ASF in the Czech Republic: current situation and control measures

Transcription

1 NOTE; the use of the content presented here is subjected to the author s permission ASF in the Czech Republic: current situation and control measures Petr Vaclavek NRL for ASF, State Veterinary Institute Jihlava Workshop on Laboratory Diagnosis and Control of CSF and ASF, May 2018

2 STATE VETERINARY INSTITUTE JIHLAVA National reference laboratory for CSF and ASF

3 First occurrence of ASF virus in wild boar in the Czech Republic First ASF positive carcass location: Příluky, Zlín district Date: 26th June 2017 Way of ASF introduction? GPS N, E Zlín

4 First ASF case in the Czech Republic Zlín city - inhabited area 1 st WB carcasses found nearby the local hospital

5 Molecular characterisation of the Czech and others Eastern European ASFV isolates from years COUNTRY YEAR P72 GENOTYPE CVR SUBTYPING IGR I73R-I329L SUBTYPING IGR MGF SUBTYPING Georgia 2007 II CVR1 IGR-1 MGF -1 Armenia 2007 II CVR1 IGR-1 MGF -1 Azerbaijan 2008 II CVR1 IGR-1 MGF -1 Russia II CVR1 IGR-1 MGF -1 Federation II CVR1 IGR-1 + IGR-2 MGF -1 + MGF -2 Ukraine 2012, 2015 II CVR1 IGR-2 MGF -1 Belarus 2013 II CVR1 IGR -2 MGF II CVR1 IGR-2 MGF -1 Estonia CVR II CVR1SNP* + IGR-2 MGF -1 CVR 2 Latvia II CVR1 IGR-2 MGF -1 Lithuania II CVR1 IGR-2 MGF II CVR1 IGR-2 MGF -1 Poland 2016 II CVR1 IGR-2 MGF -1 + MGF II CVR1 IGR-1 + IGR-2 MGF -1 Moldova 2016 II CVR1 IGR-2 MGF -1 Czech Republic 2017 II CVR1 IGR-2 MGF -1 Carmina Gallardo, CISA-INIA Romania 2017 II CVR1 IGR-2 MGF -1

6 Demarcation of the infected area and the buffering zone In accordance with the Council Directive 2002/60/EC the whole District Zlín has been declared as an infected area (1 034 km 2 ), 37 municipalities, 89 hunting grounds infected area red zone Infected area green zone buffering zone intensive hunting highway SVI Praha (Prague) Poland Czech Republic SVI Jihlava NRL for ASF SVI Olomouc rendering plant Germany Austria Slovakia

7 Sampling: sample types and quality clotted blood = first choice sample organs - compulsory in PART II. (spleen, bone marrow, kidney, lung, tonsils etc.) PART I. and buffering zone only blood

8 DNA extraction MagNA PURE LC a MagNA PURE 96 (Roche) + compatible commercial extraction kits MagNA Pure LC Total Nucleid Acid Isolation Kit MagNa Pure 96 DNA and Viral NA Small Volume Kit (Roche)

9 PCR methods vs. laboratory capacity The ability to integrate qpcr into automated platforms increases sample throughput and decreases the potential for cross-contamination. = fast, sensitive, quantitative, closed systems E.g. enhancement of lab capacity due the robotic extraction of DNA: MagNA PURE (Roche) 32/96 samples/1-2 hr QIA Symphony (Qiagen) 96 samples/3 hr + increase of Real-Time PCR systems: 4x CFX96 Touch Real-Time PCR (BIO-RAD) = 4 x 96 samples/70 min.

10 Approximate capacity of Czech NRL for ASF (samples tested/1day) Real Time PCR ELISA Ab blood tissue blood standard mode crisis mode

11 Approximate total capacity of all Czech SVI laboratories (samples tested/1day) Real Time PCR ELISA Ab blood tissue blood standard mode crisis mode

12 ASF DIAGNOSTICS TESTS used in CZECH LABORATORIES ANTIBODY DETECTION TECHNIQUES TEST TYPE REFERENCE ELISA test INGEZIM PPA Compac blocking ELISA ID Screen Indirect ELISA ID Screen Competition ELISA SVANOVIR ASFV-Ab indirect ELISA - INGENASA ID.VET ID.VET Svanova IPT test Indirect immunoperoxidase test (IPT) Gallardo et al DETECTION of the ASF VIRUS GENOME by PCR TEST TYPE REFERENCE Conventional PCR Real Time PCR OIE conventional PCR Agüero et al UPL Real-time PCR (UPL Probe) Fernandez et al Taqman Probe (OIE - Real Time PCR) King et al Zsak et al. 2005

13 Active and passive surveillance in the in buffering zone with the intensive hunting hunted WB: all ASF negative 698 foud dead: all negative 26 May 2018 financial reward for each hunted WB > 100 WB WB WB WB 5-9 WB 1-4 WB high-risk area infected area highway

14 ASF in Czech Republic: current epidemiological situation from 26 June 2017 to 26 May 2018: In total 248 cases of ASF in wild boar population the total number of positive cases in found dead WB: 214 (virus and/or antibodies) the total number of positive cases in hunted WB: 34 (virus and/or antibodies) Total number of ASF tested wild boar Part II: found dead 380 / 214 positive (56.3%) hunted 1852 / 34 positive (1.8%) Part I: found dead 152 / no positive hunted 9145 / no positive NO OUTBREAK IN DOMESTIC PIGS! PART II. PART I. 40 km

15 WB positive cases: virology / serology Positive ASF results: 26 June May 2018 WB Found dead both PCR and ELISA (IPMA) positive only PCR positive only ELISA (IPMA comfirmed) positive Total positive cases Hunted TOTAL Wild boars ASF Virus (PCR) ASF antibodies (ELISA, IPMA) Found dead Hunted TOTAL Recovering survivors piglets / adults (1:1) 6,7 %??? chronic infection? virus carriers? In 2017/2018: 606 (74%) found dead WB a 8769 (85%) hunted WB (PART I. and II.) were tested for presence of specific antibodies (ELISA, IPMA) against ASFV in the CR (8 April 2018)

16 ASF measures for domestic pigs in Part II ban on keeping of pigs in backyard farms in the infected area enhanced passive surveillance in pig farms - farmers must report all dead pigs in the infected area (all cases are tested for ASF) movement of pigs only with authorisation issued by the RVA for Region Zlín. ban on feeding with fresh grass, ban on straw bendding official controls in pig farming continue in accordance with Commission Implementing Decision 2014/709/EU. Targeted for BIOSECURITY. Density of domestic pigs in the Czech Republic (per 100 km 2 ) farms pigs Czech Republic Zlín region infected area (district Zlín)

17 ASF measures in wild boar in Part II enhanced passive surveillance of WB found dead (motivated searching of carcases) ban on hunting (any species, any hunting system) and later hunting of WB was allowed but only by individual hunting and trapping (selected and trained hunters) ban on feeding (only baiting allowed) ban of entrance for the general public into the infected area all hunted and found dead WB must be disposed of in the rendering plant (+ tested for ASF) unharvest fields left to provide food and shelter for wild boars financial rewards: each finding of dead wild boar each hunted young wild boar (up to 50 kg) each hunted adult wild boar (over to 50 kg) compensation for piglet 39, one-year old 78, adult saw 125 for hunted wild boar disposed of in the rendering plant.

18 Wild boar density in Europe (FAO/ASFORCE, May 2015)

19 The density of wild boar population in the Czech Republic (per 100 km2) Hunting year Game bags hunted animals: 1-4 / km2 real WB density?? = 1,5-2x higher the population doubles every 10 years motivated hunting in the whole country 10-12/2017 (38 EURO per hunted animal) 2x higher population in 10 years

20 Electric fences around the infected area ČTK Luděk Ovesný - MAFRA

21 Scented fences around the infected area synthetic foam with 3-Methylbutanoic acid (isovaleric acid) imitation of typical predators smell /odour strong pungent cheesy or sweaty smell it is a major component of the cause of unpleasant foot odour product: Pacholek koncentrát B, Ekoplant, s.r.o.

22 Fences around the high risk zone Area: 57 km 2 Perimeter: cca 30 km electric fence (voltage V) scent fence

23 Passive surveillance: wild boars found dead high risk area (fenced area) inside the infected area 21 May 2018 Fenced area total negat. posit. prevalence in ,7% out WB density in the fenced area: more than 520 (found dead+hunted) WB / 57 km2 = 9.1 WB per 1 km2 YEARLONG HOMERANGE ASFV escape from the fenced area (December 2017) REASON? Rut season? Hunting?

24 The speed rate of disease expansion DIAMETER 11 KM / 11 MONTHS = SLOW speed = Ø 0,5 km/ 1 month despite the high WB density (8-10 WB / km2) based on the estimated date of death of WB found May 2017 June July August October December

25 Higher risk area (fenced area) - unharvested fields left 115 hectares of unharvested fields (rape, maize and wheat) were left for wild boars providing both food and shelter

26 Enhanced passive surveillance of WB found dead Motivated or/and organised searching of carcasses very inaccessible terrain dense vegetation

27 Increased passive surveillance of dead WB motivated searching for carcases

28 Carcass removal time Estimated age of carcasses (days) vs. lab result Estimated incidence DAYS DAYS PCR result: positive negative

29 Research Institute of Agricultural Engineering, p.r.i. Alternative methods of localization of WB carcasses e.g. by thermovision

30 26 January 2018 Active surveillance in the infected area individual hunting by trained hunters hunting by police snipers hunted WB tested RED ZONE 607 (31 posit. = 5.1%) GREEN ZONE 1711 (0 posit.)

31 Hunting by police snipers in the infected area individual hunting by Police snipers (Elite Squad, Police Special Unit, Airport snipers ) started from 16 October 2017 (3 days a week during 10 weeks) in total 157 WB hunted - 8 positive for ASF snipers trained for biosecurity during hunting organization and coordination by RVA and by regional hunters over night hunting (18:00 6:00) mobile thermovision used snipers with silencers all carcases collected and rendered

32 Weekly incidence in relation to hunting measures number of WB found dead (441) part I. and II. number of ASF PCR positive found dead WB (8 April 2018) police snipers week 34: hunting by trapping week 37: hunting by Police snipers week 42-52/2017 and week 4-5/2018 individual hunting by trained hunters week 51: first positive WB cases outside fenced area

33 Carcasses age date of finding vs. date of death (estimated) Snipers activity Snipers activity

34 Disposal of hunted wild boars from the infected area in selected rendering plant Positive BIOSECURITY measure, BUT influence the sample quality = LAB RESULT

35 Trapping of wild boars 32 traps in the area cage traps with sensors and cameras subsidies for traps = Fenced area total trapped negat. posit. prevalence in % out

36 What we learned from our small outbreak? The best rated measures (effectiveness and practicality): motivated and active passive surveillance fast systematic removal of carcases ban on driven hunting (despite public/hunters opposition and political pressure) hunting biosecurity - ALL WB to rendering plant from PART II. and I. disposal of hunted wild boars from the infected area hunters motivation (financial rewards and compensation) Has to be analysed (questionable): hunting in the infected area (snipers) unclear impact unharvested fields in infected area Has to be enhanced: communication with hunters education /awareness intensive media campaign effective decreasing of WB population out of the infected area biosecurity measures on commercial farms

37 Conclusions last ASF infectiones between WB are dated (based on day of death estimation) to the January 2 last ASF positive carcasses were found on 15th April scull and bones (estimated dead is about 4-5 months ago) the uniqueness of the Czech outbreak is the local occurrence in a small solitary area without direct connection to the affected localities in the neighbouring countries incomparable with the situation in Baltic countries and in Poland pioneering authorities with some alternative measures - the crucial fact? it's easy to be wise after the event. / After the battle, everyone is a general but on the Czech example it looks that it is possible to sort out such a small outbreak predicted evolution of Czech outbreak now? there are new WB in fenced area (last week 2 fresh carcases were found) we ll let you know in July

38 Acknowledgements Colleagues from other NRLs: Pavel Bartak Pavel Vodrazka Marcela Kostkova Katerina Mikulaskova Ondrej Stepanek Bronislav Simek, etc. Petr Satran Tomas Jarosil Frantisek Svoboda Petr Kucinsky Frantisek Mahdalik Hugo Merta, etc. Imbi Nurmoja Siiri Põldma Marianna Dobroštana Svetlana Cvetkova Rūta Zagrabskaitė Grzegorz Wozniakowski Marius Masiulis, etc. Marisa Arias Carmina Gallardo Raquel Nieto, etc.

39 Thank you for your attention. 39