Trop Anim Health Prod (2009) 41:421–425
DOI 10.1007/s11250-008-9225-3
SHORT COMMUNICATION
Antibody prevalence against respiratory viruses
in sheep and goats in North-Western Turkey
Kadir Yeşilbağ & Burak Güngör
Accepted: 14 August 2008 / Published online: 28 August 2008
# Springer Science + Business Media B.V. 2008
Abstract Respiratory viruses may infect both small adenoviruses are the possible common reason of
and large ruminant species, and can be transmitted respiratory diseases in small ruminants in the region.
among those of species. Present study reports pres-
ence and serological distribution of bovine respiratory Keywords Respiratory viruses . BRSV. PI-3 .
viral infections in sheep and goats in Marmara region BVDV. BHV-1 . BAV-1 . BAV-3 . Sheep . Goat
of Turkey. Total of 388 sera, 228 from sheep and 160
from goats collected from 4 provinces were analysed.
Neutralising antibodies specific to BVDV, BHV-1, Introduction
BRSV, PI-3, BAV-1 and BAV-3 were investigated.
Among 388 serum samples 32.1% were positive for Respiratory virus infections lead to economical losses
BVDV, 23.0% for BHV-1, 72.9% for BRSV, 13.2% both in large and small ruminant species. It is thought
for PI-3, 86.0% for BAV-1 and 93.0% for BAV-3. that such infections are common around the world.
There were significant differences observed between Previous investigations demonstrated that bovine
seroprevalence rates detected in neighbouring prov- respiratory syncytial virus (BRSV), bovine parain-
inces. Prevalence of BVDV specific antibodies was fluenza virus 3 (PI-3), bovine herpesvirus 1 (BHV-1),
extremely higher (p=0.0009) in sheep, however, bovine viral diarrhoea virus (BVDV) and bovine
BHV-1 (p=0.0001) and PI-3 (p=0.0038) were more adenoviruses (BAV) are the main viral agents includ-
prevalent in goats. BRSV antibody prevalence was ed in respiratory infections of ruminant species. Other
closely related to data obtained from cattle. This study viruses e.g . rhinoviruses, reoviruses, enteroviruses
demonstrates that, like in cattle herds, BRSV and and bovine coronavirus may also detected in respira-
tory infections. Although these viral agents play a
K. Yeşilbağ (*) :B. Güngör preliminary role, they generally contribute with other
Department of Microbiology, Virology, infectious agents like bacteria and Mycoplasma spp.
Uludağ University Faculty of Veterinary Medicine, (Valarcher and Haaglund 2006). Maedi and pulmo-
Görükle campus, nary adenomatosis are specific diseases of sheep that
16059 Bursa, Turkey
e-mail: kyesilbag@uludag.edu.tr their symptoms localised in the lungs are developed in
a long period of time.
Present address: In principle those of viruses infect both small and
B. Güngör large ruminants and, moreover, can be cross-transmitted
Ministry of Agriculture and Rural Affairs,
Branch of Muş Province, among animal species. Thus in the areas where above
Muş, Turkey mentioned infections are detected, an infection rate of
422 Trop Anim Health Prod (2009) 41:421–425
these viruses in small ruminants is expected. In a recent propagation and neutralization steps of other viruses.
study we investigated the seroprevalence of bovine Cell cultures were grown in Dulbecco’s MEM
respiratory viruses including BVDV, BHV-1, BRSV, PI- supplemented with 10% foetal calf serum (FCS). Cell
3, BAV-1 and BAV-3 in non-vaccinated cattle popula- lines and FCS were pre-tested to be free from
tion of Marmara region, north-western Turkey. Results indigenous BVDV contamination.
showed the presence of all the infections in the region
(Yeşilbağ and Güngör 2008). This region is one of the Serological examinations
main dairy production areas of the country, but goat
and sheep production is also at a satisfactory level. Serological screening was performed using a VNT as
Hence, as a second step, seroprevalence and distribu- previously described (Yeşilbağ and Güngör 2008).
tion of respiratory viruses in small ruminant popula- This assay is serotype specific and sensitive for
tions in Marmara region of Turkey was investigated in serological screening of viral infections. Serum
this study. samples were pre-diluted to be used in VNT in
fallowing dilutions: 1:2 for BHV-1 and BRSV; 1:5
for BVDVand PI-3 and 1:10 for BAV serotypes. Fifty
Materials and methods microlitres of diluted serum sample was mixed with
an equal volume of virus suspension (100TCID50) in
Animals and sample collection 96-well microtitre plate wells as duplicates and
consequently incubated in a 5% CO2 atmosphere at
Serum samples used in this survey were collected 37°C. Two hours incubation was applied for BHV-1
between April 2004 and October 2005 from 4 while it was 1 hour for the other viruses. Thereafter,
provinces namely Bursa, Balıkesir, Bilecik and MDBK cell suspension (3×105 cells/ml) was added
Tekirdağ located in Marmara region. There were a into each well in a volume of 50 μl. In the VNT
total of 388 samples, 228 from sheep and 160 from performed for anti-BRSV antibodies, Bel-26 cell
goats analysed (Table 1). All the sampled animals cultures were previously prepared in 96-well plates.
were older than 1 year old and randomly selected After incubation of virus suspension with diluted
from privately owned small capacity family farms serum samples in a transfer plate; cell culture medium
including sheep and goats together. In most cases in 96-well plates was removed and 100 μl of virus-
there were also some cows in the establishment. serum mixture was inoculated onto cell culture at
There was no clinical disorder recorded during the about 50% of confluence. Test results were evaluated
sampling, and no vaccination program had been by inverted light microscope after 5-7 days of
applied against viruses examined in this study. incubation in 5% CO2 atmosphere at 37°C. All the
Serum samples were heat inactivated at 56°C for viruses used in present study are in cytopathogenic
30 min and stored at –20°C until testing. nature, thus, inhibition of virus growth indicated by
non-destructed monolayers of cell cultures was
Viruses and cell cultures evaluated as indicator of virus neutralization. For
scoring a sample as positive for the investigated
Major respiratory viruses causing infection in cattle, antibodies, both wells used for the same sample were
sheep and goats were used in this study. NADL strain asked to be free of cytopathogenic effect.
of BVDV, Cooper strain of BHV-1 and Atue strain of
BRSV were used in virus neutralization test (VNT). Statistical analyses
BVDV, BHV-1, PI-3, BAV serotype 1 and BAV
serotype 3 strains were originated from Department of Statistical significance of differences in seropreva-
Virology at Ankara University Faculty of Veterinary lence values between locations were analysed using
Medicine, Ankara- Turkey while BRSV strain was Chi-square analysis and Fischer’s exact test where it
supplied from Institute for Virology at Justus-Liebig is appropriate (GraphPad InStat V2.02) (p=0.05).
University Faculty of Veterinary Medicine, Giessen- Same analyses were also applied to carry out the
Germany. BRSV was propagated in Bel-26 diploid significance among the seroprevalence values
cell line, however MDBK cell line was employed for detected against different viruses in a given location.
Trop Anim Health Prod (2009) 41:421–425 423
Table 1 Seropositivity rates of viruses in sheep and goat populations detected according to locations
Location Number of animals Seropositivity rates (%)
BVDV BHV-1 BRSV PI-3 BAV-1 BAV-3
Sheep Bilecik 33 46.3 9.0 63.6 6.0 81.6 93.9
Balıkesir 88 47.0 9.5 65.0 11.4 86.5 82.7
Bursa 95 29.7 9.47 78.8 6.4 89.3 96.8
Kırklareli 12 33.3 16.6 * 16.6 50.0 100.0
Avarege (sheep) 228 38.8% 9.8% 71.5% 8.8% 85.0% 91.9%
Goats Bilecik 89 28.7 30.2 86.2 12.1 89.0 93.8
Balıkesir 59 13.7 55.9 78.4 31.0 84.2 94.9
Bursa 12 16.6 8.3 66.6 16.6 91.6 100.0
Avarage (goats) 160 21.6% 38.2% 74.7% 19.7% 87.4% 94.7%
Total 388 32.1% 23.0% 72.9% 13.2% 86.0% 93.0%
* : not evaluated due to insufficient volume of samples
Results Discussion
Serological results are summarised in Table 1. Prev- Cross transmission of respiratory viruses between
alence of BVDV specific antibodies was significantly small ruminants and bovines is a crucial aspect of
higher (p=0.0009) in sheep, however, BHV-1 (p= disease control and epidemiology. Presence and
0.0001) and PI-3 (p=0.0038) were more prevalent in distribution of bovine major viral respiratory diseases
goats. There was no significant difference in seropre- in Marmara region of Turkey have been recently
valence values obtained for the other viruses between reported (Yeşilbağ and Güngör 2008). Present study
animal species (p>0.05). aimed to demonstrate distribution of respiratory
No significant differencewas detected between BHV- viruses in small ruminants living in small capacity
1 and PI-3, and between BAV-1 and BAV-3 antibody family farms through antibody studies.
prevalence in sheep while extremely significant differ- Detection of antibodies against all of the investi-
ences (p≤0.0026) were observed between other viruses. gated viruses in all the locations indicates wide
That situation also existed in goat population with one
exception that there was no significant difference
between BVDVand PI-3 (p>0.05).
160
In general consideration including both sheep and
goat samples, insignificant difference was recorded for 140
BVDV, BRSV and PI-3 among analysed locations. For
120
BHV-1, the most important difference (p=0.0003) was
detected between Bursa and Balıkesir where the neigh- 100
bouring provinces are. An important difference for BAV-
80
3 was also observed between those of provinces (p=
0.0096). There were also significant differences in 60
serological distribution of BAV-1 in given locations.
40
Among 388 animals, 14 (3.6%) were free of
antibodies to all of tested viruses, while only 2 20
samples were positive for antibodies to all the viruses.
0
Most commonly, antiviral antibodies were detected 0 1 2 3 4 5 6
against 2 viruses in the same animal (n=134) which
was followed by triplet detection of antibodies in 120 Fig. 1 Multiple seropositivity numbers among 388 sheep and
goats against respiratory viruses. Number of animals is shown
animals. Total of 36 animals were found seropositive on the left (y-axis). X -axis shows the number of animals
for 1 virus (Fig. 1). seropositive against 1, 2, 3, 4, 5 or 6 viruses
424 Trop Anim Health Prod (2009) 41:421–425
distribution of these infections in small ruminants as significantly high prevalence, while there was no
well as cattle population in the region (Yeşilbağ and significant difference between seroprevalence of both
Güngör 2008). Some of these agents are known to be viruses. The close relationship detected between the
transmitted between animal species, thus small rumi- seropositivity rates of BAV-1 and BAV-3 is possibly
nants may be a source of respiratory infections for due to belonging to the same serological group (group I)
cattle. This situation is important especially for small of bovine adenoviruses.
capacity family farms in which both large and small According to multiple detection rates of viral
ruminant species are kept together. antibodies, double combination of respiratory viruses
Wide distribution of pestivirus infection in small was very common which was followed by triplet
ruminants in different territories was reported (Graham combination (Fig. 1). This situation is related to
et al. 2001; Berriatua et al. 2006; Krametter- multi-agent nature of respiratory viral infections as in
Froetscher et al. 2007). There is data on the presence cattle (Autio et al. 2007). There were no epidemio-
of BVDV infection in sheep in Turkey. Present study logical clusters of this multiple infections detected
shows that BVDV is also common in small ruminants according to locations.
in Marmara region. Consistent with studies in Austria Results of this survey carry out that respiratory
(Krametter-Froetscher et al. 2006, 2007), significantly virus infections are widely distributed in small
high prevalence was detected in sheep than in goats. ruminants in Marmara region, north-western Turkey.
This situation can be related to susceptibility of Control of BVDV and BHV-1 is crucial for cattle
animal species to the agent. production. Small ruminants may interfere these
BHV-1 may also infect small ruminants in natural attempts. Due to correlation between serological
and experimental conditions. Detecting 23.0% of prevalence rates of BRSV, sheep and goat population
small ruminants infected with BHV-1 shows that may also attributed to be source of infection for
those of animal species pose a risk of virus spread to bovines. Virological studies may be more satisfacto-
cattle population. Thus, a potential risk is generated rily to clarify the role of these viruses on clinical cases
for the success of BHV-1 control & eradication of respiratory disease in small ruminants.
programs in dairy herds in the region. In accordance
with previous reports (Yeşilbağ et al. 2003, Yeşilbağ
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