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Prevelance of Peste Des Petits Ruminants Virus In PREVELANCE OF PESTE DES PETITS RUMINANTS VIRUS IN THE BLUE NILE STATE By Raja Eltahir Haj Omer B.V.Sc. (1996) University of Khartoum Supervisor Professor Abdel Rahim El Sayed Karrar co-Supervisor Dr. Yahia Hassan Ali A thesis submitted to the University of Khartoum in partial fulfillment of the requirements for the Degree of Master of Veterinary Medicine (M.V.M) Department of Medicine, Toxicology and Pharmacology Faculty of Veterinary Medicine University of Khartoum 2011 ﺑﺴــﻢ اﷲ اﻟﺮﺣـﻤـﻦ اﻟﺮﺣـﻴﻢ ﻗﺎل ﺗﻌﺎﻟﻰ: (ﺳـﺒﺤـﺎن اﻟﺬي ﺳـﺨﺮ ﻟﻨﺎ هﺬا وﻣﺎ آـﻨﺎ ﻟﻪ ﻣـﻘﺮﻧﲔ) ﺻﺪق اﷲ اﻟﻌﻈﻴﻢ ﺳﻮرة اﻟﺰﺧﺮف - ﺁﻳﺔ (13) LIST OF CONTENTS Page Dedication…………………………………………………………… ix Acknowledgements…………………………………………………. iix English Summary…………………………………………………….. x Arabic summary ……………………………………………………. xi List of Contents……………………………………………………… i List of Tables ……………………………………………………….. vi List of Figures………………………………………………………. vii Introduction…………………………………………………………….. 1 iii Page CHAPTER ONE: LITERATURE REVIEW 1.1. Definition………………………………………………………………. 4 1.2. Etiology………………………………………………………………… 5 1.2.1. Virus structure…………………………………………………………... 5 1.2.2. Relationship between the PPR virus and rinderpest virus………………. 6 1.3. Host Range and species variation………………………………………… 6 1.4. Historical Background……………………………………………………. 7 1.5. Epidemiology of PPR…………………………………………………… 9 1.5.1 Geographical Distribution…………………………………………….. 9 1.5.2. Transmission…………………………………………………………….. 10 1.5.3. Lineages of PPRV……………………………………………………… 11 1.5.4. Morbidity and Mortality……………………………………………… 12 1.6 Pathology………………………………………………………………… 13 1.6.1 Gross lesions…………………………………………………………… 13 1.6.2 Microscopic lesions (histopathology)………………………………… 14 1.7. Clinical Signs…………………………………………………................. 14 1.7.1. Per acute Syndrome………………………………………………… 15 page 1.7.2. Acute Syndrome…………………………………………………….. 15 1.7.3. Sub acute Syndrome………………………………………………… 17 1.8. Resistance and immunity……………………………………………… 17 1.8.1 Innate and passive immunity………………………………………… 17 1.8.2 Active immunity…………………………………………………….. 17 1.8.3 Vaccination………………………………………………………….. 18 1.9. Diagnosis of PPR…………………………………………………….. 19 1.9. 1. Laboratory Diagnosis……………………………………………… 19 1.9.1.1 Serological tests…………………………………………………… 20 1.9.2. Differential Diagnosis………………………………………............. 22 1.9.3. Control of PPR……………………………………………………… 26 CHAPTER TWO: MATERIALS AND METHODS page 2. Material and Methods………………………………………………….. 29 2.1. Area of the Study ……………………………………………………… 29 2.2. Data collection………………………………………………………. 31 2.2.1. Collection of samples……………………………………………… 33 2.2.2. Material for samples collection…………………………………… 33 2.2.3. Sampling Procedure………………………………………………. 34 Page 2.3. Competitive ELISA System for detection of specific antibodies to PPR virus…………………………………….... 35 2.3.1. Materials………………………………………………………….. 35 2.3.2. Reagents………………………………………………………….. 36 2.3.3. Protocol for competitive ELISA………………………................. 37 2.4. Immunocapture ELISA for detection of antigen………………….. 41 2.4.1. Materials………………………………………………………… 41 2.4.2. Protocol of Immunocapture ELISA…………………………….. 41 CHAPTER THREE: RESULT Page 3.1 Epidemiology of PPR in Blue Nile state………………………… 43 3.2. The prevalence of PPR in ruminants in Blue Nile State……......... 44 3.3 The PPR Sero-prevalence in different species……………………. 44 3.4. The PPR sero-prevalence in different locations…………………. 44 3.5 Prevalence of PPR Antigen in the different species……………… 45 CHAPTER FOUR: Page DISCUSSION………………………………………………………… 58 Conclusions and Recommendation ……………………………………. 61 References………………………………………………………………. 63 LIST OF TABLES Page Table (1) The Number of the Serum samples collected from different species of animals at Blue Nile State during(2009-2010)……………….. 32 Table (2) Number of Animals vaccinated against PPR in Blue Nile State during(2009 -2010)………………………………………………………. 45 Table (3) Seroprevalence of PPR in the different localities in Blue Nile state, Sudan detected using cELISA during 2010…………. 46 Table (4) Result of cELISA for the detection of PPR antibodies in the different species in Blue Nile state (2010)………………………… 47 Table (5) Determination of PPR Antibodies using cELISA in the different locations in Blue Nile state, Sudan (2010)………………………………… 48 Table (6) Seroprevalence of PPR determined using cELISA in different species and localities in Blue Nile state during (2010)…......... 49 LIST OF FIGURES Page Figure (1) Geographic distribution of PPRV lineages (Dhar et al., 2002)…………………………………………......................... 10 Figure (2) Area of the Study ………………………………………………. 30 Figure (3) ELISA Plate showing positive and negative serum samples……………………………………. 40 Figure (4) ELISA Plate showing Protocol of Immunocapture ELISA………………………………………………. 42 Figure (5) Seroprevalence of PPR in the different localities in Blue Nile state during (2010)…………………………….. 50 Figure (6) PPR antibodies detected using cELISA in the different species in Blue Nile state (2010)………………………. 51 Figure (7) Seroprevalence of PPR determined using cELISA in the different locations in Blue Nile State, Sudan during (2010)……………………… 52 Figure (8) Prevalence of PPR Antigen in the different species…………………………………………………… 53 Page Figure 9 Erosive stomatitis involving: the inside of the lower lips and adjacent gum ……………………………………………………………….. 54 Figure10:Serous nasal discharge becoming mucopurulent and resulting, at times, in a profuse……………………………………………………………… 55 Figure11:Catarrhal exudate which crusts over and occludes the nostrils……………………………………………………………….. 56 Figure 12:Severe non-haemorrhagic diarrhea, Congestion of conjunctiva, crusting on the medial canthus and sometimes profuse catarrhal conjunctivitis……………………………………………………..... 57 DEDICATION I dedicate this work to: The soul of my father My dearest mother My great husband Ammar My lovely sons Ahmed Aymen Awab My dearest Brothers and Sisters With my best wishes ACKNOWLEDGEMENT First of all my thanks to Allah for giving me health and strength to complete this study. I owe my sincere Gratitude and thanks to my supervisor Professor. Abedel Rahim El Sayed Karrar for his guidance, advice, attention, kindness and unlimited help. My kind regards and thanks to my co- supervisor Dr. Yahia Hassan Ali, Dr. Intisar Kamil Saeed, Department of Virology especially Dr.Mahasin Elnour the Central Veterinary Research Laboratory (CVRL) Soba, for offering me space and facilities for work, and for performing ELISA. My thanks to IAEA Research contract for funding. The help and financial support of Ministry of Animal Resource and Fishers is very much appreciated. I extend my affable thanks to Animal health and Epizootic Disease Control General Directorate for his unreserved help. I would like to express my thanks to the staff members of the Blue Nile research laboratory (Abed Allah, Awad, Alawy) for giving a valuable support in sample collection. My full thanks to my colleague Elzain Basher for his helping in analysis of data. I am grateful to my mother, sisters, brothers, and sons with thankfulness, for their encouragement, patience and greatest help. Lastly I am really falling short of words to express my gratitude to my dearest husband Dr. Ammar Ismaiel for his moral support and blessing me with more than that I asked for. Prevalence of Peste des Petits Ruminants Virus In Blue Nile State A Thesis Submitted in partial fulfillment of the requirements of University of Khartoum for the Degree of Master of Veterinary Medicine Raja Eltahir Haj Omer B.V.Sc. (1996) Summary This study was carried out to investigate the epidemiology of peste des petits ruminants (PPR) in the Blue Nile State, Sudan, during 2009-2010, through collection of epidemiological data and antigen and antibody detection. Two methods were adopted to achieve this aim ; namely, collection of data from Veterinary services records and serological examination of sheep, goats, camels and cattle, using competitive enzyme- linked immunosorbent assay (c-ELISA) for PPR antibody detection and immunocapture ELISA (ic-ELISA) for(PPR antigen detection54. Out of 1037 serum samples collected from sheep , goats, camels and cattle in six Localities and tested for PPR virus antibodies using c-ELISA, 577 (55.64%) were positive. Among these, antibodies against PPR virus were detected in 83 samples(92.22%) from Baw Locality, 84 (63.16%) from Damazin locality, (84.38%) from Geissan Locality, 80(54.42%) from Rosirais Locality, 178 (43.84%) from Tadamon Locality and 98 (49.75%) from slaughterhouse. Throughout animal species, the highest antibodies titter of PPR was found to be in 206 samples(79.54%) of goats followed by sheep272(54.62%),cattle81 (40.50%) and camels 18 (22.50%). Out of 303 lung samples collected from sheep, goats, camels and cattle in slaughterhouse and tested for PPR antigen, using ic-ELISA only six sheep samples were positive. The results showed that PPR is an endemic disease in the Blue Nile State. ﻣﻌﺪل اﻧﺘﺸﺎر ﻓﻴﺮوس ﻣﺮض ﻃﺎﻋﻮن اﻟﻤﺠﺘﺮات اﻟﺼﻐﻴﺮة ﻓﻰ وﻻﻳﺔ اﻟﻨﻴﻞ اﻻزرق رﺳﺎﻟﺔ ﻟﻨﻴﻞ درﺟﺔ اﻟﻤﺎﺟﺴﺘﻴﺮ ﻓﻲ اﻟﻄﺐ اﻟﺒﻴﻄﺮي ﺟﺎﻣﻌﺔ اﻟﺨﺮﻃﻮم ﺑﻮاﺳﻄﺔ رﺟﺎء اﻟﻄﺎهﺮ ﺣﺎج ﻋﻤﺮ ﺑﻜﻼرﻳﻮس اﻟﻌﻠﻮم اﻟﺒﻴﻄﺮﻳﺔ ﺟﺎﻣﻌﺔ اﻟﺨﺮﻃﻮم (1996) اﻟﻤــﺴﺘﺨـﻠـﺺ ﺃﺠﺭﻴﺕ ﻫﺫﻩ ﺍﻟﺩﺭﺍﺴﺔ ﻟﻠﺘﺤﻘﻕ ﻤﻥ ﻭﺒﺎﺌﻴﺔ ﻤﺭﺽ ﻁﺎﻋﻭﻥ ﺍﻟﻤﺠﺘﺭﺍﺕ ﺍﻟﺼﻐﻴﺭﺓ ﻓﻲ ﻭﻻﻴﺔ ﺍﻟﻨﻴل ﺍﻻﺯﺭﻕ ﻓﻲ ﺍﻟﻔﺘﺭﺓ ﻤﻥ 2009- 2011ﻡ ﻤﻥ ﺨﻼل ﺠﻤﻊ ﺒﻴﺎﻨﺎﺕ ﻋﻥ ﻭﺒﺎﺌﻴﺔ ﺍﻟﻤﺭﺽ ﻓﻲ ﺍﻟﻭﻻﻴﺔ ﻭﻤﺩﻯ ﺇﻨﺘﺸﺎﺭ ﺍﻷﺠﺴﺎﻡ ﺍﻟﻤﻀﺎﺩﺓ ﻭﺍﻷﻨﺘﻴﺠﻴﻨﺎﺕ ﻓﻲ ﺍﻟﺤﻴﻭﺍﻨﺎﺕ ﺍﻟﻤﺼﺎﺒﺔ. أﺳﺘﺨﺪﻣﺖ ﻃﺮﻳﻘﺘﺎن ﻹﻧﺠﺎز هﺬﻩ اﻟﺪراﺳﺔ هﻤﺎ: ﺠﻤﻌﺕ ﺒﻴﺎﻨﺎﺕ ﻤﻥ ﺴﺠﻼﺕ ﺍﻟﺨﺩﻤﺎﺕ ﺍﻟﺒﻴﻁﺭﻴﺔ ﻭﻓﺤﺹ ﺴﻴﺭﻭﻟﻭﺠﻲ ﻟﻌﻴﻨﺎﺕ ﻤﻥ ﺃﻨﺴﺠﺔ ﺍﻟﻀﺄﻥ ﻭ ﺍﻟﻤﺎﻋﺯ ﻭﺍﻹﺒل ﻭﺍﻷﺒﻘﺎﺭ ﻤﺴﺘﺨﺩﻤﻴﻥ ﻓﻲ ﺫﻟﻙ ﺇﺨﺘﺒﺎﺭ ﺍﻻﻟﻴﺯﺍ( ﻟﻠﻜﺸﻑ ﻋﻥ ﺍﻻﺠﺴﺎﻡ ﺍﻟﻤﻀﺎﺩﺓ) ﻭﺇﺨﺘﺒﺎﺭ ﺍﻻﻟﻴﺯﺍ ﺍﻟﻤﺘﻁﻭﺭﺓ ﻟﻠﻜﺸﻑ ﻋﻥ ﺍﻷﻨﺘﺠﻴﻨﺎﺕ . ﻓﺤﺼﺕ1037 ﻋﻴﻨﺔ ﺴﻴﺭﻡ ﻤﻥ ﺍﻟﻀﺄﻥ ﻭﺍﻟﻤﺎﻋﺯ
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