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Marek’s disease Dr Venugopal Nair Institute for Animal Health, Compton, Berkshire, RG20 7NN, UK Tel.: (44 1635) 578411, Fax: (44 1635) 577263) [email protected] http://www.iah.ac.uk/ Summary of general activities related to the disease 1. Test(s) in use/or available for the specified disease at your laboratory Following tests have been developed and used in connection with the research programmes, although occasionally these tests are used for diagnostic purposes of the disease in the field. 1. 2. 3. 4. 5. 6. 7. 2. Serotyping of MDV 1, 2 & 3 isolates Agar gel precipitation tests for antigen and antibody PCR tests for detection of MDV as well as for quantitation of virus load In situ hybridisation tests for viral nucleic acid Flow cytometry to identify the phenotype of infected cells Fluorescent/confocal microscopy to detect virus/viral antigens ELISA tests to detect antibodies. Production and distribution of diagnostic reagents The diagnostic reagents are produced mostly for research purposes, but are usually available on request for diagnostic purposes. Activities specifically related to the mandate of OIE Reference Laboratories 3. International harmonisation and standardisation of methods for diagnostic testing or the production and testing of vaccines Standard reagents are not prepared since written standards for vaccine productions exist. Contributed chapters on various aspects of Marek’s disease including diagnostic testing and vaccine production to several text books (see list of publications). 4. Preparation and supply of international reference standards for diagnostic tests or vaccines Field Samples including blood and feather samples were sent from different countries for the detection of Marek’s disease virus either by PCR or by quantitative PCR for the accurate measurement of MDV genome copy numbers. 5. Research and development of new procedures for diagnosis and control The main focus of research of the group at the Institute for Animal Health, Compton is to understand the molecular mechanisms of MD pathogenesis with an aim to develop more effective diagnostic tests and vaccines. Quantitative PCR tests for the accurate measurement of MDV genome copy numbers in the feather samples developed by the group has been applied to field samples to measure the efficacy of the vaccination and also to monitor the correlation between virus load and the protective index. Annual reports of OIE Reference Laboratories and Collaborating Centres, 2007 1 Marek’s disease 6. Collection, analysis and dissemination of epizootiological data relevant to international disease control Field samples sent for diagnosis from different parts of the UK and abroad were tested for MDV diagnosis. Dr. Venugopal Nair visited different parts of India to hold discussions with scientists and different animal health professionals for ways of improving the control of the disease. 7. Provision of consultant expertise to OIE or to OIE Member Countries Continued to provide technical advice on different aspects of Marek’s disease control and diagnosis to personnel from several countries including the UK, India, Nepal, Poland and France. 8. Provision of scientific and technical training to personnel from other OIE Member Countries Presented the lecture on Marek’s disease to the participants of the Poultry Health Course held at the Institute for Animal Health during 26 FEBRUARY – 8 MARCH 2007. The meeting was attended by participants from many countries. Ms. Katarzyna Krol, Research Scientist from National Veterinary Research Institute, Pulawy, Poland joined the group for one-month training with funds from the British Council. Ms. Sabina Malik, University of Reading joined on the 19 th June 2007 as part of the summer studentship (10 weeks) to work on Molecular diagnosis of Marek’s disease. Melissa Singh, MSc student from started the 6-week research project under the supervision of Dr. Baigent as part of the Control of Infectious Diseases in Animals programme run by the Royal Veterinary College, London 9. Provision of diagnostic testing facilities to other OIE Member Countries Samples received from UK were tested for MDV by PCR, quantitative PCR or by the method of virus isolation in cell culture. There were restrictions on importing materials from other countries due avian influenza 10. Organisation of international scientific meetings on behalf of OIE or other international bodies Several Members of the group participated in the XV Congress of the World Veterinary Poultry Association September 10-15, 2007 Beijing, China 11. Participation in international scientific collaborative studies Collaboration in research on MD was continued with various leading laboratories in Europe, USA, India, Japan, China and Australia. As part of the collaboration, Dr. Venugopal Nair visited India to discuss about further collaborations on the control of Marek’s disease virus. A research programme on UK-China Collaborative Research Platform on Avian Oncogenic Virus diseases was initiated in April 2007 jointly with Prof. Zhizhong Cui, Shandong Agricultural University, Taian, China. The first meeting of members of the UK group and Prof. Zhizhong Cui’s group (Shandong University) to start the collaborative projects under the China Partnering Award was held during September 10-15, 2007 in Beijing. Dr. Steve Walkden-Brown, University of New England, Armidale, New South Wales, Australia joined the group on sabbatical as part of the Underwood Fellowship. 2 Annual reports of OIE Reference Laboratories and Collaborating Centres, 2007 Marek’s disease 12. Publication and dissemination of information relevant to the work of OIE (including list of scientific publications, internet publishing activities, presentations at international conferences) Presentations at international conferences and meetings Dr. Baigent gave a talk on ‘Marek’s disease: A virus evolving to evade vaccines’ as part of the MSc in Veterinary Epidemiology and control of infectious diseases in animals at the Royal Veterinary College on 19-1-07 Dr. Nair gave a talk on ‘Marek’s disease oncogenicity: Molecular insights’ to Oxford University Pharmacology MSc students at Compton on 9 February 2007. As part of this, the students will be offered short term research projects during this summer as part of their MSc course. Dr. Nair invited to give a talk on ‘Marek’s disease – A natural disease model for virus-induced lymphoma’ at the ESF-Wellcome Trust Conference on Animal Biotechnology and its Applications to Animal and Human Health at Sanger Centre, Hinxton on 14-16 June 2007. Dr. Yongxiu Yao gave a talk on the Identification of microRNAs of avian herpesviruses at the XV Congress of the World Veterinary Poultry Association on September 10-15, 2007 Beijing, China. Dr. Hongtao Xu gave a talk on the Analysis of MicroRNA Expression Profiles of RB-1B and CVI988/Rispens in vitro by Real-time PCR at the XV Congress of the World Veterinary Poultry Association on September 10-15, 2007 Beijing, China. Dr. Yuguang Zhao gave a talk on the Towards Developing Better Vaccines for Marek’s Diseases Virus by Genome Manipulation of Infectious Bacterial Artificial Chromosomes (iBAC) at the XV Congress of the World Veterinary Poultry Association on September 10-15, 2007 Beijing, China. Scientific publications in peer-reviewed journals 1. Stephen J. Spatz, Yuguang Zhao, Lawrence Petherbridge, Smith, L. P, Susan J. Baigent and Venugopal Nair (2007) Comparative sequence analysis of a highly oncogenic but horizontal spread-defective clone of Marek's disease virus. Virus Genes, 35(3): 753-66. 2. Keith W. Jarosinski, Neil G. Margulis, Jeremy P. Kamil, Stephen J. Spatz, Venugopal K. Nair and Nikolaus Osterrieder (2007) Horizontal transmission of Marek’s disease virus requires US2, the UL13 protein kinase, and gC. Journal of Virology, 81(19): 10575-87 3. Susan J. Baigent, Lorraine P. Smith, Richard J. W. Currie, Venugopal K. Nair (2007) Correlation of MDV vaccine virus genome load in feather tips with protection, using an experimental challenge model. Avian Pathology, 36, 467-474. 4. Yao, Y., Zhao, Y., Xu, H., Smith, L. P., Lawrie, C. H., Sewer, A., Zavolan, M. & Nair, V. (2007) Marek’s disease virus type 2 (MDV-2)-encoded microRNAs show no sequence conservation to those encoded by MDV-1. Journal of Virology, 81, 7164-7170. 5. Stephen J. Spatz, Lawrence Petherbridge, Yuguang Zhao and Venugopal Nair (2007) Comparative full-length sequence analysis of oncogenic and vaccine (Rispens) strains of Marek’s disease virus Journal of General Virology, 88, 1080–1096. 6. Katarzyna Król, Sue Baigent, Venugopal Nair, Wojciech Kozdruń & Elżbieta Samorek–Salamonowicz (2007) Comparison of Standard Polymerase Chain Reaction (PCR) with Real-Time PCR for Meq gene of Marek’s Disease Virus. Bull Vet Inst Puławy, 51, 3-7. 7. Butter C, Staines K, Baaten B, Smith L, Davison TF (2007) Route of challenge is critical in determining the clinical outcome of infection with a very virulent oncogenic herpesvirus, Marek's disease virus. Avian Pathology, 36, 93-9. _______________ Annual reports of OIE Reference Laboratories and Collaborating Centres, 2007 3