African swine fever situation in Romania

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African swine fever situation in Romania Standing Group of Experts on African swine fever in the Baltic and Eastern Europe region (under the GF-TADs umbrella) Eighth meeting (SGE ASF8) Chișinău, Moldova, 20-21 September 2017

T H E N A T I O N A L S A N I T A R Y V E T E R I N A R Y A N D F O O D S A F E T Y A U T H O R I T Y O F R O M A N I A Clinical suspicion On 27.07.2017 one pig was found dead in a backyard holding with 4 pigs, in Satu-Mare Municipality, Satu- Mare County, near to the border with Ukraine. Sample was first sent to the SVL Satu-Mare, was examined, and found positive. Sample was sent further to NRL for investigation by an acredited method on 28-th July. On 28-th July, based on preliminary positive results, and in view to contain the virus, took place more activities: - Preliminary epidemiologic investigation which established another possible contact holding with three pigs, in the neighborhood; - In the first backyard, were found: meat coming from 2 another sows slaughtered at home in the previous evening, and refrigerated; one live fattening pig with fever; none of these pigs were identified. - In the contact holding were found: one dead boar, and two sows without clinical signs; none of these pigs were also identified. - All the pigs mentioned before and found in life, in both backyards were sampled for diagnostic purposed and culled; - Took place the disposal by burning and burying on site of all culled pigs, and of the confiscated meat; - Took place disinfections of shelters, premises, vehicle and tools in both backyards; - Were established movement restrictions on live animals of other species found in both backyards.

On 31-th July and 1-st August the Local Disease Control Centre Satu-Mare was met, was activated the Contingency Plan, and were issued more Decisions for adopting the ASF Control Plan and restriction zones. Were also discussed the next activities to be followed in the restriction area, as follows: census, surveillance activities, checks in traffic and routes, resources of personell, equipments and founds etc. Informations were sent to the NSVFSA, to all commercial holdings in the county, and to the local veterinary competent authorities in the neighboring counties. All farmers, hunters, rangers etc. were informed by press releases about the disease confirmation and about the measures to be followed. On 1-st August was also met the National Disease Control Centre and issued a Decision with the necessary measures to be adopted at Government level.

ASF Confirmation: On 31-th July the NRL has confirmed the diagnostic of ASF for the first sample which was also considered the primary outbreak. The primary disease outbreak was notified to the OIE and to the EU Commission by ADNS system. The Commission and the Member States were informed about the confirmation. On 1-st August was confirmed the ASF for the rest of samples coming from the both backyards, and sampled on 28-th July. The secondary outbreak was notified the OIE and to the EU Commission. The Commission and the Member States were also informed about these confirmations. The National Reference Laboratory sent the samples to the Community Reference Laboratory.

Outbreaks coordinates: Lat: 47.749330 Long: 22.843265 The restriction zones was established under the provisions of the Council Directive 2002/60/EC: - a 3 km protection zone around the outbreaks; - a more larger than 10 km surveillance zone. In some part of it, the distance from the outbreak (West, East, North-East and North), exceeds with 6, 20, 36 and 13 km the legal limit of 10 km surveillance zone. They were approved by the CI Decision 2017/1416 and are enforced to 15-th October 2017

First pig found dead Lesion: Enlarged spleen Temperature in live pig

ACTIVITIES FOLLOWED In the protection zone: Census and clinical inspections: 100 backyard holdings, with 705 pigs, inspected twice No commercial farm inside the protection zone, only backyards; Identification of all pigs found unidentified; In the surveillance zone: Census and clinical inspection: 6217 backyard holdings with 26473 pigs, inspected twice; 4 commercial farms inside the restriction zone with 13664 pigs inspected daily; Surveillance activities High awareness level maintained amongst farmers, hunters, rangers, veterinarians, transporters. So far, the ASF situation is stable, the virus seems to be contained, any new outbreak was confirmed meantime.

Holdings and pigs in the restriction zone; clinical inspections since 1-st august Crt. no. Restriction zones No. of pig holdings No. pigs No. of clinical inspections 1 Protection zone 3 km (Satu-Mare area) 2 Surveillance zone more than 10 km (localities specified in the Annex of the Decision 2017/1416/CE) 101 709 1414 6217 nonprofessional holdings 4 commercial holdings 26473 54656 13664 Each day, all pigs 3 Total 6221 40137 54656 + pigs in commercial holdings each day

Surveillance in non-professional holdings since 1-st august Protection zone Surveillance zone Satu-Mare, outside the restriction zones Satu-Mare total Blood on EDTA Organs Blood on EDTA Organs Blood on EDTA Organs Blood on EDTA Organs 23 1 18 18 22 10 63 29

Surveillance in commercial farms all-over the county Satu-Mare since 1-st august Commercial holdings Protection zone Surveillance zone Satu Mare County outside restriction zone Satu-Mare Total Type of samples Blood- EDTA Orga ns Blood- EDTA Organs Blood- EDTA Organs Blood- EDTA Organs Total 0 0 176 74 41 26 217 100

Active surveillance in Satu-Mare, wild boars, in August 2017. No passive surveillance. No. hunting grounds 22 Under 1 year age Hunted wild boars tested by PCR 1-2 years age Older than 2 years Total 2 27 31 60

Conclusions following the epidemiological investigations: The virus could be introduced into the primary outbrek in the period 21-24-th July 2017 but sources were not possible to be traced. The source of virus could be in meat, or meat products fraudulent introduced in Romania from Ukraine, having regard the bordering of the Satu-Mare county with Ukraine where the ASF evolves both in domestic pigs and wild boars. Way of ASF introduction: most probably swill feeding. Tracing forward: infection in the secondary outbreak, already confirmed and notified on 1-st August 2017, was introduced by a boar owned illegaly, in mating purposes, and which were present from 21 to 24-th July 2017 in the backyard which constituted the primary outbreak. Tracing back: undetermined yet, only supposed to be in Ukraine.

The ASF infection timeline Holding Owners 10.iul 11.iul 12.iul 13.iul 14.iul 15.iul 16.iul 17.iul 18.iul 19.iul 20.iul 21.iul 22.iul 23.iul 24.iul 25.iul 26.iul 27.iul Butea Petru- RO1364921369 (unidentified animals, they appeared registered on the last location - Vanyl holding from where they were bought at 2 month old, in January 2017); primary outbreak. 3 sows of 9 month old, 1 fattening pig of 9 month old. The period from 22-24 July could be considered the introduction of the ASF virus, when the boar come back in the holding of Mr. Butea. This is, also, the incubation period for Mr. Butea holding. One sow shows clinical sigs. The night of 26/27 July the sow has died. clinical suspicion in the dead sow, the sample were sent to NRL for diagnostic;the 2 other sows were slaughtered at home due to clinical sigs: apathy, fever, lost of appetite Boros Florin - unregistered hoding, unidentified pigs; secondary outbreak. 2 sows of 24 moth old and 1 boar. the boar of Mr. Boros spent the period form 11-th-July to 20-th July in the holding of Mr. Butea for mating. All animals were healty the boar was returned in the origine holding for mating the sow of Ms. Pop Incubation period in Mr. Boros holding and first clinical signs in boar The boar expressed clinical signs Pop Elisabeta - holding RO1364920986 (considered as possible contact holding)- one adult sow, and 15 vietnameese pigs of different ages. the adulte sow was transfered in Mr. Boros holding in mating purposes and come back in the same evening

Conclusions of the European Union Reference Laboratory for african swine fever (EURL-ASF), INIA-CISA The presence of ASF has been confirmed throughout ASFV genome and antibody detection in the four domestic pigs received from the outbreaks occurred in Romania, in Satu-Mare on 28-th July 2017. The ASF virus has been isolated from the four domestic pigs showing the characteristic haemadsorbing pattern. The preliminary results suggest that, on the basis of the different-sized PCR products of the two intergenic regions (IGRI73R-I329L and MGF) and the CVR within the ASFV genome, the domestic pigs ROMANIA ASF viruses belong to the p72 genotype II, CVR-1, IGR-2 and MGF1 variants. These are the variants mostly circulating within the EU countries as well as described in Moldova (2016), Ukraine (2012, 2015), Belarus (2013) and in certain areas of the Russia Federation (from the data available at the EURL).

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