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Self-declaration by Romania on regaining its Bluetongue free status

Self-declaration submitted to the OIE on 26 February 2018 by Dr Marius GRIGORE, Delegate of Romania to the OIE, Director of Animal Health Directorate of NSVFSA (National Sanitary Veterinary and Food Safety Authority), Romania

I. The Bluetongue situation in Romania

On the basis of the information provided to the World Animal Health Information System (WAHIS), Romania had been free of Bluetongue virus (BTV) infection until August 2014, when the infection was confirmed in cattle in Buzau county. Through a monitoring and an eradication programme of bluetongue in place, the last outbreak of bluetongue was closed on 4 January 2016. Surveillance has been in place in accordance with Articles 8.3.14 to 8.3.17. of the Terrestrial Animal Health Code (Terrestrial Code) and no case of bluetongue was reported in Romania in the last two years.

II. Surveillance and control measures of Bluetongue in Romania

In Romania, the estimated number of ruminants at risk for bluetongue is the following:

Specie Number

Bovine 2,559,990

Sheep 12,564,438

Goats 1,853,131

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Since 2000, the surveillance of bluetongue in Romania is carried out in the frame of the annual programme for the surveillance, prophylaxis and control of animal diseases, of zoonotic diseases and environment protection. Until 2014, the objectives of bluetongue surveillance programme were the following:

- early detection of any evidences of bluetongue occurrence in Romania; - urgent and efficient intervention in case of a bluetongue outbreak; - establishing the areas with BTV vectors presence; - identifying risk zones for bluetongue in Romania; - complying with the requirements of the Terrestrial Code concerning freedom from

bluetongue.

On 22 August 2014, bluetongue infection was confirmed in cattle with clinical signs in Buzau County. The serotype identified on Romanian territory was BTV 4, as reported in WAHIS on 3 September 2014 and confirmed by the OIE Reference Laboratory in Pirbright on 1 October 2014. Following the evolution of the BTV infection starting from 2014, a monitoring and eradication programme of the disease was implemented in Romania aiming to demonstrate the absence of new infected cases of serotype BTV 4 and to detect new serotype incursions in Romania. The monitoring programme has three components: serological, virological and entomological.

Passive surveillance Active surveillance

- verification of the documents; - serological survey

- inspection of animals belonging to domestic or wild species, susceptible to the BT virus, existing in Romania.

- monitoring of culicoid vectors, identifying the genus and the species, with mobile or permanent black light traps (South Africa type), on the whole territory of the country, during an entire year.

*Details of surveillance performed in the annual frame programme is available in Annex 1.

The surveillance approach is performed on sentinel animals considering the vector activity. Vaccination is also in place on a voluntary basis at the request of an owner. During the years 2014 and 2015, the serological surveillance was performed monthly, from April to November.

Table 1. Results of Bluetongue testing in the frame of the surveillance programme, Romania, 2014 -2015.

Test performed Total number of tests Number of positive tests

2014 2015 2014 2015

Competitive/blocking ELISA 30647 48971 6975 5843

Real Time RT-PCR 6713 1312 5985 43

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In 20141, there were 1.885 outbreaks detected with 5.985 cases, while this frequency decreased to 30 outbreaks and 43 cases of BTV 4 in 2015 (See annex 2 for outbreak distribution by County). The last outbreak of BT was notified to the OIE on 24 November 2015.

III. Surveillance of BTV infection during the past two years

Surveillance of BTV infection in ruminants

The bluetongue monitoring programme is a fulfilment of the OIE - Terrestrial Code Articles 8.3.14-8.3.17 provisions, namely demonstrating the absence of certain serotypes of BTV in Romania and determining the seasonally vector free period (See Annex 1 for a detailed activity on surveillance).

Figure 1. Geographical distribution of BT outbreaks, Romania, 2014

1 Figures reported through Animal Disease Notification System (ADNS) in 2014. WAHIS figures are slightly different due to the fact, to allow timely notification, some outbreaks were merged and reported together.

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Figure 2. Geographical distribution of BT outbreaks, Romania, 2015

In case of clinical suspicions, samples are gathered and sent to the county sanitary veterinary and food safety laboratory in order to be tested by ELISA (whole blood with or without EDTA from live animals and organ fragments -spleen, lymph nodes, lung- from dead animals). The animals with negative results to ELISA test are tested once again after five days by ELISA test. For the animals with positive results to ELISA test, the suspicion is notified. They are further tested by PCR on EDTA blood samples. The confirmation of disease is made by real time RT-PCR test.

The serological monitoring includes sentinel animals, initially seronegative, unvaccinated, older than four months’ age (first cattle and then sheep and goat) in target localities established into epidemiological units represented by quadrates of 50/50 kilometres (km) respectively 2500 square km, calculated in relation with the total area of each county (See details of sentinel selection in annex). Each epidemiological unit has a target locality. Identification of target localities is performed by county or Bucharest sanitary veterinary and food safety directorates, by overlapping the square grid over the map area of the county, so that the distance between the target localities should be at least 50 km (Figure 3); the lists of target localities were sent for approval to NSVFSA.

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Figure 3. Active surveillance of Bluetongue, Epidemiological units of 50/50 km, Romania

The number of samples for each target locality (see Annex 3) is calculated to detect disease with a prevalence of 5% and a confidence of 95%. Prevalence and confidence figures are calculated for each target locality. The serological survey is implemented in June, September and November. The testing methods are: competitive/blocking ELISA, virus neutralisation, real time RT-PCR and PCR typing. The sentinel animals seroconverted during June – November testing are regarded as suspicions; the county sanitary veterinary and food safety directorate representatives carry out report of the suspicion notification and send it to National Sanitary Veterinary and Food Safety Authority, and also the animals are further tested by PCR on EDTA blood samples at the designated laboratories.

Until the confirmation of the first outbreak in each county, the blood samples on EDTA are sent to the National Reference Laboratory (NRL) for bluetongue for the confirmation of the disease and in order to determine the viral serotype by PCR typing. After the confirmation of the first outbreak in the county, the blood samples on EDTA are tested at the designated laboratories. All EDTA blood samples are tested in designated county veterinary laboratories and Institute for Diagnosis and Animal Health (IDAH) which is the NRL for bluetongue. Virological and molecular testing is conducted to confirm the disease in case of suspicions (clinical cases, following seroconversions).

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Figure 4. Target localities for active surveillance, Romania, 2016

Figure 5. Target localities for active surveillance, Romania, 2017

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Following the testing by PCR of the clinical suspicions and the seroconverted sentinels, no positive results were obtained since January 2016.

Table 3. Bluetongue testing in ruminants, annual frame programme, Romania, 2016

Sampled animals Competitive/ blocking ELISA

No. of positive tests

Real Time RT-PCR

No. of positive tests

Clinical suspicions 51 10 12 0

Sentinels 35936 3890* 342 0

Pre-movement tests 232542 1851 19872 0

* out of which 351 represent seroconverted sentinels during June-November and they were further tested by

PCR

Table 4. Bluetongue testing in ruminants, annual frame programme, Romania, 2017

* out of which 220 represent seroconverted sentinels during June-November and they were further tested by

PCR

Table 5. Virus neutralization testing on seroconverted sentinels, Romania, 2016 - 2017

Year VNT BTV 4 No. of positive tests VNT BTV8 No. of positive

tests

2016 124 101 99 0

2017 81 65 65 0

Following the evolution with BTV infection, since 15 October 2014 the entire territory of Romania is under animals movement restriction in regard to BTV4, which considers the following measures: ruminants from Romania moved to other EU Member States territory or third countries territory showed no clinical sign of bluetongue, testing by ELISA or PCR or having vaccinated animals against BTV serotype 4, depending by the BTV vector activity period and the conditions imposed by the country of destination.

Monitoring of culicoid vectors

Entomological surveillance has the following objectives:

- - establishing of the beginning and the end of the vector free period; - - identification of culicoid species and genera; - - establishing of the specific composition and the vectorial abundance.

Based on the entomological investigations performed, risk maps for bluetongue virus are presented below (see Annex 4 for details of the monitoring methodology). The following maps show the areas were competent vectors from Culicoides obsoletus and Culicoides pulicaris exist up to the altitude of 500 m.

Sampled animals Competitive/ blocking ELISA

No. of positive tests

Real Time RT-PCR

No. of positive tests

Clinical suspicions 4 2 4 0

Sentinels 35603 2510* 216 0

Pre-movement tests 173742 1190 101449 0

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Figure 6. Environmental conditions for culicoid vectors. C. obsoletus and C. pulicaris (up to 500 mts. altitude), Romania ,2012.

Figure 7. Risk areas for bluetongue, Romania, 2012

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Table 6. Results of entomological surveillance, Bluetongue surveillance programme, Romania, 2014 – 2017

Year Number of culicoids

C. obsoletus C. pulicaris C. nubeculosus C. dewulfi

2014 77397 28881 1113 1

2015 131308 66079 356 0

2016 93437 40988 1090 1

2017 17808 9631 174 0

IV. Measures for maintenance of status

Early detection system. Bluetongue is a compulsory notifiable disease on the entire territory of Romania. The notification is carried out according to the national Order 77/2005 for the approval of the Sanitary Veterinary Norm regarding the notification of animal diseases, with all subsequent amendments, published in the Official Journal of Romania, no. 964/31 October 2005; this order represents the official transposition of Council Directive 82/894/EEC on the notification of animal diseases within the European Community.

According to the legislation in force, the animal’s owner has the obligation to notify any alteration of the health status of the animals or any case of mortality registered, to the empowered free practice or official veterinarian.

The empowered free practice veterinarian has the obligation to communicate to official veterinarian any situation in which a transmissible animal disease is suspicioned. The official veterinarian communicates, by any means, the suspicion of disease to the county sanitary veterinary and food safety directorate (local sanitary veterinary competent authority). When, following inspection there is a suspicion of bluetongue disease, samples are gathered and sent in order to verify the suspicion (whole blood and/or EDTA from live animals and organ fragments (spleen, lymph nodes, lung) from dead animals.

In the case of the animals coming from other countries, clinical inspection and verification of relevant data, including sanitary-veterinary documents and other documents accompanying susceptible animal consignments coming from other member states of the European Union or third countries, before unloading at destination are performed, according to the NSVFSA and Agricultural and Rural Development Ministry Order no. 129/566/2007 approving sanitary veterinary norm regarding veterinary and zootechnical controls applicable to intra-community trade with certain live animals and animal products (See Annex 1 for details on clinical surveillance). Following European intra-community trade, serological ELISA testing is implemented (samples collected from 1% of the animals, but no less than 5 samples/batch) for ruminants coming into Romania. For ruminants coming from third countries all animals are tested. The animals sampling is performed by the authorized private veterinarian under the supervision of the official veterinarian. The samples are collected during the first 5 - 7 days after entering Romanian territory.

Vaccination. Vaccination against BTV serotype 4 of the susceptible animals is applied on a voluntary basis, at the owner’s (keeper’s) request. Inactivated vaccine (only with serotype 4) is used and is administered according to the protocol of the vaccine manufacturer. Vaccination is carried out by private veterinarians, under the surveillance and official control of the local sanitary veterinary and food safety directorate (Annex 5 lists complementary legislation in place).

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Table 7. Vaccinated animals under the surveillance and official control, Romania, 2014-2017

Year Number of vaccinated animals*

bovine animals ovine animals

2014 3794 0

2015 59832 8332

2016 74538 153143

2017 70166 28828

*indigenous animal population and for trade-movement purposes

V. Conclusions

Considering that:

Prior to the confirmation of the first case in August 2014, Romania had been free from bluetongue in ruminants;

The last outbreak of bluetongue serotype 4 was reported on 24 November 2015 and the event was closed on 4 January 2016:

Based on the surveillance programme, in accordance with Chapter 1.4. and Articles 8.3.14 to

8.3.17 of the Terrestrial Code, no evidence of infection with BTV in the country during the past

two years was demonstrated in Romania.

The Delegate of Romania declares that the country regained its bluetongue free status with effect

from 04 January 2018 in accordance with Article 8.3.3. of the OIE Terrestrial Code (2017).

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VI. Annexes

Annex 1

Passive surveillance includes:

- verification of relevant data, including sanitary-veterinary documents and other documents accompanying susceptible animal consignments coming from other member states of the European Union or third countries, before unloading at destination, according to the NSVFSA and Agricultural and Rural Development Ministry Order no. 129/566/2007 approving sanitary veterinary norm regarding veterinary and zootechnical controls applicable to intra-community trade with certain live animals and animal products;

- evaluation of health status of ruminants (cattle, sheep and goat) performed by the owners or their legal representatives and reporting of all clinical signs to authorized private or official veterinarians;

- clinical inspection of susceptible ruminants (domestic and wild) in Romania as follows:

- all ruminants entering into Romania following intra-community trade and all ruminants imported from third countries, during waiting period; the inspection is performed by the official veterinarian;

- indigenous ruminants slaughtered in authorized slaughterhouses, ruminants entering into Romania following intra-community trade or imported from third countries destined for slaughtering; the inspection is performed by the official veterinarian of the slaughterhouse;

- wild ruminants hunted under natural condition (with lesions following external inspection) or hunted because they show clinical signs of disease; it is compulsory for the wildlife officers of the hunting areas to notify the authorized private veterinarian regarding any alteration of the health status of wild animals or any lesions noticed in dead animals;

When, following inspection there is a suspicion of bluetongue disease, samples are gathered and sent to county sanitary veterinary and food safety laboratory or IDAH in order to verify the suspicion (whole blood with or w/o EDTA from live animals and organ fragments (spleen, lymph nodes, lung) from dead animals.

Active monitoring includes:

Serological monitoring

The selection of sentinel animals to be included in the monitoring programme is implemented between January - February, or the latest until the end of the vector free period, using the competition/blocking ELISA to determine if they are seronegative.

Sampling, both for sentinel selection and subsequent testing is performed by the authorised private veterinarian under the supervision of the official veterinarian. The animals with seroconversion during sentinel selection are eliminated from testing, and other animals are tested until completion of the necessary number of seronegative animals for the monitoring programme.

The selection of the sentinel animals is carried out only from the non-commercial holdings; their owners are informed about the „sentinel” status of theirs animal and their obligations.

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In addition to the necessary number of the sentinels, the group that will serve to replace the sentinels that are excluded from the programme of testing for some reason (sold by their owner, emergently culled, etc.). During the rest of the monitoring period, no other sentinels are selected apart from the initial number +10%. In case there are difficulties in selecting the sentinel animals in the target localities, the central authority may approve the selection of sentinels in localities around the target locality initially selected.

During the sampling activities in June, September and November, both blood samples for serological (ELISA) and virological (PCR) tests are collected.

The sentinel animals seroconverted during June – November testing are regarded as suspicions; the county sanitary veterinary and food safety directorate representatives carry out report of the suspicion notification and send it to National Sanitary Veterinary and Food Safety Authority, and also the animals are further tested by PCR on EDTA blood samples at the designated laboratories.

If within the county at least one outbreak of bluetongue is already declared, and if the target locality is situated within the 20 km radius area around this outbreak, then this locality is excluded from testing.

From sentinel animals with seroconversion, serum samples are sent to the National Reference Laboratory for bluetongue in order to determine the viral serotype by virus neutralisation (VNT). Maximum three positive serum samples/testing month/county are sent for VNT.

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Annex 2

Figure 8. Number of outbreaks of Bluetongue per county, Romania, 2014

Figure 9. Number of outbreaks of Bluetongue per county, Romania, 2015

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Annex 3

Table 7. Sampling model in June, September and November by counties, Romania.

No. County Total surface of

the county/km2

No. of target

localities

Total no. of

samples/month

Total no. of

samples/period

1. Alba 6242 3 177 1062

2. Arad 7754 4 236 1416

3. Argeş 6862 3 177 1062

4. Bacău 6621 3 177 1062

5. Bihor 7544 3 177 1062

6. Bistriţa Năsaud 5355 3 177 1062

7. Botoşani 4986 2 118 708

8. Brăila 4766 2 118 708

9. Braşov 5363 3 177 1062

10. Buzău 6103 3 177 1062

11. Caraş Severin 8514 4 236 1416

12. Călăraşi 5088 2 118 708

13. Cluj 6674 3 177 1062

14. Constanţa 7071 3 177 1062

15. Covasna 3710 2 118 708

16. Dâmboviţa 4054 2 118 708

17. Dolj 7414 3 177 1062

18. Galaţi 4466 2 118 708

19. Giurgiu 3526 2 118 708

20. Gorj 5602 3 177 1062

21. Harghita 6639 3 177 1062

22. Hunedoara 7063 3 177 1062

23. Ialomiţa 4453 2 118 708

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24. Iaşi 5476 3 177 1062

25. Ilfov 1583 1 59 354

26. Maramureş 6304 3 177 1062

27. Mehedinţi 4933 2 118 708

28. Mureş 6714 3 177 1062

29. Neamţ 5896 3 177 1062

30. Olt 5498 3 177 1062

31. Prahova 4716 2 118 708

32. Satu Mare 4418 2 118 708

33. Sălaj 3864 2 118 708

34. Sibiu 5432 3 177 1062

35. Suceava 8553 4 236 1416

36. Teleorman 5790 3 177 1062

37. Timiş 8697 4 236 1416

38. Tulcea 8499 4 236 1416

39. Vaslui 5318 3 177 1062

40. Vâlcea 5765 3 177 1062

41. Vrancea 4857 2 118 708

42. Mun. Bucureşti 228 1 59 354

Total 238411 114 6726 40356

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Annex 4

During 2017, entomological surveillance was implemented as follows:

– weekly catches with fixed traps placed in every county (1 fixed trap/county) in March and November or until relevant data is obtained, in order to determine the beginning and the end of the vector free period;

- monthly catches with fixed traps placed in every county (1 fixed trap/county) during the vector season, in order to establish the spatial and temporal abundance of culicoid vectors;

- monthly catches with fixed traps (1 fixed trap/county), inside shelters, during the vector free period.

The traps are placed only in the target localities where serological monitoring is in place.

For each trap collection point latitude, longitude and altitude coordinates will be recorded with GPS devices.

During the previous years, the entomological surveillance was performed by weekly catches with both permanent mobile traps during the vector activity period and monthly catches in the vector free period.

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Annex 5

The legal basis, in order to support the declaration of bluetongue free country status, represents the community legislation transposed into the national legislation, as follows:

• Transposed community legislation which establishes the strategy of the surveillance and control of bluetongue disease:

- Order no. 32/2006 for the approval of the Sanitary Veterinary Norm that establishes specific provisions for the control and eradication of bluetongue, with subsequent amendments; this Order represents the official transposition of the Council Directive 2000/75/EC laying down specific provisions for the control and eradication of bluetongue;

- Government Decision no. 1156/2013 for the approval of sanitary-veterinary actions foreseen in the Strategic programme for the surveillance, prevention and control of transmissible animal diseases from animals to humans, animal protection and environment, of identification and registration of, bovines, swine, sheep, goats and equidae, with subsequent amendments;

- Order no. 35/2016 for the approval of the applying methodological norms of Strategic programme for the surveillance, prevention and control of transmissible animal diseases from animals to humans, animal protection and environment, of identification and registration of, bovines, swine, sheep, goats and equidae, with subsequent amendments;

Transposed community legislation regarding the notification of transmissible diseases:

- Order 77/2005 for the approval of the Sanitary Veterinary Norm regarding the notification of animal diseases, with all subsequent amendments; this Order represents the official transposition of the Council Directive 82/894/EEC on the notification of animal diseases within the Community;

Transposed community legislation which establishes the identification of ruminants and the authorisation of holdings:

- Government Emergency Ordinance no 113/2002 regarding the identification and registration of bovine animals in Romania;

- Government Emergency Ordinance no 23/2010 regarding the identification and registration of swine, sheep and goats and for amending of other regulations, approved by the Law 191/2012 for approval of the Government Emergency Ordinance no 23/2010 regarding the identification and registration of swine, sheep and goats and for amending of other regulations;

- Order no. 16 of 16th March 2010 on the approval of „sanitary veterinary Norm regarding the procedure for the sanitary-veterinary registration/authorization of establishments/ assembly centre/holdings of origin and of means of transport in the field of animal health and welfare, of the establishments involved in the storage and neutralization of animal by products which are not intended for human consumption and of processed products”, published in the Official Journal of Romania, Part I, no. 194 of 26 March 2010, with further amendments;

- Commission Regulation (EC) no. 1760/2000 establishing a system for the identification and registration of bovine animals and regarding the labelling of beef and beef products and repealing Council Regulation (EC) No 820/97;

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- Commission Regulation (EC) no. 911/2004 of April 29, 2004 implementing Regulation (EC) No 1760/2000 of the European Parliament and of the Council as regards ear tags, passports and holding registers, with further amendments;

- Council Regulation (EC) no. 21/2004, of December 17, 2003 establishing a system for the identification and registration of ovine and caprine animals and amending Regulation (EC) No 1782/2003 and Directives 92/102/EEC and 64/432/EEC, with further amendments;

- Order 40/2010 on the approval sanitary veterinary Norm regarding the implementation process of identification and registration of swine, bovine, sheep and goat, published in the Official Journal of Romania, no. 286/30 April 2010, with further amendments.

• Transposed community legislation which establishes the intracommunity trade in animals:

- The NSVFSA Order no. 61/13.03.2006 for the approval of sanitary veterinary Norm regarding the animal health problems which affect the intracommunity trade in animals from bovine and swine species, which transposes the Council Directive 64/432/EEC regarding the animal health problems which affect the intracommunity trade in animals from bovine and swine species, with subsequent amendments;

- Commission Regulation (EC) No 1266/2007 on implementing rules for Council Directive 2000/75/EC as regards the control, monitoring, surveillance and restrictions on movements of certain animals of susceptible species in relation to bluetongue.

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