Bluetongue Virus Serotype 8 Infection Research Articles

Transmission of Bluetongue Virus Serotype 8 by Artificial Insemination with Frozen-Thawed Semen from Naturally Infected Bulls.

Transmission of bluetongue (BT) virus serotype 8 (BTV-8) via artificial insemination of contaminated frozen semen from naturally infected bulls was investigated in two independent experiments. Healthy, BT negative heifers were hormonally synchronized and artificially inseminated at oestrus. In total, six groups of three heifers received semen from four batches derived from three bulls naturally infected with BTV-8. Each experiment included one control heifer that was not inseminated and that remained BT negative throughout. BTV viraemia and seroconversion were determined in 8 out of 18 inseminated heifers, and BTV was isolated from five of these animals. These eight heifers only displayed mild clinical signs of BT, if any at all, but six of them experienced pregnancy loss between weeks four and eight of gestation, and five of them became BT PCR and antibody positive. The other two infected heifers gave birth at term to two healthy and BT negative calves. The BT viral load varied among the semen batches used and this had a significant impact on the infection rate, the time of onset of viraemia post artificial insemination, and the gestational stage at which pregnancy loss occurred. These results, which confirm unusual features of BTV-8 infection, should not be extrapolated to infection with other BTV strains without thorough evaluation. This study also adds weight to the hypothesis that the re-emergence of BTV-8 in France in 2015 may be attributable to the use of contaminated bovine semen.

Economic Impact of a Bluetongue Serotype 8 Epidemic in Germany.

Background and Objectives: Germany was affected by Bluetongue virus serotype 8 (BTV-8) from 2006 to 2009 and recorded new cases since December 2018. We assessed the economic impact of the epidemic from the first cases in 2006 until 2018. Direct costs include production losses, animal deaths, and veterinary treatment. Indirect costs include surveillance, additional measures for animal export, disease control (preventive vaccination and treatment with insecticides), vector monitoring, and administration.Methodology: To estimate the financial impact of BTV-8 on different species and production types at the animal level, we performed a gross margin analysis (GMA) for dairy and beef cattle, and sheep. To estimate the impact on the national level, we used a modified framework described by Rushton et al. (1) and applied a methodology described by Bennett (2). Both the GMA and the economic model on national level were implemented in Excel and the Excel Add-in @Risk. The tools, which are widely applicable, also for other diseases, are made available here.Results: The financial impact of a BTV-8 infection at the animal level was estimated at 119–136 Euros in dairy cattle, at 27 Euros in beef cattle, and at 74 Euros in sheep. At the national level, the impact of the BTV-8 epidemic ranged between 157 and 203 million Euros (mean 180 million Euros). This figure consisted of 132 (73%) and 48 (27%) million Euros for indirect and direct costs. Indirect costs included 89 million Euros (67%) for vaccination, 18 million Euros (14%) for insecticide treatment, 15 million Euros (11%) for diagnostic testing of animals dispatched for trade, 8 million Euros (6%) for monitoring and surveillance, and 3 million Euros (2%) for administration. The highest costs were induced by a compulsory vaccination campaign in 2008 (51 million Euros; 28% of the total costs) and the disease impact on cattle in 2007 (30 million Euros; 17%).Discussion: We compare the outcome of our study with economic analyses of Bluetongue disease in other countries, and discuss the suitability of GMA and the developed tools for a wider application in veterinary economics.

Schmallenberg virus infection of ruminants: challenges and opportunities for veterinarians.

In 2011, European ruminant flocks were infected by Schmallenberg virus (SBV) leading to transient disease in adult cattle but abortions and congenital deformities in calves, lambs, and goat kids. SBV belonging to the Simbu serogroup (family Bunyaviridae and genus Orthobunyavirus) was first discovered in the same region where bluetongue virus serotype 8 (BTV-8) emerged 5 years before. Both viruses are transmitted by biting midges (Culicoides spp.) and share several similarities. This paper describes the current knowledge of temporal and geographical spread, molecular virology, transmission and susceptible species, clinical signs, diagnosis, prevention and control, impact on ruminant health, and productivity of SBV infection in Europe, and compares SBV infection with BTV-8 infection in ruminants.

Survey of bluetongue virus infection in free-ranging wild ruminants in Switzerland

BackgroundIn 2006, bluetongue virus serotype 8 (BTV-8) was detected for the first time in central Europe. Measures to control the infection in livestock were implemented in Switzerland but the question was raised whether free-ranging wildlife could be a maintenance host for BTV-8. Furthermore Toggenburg orbivirus (TOV), considered as a potential 25th BTV serotype, was detected in 2007 in domestic goats in Switzerland and wild ruminants were considered a potential source of infection. To assess prevalences of BTV-8 and TOV infections in wildlife, we conducted a serological and virological survey in red deer, roe deer, Alpine chamois and Alpine ibex between 2009 and 2011. Because samples originating from wildlife carcasses are often of poor quality, we also documented the influence of hemolysis on test results, and evaluated the usefulness of confirmatory tests.ResultsTen out of 1,898 animals (0.5%, 95% confidence interval 0.3-1.0%) had detectable antibodies against BTV-8 and BTV-8 RNA was found in two chamois and one roe deer (0.3%, 0.1-0.8%). Seroprevalence was highest among red deer, and the majority of positive wild animals were sampled close to areas where outbreaks had been reported in livestock. Most samples were hemolytic and the range of the optical density percentage values obtained in the screening test increased with increasing hemolysis. Confirmatory tests significantly increased specificity of the testing procedure and proved to be applicable even on poor quality samples. Nearly all samples confirmed as positive had an optical density percentage value greater than 50% in the ELISA screening.ConclusionsPrevalence of BTV-8 infection was low, and none of the tested animals were positive for TOV. Currently, wild ruminants are apparently not a reservoir for these viruses in Switzerland. However, we report for the first time BTV-8 RNA in Alpine chamois. This animal was found at high altitude and far from a domestic outbreak, which suggests that the virus could spread into/through the Alps. Regarding testing procedures, hemolysis did not significantly affect test results but confirmatory tests proved to be necessary to obtain reliable prevalence estimates. The cut-off value recommended by the manufacturer for the screening test was applicable for wildlife samples.

Effect of an inactivated bluetongue serotype 8 vaccine on semen quality in rams

The aim of this study was to determine whether a single dose of an inactivated bluetongue virus serotype 8 (BTV-8) vaccine altered semen quality in rams. Twenty sexually mature rams were assigned to three experimental groups: two groups of four animals were vaccinated and a third group of four animals was unvaccinated. The first group included rams with a history of natural BTV-8 infection in 2007 and the second and third groups included BTV-8 naïve rams. Semen was collected prior to vaccination and for 4 months post-vaccination. There were no significant differences in semen quality traits, including motility and concentration of spermatozoa, and percentages of living, normal dead and abnormal dead spermatozoa, between vaccinated and unvaccinated groups, or over time (P>0.05). The BTV-8 vaccine tested in this study did not appear to have any adverse effect on semen quality in rams.

Experimental infection of South American camelids with bluetongue virus serotype 8

Bluetongue (BT) is an infectious, non-contagious disease of wild and domestic ruminants. It is caused by bluetongue virus (BTV) and transmitted by Culicoides biting midges. Since 1998, BT has been emerging throughout Europe, threatening not only the naïve ruminant population. Historically, South American camelids (SAC) were considered to be resistant to BT disease. However, recent fatalities related to BTV in captive SAC have raised questions about their role in BTV epidemiology. Data on the susceptibility of SAC to experimental infection with BTV serotype 8 (BTV-8) were collected in an animal experiment. Three alpacas ( Vicugna pacos) and three llamas ( Lama glama) were experimentally infected with BTV-8. They displayed very mild clinical signs. Seroconversion was first measured 6–8 days after infection (dpi) by ELISA, and neutralising antibodies appeared 10–13 dpi. BTV-8 RNA levels in blood were very low, and quickly cleared after seroconversion. However, spleens collected post-mortem were still positive for BTV RNA, over 71 days after the last detection in blood samples. Virus isolation was only possible from blood samples of two alpacas by inoculation of highly sensitive interferon alpha/beta receptor-deficient (IFNAR −/−) mice. An in vitro experiment demonstrated that significantly lower amounts of BTV-8 adsorb to SAC blood cells than to bovine blood cells. Although this experiment showed that SAC are generally susceptible to a BTV-8 infection, it indicates that these species play a negligible role in BTV epidemiology.

Viral RNA load in semen from bluetongue serotype 8-infected rams: Relationship with sperm quality

This study investigated if viral RNA was detectable in the semen of rams clinically infected with bluetongue virus serotype 8 (BTV-8) by RT-qPCR, and to what extent the amount detected may be predictive of sperm quality. Semen samples were collected on six occasions from 93 BTV-8 infected rams involved in two longitudinal (n=12 and 27, respectively) and one cross-sectional (n=54) field study. Semen quality was assessed in terms of mass motility, concentration of spermatozoa, percentage of living and dead spermatozoa as well as cytological features. An overall semen quality score (SQS) was established. Depending upon the studied population, BTV RNA was detected in 75-100% of semen samples at initial testing 25-57 days post-observation (DPO) of clinical signs, and was detectable up to 116 DPO in a proportion of rams undergoing repeated sampling. Semen quality variables were significantly altered following natural BTV-8 infection and correlated with the amount of BTV RNA present. The SQS did not return to normal when virus was no longer detectable, suggesting that clearance of BTV precedes full recovery of sperm quality. In conclusion, viral RNA may be transiently recovered from the semen of BTV-8 affected rams and may serve as an indicator in predicting ram breeding potential following natural infection.

Spatial analysis of bluetongue cases and vaccination of Swiss cattle in 2008 and 2009

Bluetongue (BT) is a vector-borne viral disease of ruminants. The infection is widespread globally with major implications for international animal trade and production. In 2006, BT virus serotype 8 (BTV-8) was encountered in Europe for the first time, causing extensive production losses and death in susceptible livestock. Following the appearance of BTV-8 in Switzerland in 2007, a compulsory vaccination programme was launched in the subsequent year. Due to social factors and difficulties to reach animals on high pasture, the regional vaccination coverage varied across the country in both 2008 and 2009. In this study, the effect of vaccination on the spatial occurrence of BTV-8 and the associated relative disease risk in Switzerland in 2008 and 2009 were investigated by a spatial Bayesian hierarchical approach. Bayesian posterior distributions were obtained by integrated nested Laplace approximations, a promising alternative to commonly used Markov chain Monte Carlo methods. The number of observed BTV-8 outbreaks in Switzerland decreased notably from 2008 to 2009. However, only a non-significant association between vaccination coverage and the probability of a spatial unit being infected with BTV-8 was identified using the model developed for this study. The relative disease risk varied significantly across the country, with a higher relative risk of BTV-8 infection in western and north-western Switzerland where environmental conditions are more suitable for vector presence and viral transmission. Examination of the spatial correlation between disease occurrence, control measures and associated ecological factors can be valuable in the evaluation and development of disease control programmes, allowing prioritisation of areas with a high relative risk of disease.

The effect of bluetongue virus serotype 8 on milk production and somatic cell count in Dutch dairy cows in 2008

The effect of bluetongue virus serotype 8 (BTV-8) infections was quantified on milk production and udder health. From July 2008 to December 2008, 1,074 seronegative cows in 15 herds that were not vaccinated against BTV-8 were tested every 3 wk for BTV-8 antibodies. Sampling stopped when cows seroconverted. Test-day records were provided and 3 traits were defined to evaluate the effect of BTV-8 on milk production and udder health: 1) the difference between observed and predicted fat- and protein-corrected milk production; 2) the natural logarithm of the somatic cell count (lnSCC); and 3) the occurrence of a new high SCC. In the default model, the variables were assumed influenced by BTV-8 when the test-day record of the seroconverted cow was taken within 30 d before seroconversion, thus, in the period in which the cow was infected. In sensitivity analyses, the time intervals were varied in which BTV-8 was assumed to affect milk production and udder health. During the study, 185 cows (17%) had a subclinical infection and seroconverted and 77 had a test-day result within 30 d before seroconversion. In this period, in cows that seroconverted, the fat- and protein-corrected milk production was 52 (95% confidence interval: 26 to 77) kg less than in the period before and after seroconversion and was 51 (95% CI: 26 to 76) kg less than in cows that remained seronegative. When the time interval was increased to within 42 d before seroconversion, the milk production in BTV-8-seroconverted cows decreased by 61 (95% CI: 28 to 94) kg compared with the period before and after seroconversion and decreased by 59 (95% CI: 27 to 92) kg compared with cows that remained BTV-8 seronegative. No significant effect of BTV-8 was found on SCC and odds for a high SCC. Subclinical BTV-8 infection in dairy cattle results in a decreased milk production.

155 EMBRYONIC APOPTOSIS AFTER BTV-8 INFECTION IN BOVINE HATCHED IN VITRO PRODUCED BLASTOCYSTS

The recent blue tongue virus serotype-8 (BTV-8) epidemic in central Western Europe has been associated with field fertility problems (De Clercq et al. 2008 Transbound. Emerg. Dis. 55, 352–359). Previous research clearly showed that in vitro produced bovine hatched blastocysts are susceptible for BTV-8 infection (Vandaele et al. 2010 Reprod. Fertil. Dev. 22, 254 abst.). The aim of the present study was to investigate the effect of a BTV-8 infection on the occurrence of apoptosis in embryos in order to gain a clear insight into the role BTV-8 might play in early embryonic death. Immature bovine oocytes were matured and fertilized in vitro. Presumed zygotes were denuded 24 h post-insemination and cultured in 50-μL droplets of modified SOF medium with 10% fetal calf serum (tested negative for BTV antibodies) at 38.5°C in 5% CO2, 5% O2, and 90% N2. At 7 days post-insemination (dpi), blastocysts were grouped to enhance hatching. At 8.5 dpi, 4 to 8 hatched embryos were placed in 800 μL of minimum essential medium (MEM), containing 103.8 to 104.9 50% tissue culture infectious doses (TCID50) of BTV-8 (Bel 2006/3, VAR, Brussels, Belgium) and incubated for 1 h at 38.5°C in an atmosphere of 5% CO2 in air. Two groups of hatched control embryos were incubated under the same circumstances in 800 μL of SOF and 800 μL of MEM, respectively. After inoculation, embryos were washed according to IETS guidelines with the exception that they were not ZP-I and cultured in new SOF. At 48 and 72 h post-inoculation (hpi), half of the embryos of each group were fixed in 4% paraformaldehyde for 12 to 24 h and subsequently stained for BTV-8 and apoptosis with a double immunofluorescent staining using a BTV-8 monoclonal antibody (8A3B.6, ID-Vet, Montpellier, France) in combination with TUNEL (Roche Diagnostics, Mannheim, Germany). All mock-inoculated embryos were negative for BTV-8 virus antigen. At 48 and 72 hpi, respectively, 45 and 38% of the embryos were BTV-8 positive in all embryonic cells, and all remaining embryos had at least some BTV-8 blastomeres. The overall apoptotic cell ratio in infected embryos (17.9 ± 1.14% at 48 hpi and 15.3 ± 1.16% at 72 hpi) was significantly higher than in noninfected, mock-inoculated embryos (10.4 ± 0.57% at 48 hpi and 4.3 ± 0.30% at 72 hpi) (P < 0.01). Furthermore, total cell number was substantially lower in infected embryos (103 ± 14.2 at 48 hpi and 120 ± 17.5 at 72 hpi) compared with noninfected, mock-inoculated embryos (258 ± 47.4 at 48 hpi and 348 ± 64.1 at 72 hpi) (P < 0.05). This study indicates that embryonic apoptosis after BTV-8 infection results in embryonic arrest and early embryonic death and thus might be involved in herd fertility problems during a BTV epidemic. The first author is supported by Research Foundation Flanders (Grant number G.0210.09).

Mortality attributable to bluetongue virus serotype 8 infection in Dutch dairy cows

In 2007, bluetongue virus serotype 8 (BTV-8) re-emerged in the Netherlands and a large number of farmers notified morbidity and mortality associated with BTV-8 to the authorities. All dead cows in the Netherlands are registered in one of the three age classes: newborn calves <3 days, calves 3 days to 1 year, and cows >1 year. These registrations result in a complete data set of dead cattle per herd per day from 2003 until 2007. In this study, the mortality associated with BTV-8 for the Dutch dairy industry was estimated, based on this census data. Default, mortality associated with BTV-8 was estimated for the confirmed notification herds. Moreover, an additional analysis was performed to determine if mortality associated with BTV-8 infection occurred in non-notification herds located in BTV-8 infected compartments. A multivariable population-averaged model with a log link function was used for analyses. Separate analyses were conducted for the three different age groups. Confirmed notification herds had an increased cow mortality rate ratio (MRR) (1.4 (95% CI: 1.2–1.6)); calf MRR (1.3 (95% CI: 1.1–1.4)); and newborn calf MRR (1.2 (95% CI: 1.1–1.3)). Furthermore, in non-notification herds in BTV-8 infected compartments, mortality significantly increased 1.1 times (95% CI: 1.1–1.1) in cows, 1.2 times (95% CI: 1.2–1.2) in calves and 1.1 times (1.1–1.1) in newborn calves compared with BTV-8 non-infected months. Using objective census data over a 5-year period, the MRRs indicated increased mortality associated with BTV-8 infection not only in herds of which the farmer notified clinical signs but also in non-notification herds in infected compartments.

Bluetongue virus serotype 8 (BTV-8) infection reduces fertility of Dutch dairy cattle and is vertically transmitted to offspring

In 2007, BTV-8 re-emerged for the second year in the Netherlands and caused morbidity and increased mortality in cattle herds. In addition, cattle farmers reported reduced fertility in their cows. For this study, fifteen herds that were not vaccinated were selected. These were matched to 10 vaccinated herds by geographic region. At the start of the study, in July 2008, all cattle in the non-vaccinated herds >1 year old were sampled. All seronegative cows entered the study program and blood samples from these cows were tested for antibodies against BTV-8 in an ELISA. Cows were sampled at intervals of three weeks and sampling was stopped once a cow tested seropositive. Sampling ceased in all remaining cows in December 2008. Newborn calves originating from infected dams or from vaccinated dams were tested by PCR for BTV-8. Fertility data were obtained from the Royal Dutch Cattle Syndicate (CRV). Multi-level generalized latent and linear models were used for analyses. In 2008, 185 (17.2%) out of 1,074 initially seronegative non-vaccinated cattle seroconverted and were assumed to be infected with BTV-8. Infected cows were 5 (95% CI: 1.9–14.3) times more likely to return for insemination within 56 days after first insemination. In addition, these cows needed 1.7 (95% CI: 1.4–2.0) times more inseminations for an assumed pregnancy, and needed 2.5 (95% CI: 2.4–2.6) times more days between first and last insemination compared to the period prior to seroconversion and compared to cows not infected by BTV-8 in 2008. No association between BTV-8 infection and the chance to abort between 100 and 260 days after last insemination was found. In total, 48 calves originating from infected cows were tested by PCR for the presence of BTV-8. Ten (20.8%) out of these 48 calves were born PCR-positive. None of 256 calves from vaccinated dams tested PCR-positive. Further, cows infected during the second half of gestation had a 15.5 times (95% CI: 1.3–190.4) higher chance of a PCR-positive newborn calf compared to cows infected in the first half of gestation. This study showed that BTV-8 has a negative effect on fertility of dairy cattle.

Virological and pathological findings in Bluetongue virus serotype 8 infected sheep

Twenty-seven sheep of the four most common Swiss breeds and the English breed Poll Dorset were experimentally infected with a northern European field strain of bluetongue virus serotype 8 (BTV-8). Animals of all breeds developed clinical signs, viremia and pathological lesions, demonstrating that BTV-8 is fully capable of replicating and inducing bluetongue disease (BT) in the investigated sheep. Necropsy performed between 10 and 16 days post-infectionem (d.p.i.) revealed BT-typical hemorrhages, effusions, edema, erosions and activation of lymphatic tissues. Hemorrhages on the base of the Arteria pulmonalis and the left Musculus papillaris subauricularis were frequently present. Histology confirmed the macroscopical findings. Using a score system, clinical manifestation and pathology were found to be significantly related. Furthermore, clinical signs and fever were shown to be indicative for the concurrent presence of high amounts of viral ribonucleic acid (RNA) in blood. Spleen, lung, lymph nodes and tonsils from all animals were analyzed regarding viral RNA loads and infectivity using real-time reverse transcriptase PCR (rRT-PCR) and virus isolation in cell culture, respectively. The highest amount of viral RNA was detected in spleen and lung and rRT-PCR revealed to be a more sensitive method for virus detection compared to virus isolation. A long-term follow-up was performed with three sheep showing that BTV-8 viral RNA in blood was present up to 133 d.p.i. and in certain tissues even on 151 d.p.i. No significant breed-related differences were observed concerning clinicopathological picture and viremia, and the Swiss sheep were as susceptible to BTV-8 infection as Poll Dorset sheep, demonstrating a remarkably high virulence of BTV-8 for indigenous sheep breeds.

Serological Survey of Bluetongue Virus Serotype-8 Infection in South American Camelids in Switzerland (2007-2008)

Outbreak of bluetongue virus serotype-8 (BTV-8) infection in domestic ruminants in Northern Europe. To investigate the South American camelids' (SAC) susceptibility to BTV-8 infection, their role in the epidemiology of the disease, and the use of currently available serological screening tests in SAC in an endemic region. Three hundred and fifty-four unvaccinated and 27 vaccinated SAC (170 llamas, 201 alpacas), ranging in age from 1 month to 17 years between June and August 2008. The SAC originated from 44 herds throughout the country, representing 10% of the Swiss SAC population. Prospective, observational study of a convenience sample of SAC. Serum samples were analyzed with 2 serological screening tests. When results diverged, a 3rd ELISA was carried out for confirmation (ID Screen Bluetongue Competition ELISA kit). All sera from the 354 unvaccinated animals were negative in the endemic region. Reliable seroconversion was observed after administration of 2 doses of vaccine. This study suggests a low susceptibility of SAC to BTV-8 despite the presence of the virus in the cattle and small ruminant population, indicating that SAC do not play a major role in the epidemiology of BTV-8. Furthermore, these results indicate that commercially available serological tests for BTV-8 can be used in SAC.

192 SUSCEPTIBILITY OF BOVINE-HATCHED BLASTOCYSTS TO BLUETONGUE VIRUS SEROTYPE 8 INFECTION

In 2006 and 2007, Bluetongue virus serotype 8 (BTV-8) caused devastating outbreaks in Northern Europe; the outbreaks were controlled in 2008 and 2009 by an international vaccination policy. Remarkably, BTV-8 differs from other serotypes in that it spread transplacentally (De Clercq K et al. 2008 Transboundary and Emerging Diseases 55, 352-359). Apart from the transplacental spreading, a significant increase in the incidence of abortions was reported in Belgium (Meroc E et al. 2009 Transboundary and Emerging Diseases 56, 39-48). The aim of the present study was to investigate the susceptibility of bovine-hatched, in vitro-produced blastocysts to BTV-8. A total of 1390 immature bovine oocytes were matured and fertilized in vitro. Presumed zygotes (n = 1148) were denuded 24 h post-insemination and cultured in 50-μL droplets of modified synthetic oviduct fluid (SOF) medium with 10% fetal calf serum (tested negative for BTV antibodies) at 39.0°C in 5% CO2, 5% O2, and 90% N2. At 7 days post-insemination (dpi), blastocysts were grouped to enhance hatching. For virus incubation, BTV-8 Bel 2006/2 from Veterinary and Agrochemical Research Centre (VAR, Brussels, Belgium) was used. At 8.5 dpi, hatched embryos were placed in 800μL of minimum essential medium (MEM) containing 103.8 50% tissue culture infectious doses (TCID50) of BTV-8 and incubated for 1 h at 39°C in an atmosphere of 5% CO2 in air. At the same time, 2 groups of hatched control embryos were incubated under the same circumstances in 800 μL of SOF and 800 μL of MEM, respectively. After infection, all embryos were washed according to IETS guidelines with the exception that they were not zona pellucida intact and cultured in new SOF. At 48, 60, 72, and 96 h post-infection (hpi), one-fourth of the embryos of each group were fixed in 4% paraformaldehyde for 12 to 24 h and subsequently stained for BTV-8 with double immunofluorescent staining using a BTV-8 monoclonal antibody (8A3B.6, ID-Vet, Montpellier, France). All control embryos (CTRL and MEM) were negative for BTV-8 virus antigen at all time points. At 48 hpi, only 1 out of 7 infected embryos was positive for virus antigen (in all its cells). At 60 hpi, all remaining embryos (n = 6) were negative, whereas at 72 hpi and 96 hpi all embryos had 25% to 100% BTV-8-positive cells (n = 6 at 72 hpi and n = 7 at 96 hpi). Furthermore, 1 embryo at 72 hpi and 2 embryos at 96 hpi showed morphological signs of degeneration. This study has showed for the first time that hatched in vitro-produced blastocysts are susceptible for BTV-8 virus infection and replication in vitro. The relatively long time between virus infection and detection of viral antigen is in accordance with the slow replication cycle of the virus. Further research is ongoing to investigate the importance of BTV-8 infection in early embryonic death. The first author is supported by Research Foundation-Flanders.

Epidemiologic characteristics of bluetongue virus serotype 8 laboratory-confirmed outbreaks in The Netherlands in 2007 and a comparison with the situation in 2006

A major epidemic of bluetongue virus serotype 8 (BTV-8) occurred in Western Europe in 2006. During 2007 it became evident that BTV-8 had survived the winter and a re-emerging epidemic quickly developed. The objective of this study was to describe the severity and clinical impact of the BTV-8 epidemic in 2007 in The Netherlands in laboratory-confirmed outbreaks and to compare this with the situation in 2006. The relative frequency of clinical signs in BTV-8 affected sheep flocks and cattle herds in 2007 and 2006 was similar. The most prominent changes were a higher proportion of sheep flocks with lameness and a much higher proportion of cattle herds reporting a decrease in milk yield in 2007. BTV-8 associated morbidity and mortality incidence rates in sheep flocks and cattle herds were significantly ( P < 0.001) higher in 2007 than in 2006. Both in sheep flocks and cattle herds, BTV-8 associated case fatality was significantly ( P < 0.001) lower in 2007, which was probably due to better medical treatment of sick animals. There were significantly ( P < 0.001) more fertility problems associated with BTV-8 infection in outbreak cattle herds in 2007 compared to 2006.

Transplacental and oral transmission of wild-type bluetongue virus serotype 8 in cattle after experimental infection

Potential vertical transmission of wild-type bluetongue virus serotype 8 (BTV-8) in cattle was explored in this experiment. We demonstrated transplacental transmission of wild-type BTV-8 in one calf and oral infection with BTV-8 in another calf. Following the experimental BTV-8 infection of seven out of fifteen multi-parous cows eight months in gestation, each newborn calf was tested prior to colostrum intake for transplacental transmission of BTV by RRT-PCR. If transplacental transmission was not established the calves were fed colostrum from infected dams or colostrum from non-infected dams spiked with BTV-8 containing blood. One calf from an infected dam was born RRT-PCR positive and BTV-specific antibody (Abs) negative, BTV was isolated from its blood. It was born with clinical signs resembling bluetongue and lived for two days. Its post-mortem tissue suspensions were RRT-PCR positive. Of the seven calves fed colostrum from infected dams, none became infected. Of the six calves fed colostrum from non-infected dams spiked with infected blood, one calf became PCR-positive at day 8 post-partum (dpp), seroconverted 27 days later, and remained RRT-PCR and Abs positive for the duration of the experiment (i.e., 70 dpp). This work demonstrates that transplacental transmission in late gestation and oral infection of the neonate with wild-type BTV-8 is possible in cattle under experimental conditions.

Blauzungenkrankheit bei Schweizer Schafrassen: Klinische Symptome nach experimenteller Infektion mit dem BTV-Serotyp 8

Clinical disease of bluetongue (BT) in sheep may differ depending on breed, age and immunity of infected sheep and may also vary between serotype and strain of BT virus (BTV). Since there are no data available on the susceptibility of Swiss sheep breeds for BT, we performed experimental infection of the 4 most common Swiss sheep breeds and the highly susceptible Poll Dorset sheep with the BTV serotype 8 (BTV-8) circulating in Northern Europe since 2006. Clinical signs were assessed regarding severity, localisation, progression and time point of their appearance. The results clearly show that the Swiss sheep breeds investigated were susceptible to BTV-8 infection. They developed moderate, BT-characteristic symptoms, which were similar to those observed in Poll Dorset sheep. Regardless of breed, the majority of infected animals showed fever, swelling of the head as well as erosions of the mouth and subcutaneous haemorrhages.