ISOLATION AND ANTIBIOTIC SENSITIVITY PROFILE OF SALMONELLA TYPHI FROM STOOLS OF PATIENTS IN UMUAHIA

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ABSTRACT

 

Fifty (50) stool samples and corresponding venous blood specimens were collected from patients who presented for widal test diagnosis of typhoid fever in five Hospitals or Clinics in Umuahia. The stool samples were cultured on Salmonella  Shigella agar  (SSA) and the sera from the blood  specimens were used to screen for antibodies to febrile antigens of Salmonella typhi and Salmonella paratyphi A, B, and C. Out  of the 50  stool samples cultured, 33(73.33%) produced  significant growth on SSA. The isolates were characterized by colonial morphology, Gram staining reaction and Biochemical tests and were provisionally identified as Salmonella specie. The antibiotics sensitivity profile of the 33 isolates was done using the Kirby-Bauer disc agar diffusion technique. All the 33(100%) isolates were sensitive to Nitrofurontoin, 27(81.8%) were sensitive to Gentamicin, 22(66.7%) to Ciprofloxacin and 19(57.5%) to Ofloxacin, while one (3.30%) isolates was sensitive to Cefuroxime, all the isolates were resistant to Augmentin, Ceftazidime and Ceftriaxone. Comparison of culture widal test results revealed that only 7(21.2%) of the culture positive cases had significant antibody titres (> 1:80) to Salmonella febrile antigens. The result of this study show that there is a high level of antibiotic resistance among Salmonella  isolates and that widal test detected less than 50% of cultured positive cases. The results of this work suggest there is a need for culture and sensitivity as the basis for diagnosis and treatment of typhoid disease in the study area.





TABLE OF CONTENTS

Title page i

Certification ii

Dedication iii

Acknowledgements iv

Table of Contents                                                                                                          v

 .List of Table         viii

 List of Figure ix

Abstract x

CHAPTER ONE

1.1 INTRODUCTION 1

1.2 Aims and objectives   4

CHAPTER TWO

2.0 LITERATURE REVIEW 5

2.1 Prevalence / incidence 5

2.2 Isolation and identification of Salmonella 7

2.3 Culture 8

2.4 Anti-microbial resistance 8

2.5 Widal agglutination test 11

CHAPTER THREE

3.0 MATERIALS AND METHODS 13

3.1 Study area 13

3.2 Sample collection 13

3.3 Media preparation 14

3.4 Isolation and identification of salmonella species 14

3.5 Culture 14

3.6 Widal tests 14

3.7 Gram Staning 15

3.8 Anti microbial Susceptiblity testing 15

3.9 Biochemical tests 16

CHAPTER FOUR

4.0 RESULTS 18

CHAPTER FIVE

5.0 DISCUSSION, CONCLUSION AND RECOMMENDATION 27

5.1 Conclusion 29

5.2 Recommendation 30

References 31

Appendix 37

LIST OF TABLES

 

Table                        Title Page

 

1: Comparison of culture and widal tests for diagnosis of typhoid

 among the study population 21

2: Antibiotic sensitivity profile of the isolates 22

3: Diameters of zone inhibition of various antimicrobial agents against

 the isolates (stated interpretative reference range in brackets) 23

 

 

 

 

 

 

 

 

 

 

LIST OF FIGURE

 

Figure

                                       Title

     Page

1

Prevalence of Salmonella spp in stool sample in Umuahia

      20

2

In vitro Antimicrobial Susceptibility profile of 33 Salmonella isolate

      26

 

 

 

 

 

 

 

 

 

 

 

 


 

 

 

 

 

 

 

 

CHAPTER ONE

1.1 INTRODUCTION

Salmonellae, a genus belonging to the family of Enterobacteriaceae. Comprises a large and complex group of human pathogen that have been long associated with a wide spectrum of infectious diseases, including typhoid fever and non typhoidal. Is one of pathogen that causes death in infants and immune compromised and aged individual through out the world. These organisms are transmitted through food, water, environmental contacts, pet (animals) and from person to person, with morbidity rates in developing countries 3 to 6 higher than in developed countries.

(Guerrant, 2001).  

Among the enteric pathogens, Salmonella is a particular concern as the cause of enteric fevers, food poisoning and Gastroenteritis. Although more prevalent in developing countries, Salmonella, with its more than 2500 different serotypes is a leading cause of food borne infections worldwide. Two hundred million to more than one billion cases of diarrhea result worldwide due to Salmonella every year  leading to 3 million deaths (Goburn, et al., 2007), and this occurs mostly in developing countries especially in children and young adult. (Kasper, et al., 2005; Niyogi, 2005; Khatun, et al., 2011). Typhoid and paratyphoid fevers (enteric fevers) are acute systemic infections caused mainly by the bacterium Salmonella enterica serotype typhi and other serotypes of Salmonella paratyphi A, B. and C (Buckle et al., 2012). It continues to be a global health problem, especially in the tropics and sub-tropical countries. Over 27 million person suffer from this disease annually (Buckel et al., 2012). Salmonella 

causes mild to severe forms of intestinal tract infections. It can cause self-limited and the more severe form of systemic typhoid fever (Kasper et al., 2005; Goburn et al., 2007). These infections are prevalent in developing countries where lack of clean water supply, lack of proper sewage disposal system and flies aggravate the spread of the disease (Kasper, et al., 2005). Salmonella infections can be divided into two major groups of clinical importance; Typhoidal salmonellosis (Salmonella typhi and Salmonella paratyphi) and non Typoidal salmonellosis (all Salmonella serovars) Abdullahi, 2010). Human Salmonellosis is usually self-limiting, and antimicrobial treatment is seldom required. Reports have indicated that antimicrobial treatment for uncomplicated Gastroenteritis does not reduce the severity or duration of symptoms; in contrast, it may prolong faecal excretion at convalescence and result in the emergence of resistant strains (Abdullahi, 2010). Nevertheless, if spread beyond the intestine occurs, effective antimicrobial treatment is essential, and the knowledge of the likelihood of resistance to commonly available drugs could be of considerable value to the clinicians.

Humans spread Salmonella mainly through the stool. Food borne illness among the people and the transmission can occur when food and water are contaminated with stool or through direct faecal – oral route. Human stool acts as an important reservoir of Salmonella serovars that are grouped based on their cell surface antigen. Species isolated from human stool are Salmonella typhi, S. paratyphi A, S. typhimurium, S. wrothington, and S. enteritidis (Kumar, et al., 2009). Salmonella is a worldwide issue in public health sector. People most at risk for serious complications due to food poisoning include older adults, pregnant women, infant and people who have compromised immune systems. Salmonellosis is manifested clinically in all hosts by one of three major syndromes, acute systemic infection, acute enteritis or chronic enteritis (Dworkin, et al., 2001). Symptoms usually include headache, nausea, vomiting, fatigue, gastroenteritis, abdominal cramps, bloody diarrhea with mucus and sometimes reactive arthritis (Reiters syndrome) (Dworkin, et al., 2001). Following dehydration with renal insufficiency and death may occur.

 

Drug resistance is the decreased sensitivity or the complete insensitivity of microorganisms to drugs that cause cell death or inhibition of growth (Sharma, et al., 2005). Microbial resistance of Salmonella is an emerging global challenge, especially in developing countries where there is an increased misuse and abuse of antimicrobial agents in humans and animals (Kasper, et al., 2005). In most developing countries, laboratory investigations of Salmonella are diagnostic challenges due to the lack of adequate facilities that enable culture and antimicrobial susceptibility testing. As a result, there is limited awareness of the prevalence of infections and antimicrobial resistance (Cook, et al., 2003; Sharma, et al., 2005). Also, the injudicious use of antibiotics by patients and physicians like in many developing countries has led to increased antibiotic resistance, and in turn reduced therapeutic efficacy in these countries (Okeke, et al., 2007; Asrat, 2008). Antimicrobial resistance has complicated the selection of antibiotics for the treatment of enteric bacterial pathogens, particularly to commonly used antimicrobial agents such as Ampicillin, Tetracycline and Trimethoprim – Sulfamethoxazole (ISenbarger, et al., 2002).  Examining the antibiotic susceptibility pattern of pathogens is important towards folioing treatment to the ever changing resistance patterns and distribution of pathogenic bacteria. Hence, there is a great need susceptibility pattern of different bacterial agents which cause enteric infections in order to introduce effective treatment for gastrointestinal illness.

1.2 Aims and objectives  

1. To access the frequency of isolation of Salmonella species from stool samples of people undergoing widal test in Umuahia.

2. To determine the antimicrobial susceptibility profile of the isolates.

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