ISOLATION AND ANTIBIOGRAM OF SALMONELLA SPP FROM POULTRY FEEDS AND FECAL DROPPINGS FROM SELECTED FARMS IN UMUDIKE, UMUAHIA.

ABSTRACT

The present study was aimed at the isolation and antibiogram study of Salmonella spp from various poultry feeds (starter, grower and layer feeds) and fecal droppings in some selected poultry farms in Umudike. 144 samples (72 feed samples and 72 fecal dropping samples) were used for the study and all  the  samples were cultured on Salmonella-Shigella agar and the organisms were identified using biochemical tests and by Gram staining, the microbial load of the organisms were determined by carrying out 5 fold serial dilution of the samples. The biochemical results and Gram staining results showed that all the isolates from the samples were Salmonella spp. The prevalence of Salmonella was highest in the poultry droppings than in the poultry feeds. The highest number of Salmonella spp occurred in the feed samples for growers (37.5%) while the lowest occurred in the starters (20.8%) while the highest occurrence of Salmonella in fecal droppings was 66.67%. The highest TVC of Salmonella isolates in the feeds were 118.67± 1.15 x 10⁵Cfuml^-1 while the highest TVC in the fecal droppings was 145.00±6.56x10⁵ Cfuml^-1.The antimicrobial testing showed that the isolated Salmonella spp were highly sensitive to Gentamycin, Ciprofloxacin and Ofloxacin, moderately sensitive to Erythromycin and Streptomycin but highly resistant to Augmentin and Ampicillin.  The present study indicates that Salmonella spp are more prevalent in the fecal droppings of poultry birds than in the feeds of poultry birds. The study also indicates that Ciprofloxacin and Ofloxacin can be used as first line therapies for Salmonella gastroenteritis from poultry sources.

TABLE OF CONTENTS

Title Page                                                                                                                            i

Certification                                                                                                                                     ii

Dedication                                                                                                                          iii

Acknowledgements                                                                                                                        iv

List of Tables                                                                                                                                    ix

List of Figures                                                                                                                                  x

Abstract                                                                                                                                                xi                

CHAPTER ONE: INTRODUCTION

1.1 Background to the study                                                                                        1

1.2 Aim of the study                                                                                                    3

1.3 Objectives of the study                                                                                          3

CHAPTER TWO: LITERATURE REVIEW

2.1 General overview of Salmonella                                                                                  4

2.1.1 History of Salmonella                                                                                        4

2.1.2 Taxonomy and Nomenclature of Salmonella                                                     4

2.2 Pathogenesis of Salmonella                                                                                 5

2.3 Epidemiology of Salmonella contamination in poultry products                          6

2.4 Salmonella Contamination                                                                                           7

2.4.1 Sources of Salmonella                                                                                        8

2.4.2 Clinical manifestation of Salmonella                                                                 9

2.5 Clinical prevalence of Salmonella spp among poultry products                           9

2.6 Chicken as a reservoir of Salmonella                                                                          11

2.7 Isolation of Salmonella from Chicken                                                                        12

2.7.1 Isolation of Salmonella from Poultry fecal droppings                                             14

2.8 Characterization of Salmonella                                                                             14

2.8.1 Cultural characteristics of Salmonella spp                                                         14

2.8.2 Biochemical characteristics of Salmonella                                                              15

2.5 Antibiogram study of Salmonella                                                                               16

CHAPTER THREE: MATERIALS AND METHODS                                     

3.1 Description of study area                                                                                      18

3.2 Sample collection and Sample sizes                                                                     19

3.3 Transportation of Samples                                                                                    19

3.4 Preparation of Media                                                                                                    22

3.5 Determination of microbial load                                                                                  22

3.6 Isolation of Salmonella from Poultry feeds                                                                 23

3.7 Isolation of Salmonella from Poultry fecal droppings                                                 23                                                                 3.8 Identification of the Isolated organisms                                                               23

3.8.1 Gram staining                                                                                                     23

3.8.2 Biochemical test of the isolates                                                                          24

3.8.3 Methyl Red test                                                                                                  24

3.8.4Voges-Proskeaur test                                                                                                  24

3.8.5 Catalase test                                                                                                               24

3.8.6 Oxidase test                                                                                                               24

3.8.7 Citrate test                                                                                                          25

3.9 Antibiogram study of the Salmonella isolates                                                     25

CHAPTER FOUR: RESULT

4.1 Result                                                                                                                       26-37

CHAPTER FIVE: DISCUSSION, CONCLUSION AND RECOMMENDATION

5.1 Discussion                                                                                                          37

5.2 Conclusion                                                                                                                40

5.3 Recommendation                                                                                                      40

 References

LIST OF TABLES

1.   Study design                                                                                               18

2.   Biochemical reaction of the Salmonella spp isolated from all the tables                  28                                                                           

3.                     The frequency of Salmonella spp in the various feed samples in NRCRI and      MOUAU poultry farms from April to July 2016.                                                29

4.              The frequency of Salmonella spp in fecal dropping samples in NRCRI and MOUAU poultry farms from April to July 2016.                                  30

5.              Microbial load for Starter feeds and droppings                                                    35

6.              Microbial load for Grower feeds and droppings                                                  36

7.              Microbial load for Layer feeds and droppings                                                    37

LIST OF FIGURES

1. Study design                                                                                                              18

2. Antibiotic sensitivity profile of Salmonella isolates  from Amobyng poultry feeds and fecal droppings collected from MOUAU                                                               31

3.  Antibiotic sensitivity profile of Salmonella isolates from Top poultry feeds and fecal droppings collected from NRCRI                                                                               32

4. Antibiotic sensitivity profile of Salmonella isolates  from vital poultry feeds and fecal droppings collected from NRCRI                                                      33

5. Antibiotic sensitivity profile of Salmonella isolates  from Amobyng poultry feeds and fecal droppings collected from NRCRI                                         34

CHAPTER ONE

INTRODUCTION

1.1  BACKGROUND TO THE STUDY

Salmonella are rod shaped gram negative bacteria of the Enterobactericeae family (OIE, 2006). The genus consist of two (2) specie; Salmonella enterica and Salmonella bongori, and are non spore forming, predominantly motile Enterobacteria with cell diameters between approximately 0.7 and 1.5 um lengths from 2-5um and peritrichous flagella (flagella that are all around the cell body). They are chemotrophs, obtaining their energy from oxidation and reduction reactions using organic sources. They are also facultative anaerobes capable of surviving with or without oxygen. More than 2300 serotypes exist based on 67 “O” antigen (for non motile species) and the numerous antigens (for motile species) recognized thus far (Blood et al., 2003).

The development of the poultry industry was described as the fastest means of bridging the protein deficiency gap in most developing nations.  This has concomitantly required trade in hatching eggs, day old chicks, feed concentrates and additives from various controlled and uncontrolled, local and international sources (Al-Nakhli et al., 1999; Gene, 2006). Poultry immediate micro environment includes the housing, the feed they consume, climatic and management factors which are reported to have effects on the performance of the birds (Apantanku, 2006). From various studies conducted, poor performance, feeds and feeding, diseases and pests have been tagged major constraints to poultry production in Nigeria. Infections due to Salmonella spp. continue to cause global concern of morbidity and mortality in human and animals as well as significant economic losses. Salmonellosis occurs mainly by a fecal-oral route through the consumption of contaminated feed and water (WHO, 2010).

Sources of Salmonella infections into poultry farms include contaminated feed  and feed ingredients, water, equipments, personnel, rodents and hatchery related unhygienic activities (Ricardo and Brazil, 2012; Roth, 2012).

Salmonella is a primary inhabitant of the gastro intestinal tract and is recognized as one of the most common causes of food borne infections and human death (mead et al., 1999). Salmonella is also reported to cause widespread food borne zoonotic infections in both developed and developing countries (Ostean, 1999). The united state centre for disease control and prevention, reported the implication of human Salmonella infection outbreaks with poultry, accounting for more than 50% of such outbreaks (Roth, 2012).

There is an increase in the number of resistant strains of Salmonella isolated from poultry and poultry environment, and this has been attributed to the widespread use of antibiotics in human and veterinary medicine (Anthony et al., 2001). Salmonella isolates from poultry and fed with antibiotics as growth hormones, could exhibit resistance to antibiotics. Collington (2003) reported the successful reduction in prevalence of antibiotic resistant strains of Salmonella due to stringent controls of antibiotics and surveillance of antibiotic resistance pattern in populations.

However, such resistance patterns are not needed in developing countries like Nigeria. This resistance occurs due to the indiscriminate and continuous use of sub therapeutic doses of antibiotics. This study was designed to select the appropriate antibiotics against Salmonella determining the resistance and sensitivity patterns of the isolates with the view to make people more prudent in the use of antibiotics.

1.2 AIM OF THE STUDY

The aim of the study to isolate and identify Salmonella spp. from various feed samples and fecal droppings from some selected farms in Umudike and a further assessment of their sensitivity patterns to different standard antibiotics.

1.3 OBJECTIVES OF THE STUDY

In view of the above considerations, this project work was undertaken within the following objectives.

i.               To isolate and identify Salmonella from the various test feed samples and fecal droppings.

ii.              To determine the prevalence of Salmonella spp in the various feed samples and fecal droppings.

iii.            To determine the microbial load of the various feed samples and fecal droppings.

iv.            To study the susceptibility of the Salmonella isolates to standard antibiotics.

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