TCS Tree Based on L-Fragment

TCS tree based on L-fragment

Kosti village 11 nt changes 12LPN1 (IP 1)

4 nt changes TUR/18/2010 BUL/1/2010 35 nt changes (Gümüşhane) (wild boar) 09/07/2010 30/12/2010 12LPN3 (IP 4) 14 nt changes Rezovo village

putative common ancestor of the Bulgarian viruses

All nucleotide substitutions are unique Neighbor-joining tree based on Leader+P1 TUR36/2010

TUR18/2010

BUL 12LPN3 (Rezovo)

979

BUL 12LPN4 (Rezovo)

1000 BUL 12LPN1 (Kosti)

767

BUL 12LPN2 (Kosti)

BUL/12010 (wild boar) 0.001 Conclusions

• The FMD outbreaks in Kosti and Rezovo villages are unlikely to be directly linked. • Both have a common ancestor close to the virus from wild boar. • The long branches between the putative common ancestor and each of the two outbreaks examined (IP1 and IP4) suggest possible intermediate hosts (either wildlife or domesticated animals). FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 Every think started from ….

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 FAO-EU FMD /EC/OIE Tripartitt Group Meeting,Plovdiv, Bulgaria, 25 March,2011 One shot wild boar

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 One shot wild boar

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 Not FMD in wild boar

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 What tests we used?

Аg FMDV LFD FMFV Ag ELISA WRL FMDV Svanova One- step rRT-PCR (Reid, S. et all, 2002,) (Callahan et all., 2002) FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 Chronology • 29th December 2010 –wild boar with lesions in the feet, shot 2 km from the Turkish border; (42° 00‘409"N; 27°46'49.33"E) • 3th Jan – the sample sent to NRL • 4th Jan 2011 – Wild boar positive for - FMDV type O, • 9th Jan – 12 goats, 16 sheep,1 bovine and 8 pigs from the village of Kosti confirmed positive for NSP (42° 3'32.01"N; 27°46'49.33"E) FMDV type O • 15th Jan – clinical signs of FMD in bovines in the village of Rezovo; • 17th Jan – FMDV type O - Rezovo village, diagnosis confirmed by NRL – start of killing and burial on the spot

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 Chronology 31 January 2011 –FMDV type O in Gramatikovo 15 th February 2011, 10 h am. Samples sent to WRL Pirbright, UK . All confirmed O-type FMDV 18-th March 2011 samples from suspected new outbreak in village G.Bukovo arrived at the NRL – 22h am. 19-th March FMDV type O confirmed G.Bukovo 22-th March 2011 samples sent to WRL Pirbrihgt, UK 22-th March 2011 started culling of animals in village G.Dabovo 22-th March 2011 samples from suspected new outbreak in village Granichar arrived at the NRL 23-th March 2011 FMDV type O confirmed Granichar Village

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 SEQUENSING ANALYZIS AT WRL UK

• The analysis made from the samples by the OIE Reference Laboratory Institute for Animal Health, Pirbright, UK, indicate this FMDV is a serotype O, Topotype ME-SA, Strain PanAsia-2ANT-10 which has more than 99.5% affinity with samples isolated in 2010 in Iran and Turkey.

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 The sub-lineage of the O-PanAsia strain was identified in Pakistan and Iran and named O- Pan-Asia-2 and become widespread in the Middl East A number of distinct sub-lineages have recently evolved and have been named as follows: • PanAsia-2YAZ-09 • PanAsia-2FAR-09 • PanAsia-2ANT-10 • PanAsia-2BAL-09 • PanAsia-2SAN-09 • PanAsia-2PUN-10 • PanAsia-2Kosti-10 FAO-EU FMD /EC/OIE Tripartite Group Meeting, P lovdiv, Bulgaria, 25 March,2011 FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 © Daniel Dalet / d-maps.com 500 km

300 mi

Closed Sequences of Isolates of FMDV О-Pan Asia 2010 in Turkey and BG

O/BUL/1/2010 (Burgas 30/12/2010) O/TUR/926/2010* (Bursa 26/07/2010) Bulgaria O/TUR/1086/2010* (Antalya 16/08/2010) O/TUR/840/2010* (Agri 15/07/2010) O/TUR/868/2010* (Eskisehir 20/07/2010) Kastamonu O/TUR/1003/2010* (Sivas 10/08/2010) Gümüşhane O/TUR/18/2010 (Gumushane 09/07/2010) Bursa Eskişehir O/TUR/883/2010* (Kastamonu 23/07/2010) Sivas Ağrı O/TUR/1094/2010* (Giresun 11/08/2010) Turkey Iran Antalya O/TUR/35/2010 (Erzincan 13/08/2010) O/TUR/153/2010* (Gaziantep) O/TUR/154/2010* (Gaziantep) O/IRN/94/2010 (West Azerbaijan 15/04/2010) O/IRN/92/2010 (West Azerbaijan 14/04/2010) 100 O/AFG/59/2010 (Samangan 14/05/2010) 97 O/PAK/36/2010 (Gilgit-Baltistan 29/07/2010) O/TUR/36/2010 (Kocaeli 13/08/2010) O/UKG/12/2001 (AJ311724) O1/Manisa/TUR/69 (AJ251477) FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 0.01 • The closest sequences of these strains isolated during 2010 from 7 different provinces in Turkey was differed by only a single nucleotide from each other and from the Bulgarian FMDV sequence (isolated January 2011 from wild boar in Kosti village Burgas district)

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 TCS tree based on L-fragment

11 nt changes Kosti village 12LPN1 (IP 1)

4 nt changes TUR/18/2010 BUL/1/2010 35 nt changes (Gümüşhane) (wild boar) 09/07/2010 30/12/2010 12LPN3 (IP 4) 14 nt changes Rezovo village

putative common ancestor of the Bulgarian viruses All nucleotide substitutions are unique Neighbor-joining tree based on Leader+P1

TUR36/2010

TUR18/2010

BUL 12LPN3 (Rezovo)

979

BUL 12LPN4 (Rezovo)

1000 BUL 12LPN1 (Kosti)

767

BUL 12LPN2 (Kosti)

BUL/12010 (wild boar) 0.001 Conclusions made at WRL

O/BUL/2010 (wild boar) 73 O/Kosti/2/2011 99 O/Kosti/1/2011 BUL isolates 2010/ 2011

100 O/Rezovo/3/2011 95 O/Rezovo/4/2011 71 O Iran 1 2010 (HQ450645) 98 O/PAK/3/2008 (FJ798192) 39 O/PAK/70/2007 (FJ798188) O/BUL/1/96 (EU553839) 100 99 O/TUR/1/96 99 O/TUR/2/2005 FJ561319 O/TUR/7/2003 (DQ164991)

100 O IRN/1/2000 (DQ164892) 74 O/IRQ/30/2000 (DQ165057) 78 O/Konya/TUR/512/10/99 (DQ296515) 96 O/UKG/11/2001 (DQ404180) O1/Manisa/Turkey/69 (AJ251477) O/BUL/1/91 (EU553837) 100 O/BUL/1/93 (EU553838

0.01 Neighbor-joining tree, showing the relationships between FMDV isolates in Bulgaria 2010 – 2011,

77 O/BUL/2010 100 Rezovo/3 BUL isolates 2010/ 2011

71 Kosti/1 O/ISL/PAK/L1573/2009 (HQ113232) 100 O/PAK/48/2008 (GU384683) O/UKG/11/2001 (DQ404180) O Campos/Bra/58 (AJ320488)

0.01 Decision voted at the SCoFCAH meeting on 12th Jan Abd r

Shumen Varna

Sliven Burgas

Yambol

Haskovo Annex I

Kardjali Annex II

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Additional regions to Annex II Bulgaria, 25 March,2011 ------

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 bovines 194 Kosti – the village of the bones sheep 117 goats 149 pigs 72

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 Rezovo – the most southerly point on the Bulgarian Black sea coast

bovines 92

sheep 68 goats 9 pigs 28

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 21 Turkish buffalos

FMD positives:

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 LPBL ELISA and Mono-O-type ELISA Prionics Results from Buffaloes Sera

• Linear end-point titration mono O-type

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 TURKISH BUFFALOES - SEROLOGY RESULTS FMDV O, A and Asia-1 LIQUID PHASE BLOCKING ELISA (IAH)

O-type LPBL A-type LPBL ASIA-1 LPBL Buffaloes ELISA ELISA ELISA

1 (>1:256) (1:16) (1:32)

2 (1:256) (-) (1:16)

3 (>1:256) (1:16) (1:64)

4 (>1:256) (1:64) (1:64)

5 (>1:256) (1:16) (1:32)

6 (1:256) (1:16) (1:32)

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 TURKISH BUFFALOES - SEROLOGY RESULTS FMD MONO-O-TYPE PRIONICS LINEAR TITRATION ELISA RESULTS

Buffaloes Mono - O – type Priocheck Linear Titration

1 1:1024

2 1:256

3 1:1024/1:2048

4 1:1024

5 1:1024

6 1:256

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 FMDV MONO-O-TYPE PRIONICS LINEAR TITRATION ELISA and rRT- PCR RESULTS Mono - O - type Buffaloes Prionics linear One Step r RT-PCR titration

1 1:1024 neg 2 1:128 neg 3 1:1024 neg 4 1 : 64 neg 5 1:256 POS (ct 31.58) 6 1:256 neg 7 1:128 neg 8 1:128 neg

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 CATTLE AND BUFFALOES VIROLOGY RESULTS FROM WRL IAH PIRBRIGHT, UK

SPECIES EAR TAG LOCATION PCR R CELL RESULT CULTURE (ct value) ISOLATION

BUFFALO-TR 3953-1887 REZOVO 33.14 POS NVD BUFFALO-TR NO EAR TAG REZOVO 31.58 POS NVD CATTLE NO EAR TAG REZOVO 19.34 POS FMDV - O CATTLE NO EAR TAG REZOVO 18.27 POS FMDV - O CATTLE NO EAR TAG KOSTI 16.86 POS FMDV - O CATTLE NO EAR TAG KOSTI 20.47 POS FMDV - O

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 TURKISH BUFFALOES - SEROLOGY RESULTS FROM WRL IAH PIRBRIGHT, UK

Bufalloes Type –O / Type - A / Type – Asia 1 FINAL RESULT VNT VNT VNT

1 POS / 512 POS / 32 POS / 128 POS 2 POS / 128 POS / 40 POS / 16 POS 3 POS / 90 NEG /< 8 NEG / 8 POS 4 POS / 356 POS / 64 POS / 64 POS 5 POS / 178 POS / 178 POS / 45 POS 6 POS / 256 POS / 45 POS / 45 POS 7 POS / 355 POS / 355 POS / 64 POS FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 rRT-PCR Results from last outbreak investigated epithelium smaples

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 rRT-PCR Results from last suspicion investigated in village St. Karadjovo-16.02.2011

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 rRT-PCR Results from last outbreak investigated epithelium smaples

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 Cattle from St. Karadjovo village

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 Detection of FMDV by rRT-PCR in serum samples from viraemic animals

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 Samples taken for FMDV surveillance from 04.01.2011 to 22.03.2011 Number Wildlife of Number of Samples Animals Villages Municipalities tested tested tested Wild Boar 4 30 Wild Goat 1 5 White tile Deer 1 14 Domestic Ruminants 50 12 24 990 East Balkan pigs 22 21 2512 Total 27 551 FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011 Thank you for the attention!

FAO-EU FMD /EC/OIE Tripartite Group Meeting, Plovdiv, Bulgaria, 25 March,2011

The beginning

 4th Jan 2011, 22:00 p.m. – Wild boar positive for FMD type O;

+ 5th Jan • Start of the measures for control and eradication of FMD according to Dir. 2003/85/EC. • Clinical and epidemiological examinations as well as blood sampling of susceptible animals in all villages located in the risk area 1st outbreak –Kosti (09.01.2011)

• 100% Stamping out in Kosti • Clinical examinations on daily basis in the settlements in the 10km zone • 100% Serological surveillance in the settlements in the 10 km zone: - 1st sampling: ended on 11th Jan (all Neg.) - 2nd: 28th Jan - 2nd Feb (Gramatikovo pos.) - 3rd: 8-9th Feb (all Neg.) - 4th: 23-27th Feb (Neg.)

bovines sheep goats pigs Settlements № herds № animals № herds № animals № herds № animals № herds № animals

1. Kosti outbreak 4 196 21 121 44 205 8 136 Settlements in the 10km surveillance zone 4. Brodilovo 3 8 26 117 47 164 3 44 5. Balgari 0 0 5 39 6 12 0 0 6. Kondolovo 0 0 3 48 1 39 0 0 7. Izgrev 0 0 1 4 0 0 1 13 3. Gramatikovo 1 1 5 192 24 111 0 0 Total 8 205 61 521 122 531 12 193 2nd outbreak –Rezovo (15.01.2011)

• 100% Stamping out in Rezovo • Clinical examinations on daily basis in the settlement in the 10km zone • 100% Serological surveillance in the settlement in the 10 km zone: - 1st sampling: 28th Jan - 2nd Feb (all Negative) - 2nd: 8-9th Feb (all Neg.) - 3rd: 23-27th Feb (Neg.)

bovines sheep goats pigs Settlements № herds № animals № herds № animals № herds № animals № herds № animals

1. Rezovo outbreak 6 87 2 68 2 8 1 29 Settlements in the 10km surveillance zone 8. Sinemorets 3 14 9 220 10 79 6 9 3rd outbreak – Gramatikovo (31.01.2011)

• 100% Stamping out in Gramatikovo • Clinical examinations on daily basis in the settlements in the 10km zone • 100% Serological surveillance in the settlements in the 10 km zone: - 1st sampling: ended on 8th Feb (all Neg.) - 2nd: 13-16th Feb (all Neg.) - 3rd: 23-27th Feb (Neg.)

Large ruminants sheep goats pigs Settlements № herds № animals № herds № animals № herds № animals № herds № animals

3. Gramatikovo 1 1 5 192 24 111 0 0

Settlements in the 3 km and 10km surveillance zone protection zone 4. Kondolovo 0 0 3 48 1 39 0 0 1. Slivarovo 0 0 3 38 3 48 3 12 2. Zabernovo 3 22 3 9 12 112 8 61 3. Vizitsa 4 57 5 84 2 21 0 0 5. Balgari 0 0 5 39 6 12 0 0 6. Kosti 0 0 0 0 0 0 0 0 Total 8 80 24 410 48 343 11 73 +

10 km 3 1

WB 2

WB Wild boar shot 1 Kosti 2 Rezovo 3 Gramatikovo Clinical examinations on daily basis and blood sampling of domestic animals in the villages out of the 10km zones

Sero-surveillance from 2 to 5 times of the susceptible animals in all villages carried out (7th Jan – 27th Feb): - 100% of the animals in the villages bordering to Turkey - All other villages - under the scheme 5% prevalence and 95% confidence Decision 2011/44 EC

Shumen Varna

Sliven Burgas

Yambol

Haskovo Annex I Kardjali Annex II Outbreak of 19th March

Sredets

Infected farm 42 km

Outbreak of 24th March

Surveillance program for FMD in wildlife Surveillance in wildlife 16 WB + 6 roe deers tested neg. so far Experimental Infection of Wild Boar and Pigs with Food and Mouth Disease Virus from Bulgaria: Preliminary Results Angele Breithaupt, Klaus Depner, Bernd Haas, Martin Beer

Two wild boar four months old were experimentally inoculated in the “bulb of the heel” with FMD virus strain isolated from a wild boar in Bulgaria in January 2011. The animals received 0.2 ml cell culture medium of the second passage of IBRS-2 cells, containing 106.5 50% TCID50 of FMD virus. One day later two wild boar of same age and two domestic pigs (8 weeks old) were placed as contact animals in the same containment room. Body temperatures and clinical signs were recorded daily during the experimental period of 4 weeks. Every two to three days blood samples for virological and serological investigations were taken from each piglet. For monitoring virus excretion nasal and saliva swabs were taken daily.

First clinical signs were noticed in the domestic pigs on day 2 post exposure (DPE). Both animals had fever (up to 41.2 °C) and poor appetite, lameness and swelling and reddening of the coronary band. Vesicles appeared on the snout and the digits. On 3 DPE pronounced symptoms were seen with numerous ruptured and unruptured vesicles affecting every feet and the snout. The pigs had to be euthanized due to animal welfare reasons.

First symptoms in wild boar were seen on 4 days post infection (DPI) and 4 DPE, respectively. The lesions started in the inoculated animals with vesicles on the dorsum of the snout and the interdigital space. During the next days (until 7DPI/DPE) several ruptured vesicles with serofibrinous in-filling were detectable on the interdigital space, the coronary band, the digits, the heel, claws and the lips. Food intake was reduced but activity was unaltered and lameness did not occur. Increased body temperature (40.5 °C) was measured between 3 and 5 DPE in contact wild boar.

In contact animals, viraemia started on 2 DPE and lasted until death (domestic pigs) or maximum until 6 DPE in wild boar. Virus excretion started 1 DPE with viral titres up to 105,75 TCID50 per mL and continued until 9 DPE in wild boar. Viral RNA in serum was detected up to 8 DPE, in saliva for at least 16 DPE (subsequent samples are under investigation). Antibodies (Priocheck® NSP) were detected on 7 DPI (needle infected wild boar) and 8 DPE (contact wild boar), respectively.

In needle infected wild boar, viraemia started on 1 DPI and lasted until 9 DPI. Virus excretion 5,25 started 1 DPE with viral titres up to 10 TCID50 per mL and continued until 12 DPI.

The most striking result of the described experiment is the discrepancy in the clinical course between domestic pigs and wild boar. While the domestic pigs had to be euthanized due to severe clinical FMD, the general condition of wild boar was less affected. The wild boar started to shed virus already one day after infection without showing any clinical signs for several days (incubation period 4 days). Although the food lesions seen in wild boar around 7DPI/DPE appeared severe, the animals were not handicapped in their movement ability. These findings suggest that an infected wild boar may survive the disease and shed FMD virus over a prolonged period (in our experiment 9 days post exposure).

FAO-EUFMD/EC/OIE Tripartite Group Meeting on control of FMD and other exotic diseases in the Southern Balkans/Aegean region 25th March 2011 Venue: Park Hotel, Plovdiv, Bulgaria

Provisional Time-Table

THURSDAY 24th (PM) FRIDAY 25th (AM) Arrival of participants. 08.30 Opening of the Tripartite meeting The participants will be met on arrival at 10.00 Coffee break Sofia International Airport, and taken by 12.30 Closing car to Plovdiv. 13.00 Lunch/Departure Thursday evening : Reception/Meal

Technical Items

Italics indicate the Country/Institution hereby requested to make a presentation

Item 1 Adoption of the Agenda.

Recap of the Recommendations of the EuFMD Executive Committee, 2nd February (K Sumption, EuFMD);

Part I : Situation reports

Item 2 FMD situation and FMD control programme in Bulgaria

2.1 Situation report (Provided by NVS, Bulgaria) including  report on surveillance activities and control measures since last meeting on 2nd February  Results of the Wildboar/wildlife surveillance conducted February-March

2.2 Epidemiological insights from FMDV genome sequencing of Bulgarian FMDV conducted at EU-CRL and at Lindholm, Denmark (Dr Georgiev, NVS, or by powerpoint and teleconference link to Pirbright/Lindholm)

2.3 Status of studies in wild boar conducted by the FLI on Bulgarian FMDV isolates

Discussion

FMD situation and FMD control programme in Turkish Thrace

2.4 Report on surveillance activities and control measures since last meeting on 2nd February (GDPC, Turkey)

Including:  serological surveillance program  Results of the Wildboar/wildlife surveillance  control measures applied in the period, and to be applied in coming months  timing of re-vaccination program  FMD situation in Anatolia

FMD situation and FMD control programme in neighbouring parts of Greece (Dr Doudonakis)

2.5 Report on surveillance activities and control measures since last meeting on 2nd February

Part II Associated scientific and technical developments

Item 3 The role of wild boar in FMDV epidemiology; issues and potential scientific study (Dr Khomenko, FAO)

Item 4 Strengthening FMD laboratory capacity in the Balkan region : potential support programme (Dr Sumption, EuFMD)

FMD CONTROL IN TURKEY

H. Haluk AŞKAROĞLU Head of Animal Health Services Department , GDCP A. Naci BULUT Head of the Diagnosis Department, Şap Institute

the EuFMD Commission Tripartite Meeting, 25March 2011 Sofia, Bulgaria Control of FMD in Turkey CURRENT SITUATION

• FMD is endemic in Anatolian Region of Turkey (Types A and O).

• Field strains circulating in Turkey are:

• A Iran 2005 (endemic in Anatolia since Nov 2005) •Sub linage A AFG-07(DEN10) New!

• New variant of Type O Panasia II (endemic in Anatolia since Oct 2006) •Sub linage O ANT-10 (2010)

• No case of Asia-1 has been reported since 2002.

• 1702 outbreaks occurred in Anatolia in 2010.

• FMD has not occurred in Thrace since 12/09/2007.

• THRACE REGION IS FMD-FREE WITH VACCINATION  Type A05 IranVaccine: Sufficiency  Vaccine strain: A Tur 2006 replaced with A22 Iraq on July 2008;  However, recent some r value results inticated that A Tur 2006 does not cover fully to field isolates?

 Type O :  Vaccine strain: O1 Manisa. covers current field isolates antigenically;

 It was detected with low r value from some isolates ,

 A decision was made to change the current strain with a homolog of circulated field virus. (O Tur-07)  Current vaccine formula contains both O Manisa and O Tur-07 (Pan Asia II) vaccine strains

3 FMD OUTBREAKS (2006-2011) 1715 1800 1557 1600 1400 1200 1000 809 800 667* 600 400 253 214 200 0 2006 2007 2008 2009 2010 2011

* 20th March DISTRIBUTION OF FMD OUTBREAKS BY PROVINCES IN 2010

L.Yellow:1-4 Yellow:5-9 L.Green:10-19 D.Green:20-29

Blue:30-39 Purple: 40-49 Red: >50 Control of FMD in Turkey

SEROTYPE DISTRIBUTION OF 2010 OUTBREAKS FMD-A; 155; 9% FMD-?; 559; 33%

FMD-A

FMD-O

FMD-?

FMD-O; 1001; 58%

Serotype Total A 155 O 1001 Unidentified(not sampled or untyped) 559 Total 1715 Comparison of FMD Situation in 2009 and 2010  Average number of animals infected per outbreak;  In 2009: 15,8

In 2010: 44,0

 Average percentage of animals infected per outbreak; In 2009: 3,2%

In 2010: 6,9% FMD Situation in 2010  Average percentage of animals infected per outbreak in some selected provinces based on TURKVET data;

 Burdur: 0,6 %

 Denizli 0,8 %

 Bursa: 1,0 %

 Konya: 3,9 %

 Ardahan: 10,8 %

 Van: 11,5 %

 Erzurum: 23,0 % Monthly Distribution on FMD Outbreaks

300 256 269 221 228 200 155 115 49 75 100 38 91 97 121 2009 17 12 18 19 15 24 2010 0 17 12 22 28 5 25 1 2 3 4 5 6 2009 7 8 9 10 11 12 2011 Disease Situation

TYPE A

275 294 41% 44% TYPE O

UNIDENTIFIED 96 (UNTYPED OR 15% NONSAMPLED)

UNIDENTIFIED (UNTYPED MONTH TYPE A TYPE O OR NONSAMPLED) TOTAL OCAK 220 76 101 397 ŞUBAT 73 22 127 222 MART 1 47 48 TOTAL 294 96 275 667 Map Distribution in timeline of DEN10/AFG-07 Genetic Diversities  Type O :  New O Pan-Asia sub-strain (Group IIA)

 Detected 4 sublineages in Turkey in 2010 (4-6% difference observed):  O SAN-09,  O TER- 08,

 O FAR-09 (exotic),  O ANT-10 (exotic).

 Type A05 Iran :  With 6-7% differrences, genetic lineage occured due to the type A in 2010:  A ARD/ERC-09,  A ARD/NEV-09,

 A AFG-07 /DEN10(exotic) NEW!

12 O PanAsia – II ANT-10 GLOBAL SITUATION

DEC-2009 JAN-2010 FEB-2010 Mar-2010 APR-2010 MAY-2010 JUN - 10 Control of FMD in Turkey

CONTROL MEASURES

• Routine control measures in case of disease outbreak (restriction, quarantine, ring vaccination, sampling, diagnosis)

• Epidemiological investigation • Active and passive surveillance • Outbreak investigation and case study • Follow-up

• Mass vaccination

• Control of animal movements

• Training and communication activities PREVENTIVE MEASURES AND SURVEILLANCE IN THRACE- 2010/2011 Preventive Measures and Surveillance in Thrace -2010  All livestock were vaccinated;  LR twice a year/SR once a year

 Animal movements  from Anatolia were controlled; execute OIE TAHC

 Check points  were established on bridges and harbours for animal movements

 Passive and active surveillance  was implemented regularly

 Regular serosurveillance was carried out Additional Measures in Kurban Festival

 Animals were imported from FMD-free countries;

 Animals from Anatolia not accepted

 All livestock were checked systematically by clinical surveillance before and after the festival

 Temporary animal markets designated for the festival were tightly checked Serosurveillance 2010 in Thrace  Serosurveillance  was carried out in order to substantiate FMD-free status with vaccination  Sera were collected  from a total of 12,758 animals in the period Sept-Nov 2010.

 The sample design was two-stage with villages selected at random and then animals within villages.

 The samples were tested using the CEDI NSP ELISA which has a specificity of 99.1% and a sensitivity of around 88%. Results of Thrace Serosurveillance  The overall initial positive rate is 0.38%.  Given the specificity of the NSP test used, this is well within the expected rate of positive test results i.e. within the expected false positive result frequency.  The distribution of weak and strong positives (45 and 3 respectively) indicates that most are weak positives, again suggestive of false positive results.  In total 48 sera were positive within the 12745 sera; 18 by Cattle and 30 by SR.  There is no clustered positivity within the individual unit indicating experienced infection ; and also there is no clustered positivity by age group.  Confirmatory tests, Western Bloting(WB) and LPBE were performed for all positive sera.  All positive sera were titrated by LPBE; there is no high titre indicated postinfectional antibody  There is no positive results by WB. Follow-up Investigation (FI)Results  FI was made for identification test results:  Epidemiological inquiry in the positive village  All animals were sampled in the positive premises  Random sampling were made in positive villages 2408 sera were collected from 45 positive villages and tested by NSP ELISA  Results 22 sera (9 from cattle and 13 from SR) were positive by NSP There is no high titre by LPBE WB testing has been pending Additional measures in Thrace in response to the outbreak in Bulgaria Measures put in place  All livestock markets have been closed,

 Animal movements stopped,

 Livestock are to be kept confined in the areas defined as risk areas.

 In the areas defined as risk areas, including the villages close to the Bulgarian border, supplementary vaccination has been implemented for all susceptible animals Additional Surveillace in Thrace in Response to the Outbreak in Bulgaria  Surveillance:  Active clinical surveillance:  Active clinical surveillance is being carried out in the villages located in borderline and forest particularly in Demirkoy with frequent examinations of animals by the district veterinary service.

 Additional manpower had been brought to the district.

 Wild boar surveillance:  An investigation has been implemented by collecting tissue samples, plain blood and sera from hunting the wild boars located in the forestry area in Thrace region Additional Surveillace in Thrace in response to the outbreak in Bulgaria(2)  Additional Serosurveillance  Serosurveillace in addition to Autumn 2010 was implemented in order to investigate probable risk introduction;  322 villages have been identified as being inside or adjacent to forested areas in Thrace  A two-stage sample design has been used with a confidence level of 97.5% for each stage.  The prevalence  Estimate at village level is 2% and for animals within a village at 5% with 97,5% confidence interval.  The NSP test sensitivity is estimated at 88%.

 147 villages and 80 animals (40+40 for LR and SR) within each unit were selected.

 Age of animals was clustered in 4-24 months

 Overall sample size is potentially 11,760. Additional Surveillace in Thrace in response to the outbreak in Bulgaria(3)  Results of Serosurveillance:  A. Wild boar Surveillance 44 wild boar were sampled:  30 Tissue Sample (Toung/Interdigital tissue)  24 Blood  26 sera were received. No virus detected from tissues and blood samples. 4 out of 26 sera were positive by NSP (WILD BOAR SERA RESULTS.xlsx) and weak ab titre by LPBE. Additional Surveillace in Thrace in response to the outbreak in Bulgaria(4)  B. Domestic Livestock Surveillance Results  10932 sera were tested by NSP ELISA (CediDiag.)  29 sera (9 and 20 from cattle and SR respectevely) were positive by NSP ELISA (prosseced data for foresty villages in Thrace Survey 2011.xlsx)

 the overall positive rate is 0,26.

 There is no strong positive and clustered results within the village and age group.  All positive sera were titrated by LPBE; there is no high titre (the highest titre 1/712 for both types) indicated postinfectional antibody  WB testing is pending  Follow up investigation will be implemented in order to asses NSP positivity.

Wild boar and FMD: issues and study opportunities

Sergei Khomenko

Wildlife and Disease Ecology Consultant EMPRES-GLEWS

Plovdiv, Bulgaria, 25.03.2011 Historical range of Sus scrofa

3 ???

1 4 3

2 FMD in wild boar

1. Caucasus 3. Europe 4. Kazakhstan 1902 1920s? 1927 1908 2011 1931 1911 1941 1917 2. Israel 1919 1987-99 1925 2007 Marek & Hutÿra, 1931; Sludskiy, 1956 Wild boar density in Russian Federation

Low High

??? What FAO is doing

• Population/harvest estimates have been collected for the whole of N Eurasia at the sub-national level; • Detailed distribution maps on the country by country basis collected; • These will be used to model wild boar densities and population dynamics in the nearest future. WB &FMD: Epi and eco notes

• Poorly documented and rarely (if ever) studied in detail in the field (mainly through modeling exercises); • Delay in clinical manifestation of 2-3 days as compared with domestic animals reported; • Always concurrent with FMD epidemics in livestock, which is typically implicated as a source of infection; • Usually domestic pigs are affected first (if there are any); • Along with wild boar FMD sometimes recorded in other wild ungulates (e.g. cases in the Caucasus, Israel). • Wild boar is unique among other ungulates as the only scavenging species (!) Occurrence of FMD (1388-1389, Farsi calendar) and wild boar range in Iran

FMD outbreaks Wild boar range

http://www.wildpigconference.com/proceedings09/karami.pdf FMD outbreaks in Turkey (2010-11)

Only 40 % are shown as remaining 60 % are yet to be georeferenced Provinces with highest density of wild boar in Turkey

FMD x Wild Boar Proposed FMD in WB surveillance project in Turkey

• Analyze retrospective FMD data for livestock (occurrence, sero-prevalence, incidence) at the level of provinces; • Define locations/facilities for sampling shot wild boar (forests along the Black Sea coast/safari or hunting clubs?); • Implement field surveillance according to the developed design and protocols. Points and consideration

• Wild boar population estimates to define sample sizes are ideally needed; • Local (Turkish) wildlife expertise to better understand and interpret results epidemiologically; • Which logistical arrangements are needed; • Cost estimates for this sampling effort; • Whether samples can/should be tested for other diseases (CSF?); • It would be good to have a look at the external parasites as well. Measures of the Greek Authorities following the confirmation of the first FMD outbreak in Bulgaria (5 Jan 2011)

Circular letter : 132619 /07.01.2011 Increased awareness – vigilance of the regional veterinary authorities (domestic-wild animals) Recording of all live animal – animal products consignments originating from Bulgaria since 1 Dec 2011 Precautionary collection of blood samples from all consignments of live animals of the susceptible species (10% from each lot) originating from Bulgaria henceforth

Circular letter : 132620 /07.01.2011 Increased awareness – vigilance among hunters Description of clinical signs among susceptible wild species and directions to follow in case of suspicion Strong recommendation to hunting associations to advise their members to avoid travelling to Bulgaria for hunting

Circular letter : 132633 /13.01.2011 Targeted sero-surveillance in areas close to Bulgaria - Turkey Serosurveillance against FMD in Greece - Modalities:

Minimum number of holdings to be sampled per region (10-20 depending the size of the region)

Samplings in holdings close to the border (within 20 km) with Bulgaria –Turkey

In islands sampling from areas that may be at risk (e.g. close to the Turkish Coast

2/3 of samples from small ruminant holdings , the rest from bovines –swines e.t.c. Serosurveillance against FMD in Greece - Results (Jan –Mar 2011)

NUMBER OF RESULTS AREA SAMPLES POSITIVE NEGATIVE PENDING SERRES 450 0 450 0 DRAMA 450 0 450 0

XANTHI 1150 0 1150 0

RODOPI 498 0 498 0

EVROS 540 0 540 0

N ORTHERN EVROS 460 0 460 0

LESVOS Island 450 0 450 0

LEMNOS Island 100 0 100 0

CHIOS Island 470 0 470 0

SAMOS Island 172 0 172 0

IKARIA Island 64 0 64 0

RODOS Island 482 0 482 0

KOS Island 223 0 223 0 TOTAL 5509 0 5509 0 1150/0 498/0 450/0 450/0 460/0 No of blood samples tested serologically for 540/0 FMD in Greece 100/0

Period: 450/0 Jan-Mar 2011

470/0

No of samples 172/0 per region:

Tested /Posive 72/0 223/0

482/0