eufmd etc4 19th-22nd october 2009 eastern anatolia …erzurum is the largest province in eastern...

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EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia Region, Erzurum, Turkey Clinical and Epidemiology Report of Investigations Executive summary 1. On October 20 th and 21 st 2009 respectively, the EUFMD ETC4 training group visited the villages of Velibaba and Ortadüzü in the province of Erzurum. The aim of the visits was to collect epidemiological and clinical data regarding potential outbreaks of FMD and take samples to confirm the presence of the disease. 2. Each village was regarded as one epidemiological unit as all animals in each village has regular direct and/or indirect contact with each other. 3. In the village of Velibaba, the oldest lesions in the examined animals were estimated to be 7 to 10 days old. Based on this finding and considering the most commonly incubation period of 3 to 5 days, it was estimated that the most likely introduction of FMD occurred between October 5 th and October 10 th 2009. 4. In the village of Ortadüzü, the oldest lesions in the examined animals were estimated to be 2 to 3 days old. Based on this finding and considering the most commonly incubation period of 3 to 5 days, it was estimated that the most likely introduction of FMD occurred between October 13 th and October 16 th 2009. 5. Samples of blood, loose epithelium from vesicle lesions, vesicle fluid and saliva samples were collected in 20 animals for immediate investigation of antibodies and virus antigen using NSP ELISA, Ag ELISA and LFD, as well as later analyses at the NRL in Ankara (ŞAP-institute). Probang samples were taken from two animals in the village of Velibaba. 6. The SVANODIP® FMDV-Ag test was used on epithelium samples from 2 animals in Velibaba and on 3 epithelium samples, 2 saliva samples and one sample of vesicle fluid from 3 different animals in Ortadüzü. This test is a rapid and simple direct LFD-test for the detection of all seven serotypes of the FMDV antigen in swab and tissue samples. All samples tested were positive. However, the samples from Ortadüzü gave a weaker band than the ones from Velibaba. A possible explanation for this is that the animals in Ortadüzü had been treated by topical application of a disinfectant that may have disrupted the antigenic properties of the virus in the lesions. 7. The Ag ELISA test was used on 2 samples from animals in Velibaba and 3 samples from animals in Ortadüzü. In both villages type O of FMD was confirmed. 8. The NSP ELISA was used on blood samples from 17 animals in Velibaba and 3 animals in Ortadüzü. Although some results may indicate a longer presence of the virus in Velibaba, the results of the analysis are for the greater part in line with the estimated time that has elapsed since the introduction of the virus in the village. In Ortadüzü, all 3 samples were already positive despite the short time that had elapsed since the introduction of the virus in this village (the animals showed lesions that were only 2 to 3 days old). One possible explanation for this positive reaction is that the animals were quite young and may have been exposed to large quantities of virus, prompting an earlier than usual immune response to NSP.

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Page 1: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

EUFMD ETC4

19th-22nd October 2009 Eastern Anatolia Region, Erzurum, Turkey

Clinical and Epidemiology Report of Investigations

Executive summary

1. On October 20th and 21

st 2009 respectively, the EUFMD ETC4 training group visited the villages of

Velibaba and Ortadüzü in the province of Erzurum. The aim of the visits was to collect epidemiological

and clinical data regarding potential outbreaks of FMD and take samples to confirm the presence of the

disease.

2. Each village was regarded as one epidemiological unit as all animals in each village has regular direct

and/or indirect contact with each other.

3. In the village of Velibaba, the oldest lesions in the examined animals were estimated to be 7 to 10 days

old. Based on this finding and considering the most commonly incubation period of 3 to 5 days, it was

estimated that the most likely introduction of FMD occurred between October 5th and October 10

th 2009.

4. In the village of Ortadüzü, the oldest lesions in the examined animals were estimated to be 2 to 3 days

old. Based on this finding and considering the most commonly incubation period of 3 to 5 days, it was

estimated that the most likely introduction of FMD occurred between October 13th and October 16

th 2009.

5. Samples of blood, loose epithelium from vesicle lesions, vesicle fluid and saliva samples were collected

in 20 animals for immediate investigation of antibodies and virus antigen using NSP ELISA, Ag ELISA

and LFD, as well as later analyses at the NRL in Ankara (ŞAP-institute). Probang samples were taken

from two animals in the village of Velibaba.

6. The SVANODIP® FMDV-Ag test was used on epithelium samples from 2 animals in Velibaba and on 3

epithelium samples, 2 saliva samples and one sample of vesicle fluid from 3 different animals in

Ortadüzü. This test is a rapid and simple direct LFD-test for the detection of all seven serotypes of the

FMDV antigen in swab and tissue samples. All samples tested were positive. However, the samples

from Ortadüzü gave a weaker band than the ones from Velibaba. A possible explanation for this is that

the animals in Ortadüzü had been treated by topical application of a disinfectant that may have disrupted

the antigenic properties of the virus in the lesions.

7. The Ag ELISA test was used on 2 samples from animals in Velibaba and 3 samples from animals in

Ortadüzü. In both villages type O of FMD was confirmed.

8. The NSP ELISA was used on blood samples from 17 animals in Velibaba and 3 animals in Ortadüzü.

Although some results may indicate a longer presence of the virus in Velibaba, the results of the analysis

are for the greater part in line with the estimated time that has elapsed since the introduction of the virus

in the village. In Ortadüzü, all 3 samples were already positive despite the short time that had elapsed

since the introduction of the virus in this village (the animals showed lesions that were only 2 to 3 days

old). One possible explanation for this positive reaction is that the animals were quite young and may

have been exposed to large quantities of virus, prompting an earlier than usual immune response to

NSP.

Page 2: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

9. The latest FMD outbreaks in the province of Erzurum in 2009 prior to the outbreaks in Velibaba and

Ortadüzü were notified in June.

10. There were many contacts, both indirect and direct, that could have introduced FMD virus into the two

villages. Among these, high risk contacts such as live animal trade, both directly and via dealers or

markets, and pasture contact with animals from other villages were regarded as the most likely routes of

introduction of FMD virus in the village.

11. It was difficult to obtain exact information on which to base assumptions about the introduction of FMD to

each village as well as the possible further spread of the virus. The information that could be retrieved

from the TürkVet system in Erzurum did not allow verifying up-to-date vaccination details for the

examined animals, nor could the system be used for contact tracing as it was not possible to obtain

detailed information on animal movements to and from each village.

12. The endemic situation of FMD in Turkey means that many different contacts may spread FMD virus. The

disease is well known by animal owners and is regarded as an ordinary feature of animal husbandry.

This means that owners are expected to recognise the signs of the disease but not necessarily feel the

need to react on them, something that may contribute to the risk of spread.

Introduction

General information

Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the

province is elevated. Depression plains are located between the mountains and plateaus. The province has

a population of about one million people. The largest city and provincial capital is Erzurum, which has a

population of about 400,000. For administrative purposes, the province is divided into 18 districts. Each

district is subdivided into villages. Each district has its Director of Agriculture while each village has its

Headman (Muhtar) with important authority in administration at village level.

Erzurum has the highest ratio of meadows and pastures in Turkey, ideal for stockbreeding. There are about

500,000 cattle in the province. Livestock breeding is based on communal grazing by cattle, sheep and goats

in individual villages. All the animals owned by different farmers share grazing land around the village from

spring to autumn. The animals are usually brought back to the barns each night. During winter, the animals

are kept in barns within the village and fed on hay. Each family has its own barn. The area knows a system

of transhumance (called ‘Yayla’ in Turkish) that is practiced during the summer months: part of the cattle is

then moved to highland pastures for grazing, usually from June to September. These normally include

neither milking cows nor un-weaned calves.

Turkey has a mandatory vaccination programme for FMD in cattle and sheep. Large ruminants are

vaccinated twice a year and small ruminants once per year. In the province of Erzurum a bivalent vaccine

was used in 2008 and 2009 to cover FMD serotypes O and A.

Despite the vaccination, FMD is considered to be endemic in the whole country except the Trace region. The

province of Erzurum has one of the highest incidences of the disease in Turkey.

Table 1 and figure 1 summarize the outbreaks in the Erzurum province that were notified to the Turkish

veterinary service in 2009. The most recent outbreak was notified in June 2009.

Table 2 and figure 2 summarize the outbreaks in the Horasan district in the eastern part of Erzurum province

Page 3: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

that were notified to the Turkish veterinary service since 2007. No outbreak had yet occurred in this district in

2009: the last outbreak dated from December 2008, following a few outbreaks in the first half of 2007.

Table 1. Outbreaks of FMD notified in the Erzurum province in 2009 (source: Erzurum veterinary service)

notification no province district village FMD type outbreak date confirmation

date

25/09/00003 Erzurum Oltu Iğdeli not known 10.01.2009 16.01.2009

25/09/00018 Erzurum Oltu Merkez-Aslanpaşa

not known 15.01.2009 27.01.2009

25/09/00047 Erzurum Yakutiye Umudum O 02.02.2009 12.02.2009

25/09/00047 Erzurum Yakutiye Umudum O 02.02.2009 12.02.2009

25/09/00057 Erzurum Aşkale Abdalcik O 24.03.2009 08.04.2009

25/09/00057 Erzurum Aşkale Abdalcik O 24.03.2009 08.04.2009

25/09/00129 Erzurum Palandöken Tuzcu not known 28.05.2009 28.05.2009

25/09/00120 Erzurum Pasinler Kevenlik O 28.05.2009 29.05.2009

25/09/00120 Erzurum Pasinler Kevenlik O 28.05.2009 29.05.2009

25/09/00121 Erzurum Oltu Çatak not known 03.06.2009 04.06.2009

Figure 1. Map of the outbreaks in the Erzurum province in 2009

Table 2. Outbreaks of FMD notified in the Horasan district since 2007 (source: Erzurum veterinary service)

notification no province district village FMD type outbreak date confirmation

date

25/07/00007 Erzurum Horasan Dalbaşi O 17.01.2007 24.01.2007

25/07/00033 Erzurum Horasan Kizlarkale A 15.02.2007 02.03.2007

25/07/00074 Erzurum Horasan Kirik O 12.05.2007 23.05.2007

25/08/00323 Erzurum Horasan Kizlarkale O 25.12.2008 30.12.2008

Page 4: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

Figure 2. Map of outbreaks in the Horasan district since 2007

Information regarding the visits made

The ETC4 team had the opportunity to visit two villages in the Erzurum province that had notified recent

suspected FMD outbreaks in their cattle to the Erzurum veterinary service.

- Velibaba (Aras) was visited on October 20th 2009. It is a village in the district of Horasan in the east of

the province. The economy of the village revolves almost entirely around farming; most of the villagers

are involved in it. There are approx. 50 households in the village that rear cattle, sheep and goats.

- Ortadüzü was visited on October 21st 2009. This village is located in the district of Erzurum, at about

10 km from the city of Erzurum. The economy of Ortadüzü also mainly revolves around farming and as

is the case in Velibaba most of the villagers are involved in it. There are approx. 80 households in the

village that rear only cattle.

During each visit the team collected information from the villagers about the village and its farming practices,

performed an investigation on both FMD diseased and apparently healthy animals of the village and made

inquiries about possible routes of introduction and spread of the disease.

The clinical examination performed on an animal included firstly a visual examination of the animal with

specific attention to hypersalivation, lameness and lesions on foot and muzzle. For most animals examined,

this first superficial examination was followed by a close and thorough inspection of the lips, gums, tongue,

dental pad, nostrils, coronary band and the interdigital space. Where indicated, the temperature of the animal

was also taken.

Sampling was performed both on clinically healthy animals and animals showing clinical signs. Samples

included blood sample, epithelium, saliva, vesicle fluid and probang samples. In most animals several

samples were taken. An overview of the samples taken is given in tables 3 and 4, attached at the end of this

document.

The information in the tables regarding individual animals and their vaccination status was retrieved from the

Turkish identification and registration system TürkVet. It is important to notice that regarding the vaccination

status of the animals examined discrepancies might be observed when the information of the tables is

compared to the information forwarded by the villagers. These discrepancies are most likely due to the fact

that the TürkVet system is not yet fully operational with regards to the vaccination status of individual

animals.

Page 5: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

Investigations and information concerning the Velibaba village

Timeline

The timeline below summarizes the major events regarding Velibaba.

15-Aug-09 25-Oct-09

20-Aug-09

return from transhumance

of animals that later on got sick

18-Oct-09

notification ofsuspicion to

veterinary service

20-Oct-09

visit byETC4 team

13-Oct-09

first observation

of clinical signs

26-Sep-09

start of possible

window ofintroduction

of FMDV

10-Oct-09

end of likelywindow of

introductionof FMDV

10-Sep-09

vaccination of village herd

5-Oct-09

start of likely

window ofintroduction

of FMDV

Clinical examination and sampling of the animals

In the Velibaba village, animals of 3 different barns were examined.

- In the first barn, all 3 animals present were examined thoroughly (cf. table 3; animals no 1 to 3 owned

by Necmettin Durgun).

- In the second barn, some 15 animals were kept for fattening or milking. Only 4 of these were

examined thoroughly (cf. table 3; animals no 4 to 7 owned by Sulahattin Aydin).

- In the third barn 9 animals were kept (cf. table 3; animals no 7 and 10 owned by Suat Aydin, animals

no 8, 9 and 11 to 13 owned by Mehmet Durak and animals no 14 to 16 owned by Sulahattin Aydin).

The villagers had reported to the team at the beginning of the visit to the village that 7 of these animals

were showing FMD clinical signs. The remaining 2 animals apparently were healthy. All 9 animals

were subjected to a thorough examination.

The visual examination of the animals present in the first 2 barns did not reveal any apparent clinical signs

typical for FMD (hypersalivation, lameness, visual FMD lesions). Neither could lesions of FMD be observed

on 7 of these animals subjected to a more thorough examination.

This was different in the diseased barn: the 7 animals reported sick showed clear hypersalivation and on

thorough examination typical FMD lesions in the mouth and/or on the feet. The nature of the lesions is

reported in table 3. The 2 non-diseased animals did indeed not show any FMD lesions.

Following the visit to the barn, one group of young animals kept on pasture close to the village was visually

examined in order to gather more information about the disease situation in the village and to complete the

enquiries. In this group of about 80 animals, several showed mild hypersalivation. No lameness was

observed. Three animals (animal no 17 owned by Muhuttin Durgun, animal no 18 owned by Mehmet Durgun

and animal no 19 owned by Yusuf Turgut; cf. table 3) were subjected to a more thorough clinical

examination. Two of the animals showed relatively small almost healed lesions on the tongue and the gums.

Page 6: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

The results of the examination of all animals and the samples taken in Velibaba are summarized in table 3.

The age of the lesions found matches the information given by the Muhtar of the village and the owners of

the different animals examined. Based on the age of the oldest lesions that were observed on the animals

examined in the village, it is likely that the clinical signs started in the village about one week prior to the visit,

around October 13th 2009. However, the nature of the lesions found on the young animals on pasture may

suggest that the virus may already have been longer present in the herd, possibly without being observed by

the villagers.

Information concerning the epidemiological unit

Velibaba is a village situated in the Horasan district in the eastern part of the province of Erzurum.

The following description can be given of this village.

- Animal population susceptible to FMD:

- About 1,000 cattle are reared in the village. The animals are of a mixed type with most

households keeping dairy cows that produce milk for own consumption (there is no selling of

milk outside the village) and fattening calves/animals that are either destined for own

consumption or sold for meat. Each household has its own barn in which its animals are stabled

at night and fattening animals are kept for a more intensive feeding. During the day, all animals

except the animals in the final stages of fattening are kept together in age groups on communal

pasture.

- About 1,200 sheep are kept for meat and dairy production. During the spring the milk is sold to

a dairy outside the village. As is the case with the cattle, the animals are traditionally reared and

graze together on communal pasture.

- The village also keeps about 50 goats.

It is impossible to separate the herd into epidemiological subunits: all animals graze together (although

somehow separated into age groups), use the same pastures and sometimes are stabled in a barn

belonging to a different owner. Since there is no effective separation between households, the entire

village is to be considered as a single epidemiological unit.

- The traditional transhumance grazing of animals on highland pasture during summer is practised by

some farmers in the village. This grazing is apparently done in herds that are composed of animals

coming from different villages, sometimes at a far-away distance from Velibaba.

- All animals observed in the village were ear-tagged. Only a few calves were seen without any

identification mark (in Turkey calves are supposed to be identified at the latest when 6 months old).

Maps of Velibaba and the bordering villages

Figures 3 and 4 show maps of Velibaba, the surrounding villages and the wider area of the Horasan district

of which Velibaba is part.

Vaccination history

All animals of the village were reported to have been vaccinated on September 10th 2009. The previous

spring vaccination occurred in March-April 2009.

Page 7: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

Figure 3. Map of Velibaba and the neighbouring villages

Figure 4. Map of the Horasan area of which Velibaba is part

Trace back information

With regard to the possible sources of infection of the disease in Velibaba, the team collected the following

information:

- The animals of the infected group in barn no 3 were brought for transhumance grazing to the

highlands at the beginning of the summer and returned back to the village on the August 20th 2009. In

the highlands they had contact with other animals coming from the vicinity of the city of Kars. It is likely

that this kind of grazing is also practiced with other animals in the village and this must be considered

as one of the possible routes of introduction of FMDV in the village. However, in view of the long time

that has elapsed between the return of the animals from the highlands and the onset of symptoms, it is

unlikely that the diseased animals in barn no 3 did indeed get infected in that way and introduced the

disease in the village.

- During the last month, several villagers had bought cattle on the Horasan market. No small ruminants

were bought recently. Given the absence of effective separation between animals of the various ages,

this contact cannot be excluded as a possible source of infection. Velibaba animals are also traded

through the Erzurum market.

Page 8: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

- Trade also has occurred through dealers although no relevant information on this subject was

forwarded to the team.

- Velibaba is along a road that joins 3 more villages. The team observed movement of vehicles, tractors

and material entering and leaving the village. No bio security measures (disinfection) were

implemented at the entrances of the village.

It may be concluded that most likely live animals, either bought on the market and through traders or having

been in contact with diseased animals on pasture, introduced the FMD virus into the village herd. No

information was issued to the team regarding a follow-up of potential trace back contacts.

Trace forward information

All routes by which the virus may have entered the village can also be considered as possible routes for

spread of the virus. Although the sick animals were taken out of the herd and stabled in the village, no

particular bio security measures were applied to prevent spread of the disease. Similar to the trace back

contacts, the most likely way of spreading the virus to other villages and herds is contact with other

susceptible animals on pastures or the movement of live animals, either through traders or the markets,

although mechanical spread through people/traders, vehicles and other equipment that have been in contact

with infected animals or material can not be excluded.

This means that all villages that receive animals from Velibaba either directly or via traders or that have

animals that have been in contact with animals from Velibaba (e.g. through the markets of Horasan and

Erzurum) are potentially at risk of infection. It was reported that more than 70 villages are located in a radius

of approx. 30-40 km around Velibaba; they all market animals, mainly through the Horasan market.

Since the history of ownership of cattle is registered in the Turkish identification and registration database

TürkVet, the holdings potentially at risk can be identified by querying the database. However, it must be

noted that the TürkVet system is not yet fully operational: at the time of the training details could not be

retrieved on the contacts (market, trader, go-betweens) that an animal might have had before arriving at its

final destination. No information was issued to the team on a follow-up of potential trace forward contacts.

Page 9: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

Investigations and information concerning the Ortadüzü village

Timeline

The timeline below summarizes the major events regarding Ortadüzü.

25-Sep-09 25-Oct-09

19-Oct-09

first observation

of clinical signs

21-Oct-09

visit byETC4 team

20-Oct-09

notification of

suspicion to

veterinary service

16-Oct-09

end of likely

window of

introduction

of FMDV

4-Oct-09

start of possible

window of

introduction

of FMDV

12-Oct-09

vaccinationby practicioner

5-Oct-09

vaccination

by vaccinationteam

13-Oct-09

start of likely

window of

introduction

of FMDV

Clinical examination and sampling of the animals

The team visited 2 barns in the village of Ortadüzü.

- 4 animals were present in the first barn. Two of these were subjected to a thorough clinical

examination (cf. table 4; animals no 20 and 21 owned by Mehmet Seferoglu).

- The second barn was not visited inside, but the – according to the owner – most clearly sick animal

was brought outside for an examination (name of the owner not known). It is not known how many

animals were further present in this stable.

At the visual examination of the 4 animals in the first barn, 3 animals showed hypersalivation. Two of these 3

were subjected to further examination. Although none had a fever, both showed typical FMD lesions on the

nostrils, gums, lips and tongue. None showed lesions on the foot. Furthermore, the largest animal of the 2

(animal no 21; cf. table 4) was also depressed and showed a weak lameness in the right front leg (despite

the absence of lesions on the foot).

The animal of the second barn showed typical clinical signs of FMD: it was lame, hypersalivating and

showed very large and obvious, recent lesions of FMD on the muzzle, nostrils, lips and gums. An inspection

of the feet also revealed a small vesicle in the interdigital space of the left rear foot.

The results of the examination of all animals and the samples taken in Ortadüzü are summarized in table 4.

The age of the lesions found matches the information given by the owners of the animals examined. Based

on the age of the oldest lesions that were observed, it is likely that the clinical signs started about 3 days

prior to the visit, around October 18th 2009.

Information concerning the epidemiological unit

Ortadüzü is a village at about 10 km from the provincial capital Erzurum.

The following description can be given of this village.

- Animal population susceptible to FMD:

- About 1,100 cattle are reared in the village. As was the case with the Velibaba village, the

Page 10: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

animals are of a mixed type with most households keeping dairy cows that produce milk and

fattening calves/animals that are either destined for own consumption or sold for meat. Different

to Velibaba, the milk is sold to a dairy plant that collects the milk once a day by lorry.

- Neither sheep nor goats are reared in the village.

Again, it is impossible to separate the herd into epidemiological subunits: all animals graze together

(although somehow separated into age groups), use the same pasture land and return at night to the

village to be stabled. Thus, there is no effective separation between households and the entire village

is to be considered as a single epidemiological unit.

- All animals seen were ear-tagged.

Maps of Velibaba and the bordering villages

Figures 5 and 6 show maps of Ortadüzü and the surrounding villages.

Figure 5. Map of Ortadüzü and the neighbouring villages

Figure 6. Detailed map of Ortadüzü

Page 11: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

Vaccination history

According to the villagers, the animals of the village were vaccinated in the first half of October 2009. Most

animals were vaccinated by the official vaccination team 2 weeks prior to visit. Some households had their

animals vaccinated by the veterinarian only a week prior to the visit. Although at first glance this information

might raise a concern about the safety of the vaccines used, this was immediately denied by the villagers

themselves who seemed very well informed about the disease and were convinced of the safety of the

vaccines.

Trace back

Based on the information provided by the villagers, the following sources of infection are possible:

- The villagers mentioned pasture contacts with animals from the neighbouring village as the most likely

source of infection. Animals of this second village and of the 2 traders living in this village graze on the

same or adjacent pastures as the Ortadüzü animals. Direct contact between the animals has most

likely occurred.

- Frequent trade of animals (belonging to any category: cows, calves, etc.) occurs with bordering

villages.

- Trade of animals of the Ortadüzü village mainly passes through the Erzurum market (both selling and

buying of animals).

- Animals belonging to several households in the village and that were sent for transhumance grazing in

the highlands returned to the village on September 20th 2009. It is not known whether they had contact

in the highlands with animals from other villages or other parts of the province.

- A vehicle of a dairy plant enters the village once a day to collect milk. Furthermore, several vehicles

were observed entering and leaving the village. As was the case in the Velibaba village, no particular

biosecurity measures (disinfection) existed at the entrances of the village.

A follow up of these contacts was not yet performed at the moment of the visit.

Trace forward

The same contacts as mentioned in the trace back tracing can be considered as possible contacts through

which the infection might have spread, namely:

- trade of animals by villagers to other villages;

- trade of animals by dealers;

- trade of animals through the Erzurum market;

- pasture contacts between animals of Ortadüzü village and those of neighbouring villages;

- pasture contacts during transhumance (less likely type of contact);

- indirect contacts through vehicles or other material leaving Ortadüzü.

In view of the recent vaccination of the village herd, iatrogenic spread through either the vaccination team or

the private veterinarian can also not be excluded as a possible way of spreading the FMDV further.

Page 12: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

Results of the laboratory analysis

The results of the analysis are summarized in tables 5 and 6.

Three different tests were performed on the samples: the NSP (non-structural protein) ELISA, the LFD

(lateral flow device) test and the antigen ELISA:

- The LFD and the Ag ELISA can be used to detect virus antigen in the epithelium, vesicle fluid or

saliva.

- The NSP ELISA is used to detect antibodies to non-structural proteins of FMD virus triggered by

infection but not by vaccination with purified vaccines as the ones used in the Turkish vaccination

programme.

The outcome of all analysis is given as a qualitative value (positive or negative).

Some of the LFD results were very weak. A likely explanation is that the animals in Ortadüzü had been

treated by topical application of a disinfectant and that this may have disrupted the antigenic properties of the

virus in the lesions. However, since the LFD is not a quantitative assay, no definitive conclusions can be

drawn from the weakness of the signal. Furthermore, in this village all 3 samples tested with the NSP ELISA

were already positive despite the short time that had elapsed since the introduction of the virus in this village

(the animals showed lesions that were only 2 to 3 days old). One possible explanation for this positive

reaction is that the animals were quite young and may have been exposed to large quantities of virus,

prompting an earlier than usual immune response to NSP.

Table 5. Results of the laboratory investigation of the samples taken in the Velibaba village

animal ID LFD NSP ELISA Ag ELISA clinical signs

conclusions2

251750705 not

performed -

not performed

- not infected or incubating

not identified not

performed -

not performed

- not infected or incubating

not identified not

performed +

not performed

- either maternal antibodies or old infection (recovered)

259615563

not performed

+ not

performed - old infection (recovered)

2517504493

not performed

- not

performed - not infected or incubating

251567302 not

performed +

not performed

+ (6-7 d) infected

251637385 not

performed +

not performed

+ (5-6 d) infected

251637386 +1 +

not yet known

+ (7 d) infected

251567301 +1 + type O + (5-6 d) infected

251637375 not

performed +

not performed

+ (7 d) infected

251637374 not

performed -

not performed

+ (7d) probably infected but not yet seroconverted

251637383 not

performed +

not performed

+ (7 d) infected

251567366 not

performed -

not performed

- not infected or incubating

251567365 not

performed -

not performed

- not infected or incubating

2515673643

not performed

- not

performed - not infected or incubating

Page 13: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

251671046 not

performed -

not performed

+ (7-10 d) probably infected but not yet seroconverted

not identified not

performed -

not performed

+ (7-10 d) possibly infected but not yet seroconverted or lesion due to other causes

1 epithelium

2 conclusion based on references in table 7

3 animals are related: 251750449 and 251671046 are the calves of 25971556 (source: TürkVet)

Table 6. Results of the laboratory investigation of the samples taken in the Ortadüzü village

animal ID LFD NSP Ag ELISA clinical signs conclusion3

251532462 +1

+ inconclusive + (2-3 d) infected

251532461 +1 + type O + (2-3 d) infected

251759948 +2 + - + (2-3 d) infected

1 epithelium and saliva

2 epithelium and vesicle fluid

3 conclusion based on references in table 7

Table 7. Progression of disease as related to expected signs of disease and diagnostic detection

age of lesions -3 -2 -1 0 1 2 3 4 5 6 7 8 9 10

expected virus excretion

expected fever

detection with PCR on blood

detection with LFD

detection with Ag ELISA

detection with NSP ELISA

Red = most likely time frame of detection Yellow = likely time frame of detection Pale yellow = less likely time frame of detection

Conclusion

In general, the results of the diagnostic analyses performed on the samples taken correspond with the

clinical picture observed in both villages. The results of the clinical examination and the diagnostic analyses

clearly demonstrate that both villages are infected with FMD virus.

The O type of FMD virus was detected by the Ag ELISA. This finding still has to be confirmed by further

virological analysis in the Turkish reference lab in Ankara.

Taking into account the most commonly incubation period of 3 to 5 days, the symptoms and age of the

lesions indicate that the Velibaba village probably was infected between October 5th and October 10

th 2009

and the Ortadüzü village between October 13th and October 16

th 2009. However, the serological results of

some animals indicate that infection (perhaps with another serotype) might have been present in the villages

earlier. In theory, the incubation period can range from 1 to 14 days.

In general, the following conclusions can be made regarding the outbreaks in the villages of Velibaba and

Page 14: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

Ortadüzü:

- FMD, being endemic in this part of Turkey, is a disease that is well known by farmers and is accepted

as being part of the day-to-day livestock breeding practices.

- The animal husbandry practices of a village like Velibaba and Ortadüzü means that each village has to

be considered as an epidemiological unit with regard to animal diseases. Once FMDV enters the

village, it will affect the whole herd of the village due to common grazing and stabling of all animals.

- Although diseased animals were stabled separately, no other particular biosecurity measures were

applied to prevent spread of the disease.

- It was difficult to obtain all the necessary information to get a complete picture of how FMD might have

spread to and from the village and what exactly had happened since the introduction.

- It was difficult to get a complete list of trace back and trace forward contacts. Regarding the animal

contacts, part of the information could be drawn from the TürkVet database.

Acknowledgements

The ETC4 team wishes to thank the FAO-EUFMD, the Turkish authorities, the Turkish veterinary service, the

Turkish national reference laboratory for FMD, the trainers of the ETC4 course and the people of Velibaba

and Ortadüzü for offering the opportunity to get to know FMD better.

Page 15: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

Table 3. Animals examined and sampled in the village of Velibaba on 20th October 2009.

clinical signs type of lesions

no animal ID species

and sex

1 age

1

lam

en

ess

fever2

sali

vati

on

foo

t3

mo

uth

4

teats

inta

ct

vesic

le

recen

tly r

up

-tu

red

vesic

le

raw

ero

ded

are

a

ulc

er

wit

h

fib

rin

ou

s s

cab

ulc

er

wit

h

fib

rosis

ind

isti

nct

bre

ak c

oro

-n

ary

ban

d samples

taken5

vacci-nation status

1

estimated age of the oldest lesions

1 251750705 bov / M 7 months - NT - - - - - - - - - - B - S not reported

2 not identified bov / M not known - NT - - - - - - - - - - B - S not reported

3 not identified bov / F not known - NT - - - - - - - - - - B - S not reported

4 25961556 bov / F 4.5 year - NT - - - - - - - - - - B 10.2008

5 251750447 bov / F 4.5 year - NT - - - - - - - - - - not sampled

not reported

6 251750449 bov / M 7 months - NT - - - - - - - - - - B not reported

7 251567302 bov / F 1.5 year - NT + - LTD - - - - + + - B not reported

6 to 7 days

8 251637385 bov / F 1.3 year + NT + C G - - - - + - B 10.2008 5 to 6 days

9 251637386 bov / F 1.3 year + NT + I LG - - + (I) - + (M) - B - E 10.2008 7 days

10 251567301 bov / F 1.5 year + NT + - MLD - - - - + - B - S - E not reported

5 to 6 days

11 251637375 bov / F 1.3 year + NT + I L - - - - - + - B 10.2008 7 days

12 251637374 bov / F 1.3 year - NT + - ML - - - - - + - B 10.2008 7 days

13 251637383 bov / F 1.3 year + NT + I LTD - - - + (I) + (T) + (D) - B 10.2008 7 days

14 251567366 bov / M 1.5 year - NT - - - - - - - - - - B 10.2008

15 251567365 bov / M 1.5 year - NT - - - - - - - - - - B 10.2008

16 251567364 bov / F 1.5 year - NT - - - - - - - - - - B 10.2008

17 251750479 bov / F 7 months - NT - - - - - - - - - - not sampled

not reported

18 251671046 bov / M 7 months - NT + - GT - - - - - + - B - P not reported

7 to 10 days

19 not identified bov / M not known - NT - - - - - - - - +? - B - P not reported

7 to 10 days

1 information retrieved from the TürkVet information system

2 NT: not tested (animals did not appear to have fever)

3 foot: Coronary band - Interdigital space /

4 mouth: Muzzle - Lips - Gums - Tongue - Dental pad /

5 samples: Blood - Saliva - Vesicle fluid - Epithelium - Probang sample

Page 16: EUFMD ETC4 19th-22nd October 2009 Eastern Anatolia …Erzurum is the largest province in Eastern Anatolia and is located on a high plateau. The majority of the province is elevated

Table 4. Animals examined and sampled in the village of Ortadüzü on 21st October 2009.

clinical signs type of lesions

no animal ID species

and sex

1 age

1

lam

en

ess

fever2

sali

vati

on

foo

t3

mo

uth

4

teats

inta

ct

vesic

le

recen

tly r

up

-tu

red

vesic

le

raw

ero

ded

are

a

ulc

er

wit

h

fib

rin

ou

s s

cab

ulc

er

wit

h

fib

rosis

ind

isti

nct

bre

ak c

oro

-n

ary

ban

d samples

taken5

vacci-nation status

1

estimated age of the oldest lesions

20 251532462 bov / M 14 months - - - - MGT - - + + - - - B - S - E 03.2009 2 to 3 days

21 251532461 bov / M 14 months - - - - MLGT - - + - - - - B - S - E 03.2009 2 to 3 days

22 251759948 bov / M 8 months + NT + I ML - + (I) + (M) - - - - B - V - E not reported

2 to 3 days

1 information retrieved from the TürkVet information system

2 NT: not tested (animals did not appear to have fever)

3 foot: Coronary band - Interdigital space /

4 mouth: Muzzle - Lips - Gums - Tongue - Dental pad /

5 samples: Blood - Saliva - Vesicle fluid - Epithelium - Probang sample