2.9 Prevention and treatment                                                                                                 11

2.9.1 Prevention                                                                                                                      11

2.9.2 Treatment                                                                                                                      12

2.10 Antibiotic susceptibility and resistance                                                                          13

2.11 Classificaion of antibiotics                                                                                             14

2.12 Mechanism of action of antibiotics                                                                                16

2.12.1 Inhibition of cell wall synthesis (Beta- lactams)                                                         16

2.12.2 Inhibition of cell function                                                                                            17

2.12.3 Inhibition of protein synthesis                                                                                     17

2.12.4 Inhibition of nucleic acid synthesis                                                                             18

2.12.5 Inhibition of other metabolic processes (anti metabolites)                                         18

CHAPTER THREE

3.0 Materials and methods                                                                                                      19

3.1 Samples and samples/study population                                                                            19

3.2 Specimen collection                                                                                                         19

3.3 Sterilization of glass wares                                                                                               19

3.4 Preparation of culture media                                                                                            19

3.5 Preparation of innoculum size                                                                                          20

3.6 Identification test                                                                                                              20

3.6.1 Gram staining                                                                                                                20

3.7 Biochemical test                                                                                                               21

3.7.1 Catalase test                                                                                                                   21

3.7.2 Indole production test                                                                                                    21

3.7.3 Coagulase test                                                                                                                21

3.7.4 Oxidase test                                                                                                                   22

3.7.5 Urease test                                                                                                                     22

3.7.6 Citrate utilization test                                                                                                    22

3.7.7 Methyl red test                                                                                                               23

3.7.8 Voges prokauer test                                                                                                       23

3.7.9 Motility test                                                                                                                   23

3.9 Antibiotics susceptibility test                                                                                           23

3.9.1 Reading of inhibition                                                                                                     24

CHAPTER FOUR

4.0 Results                                                                                                                              25

CHAPTER FIVE

5.0 Discussion, conclusion and recommendation                                                                  31

5.1 Discussion                                                                                                                         31

5.2 Conclusion                                                                                                                        33

5.3 Recommendation                                                                                                             33

REFERENCES                                                                                              

LIST OF TABLES

Table 1: Morphological and biochemical characteristics                                                      27

Table 2: Percentage Occurance                                                                                              28

Table 3: Antibiogram of Salmonella and Shigella Isolates from Female Students                      29

Table 4: Antibiogram of Salmonella and Shigella Isolates frommale students                       30

CHAPTER ONE

1.0  INTRODUCTION

The discovery of Salmonella and Shigella is well documented in recorded history. As early as the mid 1800s scientific interest in Salmonella was initiated by the organisms potential etiological association with typhoid fever (Cunha, 2004).Salmonella and Shigella are genus of rod-shaped, gram negative motile, falcultatively anaerobic bacilli belonging to the family Enterobacteracae. (Ellermeier and Stauch2006).The bacterial genius, Salmonella is divided into two species, Salmonella bongori and Salmonella enteric. Salmonella enteric itself comprised of six subspecies; S.enterica subsp.enterica,S.enterica subsp.salamae,S.enterica susp.arizonae, S.enterica subsp.indica, S.entrica subsp.boutenae and S.enterica subsp.diarizonaeor I, ii, iiia, iiib, iv and vi respectively (Porkwollik et al.,2003) 

Of date, there exist over 2,300 serovars identified within subspecies 1. However, only a small fraction of the thousands of described subspecies 1 serovars frequently cause disease in human and domestic animals.The bacterial genus Shigella on the order hand is divided into four species which includes; Shigella dysenteriae (group A), Shigella flexneri (group B), Shigella boydii (group C) and Shigella sonnei (group D). (Robert, 1995).Shigella is transferred from person to person usually by asymptomatic carriers and via contaminated food flies, feces, fingers and water. (Robert, 1995). Infection caused by Shigella species are associated with human, no animal reservoir has been identified. Salmonella and Shigella caused mild to severe forms of intestinal tract infection. Salmonella cause self-limited gastro-enteritis and the more severe forms of systemic typhoid fever (Kasper et al., 2005). Shigella species are limited to the intestinal tract of humans and cause bacillary dysentery leading to watery or bloody diarrhea. Bacillary dysentery or shigelliosis and typhoidal salmonellosis are major diseases caused by Shigella and Salmonella and they are major health problems in developing countries which Nigeria is not an exception. Humans acquire the infection by ingesting the organism in contaminated food and water. These infections are prevalent in areas where lack of clean water supply, lack of proper sewage disposal system and flies aggravate the spread of the disease (Kasper et al., 2005). Epidemiological surveillance is an essential component in controlling Salmonella and Shigella infections (Shama et al., 2005).

Due to different factors in the last few decades, Shigella and Salmonella have become increasingly resistant to the most commonly used antimicrobials from selecting the drug for therapeutic management (Gubet al., 2012). The emergence of antibiotics resistant Shigella and Salmonella are serious problems in antimicrobial therapy globally. The incidence varies the area of isolation of these strains. The progressive increase in antibiotics resistance among these pathogens in developing countries is also becoming a critical area of concern. ( Huruyk et al., 2008).

In developing countries like Nigeria, resistance may be acquired mostly by selective pressure due to indiscriminate and misuse of antibiotics. This leads for the emergence of resistance strains of Shigella and Salmonella which may be difficult for treatment and prevention (Chessbrough, 2009). In the 1940s, Shigellosis was treated with tetracycline. In the 1970s Ampicillin was the drug of choice for the treatment of bacillary dysentery (Braud, 1976). After the pathogen began to develop resistance to Ampicillin the new drug Trimethoprime – sulphamethoxazole (TMP-SXT) was used (Ericsson et al., 1990). Even if the drug of choice in that time was TMP-SXT, in the 1980s, the Shigella species started to develop resistance to the drug. (Harnett et al., 1991). Then the emergence of resistance strains continues with any new drugs. Until about 1960, nearly all Salmonella were sensitive to a wide range of antimicrobial agents but since 1962 emergence of resistance, frequently plasmid mediated, have appeared in Salmonella worldwide. The relative importance of antibiotics resistance and the serotype, in which it occurs, differs from country to country (Coburn B, et al., 2006). Knowing the prevalence and examining the antibiotic susceptibility pattern of these pathogens is important towards tailoring treatment to the ever changing resistance patterns and distribution of pathogenic bacteria.

However, not much work has been done on the incidence and antibiotic susceptibility pattern of Salmonella and Shigella species among healthy students of MOUAU. Such a data will be necessary in order to develop rational strategies for therapy of Salmonella infection. Therefore, this study fills the knowledge gap on the prevalence and antibiotic susceptibility of the pathogenic Salmonella and Shigella among MOUAU students.

1.1       AIMS AND OBJECTIVES

The broad objective of this study is to review and ascertain the presence of Salmonella and Shigella in stool and also the occurrence of antimicrobial resistance among species of Salmonella and Shigella.The specific objectives are:

v  To isolate and identify Salmonella and Shigella in stool samples of healthy students of MOUAU.

v  To know the prevalence of Salmonella and Shigella among both male and female healthy students of MOUAU.

v  To determine antibiotic susceptibility pattern of these organisms and hence provide information on the best antibiotics to be used for treatment of Salmonella and Shigella infection.

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