Activities in 2009 Marek’s disease Venugopal Nair Institute for Animal Health, Compton, Berkshire, RG20 7NN, UK Tel.: (44-1635) 57.84.11, Fax: (44-1635) 57.72.63 E-mail:venu.gopal@bbsrc.ac.uk website: http://www.iah.ac.uk/ Summary of general activities related to the disease The Reference Laboratory continued to provide technical guidance, reagents and training to personnel from different countries on diagnosis and control of Marek’s 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. 8. 9. 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 Quantitation of vaccine virus load in samples including the dust 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. Expression analysis viral transcripts and microRNAs 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. 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 research focus 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 Annual reports of OIE Reference Laboratories and Collaborating Centres, 2009 1 Marek’s disease 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. 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. Team from IAH visited different parts of China to participate in avian tumor diseases and 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 Members Continued to provide technical advice on different aspects of Marek’s disease control and diagnosis to personnel from several countries including the UK, India, China, Poland and France. 8. Provision of scientific and technical training to personnel from other OIE Members Following personnel visited the laboratory: 1. 2. 3. 9. Dr. Anand Deshpande from Venketeswara hatcheries, Pune India visited the lab for two week training from 21st Sep 2009 Dr. N Bulbule from Venketeshwara Hatcheries visited the lab for training during 18 th -30th October 2009 Visit by Pro. K. Kumanan and Dr. A. Raja from Tamilnadu Univeristy of Veterinary & Animal Sciences (TANUVAS), India for training as part of the India Partnership Award from Oct 18 till Nov 3rd 2009 Provision of diagnostic testing facilities to other OIE Members 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 V Nair and Prof. Zhizhong Cui jointly organised the ‘UK-China Symposium on Tumor Pathology and Tumor viruses in chickens’ on 7th-8th July at Shandong Agricultural University, Taian. Following presentations were given by staff members of the OIE Reference Centre: 1. V. Nair- Avian oncogenic viruses: Major pathogens that help to unravel molecular pathways to cancer 2. Yongxiu Yao- Identification and expression of microRNAs of avian herpesviruses 3. Hongtao Xu- Role of Meq in upregulating MDV-encoded microRNAs to influence caspase-6 expression 4. Yuguang Zhao- Towards understanding Marek’s disease virus oncogenic pathways 11. Participation in international scientific collaborative studies Collaboration in research on MD was continued with various leading laboratories in Europe, USA, India, China and Australia. 12. Publication and dissemination of information relevant to the work of OIE (including list of scientific publications, internet publishing activities, presentations at international conferences) 1. 2 Presentations at international conferences and meetings V Nair, presented a paper on Marek’s disease vaccines and Pathogen diversity on 13 Nov at Oxford as part of the Vaccinology course organised by the Oxford University and Jenner Institute. Annual reports of OIE Reference Laboratories and Collaborating Centres, 2009 Marek’s disease 2. 3. 4. 5. 6. 7. 8. 9. V. Nair invited to give the keynote talk Recent advances in Marek’s disease research at AFFSA Paris as part of the Epizone meeting on 18-19th November 2009. V. Nair invited to talk at the Department of Zoology, University of Oxford, on ‘Marek’s disease: a model for virus evolution, vaccines, microRNAs and cancer’ on 19 October 2009. V. Nair invited to talk at the ‘MicroRNAs in Infection & Immunity’ meeting organised by the University of Edinburgh Sue Baigent gave a talk on “Marek’s disease: an oncogenic herpesvirus disease of chickens” to the 2nd year BSc Students from Leicester University Monday 20th April 2009 at IAH, Pirbright Laboratory. Dr. V. Nair talked to the participants on the Poultry Health Course on 27 th Feb 2009 V Nair invited to talk at the RNAi2009 conference at St. Anne’s College, Oxford on March 18th, 2009 V Nair gave invited talks on various aspects of Marek’s disease at following Institutions in China: 1. Shandong Academy of Agricultural Sciences, Jinan, Shandong (5th July 2009). 2. Shandong Agricultural University, Taian, Shandong, China (7th July 2009). 3. College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, China (14th July 2009). 4. Harbin Veterinary Research Institute, CAAS, Harbin, Heilongjiang (16th July 2009). 5. School of Veterinary Medicine, China Agricultural University, Beijing, China (17th July 2009) Dr. Sue Baigent gave a talk on the Advances in Marek’s disease at RVC16th January 2009, as part of the MSc-Control in Infectious Diseases course. Scientific publications in peer-reviewed journals 1. Tahiri-Alaoui, A., Matsuda, D., Panagoitis, P., Burman, L., Lambeth, L., Petherbridge, L., James, W., Mauro, V. & Nair, V (2009). The 5′ leader of the mRNA encoding the MDV-1 pp14 protein contains intronic IRES with allosteric properties. Journal of Virology 83:12769–12778). 2. Brown, A. C., Smith, L-P., Kgosana, L., Baigent, S. J., Nair, V. and Allday, M. J. (2009). Homodimerization of Meq viral oncoprotein is necessary for the induction of T cell lymphoma by Marek’s disease virus Journal of Virology 83:11142-11151. 3. Yongxiu Yao, Yuguang Zhao, Lorraine P Smith Michael Watson & Venugopal Nair (2009) Novel microRNAs encoded by herpesvirus of turkeys: Evidence of microRNA duplication within the cluster Journal of Virology 83:6969-6973. 4. Yuguang Zhao, Dominic Kurian, Hongtao Xu, Lawrence Petherbridge, Lorraine P Smith, Lawrence Hunt and Venugopal Nair (2009) Interaction of Marek's disease virus oncoprotein Meq with heat shock protein 70 in lymphoid tumour cells. Journal of General Virology 90:2201-2208. 5. Tahiri-Alaoui, A., Smith, L. P., Baigent, S., Kgosana, L., Lambeth, L., Petherbridge, L., James, W. and Nair, V (2009). Identification of intercistronic IRES in Marek’s disease virus immediate early gene Journal of Virology 83:5846-5853. 6. Yongxiu Yao, Lorraine P Smith, Charles H Lawrie, Nigel J Saunders, Michael Watson and Venugopal Nair (2009). Differential expression of microRNAs in Marek's disease virus-transformed T-lymphoma cell lines. Journal of General Virology 90:1551-1559. 7. Crabb JR, Walkden-Brown SW, Baigent SJ, Smith LP, Nair VK (2009) Rate of natural transmission of Marek's disease virus (MDV) to susceptible chickens in the post-challenge period: effect of vaccination with HVT and association with MDV genome count in air. Proc. Aust. Poult. Sci. Symp. 20, 184-187. 8. Luke S Lambeth, Yongxiu Yao, Lorraine P Smith, Yuguang Zhao and Venugopal Nair (2009). MicroRNAs 221 and 222 target the p27Kip1 in Marek’s disease virus-transformed tumour cell line MSB-1 Journal of General Virology, 90:1164-71. 9. Lawrence Petherbridge, Hongtao Xu, Yuguang Zhao, Lorraine P Smith, Jennifer Simpson, Susan Baigent and Venugopal Nair (2009). Cloning of Gallid herpesvirus 3 (Marek’s disease virus serotype-2) genome as infectious bacterial artificial chromosomes for analysis of viral gene functions. Journal of Virological Methods 158 (1-2):11-7. 10. Yuguang Zhao, Yongxiu Yao, Hongtao Xu, Luke Lambeth, Lorraine P Smith, Lydia Kgosana, Xiaowei Wang and Venugopal Nair (2009) A functional microRNA-155 ortholog encoded by the oncogenic Marek’s disease virus. Journal of Virology 83. 489-92. 11. Dr. Sue Baigent was awarded Dr. Bart Rispens Research Award at the WVPA at the XVI WVPA Congress, Marrakesh, Morocco, for the paper "Correlation of Marek's disease herpesvirus vaccine virus genome load in feather tips with protection, using an experimental challenge model" in AVP 36(6), 467-474. Other communications Sue Baigent (2009) Optimized Quantitation of Marek’s Disease Virus using Multiplex QPCR. INsights issue 23, 16-17. www.thermo.com/gen Annual reports of OIE Reference Laboratories and Collaborating Centres, 2009 3 Marek’s disease 13. Inscription of diagnostic kits on the OIE Register i) Did you participate in expert panels for the validation of candidate kits for inscription on the OIE Register? If yes, for which kits? No ii) Did you submit to the OIE candidate kits for inscription on the OIE Register? If yes, for which kits? Not applicable. _______________ 4 Annual reports of OIE Reference Laboratories and Collaborating Centres, 2009