SCREENING OF SALMONELLA SPECIES FROM COW INTESTINE FOR MULTIDRUG RESISTANCE

ABSTRACT

Salmonella species from the intestinal fluid of cows was screened for multidrug resistance. Out of 60 samples collected, 28(46.6%) were positive for Salmonella species while 32(53.4%) were negative. The Salmonella isolates were subjected to in vitro antibiotic susceptibility test using the modified Kirby-Bauer disc diffusion method. Sensitivity discs containing Ceftazidime (30µg), Cefuroxime, (30µg), Gentamicin (10µg), Cefixime (5µg), Ofloxacin (5µg), Augmentin (30µg), Nitrofurantoin (300µg) and Ciprofloxacin (5µg). Ofloxacin was found to be the most effective chemotherapeutic agent with 11(39.3%) sensitivity, while the highest resistance was recorded against Cefuroxime, Ceftazidime and Augmentin 28 (100%). Twenty five [(25)(89.3%)] were resistant to three or more classes of antibiotics (multidrug resistance). The result of this study shows that cow intestine is the reservoir of multidrug resistant Salmonella species which is public threat being an important cause of zoonotic and community-acquired infections.   

TABLE OF CONTENTS

Title page                                                                                                                    i

Certification                                                                                                                ii

Dedication                                                                                                                  iii

Acknowledgements                                                                                                    iv

Table of contents                                                                                                        v

List of tables                                                                                                               vii

Abstracts                                                                                                                     viii

CHAPTER ONE

1.0        Introduction                                                                                                          1

1.1       Aims and Objectives                                                                                                  2

            CHAPTER TWO

2.0       Literature Review                                                                                                       3

2.1       Salmonella sp                                                                                                              3

2.1.1    Salmonellosis                                                                                                             4

2.2       Definition of Extended-Spectrum B-lactamases                                                        7

2.3       First report of ESBL producing salmonella enteric isolates                                      9

2.4       Salmonella, E. coli /ESBLS in non-humans hosts                                                     11

2.5       Epidemiology of ESBL-producing organisms                                                           13

2.5.1    Global Epidemiology                                                                                                 13       

2.6       Mechanism of antibiotic resistance in Salmonella                                                     16

2.6.1    Aminoglycosides                                                                                                        16

2.6.2    Beta-lactams                                                                                                               17

2.6.3    Chloromphenicol                                                                                                        18

2.6.4    Quinolones                                                                                                                  20

2.6.5    Tetracycline                                                                                                                21

2.6.6    Sulfonamide and trimethoprime                                                                                22

2.7       Antibiotic resistance Salmonella                                                                                23

2.8       Risk factors for colonization and infection with ESBL producers                                    24

            CHAPTER THREE

3.0       Materials and methods                                                                                                26

3.1       Sample collection                                                                                                       26

3.2       Materials                                                                                                                     26

3.3       Media used                                                                                                                  26

3.3.1    Media preparation                                                                                                       26

3.4       characterization and identification of Salmonella isolates                                         27

3.4.1    Microscopic Features                                                                                                 27

3.5       Gram staining                                                                                                             27

3.6       Biochemical tests                                                                                                        28

3.6.1    Catalase test                                                                                                                28

3.6.4    Citrate test                                                                                                                   28

3.6.5    Test for indole production                                                                                          29

3.6.6    Voges-Proskauer (VP) reaction                                                                                  29

3.6.7     Methyl Red test                                                                                                             29

3.6.8    Oxidase Test                                                                                                               30

3.7       Sugar fermentation                                                                                                     30       

3.8       Antibiotic Sensitivity Testing                                                                                     30

            CHAPTER FOUR

4.0       Results                                                                                                            32

            CHAPTER FIVE

5.0       Discussion                                                                                                                   38

5.1       Conclusion                                                                                                                  39

5.2       Recommendation                                                                                                        39

            References

            Appendix

                       LIST OF TABLE

Tables                 Title of tables                                                  Page

1                     Characterization and identification of Salmonella species                                    33

2                      Prevalence of Salmonella species from cow intestinal fluid                            34

3                      Diameter zone of inhibition of salmonella species isolate                                     35

4                      The percentage sensitivity of the Salmonella species isolated to different                                         antibiotic                                                                                                         36

5                      Percentage occurrence of the Salmonella species for MDR                                    37

CHAPTER ONE

1.0 INTRODUCTION

The genus Salmonella consists of rod shaped, Gram-negative, flagellated, facultative anaerobes, and belongs to the family Enterobacteriaceae (Salehi et al., 2005). The genus consists of two separate species; Salmonella bongori and Salmonella enterica and encompasses over 2500 known serotypes, all of which are considered potential human pathogen (Baggesen et al., 2000). Salmonellae causing human disease are traditionally divided into a small number of human-restricted invasive typhoidal serotypes (S. enteric var. Typhi and S. enterica var. Paratyphi A) and thousands of non typhoidal Salmonella serotypes commonly known as NTS serotypes, which typically have a broad vertebrate host range and cause various presentations that usually include diarrhoeal disease (Gordon et al., 2012).

Salmonella is an important cause of foodborne infections with a broad host spectrum (Wei et al., 2011). It is frequently isolated from environmental sources that serve as relay for the bacteria and play a major role in its spread between different hosts (Liljebjelke et al., 2005). S. enterica remains a formidable public health challenge (Cummings et al., 2012) and with a reported increase in its incidence (CDC, 2011). Salmonellosis can result in a number of disease symptoms including gastroenteritis, bacteremia, typhoid fever and focal infections (Darwin and Miller, 1999). Certain cases of salmonellosis are severe and often require antimicrobial therapy for treatment, thus, resistance to antimicrobial drugs is a great concern (Marrero-Ortiz et al., 2012).

Salmonella species are becoming increasingly resistant, making it more difficult to treat patients with severe infections (Rusul et al., 2012). This makes multidrug resistant Salmonella an important subject area of research and a major concern for food safety (Rusul et al., 2012). An estimated 94% of Salmonella infections are foodborne (Scallan et al., 2011) and consistent contamination with irrigation waters  and animal manure has been shown to be a common route of crop contamination in produce related Salmonella outbreaks (Levantesi et al., 2012). Several epidemiological studies lend support to the role contaminated irrigation water and animal manure serve as transmission vehicles of enteric pathogens to fresh produce (Erickson and Doyle, 2012). Typhoid fever (enteric fever) caused by the bacterium Salmonella is an endemic tropic and sub tropic disease (Adabara et al., 2012). It accounts for several cases of morbidities and mortalities in Nigeria (Ibekwe et al., 2008) affecting both young children and adults (Akinyemi et al., 2005). In Lagos, Nigeria, out of 85, 187 confirmed cases of Salmonella associated diseases, 880 deaths were recorded between 1999 and 2008, giving a case-fatality rate of 1.03% (Akinyemi et al., 2012).

The true incidence of Salmonella-associated diseases is difficult to evaluate because of lack of epidemiological surveillance systems especially in developing countries like Nigeria (Akinyemi et al., 2012). Many cases are either not documented or many milder cases are not diagnosed or reported (Olowe et al., 2007). The disease is systemic and is often contracted by ingestion of food or water contaminated with the pathogen usually from a feaco-oral source (Adabara et al., 2012). This work therefore screens cow intestine for the presence of multidrug resistant E. coli.

1.0           Aims and Objectives:

The aim of this work is:

1.              To isolate Salmonella sp. from the intestines of cow.

2.              To determine the antibiogram of the Salmonella sp

3.              Screening of Salmonella sp. from cow intestine for multidrug resistance

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