OIE Reference Laboratory Reports

OIE Reference Laboratory Reports Activities in 2010Name of disease (or topic) for Koi herpesvirus disease which you are a designated OIE Reference Laboratory:Address of laboratory: Centre for Environment, Fisheries and Aquaculture Science (Cefas) Weymouth laboratory, The Nothe, Weymouth DT4 8UB, UNITED KINGDOMTel.: (+44-1305) 20.66.39Fax: (+44-1305) 20.66.01 e-mail address: [email protected] website: www.cefas.co.ukName of Head of Laboratory Stephen Irving (Responsible Official):Name of OIE Reference Expert: Keith WayName of writer of this report (if different from above):Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010 1 Koi herpesvirus disease Part I: Summary of general activities related to the disease1. Test(s) in use/or available for the specified disease/topic at your laboratoryTest For Specificity Total + (Number positive for KHV)PCR-KHV Viral DNA detection KHV 58 (15)PCR-HPV Viral DNA detection Cyprinid herpesviruses 50 (14)CCB cell culture at 20°C Virus isolation (Diagnosis only) Broad 16* (7)* - virus isolation on CCB cells only carried out from July to October, inclusive.2. Production and distribution of diagnostic reagents The laboratory produced : details of PCR primer sequences and protocols for the detection and confirmation of KHV.  reference cyprinid herpesvirus-1 (CyHV-1, carp pox) and CyHV-3 (KHV) isolates.  CyHV-1 and CyHV-3 (KHV) DNA.  Carp serum containing antibodies to KHV and antibody negative carp serum.Diagnostic reagents were supplied for internal use but there were no requests in 2009 from national (UK) laboratories to supply reagents. Requests were received from the following OIE member countries for supply of diagnostic reagents:Growing cultures of KHV were supplied to the Marine Institute, Galway, Republic of Ireland and the National Food and Veterinary Risk Assessment Institute, Vilnius, Lithuania. Also, a growing culture of CyHV-1 (carp pox) was supplied to MicroTechnologies, Richmond, Maine, USAKHV-infected gill tissue preserved in alcohol was supplied to Marine Institute, Galway, Republic of Ireland.Inactivated freeze-dried preparation of KHV (for PCR positive control) was supplied to National Aquatic Resources Agency, Colombo, Sri Lanka. KHV PCR protocols were provided to the Institute of Veterinary Research, Tunis, Tunisia, Bornova Veterinary Institute, Izmir, Turkey and the National Food and Veterinary Risk Assessment Institute, Vilnius, Lithuania. A KHV PCR protocol with generic CyHV primers was provided to the National Veterinary Institute, Arhus, Denmark.Part II: Activities specifically related to the mandate of OIE Reference Laboratories3. International harmonisation and standardisation of methods for diagnostic testing or the production and testing of vaccinesIn 2010 the designated expert contributed to a report on ‘KHV diagnosis and surveillance’ produced by the participants at the EPIZONE KHV workshop held at the Central Veterinary Institute (CVI) of Wageningen UR, Lelystad, The Netherlands in November 2009.2 Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010 Koi herpesvirus disease In 2010 the Cefas Weymouth laboratory participated in an EPIZONE KHV serology ring trial – a panel of carp sera was tested by seroneutralisation, ELISA and IFAT, the participants included laboratories in France, Denmark, Germany, The Netherlands, Italy, Poland, Sweden, UK (Cefas) and China.The Cefas laboratory also participated in the Annual Inter Laboratory Proficiency Test, distributed to over 30 laboratories, by the EU Reference laboratory for fish diseases in Arhus, Denmark. The proficiency test consisted of 10 ampoules containing lyophilised, virus-infected, cell culture supernatant. In 2010, for the first time, two of the ampoules contained KHV.The Cefas Weymouth laboratory began development of a KHV PCR Proficiency test in collaboration with the QA unit (VETQAS) at the Veterinary Laboratories Agency in Sutton Bonington, UK. Over 75 international laboratories including those that participated in the 2006-2008 KHV PCR ring-trials, organised by Cefas, have been invited to join the scheme. Five European laboratories, in England, France, The Netherlands, Denmark and the Republic of Ireland, have participated in a successful pilot proficiency test. All five laboratories achieved clean, correct results and encountered no problems when submitting the results on-line.4. Preparation and supply of international reference standards for diagnostic tests or vaccinesNo internationally recognised standard reference materials were supplied to other OIE Member Countries.5. Research and development of new procedures for diagnosis and control The following research is being carried out as part of Department of Environment, Food & Rural Affairs (Defra) funded projects at Cefas:1] To determine the conditions for koi herpesvirus (KHV) survival and persistence in the aquatic environment. In an extended monitoring study, carp populations that had suffered KHV outbreaks in 2006 have been shown to maintain a high prevalence of KHV-antibody positive fish for at least four years. Virus transmission was conducted successfully with carp obtained from one of these antibody-positive carp populations. Further transmission studies to assess the potential for disease spread via angling equipment have shown that, during a disease outbreak, KHV can be transmitted on contaminated keep-nets. Further studies began in 2010 to assess the risk of KHV transmission via fomites and vectors.2] To determine prevalence of KHV in susceptible species and identify KHV-vector species. Information is being collected on KHV-susceptible and vector species. Studies will also seek to confirm the susceptibility of goldfish or crucian carp x common carp hybrids to KHV. 3] To improve tests to provide accurate identification of KHV and to detect latent KHV and improve surveillance tools. Studies are continuing on variant KHV and other putative cyprinid herpesviruses to determine their impact on the reliability of detection methods and surveillance tools for KHV (see also section 6). In vivo and in vitro studies are also continuing on virus latency and reactivation. Studies of KHV latency are being enhanced by collaboration on a post-graduate project at the Marine Institute, Galway, Republic of Ireland. Results from this project will be used to improve methods for detecting latent virus. 4] Studies on the molecular epidemiology of KHVD. Tools have been developed to screen regions of polymorphism in the KHV genome. These have been used to identify an alternative marker that discriminates between vaccine and wt-KHV infections and also to identify different strains of KHV. Currently, the use of KHV genome length polymorphisms, as a tool in epidemiological investigations, is being assessed.6. Collection, analysis and dissemination of epizootiological data relevant to international disease controlA previously identified new strain (variant) of KHV was detected at a further 3 fishery sites in the UK in 2010 by PCR. This variant was first detected in the UK in 2006 with further detections, at a handful of fishery sites, inAnnual reports of OIE Reference Laboratories and Collaborating Centres, 2010 3 Koi herpesvirus disease 2007 and 2008. The variant shares 96% nucleotide identity with wild-type KHV (CyHV-3) and was detected using the generic Cyprinid herpesvirus (CyHV) DNA polymerase primer set developed at Cefas.A previously identified CyHV, that appears to be genetically distinct from CyHV-1,-2 and -3, was again detected, by PCR, at a fishery site in the UK in 2010. This putative CyHV-4 was also detected at the same site in 2008 and 2009.A real-time quantitative PCR assay (qPCR) was developed for detection of the KHV variant that shares 96% similarity with wt-KHV. Initial results with the qPCR indicate virus genome copy numbers in some tissue samples is comparable to the copy numbers of wt-KHV observed in tissues of diseased carp. Any development of sensitive assays to detect KHV in carp tissue will need to be aware of the possible presence of variants and/or other cyprinid herpesviruses. The presence of potential cross-reacting viruses could compromise the specificity of molecular- based assaysA first occurrence of KHVD at two sites in Romania was notified to the OIE in August 2010. No other member countries reported KHVD outbreaks to the OIE in 2010. The WAHID interface ( OIE World Animal Health Information Database) lists in 2010, suspected KHVD in wild carp in Canada and domestic carp in Sri Lanka and clinical KHVD demonstrated in Singapore and Malaysia. However, this information may have been included in 6 monthly or annual reports and relate to disease occurrence in 2009.Large mortalities of common carp (Cyprinus carpio carpio), suspected to be caused by KHV, were reported on ProMed Mail to have occurred in the USA, at lakes in California and New Jersey in May and a lake in Nebraska in August.Publications of disease occurrence include the first clinically apparent koi herpesvirus infection in the Czech Republic [Novotny L., Pokorova, D., Reschova, S., Vicenova, M. and 3 others (2010), Bull.Eur.Ass.Fish Pathol. 30(3), 2010, pp 85-91], a mass mortality associated with KHV in wild common carp in Canada [Garver K., Al- Hussinee L., Hawley L., Schroeder T., Edes S. and 8 others (2010), J Wildlife Diseases 46(4), pp 1242-1251] and KHV-associated mortalities in quarantined koi carp in the Philippines [Somga J., de la Pena L., Sombito C., Paner M. and 4 others (2010), Bull.Eur.Ass.Fish Pathol. 30(1), pp 2-7]. There were further published reports that confirmed Goldfish (Carassius auratus) are a vector of KHV [Bergmann SM, Lutze P., Schutze H, Fischer U. and 3 others (2010) Goldfish (Carassius auratus) is a susceptible species for koi herpesvirus (KHV) but not for KHV disease. Bull.Eur.Ass.Fish Pathol 30 (2) 74-84]. Also, a study in Germany demonstrated transmission of KHV from infected goldfish to common carp [El-Matbouli M. & Soliman H. (2010) Transmission of Cyprinid herpesvirus-3 (CyHV-3) from goldfish to naïve common carp by cohabitation. Research in Veterinary Science doi:10.1016/j.rvsc.2010.07.008].Elsewhere, a small study by the Department of Aquaculture at the Agricultural University in Szczecin, Poland reported the detection of KHV by PCR in four swan mussels (Anodonta cygnea) and one pool of five scud (Freshwater shrimp, Gammarus pulex) collected from ponds on a carp farm in Southern Poland. The farm had suffered outbreaks of KHVD between 2002 and 2007 but not in 2008. Samples were collected in November of 2007 and in August 2008 [Kielpinski M., Kempter J., Panicz R., Sadowski J., and 3 others (2010), Israeli Journal of Aquaculture (Bamidgeh) 62 (1): 28-37]. This report does not demonstrate the presence of viable virus but does imply that some invertebrate species are potential vectors of KHV.All data on KHV occurrence published in 2010 by OIE or in the scientific literature, was entered into the International Database on Aquatic Animal Diseases to add to the historical data. Access to the database is available on-line to OIE member countries via the Aquatic Animals Commission pages on the OIE web site, and the web site of the OIE Collaborating Centre for Information on Aquatic Animal Diseases at www.collabcen.net.7. Provision of consultant expertise to OIE or to OIE MembersAdvice on KHV serology provided to the National Fish Health (NAFish) Management Centre and the National Fish Health Research Centre, both in Pulau Penang, Malaysia, to the Bavarian Landesamt for Health and Food Safety, Erlangen, Germany, to the University of Veterinary Medicine, Vienna, Austria and the Lianyungang Entry-Exit Inspection and Quarantine Bureau, Lianyungang, P.R.China Advice on KHV PCR methods provided to the University of Veterinary Medicine, Vienna, Austria and advice on KHV diagnostics training to the Tai Lung Veterinary Laboratory, Hong Kong 4 Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010 Koi herpesvirus disease Information on KHV purification and on diagnostic test licensing provided to the Frederich Loeffler Institute, Griefswald, Germany.Information/advice on KHV test kits provided to Labofarm, Loudeac, France.Information on KHV diagnostic proficiency testing provided to the State Veterinary & Food Institute, Dolny Kubin, Slovakia and to the Central fish health laboratory, Ministry of Agriculture, Nir David, IsraelInformation on KHV detection in England and Wales in 2010, advice on KHV genotyping and strain similarities and information on KHV vectors provided to the Ornamental Aquatic Trade Organisation (OATA), Westbury, UK. Information on PCR and antibody screening for KHV provided to the AISA Quality and Health Management Programme, Stellenbosch, South Africa and to the Marine Institute, Galway, Republic of Ireland.Advice on CyHV detection in Mahseer carp provided to the National Fish Health Research Centre, Fisheries Research Institute, Pulau Penang, Malaysia Advice/information on KHV health status categories provided to the Veterinary Services, Kibbutz Maagan Michael, Israel and on Category 2 surveillance to the Bavarian Fish Health Service, Weiden, Germany.Advice on stress-testing for detection of KHV provided to the Marine Institute, Galway, Republic of Ireland. Information on source of CyHV-1 (Carp pox) isolates provided to the School of Veterinary Medicine, University of California, Davis, USA. Advice on KHV infection and replication in cell lines provided to the School of Life Sciences, Keele University, UK. The designated expert attended the Second Global Conference of OIE Reference Laboratories & Collaborating Centres in Paris, 21-23 June 2010 and responded to the OIE questionnaire survey on the transport of infectious animal substances by air.8. Provision of scientific and technical training to personnel from other OIE MembersNo requests were received.9. Provision of diagnostic testing facilities to other OIE MembersNo requests were received.10. Organisation of international scientific meetings on behalf of OIE or other international bodiesNo requests were received from OIE or other bodies to organise scientific meetings. 11. Participation in international scientific collaborative studiesStudies on KHV latency (PhD project) with Marine Institute, Galway, Republic of Ireland (on-going).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 meetingsOral presentations (x4) at the 8th International Symposium of Viruses of Lower Vertebrates, Santiago de Compostela, Spain, April 25-29, 2010. Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010 5 Koi herpesvirus disease K Way, C L Joiner, G Wood, K R Jeffery & N G H Taylor. Studies of antibody response and virus persistence in carp populations that have survived KHV outbreaks. P.F. Dixon, C.L. Joiner, K. Way, R.A. Reese, G. Jeney & Z. Jeney. Comparison of the resistance of selected families of common carp Cyprinus carpio (L.), to koi herpesvirus: Proteomic study. O Donohoe, K Henshilwood, K Way, D Stone & D Walls. Investigation of latency associated with koi herpesvirus infection in carp, (Cyprinus carpio). D M Stone, M C Alonso-Sanchez, P D Martin & K Way. Micro and mini satellite repeat sequences in the koi herpesvirus (KHV) genome: A potential epidemiological tool.  Scientific publications in peer-reviewed journalsN.G.H. Taylor, K. Way, P.F. Dixon, E. J. Peeler, K. Jeffrey and K.L.Denham (2010). Koi Herpesvirus (KHV): distribution and prospects for control in England and Wales Journal of Fish Diseases 33 (3) 221-230 N.G.H. Taylor, K. Way, K.R. Jeffery & E.J. Peeler. The role of live fish movements in spreading koi herpesvirus throughout England and Wales. Published online in Journal of Fish Diseases [doi:10.1111/j.1365- 2761.2010.01198.x]J. Ødegård, I. Olesen , P.Dixon, Z. Jeney , H-M Nielsen, K. Way, C Joiner, G. Jeney, L. Ardó, A. Rónyai, B. Gjerde (2010). Genetic analysis of common carp (Cyprinus carpio) strains. II: Resistance to koi herpesvirus and Aeromonas hydrophila and their relationship with pond survival. Aquaculture 304 (1-4), 7-13. Other communicationsAnnual report of reference laboratory activities on KHVD in 2009 submitted to OIE.O. Donohoe., K. Henshilwood, K. Way, D. Stone & D. Walls (2010) Investigation of latency associated with koi herpesvirus infection in carp (Cyprinus carpio). Poster presentation at Biotech Research Day 2010, Dublin City University, 29 January 2010.K. L. Denham (2010). Fish health update and a report on KHV transmission studies. Oral presentation at The Coarse Fish Meeting – Sparsholt College, UK, September 2010. N. Stinton (2010). Transmission of KHV via contaminated keep-nets and tank water. Oral presentation to the Fish Welfare Group, National Agricultural Centre (NAC), Stoneleigh Park, UK, September 2010.13. 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? NO______6 Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010

OIE Reference Laboratory Reports

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