Epidemiology of Foot and Mouth Disease in Buffaloes and Cattle Of
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Epidemiology of Foot and Mouth Disease in Buffaloes and Cattle of Punjab By Muhammad Zeeshan Nawaz M.Sc. (Hons.) A thesis submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy In Microbiology INSTITUTE OF MICROBIOLOGY FACULTY OF VETERINARY SCIENCE UNIVERSITY OF AGRICULTURE FAISALABAD, PAKISTAN 2014 0 To My Affectionate Parents I would like to dedicate this manuscript to my Honorable & Respected father Sheikh Muhammad Nawaz, who supported and encouraged me a lot at every moment during my degree program and made possible for me to complete my Doctorate. Special dedication to my mother also who never fail to remember me for a second in her prayers for my progress and success 1 CONTENTS CHAPTER TITLE PAGE ACKNOWLEDGEMENT i LIST OF TABLES ii LIST OF FIGURES iii 1 INTRODUCTION 01 2 REVIEW OF LITERATURE 04 3 MATERIALS AND METHODS 21 4 RESULTS 46 5 DISCUSSION 86 6 SUMMARY 93 7 LITERATURE CITED 95 8 ANNEXTURES 112 2 ACKNOWLEDGEMENTS If oceans turn into ink and all the woods become pens, even then the praise of Allah Almighty cannot be expressed. He, who created the universe and knows whatever, is there in it, hidden or evident and who bestowed upon me the intellectual ability and wisdom to search for its secrets. Countless salutations be upon the Holy Prophet (Salutations and peace of Allah be upon him), the city of knowledge who guided his ‘Ummah” to seek knowledge from cradle to grave. I feel highly privilege to ascribe the most and ever burning flame of gratitude and deep sense of devotion to my reverend supervisor Dr. Muhammad Arshad, Professor Institute of Microbiology, University of Agriculture, Faisalabad, for his keen potential interest, sagacious suggestions, consistent encouragement, constructive criticism, incentive teachings, polite behavior, guardian and sympathetic attitude, step to step guidance and dynamic supervision during the conduct of these investigations and in preparation of this manuscript. It gives me great pleasure in expressing my gratitude with the profound sense of benediction to Prof. Dr. Sajjad ur Rahman, Institute of Microbiology, University of Agriculture, Faisalabad, for his cooperation, scholastic guidance, immense moral support, keen interest, unerring vigor, staunch help, masterly advice and constructive suggestions. I offer my special and sincere thanks to a dignified personality Prof. Dr. Zafar Iqbal, Department of Parasitology, University of Agriculture, Faisalabad, and Prof. Dr. Iftikhar Hussain, Director, Institute of Microbiology, University of Agriculture, Faisalabad, for his valuable suggestions and guidance. Family is truing dense of my humblest obligations and my words will fail to give them credit especially to my Parents, Sisters and Wife whose prayers, love, patience, sacrifice, encouragement and spiritual inspiration was a great source of motivation for the quest of knowledge at every stage of my education. I would not be doing justice without acknowledging the invaluable moral support, excellent cooperation and constant encouragement extended to me by Saif ur Rehman, Bahar e Mustafa, Shahid Ali, Saad Zubair, Umair Hassan, Iftikhar bhai and Intezar Hussain. I fervently extended my zealous, sincerest and warmest thanks to my friends for which I could not mention, for their endless and excellent cooperation and moral support. Finally, I again thank all those whom I have mentioned and whom I could not, for helping me in completing this work, as it is customary, the errors that remain are entirely mine. At the end I gratefully acknowledge the financial support provided by Higher Education Commission (HEC) Pakistan, Indigenous Scholarship Phase VI for the completion of this work along with the Endowment Fund Secretariat (EFS), University of Agriculture, Faisalabad, Pakistan for their support and logistics. Muhammad Zeeshan Nawaz 3 LIST OF TABLES Page No. Title No. Climatic and geographic detail of three districts of Punjab selected for the 3.1 24 seroprevalence of Foot and Mouth Disease Area & species related distribution of samples for seroprevalence of Foot 3.2 28 and Mouth Disease Sex related distribution of samples for seroprevalence of Foot and Mouth 3.3 29 Disease Age related distribution of samples for seroprevalence of Foot and Mouth 3.4 30 Disease Pregnancy status related distribution of samples for seroprevalence of Foot 3.5 31 and Mouth Disease Herd type related distribution of samples for seroprevalence of Foot and 3.6 32 Mouth Disease Distribution of samples for molecular detection of Foot and Mouth Disease 3.7 35 virus Sequences for universal and serotype specific primers of Foot and Mouth 3.8 39 Disease Different experimental groups for Foot and Mouth Disease vaccines 3.9 43 evaluation Comparative seroprevalence of Foot and Mouth Disease in three districts 4.1 47 of Punjab Comparative sex related seroprevalence of Foot and Mouth Disease in 4.2 49 three districts of Punjab 4 Page No. Title No. Comparative age related seroprevalence of Foot and Mouth Disease in 4.3 50 three districts of Punjab Comparative pregnancy related seroprevalence of Foot and Mouth Disease 4.4 51 in three districts of Punjab Comparative herd type related seroprevalence of Foot and Mouth Disease 4.5 53 in three districts of Punjab Molecular detection of Foot and Mouth Disease virus by Reverse 4.6 57 Transcriptase –Polymerase Chain Reaction Serotyping of Foot and Mouth Disease virus by Indirect Sandwich-Enzyme 4.7 62 Linked Immunosorbent Assay Serotyping of Foot and Mouth Disease virus by Reverse Transcriptase – 4.8 63 Polymerase Chain Reaction Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.9 68 virus, Serotype ‘O’ in buffaloes and cattle at 2 weeks Post Vaccination Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.10 69 virus, Serotype ‘O’ in buffaloes and cattle at 4 weeks Post Vaccination Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.11 70 virus, Serotype ‘O’ in buffaloes and cattle at 6 weeks Post Vaccination Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.12 71 virus, Serotype ‘O’ in buffaloes and cattle at 8 weeks Post Vaccination Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.13 72 virus, Serotype ‘O’ in buffaloes and cattle at 10 weeks Post Vaccination Comparative Geometric Mean Titers and Cumulative Mean Titers of 4.14 73 different vaccines against serotype ‘O’ of Foot and Mouth Disease virus 5 Page No. Title No. Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.15 74 virus, Serotype ‘A’ in buffaloes and cattle at 2 weeks Post Vaccination Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.16 75 virus, Serotype ‘A’ in buffaloes and cattle at 4 weeks Post Vaccination 4.17 Indirect Hemagglutination antibody titers against Foot and Mouth Disease 76 virus, Serotype ‘A’ in buffaloes and cattle at 6 weeks Post Vaccination Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.18 77 virus, Serotype ‘A’ in buffaloes and cattle at 8 weeks Post Vaccination Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.19 78 virus, Serotype ‘A’ in buffaloes and cattle at 10 weeks Post Vaccination Comparative Geometric Mean Titers and Cumulative Mean Titers of 4.20 79 different vaccines against serotype ‘A’ of Foot and Mouth Disease virus Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.21 80 virus, Serotype ‘Asia I’ in buffaloes and cattle at 2 weeks Post Vaccination Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.22 81 virus, Serotype ‘Asia I’ in buffaloes and cattle at 4 weeks Post Vaccination Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.23 82 virus, Serotype ‘Asia I’ in buffaloes and cattle at 6 weeks Post Vaccination Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.24 83 virus, Serotype ‘Asia I’ in buffaloes and cattle at 8 weeks Post Vaccination Indirect Hemagglutination antibody titers against Foot and Mouth Disease 4.25 virus, Serotype ‘Asia I’ in buffaloes and cattle at 10 weeks Post 84 Vaccination Comparative Geometric Mean Titers and Cumulative Mean Titers of 4.26 different vaccines against serotype ‘Asia I’ of Foot and Mouth Disease 85 virus 6 LIST OF FIGURES No. Title Page No. Map of Punjab, showing three district selected for 3.1 23 seroprevalence of Foot and Mouth Disease Detection of seroprevalence of Foot and Mouth Disease with 4.1 54 Non Structural Protein- Enzyme Linked Immunosorbent Assay 4.2 Symptoms of Foot and Mouth Disease infected cattle 56 4.3 Clinical signs of Foot and Mouth Disease infected cattle 56 DNA amplified product (328 bp) of Foot and Mouth Disease 4.4 58 virus using universal primers DNA amplified product (1301 bp) of Foot and Mouth Disease 4.5 59 virus using serotype specific primers DNA amplified product (923 bp) of Foot and Mouth Disease 4.6 60 virus using serotype specific primers Detection of serotypes of Foot and Mouth Disease with Indirect 4.7 64 Sandwich Enzyme Linked Immunosorbent Assay Detection of antibody titers of Foot and Mouth Disease vaccines 4.8 67 with Indirect Haemagglutination test 7 CHAPTER – 1 INTRODUCTION Pakistan is an agricultural country and livestock have a great importance in its economy. Every 5th family in Pakistan is engaged directly or indirectly in livestock farming. It accounts for about 11.9% of total GDP and 55.4 % of agricultural value added. Total population of buffaloes and cattle in Pakistan is 33.7 million and 38.3 million respectively among which the major part (>61% of buffalo and >43% of cattle population) is residing in Punjab. Livestock is a major source of milk, meat, wool, hair, bones, fats, blood, hides and skins. Total 39945 thousand tons milk production and 1829 thousand tons beef production was recorded during the year 2012-2013 (Economic Survey of Pakistan, 2013).