EARLY DETECTION AND CONTINGENCY PLANS FOR ASF ASF REFERENCE LABORATORY Prof. José M. Sánchez- Vizcaíno jmvizcaino@visavet.ucm.es www.sanidadanimal.info
ASFV: A old friend 1978- ASF REFERENCE LABORATORY MAIN WORK: DIAGNOSIS TEST & REAGENTS EPIDEMIOLOGY-CONTROL and ERADICATION
ASF AGENDA: ASF REVIEW. RISKs FACTORS ASF EARLY DETECTION ASF CONTINGENCY PLANS FAST RESPONCE ARE WE AWARE?. ARE WE READY?
SOME ASF CHARACTERISTICS
VERY DIFFICULT AND COSTLY CONTROL AFRICAN SWINE FEVER VIRUS (ASFV) Very complex virus, big syze, large genome: 170 kb Very complex molecular structure Genetic variability Replication in macrophages NO production of neutralizing antibodies 200 nm More than 100 structural proteins 22 genotypes (VP72)
ASF PIGS (D & W) and SOFT TICKS Very resistant to inactivation in E Genetic variability
ASF PROTECTION: NO VACCINE NO INACTIVATED VACCINE NO ATENUATED VACCINE. ONLY PARCIAL PROTECTION NO RECOMBINAT VACCINE: NO TARGET GENES ANTIBODIES ARE RELATED WITH SOME TYPE OF PROTECTION AS WELL AS IN CHRONIC AND ENDEMIC ASF INFECTION Eradication without vaccine is possible but not easy
AFRICAN SWINE FEVER VIRUS (ASFV) Very resistant in the environment Clinical signs can resemble other haemorragic diseases Conditions ASFv survival time Source Variable Blood stored clinical at 4ºC signs: hiperacute 18 months subacute Iowa, 2006 Depending: isolate, route and doses of exposure Faeces in room temperature 11 days Iowa, 2006 Classical swine fever (CSF) Erysipela s Contaminated pig pens 1 month Iowa, 2006 Temperature of 56ºC 70 minutes Mebus et al. 1998 in Foreign Animal Diseases Temperature of 60ºC 20 minutes Mebus et al. 1998 in Foreign Animal Diseases ph<3.9 or ph>11.5 (serumfree media) Minutes Mebus et al. 1998 in Foreign Animal Diseases/Plowright,1994 ph 13.4 in serum free media 21 hours http://www.oie.int/esp/maladies/fiches/e_a120. htm ph 13.4 with 25% serum 7 days http://www.oie.int/esp/maladies/fiches/e_a120. htm LABORATORY DIAGNOSIS IS NEEDED
Clinical Signs: Easily Confused WITH: Classical Swine Fever Erisipelas Salmonellosis Other Septicaemic conditions PDNS
AFRICAN SWINE FEVER VIRUS (ASFV) No zoonotic Domestic pigs Both Could become Chronic European Wild Boar AFRICA: Wild Suids Innaparents Infection Low viraemia Persistently Infected Similar clinical to Domestic pigs. More resistence
ASF. Vectors: Soft ticks. Ornithodoros EUROPE: O. ERRATICUS: NO TRANSOVARIAL TRANSMISSION VIRUS: 3-5 YEARS AFRICA: The Natural History Museum, London 2005. O. MOUBATA TRANSOVARIAL (5 ninphe stages) Others: O. Sonrai, O. Porcinus, O. Verrucosus, etc. All O. tested are susceptible to ASF Virus
ASFV. Routes of Entrance:
ASF. Routes of transmission FREE AREAS: By By eating infected swill swill or or garbages Contaminated transport, By By direct contact between sick sick or or carrier and and healthy animals (exudates, (exudates, wounds,...) wounds,...) ONCE ESTABLISHED: INFECTED OR OR CARRIERS PIGS PIGS (D&W) play play an an important role role in in the the dissemination of of the the disease disease infected swill swill or or garbages freezer Wild Wild boar,ticks and and back back yard yard slurry, insects, rats...
1. Raw pork waste at airport/port 2. Movement of pork or pig product Lisbon 1957 Malta, 1978 Sardinia, 1978 Georgia, 2007 Portugal, 1960 Spain 1960 Italy, 1983 Belgium, 1985 3. Movements of infected wild boars Russia, 2007
2. Boar or pork meat of infected swine fed to pigs (movement of infected meat): South Africa 1973; Sao Tome&Principe, 1979; Ivory Coast 1996; Namibia 2004; Kenya 2007 1. Movements of infected pigs or carriers from infected neighbouring countries: Nigeria, Togo, Benin, all in 1997-98; Zambia, 2001; Kenya 2001; Ghana 2002;Tanzania 2004;Burkina Faso 2007;Kenya 2007;Tanzania 2008
AFRICAN SWINE FEVER IMPORTANCES POINTS Domestic & Wild animals affected Carrier animals Freezer High variability PROBLEMS FOR ASF CONTROL Pigs &products Clinical signs similar to other diseases No vaccine available Ticks Laboratory diagnosis needed Very resistant in the environment
Asfv, genotype II Mozambique, Madagascar, Zambia 186 Outbreaks. More 75.000 Dead pigs
ASF EPIDEMIOLOGY: Historical distribution 1957 Kenya1921
ASF EPIDEMIOLOGY: Historical distribution 1978
ASF EPIDEMIOLOGY: Historical distribution 2007 1998 2001
World globalization Economic crisis WHY IS ASF MOVING? More ASF virus in Africa Swill feeding
ASF EPIDEMIOLOGY: 4 affected countries 234 notified outbreaks +74.000 affected pigs Source: OIE, own elaboration
WILD BOARS DOMESTIC PIGS Source: OIE, own elaboration
0 10 20 30 40 50 60 70 80 2007 2008 2009 2010 OUTBREAKS BY YEAR 0 20 40 60 OUTBREAKS BY MONTH 0 5 10 15 20 25 30 35 40 45 50 May 07 Jun 07 Jul 07 Aug 07 Sep 07 Oct 07 Nov 07 Dec 07 Jan 08 Feb 08 Mar 08 Apr 08 May 08 Jun 08 Jul 08 Aug 08 Sep 08 Oct 08 Nov 08 Dec 08 Jan 09 Feb 09 Mar 09 Apr 09 May 09 Jun 09 Jul 09 Aug 09 Sep 09 Oct 09 Nov 09 Dec 09 Jan 10 Feb 10 Mar 10 Apr 10 May 10 Jun 10 Jul 10 Aug 10 Nºoutbreaks Epidemic curve ASF EPIDEMIOLOGY:
ASF EPIDEMIOLOGY: WILD BOAR DOMESTIC 19% 81% OUTBREAKS BY COUNTRY Armenia Azerbaiyán Georgia Rusia 8% 0% 26% 66%
IDENTIFICATION OF RISK FACTORS ENTRANCE IN CAUCASUS REGION: Increase of the ASF V in Africa Economic crisis Swill feeding Lack of biosecurity SPREAD: Swill feeding!! TRADITION Wild boars contact with domestic No biosecurity (ASF don t need high biosecurity) Movement of animals and products WITHOUT CONTROL ENDEMICITY: Wild boar affected Swill feeding Carriers and chronic infections NO ADVENTAGES FOR FARMES Vector? Low Biosecurity and education information to vets and farmers Delayed compensation + lack of resources veterinary service no contact with farmers inadequate connection field laboratory
EARLY DETECTION THE CHALLENGE
SOME DISEASES IN THE EU DURING THE LAST YEARS CSF FMD BT A IF IF SVD Etc
Good and rapid diagnostic tests 1 2 3 4 5 6 7 8 9 M V 257 bp- 108 bp- SAMPLES ARRIVED LATE
LACK OF EQUILIBRIUM LAB. DETECTION FIELD DETECTION
FIELD IS THE FIRST STEP: COLLABORATION: ADMINISTRATION- FARMERS
RISK FACTORS for EARLY DETECTION I No perception of the risk for entrance: No good information to farmers and Vets Legal and ilegal imports of live pigs and meat products Commercial relations with affected countries Low biosecurity practices Swill feeding Backyard systems Low education level, no information about the disease to FAMERS and Vets No information of ASF. How is ASF?
RISK FACTORS for EARLY DETECTION II Late detection caused by: Wrong surveillance program Late detection in field no formation of veterinary services and farmers Low veterinary service in the area inadequate connection field laboratory Unknown of the risk
RISK FACTORS for EARLY DETECTION III Inadequate control program: Movement of animals and products WITHOUT CONTROL Swill feeding. Difficult to changes mentality Not establish the control and surveillance area Delayed compensation + lack of resources Pig production system: Pig density Farming practices (backyard, swill feeding!!) Presence of wild boars and contact with domestic pigs Presence of ticks
ASF EARLY DETECTION NEEDED:
EVALUATING THE RISK OF ASFV ENTRANCE IN THE EU Development of a generic CUANTITATIVE risk assessment framework for the introduction of ASF in EU countries
http://www.efsa.europa.eu/en/scdocs/scdoc/1556.htm
CONCLUSIONS AND RECOMMENDATIONS ASF in the Caucasus and Russian Federation The ASFV circulating in the Caucasus and the Russian Federation is a highly virulent virus. No reduction of virulence since the first outbreak in 2007 in Georgia. ASF has spread in the TCC and in the RF since 2007; measures put in place were not sufficient to control the spread The risk of maintaining ASF and its spread within the TCC and the RF in domestic pigs is HIGH The risk of ASF introduction into the EU is moderate Risk of ASF become endemic in domestic pigs in EU is negligible (High Biosecurity), low or moderate (free range)
Wild boars: CONCLUSIONS AND RECOMMENDATIONS Very low density of wild boar, usually less than one head per km2 in Caucasus and Russian Federation. Wild boar populations are linked with those of the EU through continuous corridors. In particular, Belarus is well connected with Poland and Lithuania, and Ukraine with Poland, Romania. Wild boar in many European countries and high density populations are present in some areas, notably Germany, northern France and central Italy. Risk of ASF remaining endemic in wild boar in the TCC was considered low, and medium in RF. The likelihood of introducing the disease into the EU was moderate. Areas at risk are mainly Belarus (Poland), Ukraine (Romania), Lithuania, Latvia, Estonia. The risk of ASFV becoming endemic in wild boar in the EU is moderate
POSIBLE ENTRANCE: MODERATE Efsa,2010
FIELD TRAINING: farmer and Vets
FARMERS INFORMATION:
TV RADIO JOURNAL CAFE
http://rasve.mapa.es/publica/formacion/curso/index.html
FIELD SIMULATION
ASF DIAGNOSIS: INFECTION DISEASES/DEAD CARRIERS Ag Ag Nucleid Ac Ac VIRUS Ab Infection Clinical Carriers
ASF Laboratory DIAGNOSIS: KEY POINTS No Vaccine Available Antibodies = INFECTION No Neutralizing Antibodies Viremia for Long period of Time Antibodies Persist During Month, Even Years From 7-127 dpi. Abs good infectious marker SEVERAL TEST AVAILABLE
ASF LABORATORY DIAGNOSIS ASF LABORATORY DIAGNOSIS Identification of the Agent Haemoadsorption autorosette (HA) test Isolation in cells cultures: Haemoadsorption with peripheral blood leukocytes from infected pigs. TIME: 3 to 10 Days. ONLY R.Labs Antigen Detection. TIME 75 MIN. Direct immunofluorescent test (DIF) DIF) Low sensitivity in subacute and chronic forms Significant lack of sensitivity due to Ag-Ab Ab complex formation. Not recomended for analysis of serum and tissue t homogenated samples after first week pi. due to false negative results. r
PCR: CONVENTIONAL or REAL TIME PCR: CONVENTIONAL or REAL TIME 0 dpi 1 dpi 2 dpi 3 dpi 4 dpi 7 dpi B S B S B S B S B S B S M 257 bp- 257 bp- 257 bp- 108 bp- 1 2 3 4 5 6 7 8 9 10 11 12 13 14 C M V MOST COMMONLY USED PPC-3/4 + PPA-1/2 TIME: 5 to 6 H
ASF LABORATORY DIAGNOSIS ASF LABORATORY DIAGNOSIS Antibody Detection ELISA tests Indirect ELISA (OIE) In House ELISAs Commercial ELISA, Ingezim K3 MOST COMMONLY USED. TIME 3 H Indirect immunofluorescent test (IIF) Inmunoblotting (confirmation test)
CD-VIDEO: ASF Review and Protocols with Videos http://sanidadanimal.info/cursos/asf/
www.sanidadanimal.info
ASF CONTINGENCY PLANS. FAST RESPONCE: ADMINISTRATIVE STRUCTURE: Veterinary Service Legislation, diagnosis methodology, etc. LIVESTOCK PRODUCTION STRUCTURE: Census, farme Location, movements, wild live population. ASF CHARACTERISTICS: Risks factors, susceptible animals Vectors, clinical sings, lesions, samples to labs, route of entrance, diagnosis methods, desinfectans,..
MANUAL for a FAST RESPONCE 1. NOTIFICATION SYSTEM 2. ZONING OF AFECTED AREAS 3. BAN ON ANIMALS MOVEMENTS 4. LABORATORY CONFIRMATION 5. PROCEDURE FOR DESTRUCTIONS OF CARCASSES 6. DEPOPULATION 7. CLEANING AND DISINFECTION 8. SEROLOGICAL CONTROLS 9. STUDY WILD BOAR AND/OR VECTORS 10. SENTINEL ANIMALS 11. REPOPULATION
ASF virus is in GOOD FORM Moderate risk for the EU countries Possibility of became endemic in some countries of Europe ARE WE AWARE????? ARE WE READY?????
ASF REFERENCE LABORATORY INTERNATIONAL COOPERATION. WE ARE NOT ALONE
jmvizcaino@visavet.ucm.es