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OIE Reference Laboratory Reports Activities in 2011 Name of disease (or topic) for which you are a designated OIE Reference Laboratory: Address of laboratory: Infectious salmon anaemia Department of Pathology an Microbiology, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, PE, CANADA, C1A 4P3 Tel.: (1-902) 566-0967 Fax: (1-902) 566-0851 e-mail address: [email protected] website: http://www.upei.ca/~avc/html/oie.html Name (including Title and Position) of Head of Laboratory (Responsible Official): Dr Frederick S.B. Kibenge Professor and Chairman Name(including Title and Position) of OIE Reference Expert: Dr Frederick S.B. Kibenge Professor and Chairman Name (including Title and Position) of writer of this report (if different from above): Annual reports of OIE Reference Centres, 2011 1 Infectious salmon anaemia Part I: Summary of general activities related to the disease The Atlantic Veterinary College (AVC) is one of six faculties of the University of Prince Edward Island (UPEI), in Charlottetown, Canada. The university has approximately 4,000 full-time students. AVC offers a 4-year Doctor of Veterinary Medicine degree to Atlantic Canadian and international students, which is accredited by the Canadian Veterinary Medical Association, the American Veterinary Medical Association, and recognized by the Royal College of Veterinary Surgeons in the United Kingdom. AVC is recognized by other veterinary colleges around the world for its expertise and extensive research and service in aquaculture and fish health. AVC has a full service Veterinary Teaching Hospital and Diagnostic Services laboratories, with a Regional Diagnostic Virology Service for mammalian, avian and aquatic animal species. The OIE Reference Laboratory for ISA at AVC is located within the UPEI Aquatic Virology Collaborating Centre, one of UPEI’s six Centres of Expertise. 1. Test(s) in use/or available for the specified disease/topic at your laboratory Test For Specificity Total* Conventional RT-PCR Virus RNA Group 402 Real-time RT-PCR Virus RNA Group & viral load 445 SHK-1/ASK-2/TO/CHSE-214 cell cultures at 16oC Virus isolation Phenotype 8 Electron microscopy Virion morphology Group 2 In-situ hybridization Virus mRNA Group 0 ELISA Antibody detection Group 0 ELISA Antigen detection Group 0 Virus neutralization Antibody titre Serotype 0 DNA sequencing Nucleotide sequence Genotype 81 * Total includes all tests on surveillance, diagnostic, and research samples 2. Production and distribution of diagnostic reagents Type of reagent Fish tissues harvested from healthy fish held in quarantine ISAV cRNA for RT-PCR ISAV Proficiency panels for real time RT-PCR 2 Amount supplied nationally (including for own use) Amount supplied to other countries 10 10 vials (one gene) 10 vials (one gene) each to Taiwan and Chile 2 panels (38 vials per panel) 1 panel (38 vials) each to Taiwan and Chile Annual reports of OIE Reference Centres, 2011 Infectious salmon anaemia Part II: 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 a) Establishment and maintenance of a network with other OIE Reference Laboratories designated for the same pathogen or disease and organisation of regular inter-laboratory proficiency testing to ensure comparability of results Validation of antibody ELISA for the detection of antibodies against ISAV in fish has been initiated. Formal validation of this test in accordance with the OIE Validation Template is anticipated in the near future. b) Organisation of inter-laboratory proficiency testing with laboratories other than OIE Reference Laboratories for the same pathogens and diseases to ensure equivalence of results Organized two separate ISAV laboratory proficiency testing trials: (1) ISAV laboratory proficiency testing trial with a company in Chinese Taipei. (2) ISAV laboratory proficiency testing trial as part of the OIE Laboratory Twinning Project between AVC and Pontificia Universidad Catolica de Valparaiso, Chile. 4. Preparation and supply of international reference standards for diagnostic tests or vaccines There are no standard reference materials officially recognized by OIE. Other reference material 5. Amount available Amount supplied to other OIE Member countries ISAV antigens for antibody ELISA Prepared upon request Atlantic salmon serum (positive control) for ISAV antibody ELISA Prepared upon request ISAV Rabbit polyclonal antisera Prepared upon request ISAV cRNA for RT-PCR Prepared upon request 10 vials (one gene) each to Taiwan and Chile ISAV Proficiency panels for real time RTPCR Prepared upon request 1 panel (38 vials) each to Taiwan and Chile ISAV cDNA clones of gene segments Prepared upon request Histology slides of ISA Prepared upon request Antigens for ISAV vaccines Prepared upon request Research and development of new procedures for diagnosis and control The OIE Reference Laboratory for ISA at AVC has published results of a Ringtest on ISAV diagnostic process and TaqMan® Real-time RT-PCR of 14 participating fish diagnostic laboratories from Europe (1), South America (12) and Asia (1) (Kibenge et. al., 2011, J. Aquaculture Res. Dev., 2, 110). This Ringtest showed that there are various RNA extraction kits as well as thermocyclers that are used in the laboratory diagnosis of ISAV, resulting in poor reproducibility of the test result as a cycle threshold (Ct) value. The variation in performance of the different laboratories could result in false reporting of the fish tissue sample status. A significant factor influencing the Ct values and therefore the diagnostic sensitivity might be the thermocycler software used. Annual reports of OIE Reference Centres, 2011 3 Infectious salmon anaemia 6. Collection, analysis and dissemination of epizootiological data relevant to international disease control The OIE Reference Laboratory for ISA at AVC received 4 submissions of samples from wild fish in British Columbia-Canada. ISAV sequences were detected in some of the samples in two of the submissions, and the positive results were reported to OIE and CFIA. As a consequence, CFIA plans to sample wild fish in 2012 to test for ISAV. 7. Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the pathogen and the disease concerned The OIE Reference Laboratory for ISA at AVC participated in a CFIA-initiated Laboratory assessment. 8. Provision of consultant expertise to OIE or to OIE Member Countries Dr Fred Kibenge was expert witness for the Cohen Commission of Inquiry during the ISAV hearings on December 15-16, 2011, in Vancouver, BC, Canada. The OIE Reference Laboratory for ISA at AVC was an invited participant at the OIE Global Conference on Aquatic Animal Health Programmes: Their Benefits for Global Food Security, Panama City, Panama, 28–30 June, 2011. A presentation titled “Laboratory issues – Aquatic animal disease, diagnosis, and global trends” was made by Dr Fred Kibenge to the conference delegates. Dr Fred Kibenge was OIE Delegate to the OIE Laboratory Twinning Feedback Session, OIE Headquarters, Paris, France, 30–31 March 2011. 9. Provision of scientific and technical training to personnel from other OIE Member Countries OIE Laboratory Twinning Project between AVC and Pontificia Universidad Catolica de Valparaiso, Chile, was launched on 15 October, 2010 in Valparaiso, Chile. The first training workshop was held from 12–15 October, 2010 in Valparaiso, and included training in Taqman RT-PCR assay for ISAV. The training of two personnel from the Candidate Laboratory took place in the Parent OIE Reference Laboratory in Charlottetown, Canada, from 5– 17 June, 2011. At the end of the training program, the Candidate Laboratory was provided with a proficiency panel to try out on their own in their lab in Valparaiso, Chile. 10. Provision of diagnostic testing facilities to other OIE Member Countries No activity. 11. Organisation of international scientific meetings on behalf of OIE or other international bodies No activity. 12. Participation in international scientific collaborative studies Dr Fred Kibenge was a member of a blue ribbon team that analyzed the recovery of the Chilean salmon industry from the ISA crisis. Title of study: The recovery of the Chilean salmon industry: The ISA crisis and its consequences and lessons. Objectives: The objective of this study was to analyze and report the evolution of the ISA crisis in the Chilean salmon industry and to evaluate the recovery process from various perspectives, extracting lessons learned that might prevent or mitigate impacts of similar outbreaks in aquaculture in Chile and in other countries. 4 Annual reports of OIE Reference Centres, 2011 Infectious salmon anaemia Project Partners: This study was an initiative of the Global Aquaculture Alliance (GAA) co - sponsored by The World Bank, the Undersecretariat of Fisheries-Chile and the Chilean Salmon Industry Association (SalmonChile). The GAA brought together an International Group of Experts (Adolfo Alvial, John Forster, Frederick Kibenge, José M. Burgos, Rolando Ibarra, and Sophie St-Hilaire) to analyze the progression and recovery of the ISAV crisis in the Chilean Salmon Industry. Time frame: 1 August, 2011 –30 November, 2011. Significant outcomes: The Chilean salmon farming industry recognizes that it will take more than laws, regulations and improved practices to prevent another disease crisis. The values and attitudes of those involved in the industry are a crucial component of preventing a future problem. Cooperation between groups is essential due to the nature of the shared water bodies used by closely interconnected producers. From the ISA diagnostics perspective, there are two main lessons learned: (i) Countries should increase surveillance efforts of the aquaculture industry (farmed fish, slaughter plants, marine environment) in order to detect any significant new pathogens early, and (ii) Countries should invest in well developed capacity and technology in their diagnostic laboratories, and not rely on external labs in other countries. Mitigation of infectious salmon anaemia (ISA) by vaccination and genetic selection is a four-year project (20082012) being undertaken by Novartis Animal Health Canada Inc. Aqua Health Business, which is located in Victoria, Prince Edward Island, Canada. This project represents a partnership between Novartis, AVC/UPEI (OIE Reference Laboratory), Cooke Aquaculture Inc., New Brunswick, New Brunswick Research and Productivity Council (RPC), and the Norwegian Veterinary Institute (NVI), Norway. 13. 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 Alvial A., Forster J., Kibenge F., Burgos J.M. and Ibarra R. 2011. Chile’s Recovery from ISA. Goal 2011, Santiago, Chile, 6–9 November, 2011. http://www.gaalliance.org/update/GOAL11/goal11prog.pdf MacLellan, R.M.A., Kibenge, M.J.T. and Kibenge, F.S.B. 2011. What types of genetic changes occur to infectious salmon anaemia virus (ISAV), a fish orthomyxovirus, during mixed infections with different ISAV genotypes? 2011 Merial-NIH National Veterinary Scholars Symposium, University of Florida College of Veterinary Medicine, Gainesville, Florida, USA, 4–7 August, 2011. Kibenge, F.S.B., Godoy, M.G., and Kibenge, M.J.T. 2011. Laboratory issues – Aquatic animal disease, diagnosis, and global trends. OIE Global Conference on Aquatic Animal Health Programmes: Their Benefits For Global Food Security, Panama City, Panama, 28-30 June, 2011. OIE Scientific and Technical Review, in press. Kibenge, F.S.B. 2011. International Veterinary Medicine Laboratory Practice. Puerto Montt, Chile, 10 March, 2011, sponsored by Unversidad San Sebastian, Puerto Montt, Chile. Kibenge, F.S.B. 2011. The new finding regarding the molecular epidemiology of ISAV. Update of Important Viral Diseases in Salmon Aquaculture. Puerto Montt, Chile, January 9–16, 2011, sponsored by Aquagestion S.A., Chile. Scientific publications in peer-reviewed journals Kibenge, F.S.B., Kibenge, M.J.T., and Masaoud, E. 2011. Infectious Salmon Anaemia Virus (ISAV) Ringtest: Validation of the ISAV Diagnostic Process using Virus-spiked Fish Tissues and ISAV TaqMan® Real-time RTPCR. Journal of Aquaculture Research and Development, 2:110. doi:10.4172/2155-9546.1000110 _______________ Annual reports of OIE Reference Centres, 2011 5