ALPHAVIRUS, PRV AND PMCV SITUATION IN CHILE



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ALPHAVIRUS, PRV AND PMCV SITUATION IN CHILE Marcela Lara Head Animal Health Unit Sernapesca Trination meeting Trondheim, February 2014

Geographic Distribution of salmon production C h i l o é cultivated species - Salmo salar - O. mykiss - O. kisutch - O. tshawytscha Fresh Water Sea Water

Chilean Salmon Sea farms Sanitary Report 2013 PRODUCTIVE CONTEXT:NUMBER OF SEA FARMS Average number of sea farms in activity: 370 sea farms located in Los Lagos (43.1%), Aysén (51.3%) and Magallanes Regions (5.7%). The numbers are similar to 2012. Gobierno de Chile Ministerio de Economía Fomento y Turismo Servicio Nacional de Pesca y Acuicultura 3

Chilean Salmon Sea farms Sanitary Report 2013 PRODUCTIVE CONTEXT: BIOMASS Increased biomass post health crisis Cultivated biomass (tons): similar to 2012, where the historical maximum ocurred in october (600,000 tons) Aysén Region concentrates 56.5% of the entire biomass. Gobierno de Chile Ministerio de Economía Fomento y Turismo Servicio Nacional de Pesca y Acuicultura

Chilean Salmon Sea farms Sanitary Report 2013 PRODUCTIVE CONTEXT: BIOMASS/SPECIES Atlantic salmon represents 75.6% of the cultivated biomass, followed by rainbow trout (17.1%) and coho salmon (7.2%) Gobierno de Chile Ministerio de Economía Fomento y Turismo Servicio Nacional de Pesca

Region Atlan,c salmon Rainbow trout Coho salmon Total Los Lagos (X) 82 39 24 145 Aysén (XI) 137 31 15 183 Magallanes (XII) 21 4 25 Total 240 74 39 353

List of Diseases List EAR 1 List EAR 2 " Epizootic haematopoietic necrosis " Infectious haematopoietic necrosis virus " Infection with Aphanomyces invadans (Epizootic ulcerative syndrome " Infection with Gyrodactylus salaris " Koi herpesvirus disease " Viral haemorrhagic septicaemia virus " Spring viraemia of carp virus " Red sea bream iridovirus " Alphavirus Infectious (2008/2013) " Totivirus Infectious (2013) Free " Infectious pancreatic necrosis " Piscirickettsiosis " Renibacteriosis (BKD) " Infectious salmon anaemia virus (ISAV) (HPR 0 and HPR delete) " Sea lice " List EAR 3 PRV (purpose of collecting information) Control Program In Chile we have 3 lists of diseases

Active Surveillance Program Active surveillance system implemented in Chile - 2003, (Specific for High Risk Diseases (HRD)) Two annual visits to confirm the absence of HRD, These visits are performed by veterinarians (Certifier health condition). Since the Program's inspection, analyses has been made based on cell culture, using CHSE-214 and EPC or BF-2 cell lines. From 2009 was incorporated RT-PCR to Alphavirus and, finally, in the second semestre of 2013 was incorporated RT-PCR to Totivirus. The organs sampled for cell culture are: kidney, spleen, brain and heart. To perform the RT-PCR for SPDV and PMCV, heart samples are used. The samples are combined and consist in pools of no more than 5 fish.

SAV / PD May 2008: inclusión of PD as high risk list disease as a mandatory declaration disease.

Alphavirus 2009 included in the Official Active Surveillance Program for Hight Risk Disease 2 anual samplings in every site stocked with susceptible species. Official testing: Real time RT-PCR Hodneland & Endresen (2006) All samplings have been negative

PRV/HSMI Study - 2012 Sernapesca/Subpesca runs a study to confirm or rule out the presence of PRV/HSMI. Methods: Sampling of 28 sea sites located in X, XI and XII region. Sites selection criteria: species (Salmo salar) and average weight (>1 kg). Sampling: 15 fishes per site (n=365 samples). Analysis: RT-PCR (heart and Kidney), methods: Palacios, 2010 (HSMI). Histology: muscle, heart, piloric caeca, gill, liver, spleen and kidney. Studies, to conclude that PRV is present in the species Salmo salar in sea farming centers in X, XI and XII. Histopathology: non-specific findings

PMCV Surveillance of PMCV (Totivirus) (Listed July 2013): Official surveillance program from August 2013. Must be included in the second annual sampling. Farms already made, should make special sampling for PMCV. Diagnostic method: Real time RT-PCR Løvoll et al. 2010 (Virology Journal, 7:309) and Haugland et al. 2011 (Journal of virology, June 2011, p. 5275-5286). All samplings have been negative (15.065 fishes) Gobierno de Chile Ministerio de Economía Fomento y Turismo Servicio Nacional de Pesca y Acuicultura Marzo 2013 12

Year Type of site N of Sampled Fish Techniques 2007 2008 2009 2010 2011 2012 2013* Sea 41.852 Cellular cultura in CHSE- 214 + EPC o BF2 Freshwater 38.113 Cellular cultura in CHSE- 214 + EPC o BF2 Sea 31.312 Cellular cultura in CHSE- 214 + EPC o BF2 Freshwater 28.541 Cellular cultura in CHSE- 214 + EPC o BF2 Sea Freshwater Sea Freshwater Sea Freshwater Sea Freshwater Sea Freshwater Total general - all sample negative 18.311 14.880 17.311 15.458 14.656 17.525 18.820 15.490 20.674 15.590 685.866 * Only considers the informalon of the first days of December 2013 RT- PCR (Alphavirus) RT- PCR RT- PCR RT- PCR RT- PCR RT- PCR RT- PCR RT- PCR RT- PCR RT- PCR (Alphavirus and ToLvirus)

Official diagnosis system of high-risk disease in Chile Legal framework Obligations of laboratories (Private laboratories): Standardized work procedures. ISO 17025 accreditation Deliver periodic results of PVA, PVP, PSGR and Specific Surveillance and Control Programs ( I S A a n d Piscirickettsiosis). Immediate reporting of suspected or positive results EAR List 1 and List 2 PSEVC-ISA and PSEVC-Piscirickettsiosis Gobierno de Chile Ministerio de Economía Fomento y Turismo Servicio Nacional de Pesca y Acuicultura Marzo 2013 14

Official diagnosis system of high-risk disease in Chile Legal framework Reference Laboratories: (Resolution No. 1448 of 6/07/2011). Laboratorio del NBC Universidad Católica de Valparaíso. Laboratorio de Virología Universidad de Valparaíso Laboratorio de biotecnología y patología acuática Universidad Austral de Chile. Laboratorio de anatomía patológica Universidad de Chile. ISA PS (molecular) IPN Exóticas (PD, PMVC, VHS) PRV BKD PS (histopatología) Gobierno de Chile Ministerio de Economía Fomento y Turismo Servicio Nacional de Pesca y Acuicultura Marzo 2013 15

Next Steps Continuing with the investigation PRV in fresh water (February 2014). Implement new diagnostic techniques: Immunohistochemistry (PRV/HSMI) Training in cardiac pathologies Biosecurity Conference 2014 (Aquasur, October) - PD / CMS / HSMI

Gracias