ANTIMICROBIAL SUSCEPTIBILITY PATTERN OF GRAM NEGATIVE PATHOGENIC ORGANISMS ISOLATED FROM WATERSIDE ABATTOIR ABA, ABIA STATE

ABSTRACT

The antibiotic susceptibility pattern of six Gram negative pathogenic bacteria; Escherichia coli, Pseudomonas, Klebsiella, Salmonella, Shigella and Enterococcus was evaluated. The test organisms were subjected to antibiotic susceptibility test using the Kirby Bauer method. A multi-disc containing eight (8) antibiotics; Ceftrazidime (30ug), Cefuroxime (30ug), Gentamicin (10ug), Ceftriaxone (30ug), Ofloxacin (5ug), Ciprofloxacin (5ug), Augmentin (30ug), Nitrofurantoin (300ug), was used. The result showed all (100%) the isolates were sensitive to Ciprofloxacin and Ofloxacin while all (100%) were resistant to Augmentin. 5(83.3%) of the test organisms were resistant to  Nitrofurantoin and Ceftriaxone. Apart from Shigella  spp which was resistant to only one antibiotic (12.5%), all the others recorded resistance to two or more antibiotics. The result showed that the test organisms are multidrug resistant hence, might have chemical consequences since the organisms are also Zoonoses.

TABLE OF CONTENTS

Title page i

Certification ii

Dedication iii

Acknowledgments iv

Table of Contents v

List of Tables vii

Abstract viii

CHAPTER ONE

1.1    INTRODUCTION  1

CHAPTER TWO

LITERATURE REVIEW

2.1      Potential Bacterial Contaminants associated with Abattoir Wastewater 4

2.1.2 Escherichia coli 4

2.1.3 Salmonella 6

2.1.4 Shigella 8

2.1.5 Klebsiella 9

2.1.6 Enterobacter 10

2.1.7 Serratia 11

2.1.8 Pseudomonas 11

2.2 Antibiotic Resistant 11

2.3 Definition of ESBL’s (Extended Spectrum Beta Lactamase) 12

2.4 Resistance to -Lactams 12

2.5 Antimicrobial Susceptibility14

CHAPTER THREE

MATERIALS AND METHODS

3.1  Study Area 16

3.2 Sample Collection 16

Sterilization of Materials 16

3.4 Preparation of Culture Media 16

3.5 Identification and Confirmation of Isolates 17

Gram Staining 17

3.5.2  Motility Test 18

3.5.3 Biochemical Tests 18

3.5.4 Catalase Test 18

3.5.5 Oxidase Test 19

3.5.6 Urease Test 19

3.5.7 Indole Test 19

3.5.8 Methyl Red-Voges Proskauer Test 20

3.5.9 Citrate Utilization Test 20

3.5.10 Sugar Fermentation Tests 20

3.6  Antimicrobial Susceptibility Testing 21

3.7 Measurement of Zone of Inhibition 21

CHAPTER FOUR

4.1 Results 22

CHAPTER FIVE

DISCUSSION AND CONCLUSION

5.1 Discussion 27

5.2 Conclusion 28

5.3  Contribution to Knowledge 29

5.4 Recommendation 29

5.5 Suggestion for Further Studies 29

REFERENCES

APPENDIX

.

LIST OF TABLES

Table                                                                          Title Page                                                                                                              

1 Identification and confirmation of the Bacterial isolate from water-side abattoir effluents and river samples 23

 2  Antimicrobial susceptibility pattern of gram negative pathogenic

 isolates from Abattoir (pattern inhibition) 24

 3   Percentage susceptibility Pattern of the test Isolate 25

 4 Percentage susceptibility pattern of inhibition of the 8 antibiotics to the test organisms. 26

CHAPTER ONE

1.1    INTRODUCTION      

The characteristics of a slaughter house waste and effluent vary from day to day depending on the type of stock being processed and the method. Waste generated by abattoirs include solid waste, bones, horns and faecal components, slurry of suspended solids, fat, blood and soluble materials (Sangodoyin, 1992). These wastes from abattoir operation can be separated into solid, liquid and fat. These wastes are highly organic. The solid wastes include condensed meat, undigested ingesta, bones, horns hairs and aborted fetus. The liquid waste is usually composed of dissolved solid, blood and gut content, urine and water, while fat waste consist of fat and oil, grease which has high organic levels (Bull, 1982).

The total of waste product per animal slaughtered is approximately 35%  it’s total body weight (world bank, 1998). In an earlier study (Verheijen, 1996), found out that for every 1,000kg of carcass weight, a slaughtered cow produce 5.5kg of manure (excluding rumen content or stockyard manure) and 100kg of paunch manure (partially digested food). The weight of a matured cow varies with size, ranging from 40kg for a thin animal, 550kg for a moderate one, to 750kg  for the extremely fat one (Hammack and Gill, 2002). (Scahill, 2003) gave more detained statistics on both the live and death weight of a cow.

Potential health risks from water borne pathogens can exist in water contaminated by abattoir effluents (Cadmus, 1999), runoff from feedlots (Minner, 1966) ,dairy farm (Jazen, 1974) ,grazed pastures (Doran, 1979) and sewage sludge treated land (Dunigan, 1980).Such contamination of water bodies from abattoir wastes could be constitute significant environmental and public health hazards (Nafarnda, 2006).

Bacteria from abattoir wastes discharged into water columns can subsequently be absorb to sediments, and when the bottom stream is disturbed, the sediments releases the bacterial back into the water columns presenting long-term hazard (Sherer, 1992). Pathogens present in animal carcassesor shed in animal wastes may include Escherichia coli, Yersina enterocolitica, Campylobacter spp, Cryptosporidium parvum and Giardia lambia, others include microorganisms that has been isolated from abattoir effluent; Proteus spp, Staphylococcus aureus, Escherichia coli, Klebsiella spp, Pseudomonas sp, Aspergillus  flavus, Penicillium sp, Salmonella spp, faecal Streptococcus, Listeria monocytogenes(Cadmus, 1999).

There primary reservoir for E.coli 0157:H7 has been reported to be in healthy cattle in a study in Canada although the bacterium is also endemic to swine and sheep (Jackson, 1998). These zoonotic pathogens can exceed millions to billions per gram of feces, and may infect humans through various routes such as contaminated air, contact with livestock animals or their waste products, swimming in water impacted by animal feces, exposure to potential vectors (such as flies, mosquitoes, water fowl, and rodents), or consumption of food or water contaminated by animal wastes (Armand, 2005; Schlech, 2005).

The consequences of infection by contaminated by animal wastes can range from temporary morbidity to mortality, especially in high-risk individuals. Due to the difficulties in qualifying pathogens, indicators of fecal pollution, including coliform bacterial, Fecal coliform, Escherichia coli and Enterococci have been mentioned in lieu of overt pathogen for more than 100 years (Byamukama, 2015).     

1.2 Aim/Objectives

§ The antibiotic susceptibility pattern of gram negative organism isolated from water-side abattoir.

§ To recommend from the result the right antibiotic of choice for the treatment of these pathogenic organisms.

1.3 Scope of Antimicrobial Susceptibility

There are three test methods for carrying out/analyzing an antimicrobial susceptibility

§ Disk diffusion

§ Broth dilution

§ Agar dilution

Antimicrobial susceptibility testing methods that consistently provide reproducible and repeatable results is achieved when followed correctly.

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