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BACTERIOLOGICAL QUALITY and OCCURRENCE of SALMONELLA SPECIES and ESCHERICHIA COLI O157:H7 in ROASTED RAT (Arvicanthis Niloticus) MEAT SOLD in ZARIA, NIGERIA

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BACTERIOLOGICAL QUALITY and OCCURRENCE of SALMONELLA SPECIES and ESCHERICHIA COLI O157:H7 in ROASTED RAT (Arvicanthis Niloticus) MEAT SOLD in ZARIA, NIGERIA BACTERIOLOGICAL QUALITY AND OCCURRENCE OF SALMONELLA SPECIES AND ESCHERICHIA COLI O157:H7 IN ROASTED RAT (Arvicanthis niloticus) MEAT SOLD IN ZARIA, NIGERIA BY TERSEER IYENE ADDAI DEPARTMENT OF VETERINARY PUBLIC HEALTH AND PREVENTIVE MEDICINE, FACULTY OF VETERINARY MEDICINE, AHMADU BELLO UNIVERSITY, ZARIA, NIGERIA APRIL, 2018 BACTERIOLOGICAL QUALITY AND OCCURRENCE OF SALMONELLA SPECIES AND ESCHERICHIA COLI O157:H7 IN ROASTED RAT (Arvicanthis niloticus) MEAT SOLD IN ZARIA, NIGERIA BY TERSEER IYENE ADDAI (P15VTPH8019) A DISSERTATION SUBMITTED TO THE SCHOOL OF POST GRADUATE STUDIES, AHMADU BELLO UNIVERSITY, ZARIA IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE AWARD OF MASTERS DEGREE IN VETERINARY PUBLIC HEALTH AND PREVENTIVEMEDICINE, DEPARTMENT OF VETERINARY PUBLIC HEALTH AND PREVENTIVE MEDICINE, FACULTY OF VETERINARY MEDICINE, AHMADU BELLOUNIVERSITY, ZARIA. SUPERVISORY COMMITTEE PROF. E.C OKOLOCHA CHAIRMAN DR. B.V. MAIKAI MEMBER APRIL, 2018 DECLARATION I hereby declare that this project titled “Bacteriological quality and occurrence of Salmonella species and Escherichia coli O157:H7 in roasted rat (Arvicanthis niloticus) meat sold in Zaria, Nigeria” was done by me in the Department of Veterinary Public Health and Preventive Medicine, Ahmadu Bello University, Zaria, under the supervision of Prof. E.C Okolocha and Dr. B.V. Maikai. The information derived from the literature have been duly acknowledged in the list of references provided. No part of this work has been presented for another degree or diploma in any institution. Terseer Iyene ADDAI (Signature) (Date) i CERTIFICATION This dissertation entitled “BACTERIOLOGICAL QUALITY AND OCCURRENCE OF SALMONELLA SPECIES AND ESCHERICHIA COLI O157:H7 IN ROASTED RAT (ARVICANTHUS NILOTICUS) MEAT SOLD IN ZARIA, NIGERIA” by Terseer Iyene ADDAI meets the regulation governing the award of Master of Science in the Department of Veterinary Public Health and Preventive Medicine, Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria and is approved for its contribution to knowledge and literary presentation. Prof. E.C. Okolocha Chairman Supervisory Committee (Signature) (Date) Ahmadu Bello University, Zaria Dr. B.V. Maikai Member Supervisory Committee (Signature) (Date) Ahmadu Bello University, Zaria Prof. M. Bello Head of Department (Signature) (Date) Veterinary Public Health And Preventive Medicine. Ahmadu Bello University, Zaria Prof. S.Z. Abubakar Dean, School of Postgraduate Studies (Signature) (Date) Ahmadu Bello University, Zaria ii DEDICATION This project work is dedicated to the Lord Almighty, my parents, siblings, all my friends, and in loving memory of my father, Late Mr. J.H. Addai. iii ACKNOWLEDGEMENT My immeasurable gratitude goes to the Almighty God for the grace given to me to accomplish a part of my dream. My unquantifiable gratitude goes to my parents, Mr. J.H. Addai (Late) and Mrs. E. Addai and my siblings, Terna, Aondongu, Arita and Aondofa, for their relentless support in all ramifications throughout the course of this work. Words are not enough to express my appreciation to my sedulous supervisors and role models; Professor E.C. Okolocha and Dr. Mrs. B.V. Maikai for their guidance through every fragment of this work. My heart filled appreciation also goes to the wonderful lecturers in the Faculty of Veterinary Medicine especially in the Department of Veterinary Public Health and Preventive Medicine for tutoring and mentoring me through this program. Special thanks also to Dr. M. Chia (Department of Botany, Faculty of Life Science) and Chief M.B. Odoba (Department of Veterinary Public Health and Preventive Medicine) who gave fatherly advice and support to the work. My heart filled gratitude also goes to the staff in the Bacterial, Parasitic and Viral Zoonoses laboratories especially Mallam Mahmoud and Mallam Yahuza Maitala. I also want to thank the staff of Central laboratory, City campus, Usman Danfodio Univerisity for their technical support during the molecular analyses. I also extend my profound gratitude to the Department of Zoological, Department of Microbiology, Faculty of Life Science and to Prof. Inabo and Dr. Gadzama for their support in this project. I also want to say a very big “thank you” to Prof. J.P. Kwaga, Dr. G.N.S. Kia, Dr. Lawan for their support. Also a big ‘thank you” to Mary Akpan, Basil Akpan and Edo Akpan. To my very interesting and intelligent classmates; 2015/2016 academic session, Department of Veterinary Public Health and Preventive Medicine, it was an exciting journey and there could never be a more exciting people to learn with. I will love to appreciate everyone whose contributions made this iv work a success; space and time is not enough to acknowledge you all by your names. May the Almighty God bless you. v ABSTRACT Bacterial contamination of roasted ready-to-eat rat meat and occurrence of foodborne pathogens may be injurious to human health. In Nigeria, the incidence and multidrug resistance of Salmonella spp. and E. coli O157:H7 in human cases is on the increase and these organisms have been found associated with meat and meat products in several studies. A cross-sectional study was carried out to determine the bacteriological quality and occurrence of Salmonella spp. and Escherichia coli O157:H7 in roasted ready-to-eat rat (Arvicanthis niloticus) meat sold in Zaria, Nigeria. A total of 384 samples were examined from four purposively selected districts in the study area; Samaru, Basawa, Jushi and Sabon Gari. Total aerobic plate count was determined using spread plate technique and bacteria load determined. Isolates were screened for Escherichia coli O157:H7 and Salmonella using the conventional biochemical characterization, standardized micro-substrate (Microgen GN-ID A+B) detection kit for Gram negative bacteria and further confirmed using Rapid latex agglutination test for E. coli O157:H7 and PCR for Salmonella. Agar disc diffusion method was used to evaluate the antibacterial resistance pattern; data obtained were interpreted using clinical laboratory standards institutes (CLSI). Mean log and standard deviation of total aerobic plate counts (TAPC) were 10.01±0.27, 10.03±0.26, 10.10±0.29 and 9.9±0.29, from four sampling areas, Samaru, Sabon Gari, Jushi and Basawa. The average TAPC ranged from 12 x 109 cfu.g-1 in Samaru to 15 x 109 cfu.g-1 in Jushi sampling districts. Salmonella was isolated from 2(0.5%) and E. coli O157:H7 isolated from 5 (1.3%) of the total 384 samples. Multidrug resistance of the Salmonella and E. coli O157:H7 was observed. Of the total Salmonella isolates, 100% were found to be resistant to tetracycline, doxycycline, amoxicillin/clavulanic acid, trimethoprim/sulphamethoxazole and ampicillin, 83.3% resistant to chloramphenicol and cefixime, 67% resistant to gentamicin, 33.3% resistant to azithromycin, 17% resistant to kanamycin and cefuroxime sodium, with no isolate (0%) resistant to ciprofloxacin. Of the total E. vi coli O157:H7 isolates, 100% were found to be resistant to ampicillin and penicillin, 80% resistant to streptomycin, 60% resistant to trimethoprim/sulphamethoxazole and doxycycline, 40% resistant to chloramphenicol and gentamicin, with no isolate (0%) resistant to imipenem. Ninety-six structured questionnaires were used to assess the knowledge and practices of retailers and consumers of roasted ready-to-eat rat meat in the study area. Fifty-one percent of the consumers and 67% of the retailers of roasted ready-to-eat rat meat in Zaria, use bare hands as a means of selection in bargaining. Air drying is the major means of preservation, as documented by 52% of the retailers and 42% of the consumers. The roasted ready-to-eat rat meat is consumed either in soup or with spices with no further reheating. The high bacterial load beyond the permissible level, the occurrence of Salmonella spp. and E. coli O157:H7 and the presence of multidrug resistant strains of Salmonella and E. coli O157:H7 in roasted ready-to-eat rat meat sold in Zaria, are of serious public health significance. Therefore, there is for public health enlightenment of the consumers and retailers of roasted ready-to-eat rat meat sold in Zaria, so as to avoid bacterial contamination of the roasted rat meat. vii TABLE OF CONTENTS TITLES PAGE Title page Declaration------------------------------------------------------------------------------------------------------i Certification----------------------------------------------------------------------------------------------------ii Dedication-----------------------------------------------------------------------------------------------------iii Acknowledgement--------------------------------------------------------------------------------------------iv Abstract--------------------------------------------------------------------------------------------------------vi Table of contents--------------------------------------------------------------------------------------------viii List of Tables------------------------------------------------------------------------------------------------xiii List of Figures------------------------------------------------------------------------------------------------xv List of Plates-------------------------------------------------------------------------------------------------xvi List of Appendices-----------------------------------------------------------------------------------------xvii Abbreviations----------------------------------------------------------------------------------------------xviii CHAPTER ONE: INTRODUCTION - - - - - - - 1 1.1 Background
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