Schmallenberg virus in Poland endemic or re-emerging? A six-year serosurvey.

A novel arbovirus, called Schmallenberg virus (SBV), emerged in Europe in 2011 infecting domestic as well as wild ruminants. The virus was first detected in Poland during the 2012 vector season. In order to study the SBV post-epidemic period in Poland, over twenty-one thousand domestic ruminants (cattle, sheep, goats) were tested for SBV infection between 2013 and 2018. Samples were collected as part of the national Bluetongue virus (BTV) surveillance programme. Thirteen per cent of all samples were collected from animals between 6 months and one year of age. Overall, 37.5% of ruminants tested seropositive. The seroprevalence fluctuated yearly and was highest in 2014 and 2017; however, seroconversion was detected in younger animals throughout the study indicating continuous virus circulation during the 6-year study period. A significantly higher proportion of seropositive animals were detected among cattle and older animals. Uneven distribution of seropositive animals between provinces was identified and may be a result of different housing and breeding practices and/or meteorological conditions influencing local and regional vector abundances, rather than farm stocking densities. A small number of animals were identified as being exposed to both SBV and BTV; this is likely due to the fact that the same Culicoides species transmit these two viruses thus increasing the risk of co-exposure. Considering these results, in addition to virological and entomological studies carried out in Poland previously, it can be concluded that SBV is endemic in Poland with cyclical waves of virus circulation happening every 3-4 years.

Potential use of hematological and acute phase protein parameters in the diagnosis of acute Schmallenberg virus infection in experimentally infected calves.

The initial viraemic phase of Schmallenberg virus (SBV) infection in bovine animals is characterized by the non-specific and inconspicuous clinical signs of pyrexia (>40 °C), drop in milk yield and sometimes diarrhea. As a result, the early detection of SBV epizootics can difficult, and typically only become apparent when the congenital form of the disease is observed. The aim of the study was to describe the course of the acute phase response and haematological findings in bovine calves following experimental SBV infection. No clinical signs except for increase in rectal temperature were observed in the calves inoculated subcutaneously with a Polish strain of SBV. Viral RNA was detected in serum at 2 and 4 days post inoculation (dpi). SBV antibodies were first detected by ELISA (9-21 dpi), and subsequently by virus neutralization test (14-32 dpi). The hematological parameters showed a reduction in mid-size leucocytes (MID), and also in red blood cell count (RBC). An increase in mean corpuscular hemoglobin was also observed in SBV infected calves. No significant difference in acute phase proteins (APP) was observed between experimentally infected and control calves, suggesting limited potential as diagnostic biomarker of acute SBV infection.

Post-Epidemic Distribution of Schmallenberg Virus in Culicoides Arbovirus Vectors in Poland.

Pooled samples of female and male Culicoides midges (5146 and 332 pools, respectively) that corresponded to a total number of 124,957 specimens were collected between 2013-2017 in the vicinity of cattle barns that were distributed throughout Poland were analyzed for the presence of Schmallenberg virus (SBV) RNA. Sixty-six pools tested positive (1.2%) with mean Ct value of 34.95. The maximum likelihood estimated infection rate (MLE) was calculated at 0.53 per 1000 individuals; however, it peaked in 2016 with MLE of 3.7. Viral RNA was detected in C. obsoletus/scoticus complex, C. punctatus, and C. pulicaris pools. Moreover, viral material was present in nulliparous (virgin) Culicoides females (MLE 0.27) and for the first time reported in males (MLE 0.34), which suggests the possibility of transovarial route of SBV or virus RNA transmission, as both do not fed on host blood. The accuracy of targeted versus random SBV surveillance in Culicoides vectors was compared. The relationship between infection rate (expressed as minimum infection rate; MIR), in addition to MLE, was compared with the density of virus infected midges (DIM). In conclusion, the SBV infection rate in the vector was significantly higher in 2016 as compared to other surveillance years; this is consistent with the simultaneous increase in SBV seroprevalence (seroconversion) in ruminants during the same year.

Comparison of Schmallenberg virus sequences isolated from mammal host and arthropod vector.

Schmallenberg virus (SBV) is the member of Peribunyaviridae family, which comprises pathogens of importance for human and veterinary medicine. The virus is transmitted only between animals and mainly by biting midges of the genus Culicoides. This study was performed in order to determine SBV genetic diversity and elucidate the host-vector adaptation. All three viral segments were analysed for sequence variability and phylogenetic relations. The Polish SBV strains obtained from acute infections of cattle, congenital cases in sheep, and from Culicoides midges were sequenced using Sanger and next-generation sequencing (NGS) methods. The obtained sequences were genetically similar (99.2-100% identity) to the first-detected strain BH80/11-4 from German cattle. The sampling year and origin of Polish sequences had no effect on molecular diversity of SBV. Considering all analysed Polish as well as European sequences, ovine-derived sequences were the most variable, while the midge ones were more conserved and encompassed unique substitutions located mainly in nonstructural protein S. SBV sequences isolated from Culicoides are the first submitted to GenBank and reported.

Seroprevalence and risk factors for selected respiratory and reproductive tract pathogen exposure in European bison (Bison bonasus) in Poland.

After the complete extinction from the wild of European bison (Bison bonasus) at the beginning of the twentieth century, the worldwide species population was restored to approximately 5500 individuals, with the species however remaining endangered. Despite numerous studies on the ecology and genetics of European bison, the threats of infectious diseases have been largely unexamined. The aim of this study was to screen the exposure of the world's largest population of European bison to the pathogens, which may influence the condition and development of the endangered species. A total of 240 free-ranging and captive European bison from eight main Polish populations sampled were tested for the presence of specific antibodies against ten different viruses, bacteria or protozoan. The samples were collected from chemically immobilized, selectively culled or found dead animals. Based on serology, the exposure to bovine viral diarrhea virus (BVDV), bovine herpesvirus type 1 (BoHV-1), Mycoplasma and Brucella spp. was determined as rather accidental. Using gamma-interferon assay followed by Mycobacterium tuberculosis subs. caprae detection in tissues, diagnosis of bovine tuberculosis was made for 6 out of 78 (7.7%) bison from one captive herd. The highest seroprevalence was found for bovine adenovirus type 3 (BAdV-3) -60.2% and bovine parainfluenza type 3 (PIV-3) -34.0%, while the antibodies against bovine respiratory syncytial virus (BRSV), Toxoplasma gondii and Leptospira spp. were found in 10.4%, 10.4% and 8.7% of samples, respectively. In the multivariable statistical analysis using generalized linear mixed models (GLMMS), the risk factors for PIV-3 seropositivity included population type (free-living/captive), age and health status (apparently healthy/eliminated due to the poor condition). Higher risk of BAdV-3 seropositive result was observed in free-living female European bison. The high BAdV-3 and PIV-3 seroprevalences may suggest involvement of these pathogens in the most frequently observed respiratory disorders in European bison. Moreover, this is the first study demonstrating BAdV-3 exposure in the species.

EPIDEMIOLOGY OF SCHMALLENBERG VIRUS IN EUROPEAN BISON ( BISON BONASUS) IN POLAND.

Schmallenberg virus (SBV), an emerging arbovirus in Europe, is an important pathogen in domestic ruminants; however, its impact on free-ranging wild ruminants is not well studied. Three hundred and forty-seven serum samples collected between 2011 and 2016 from 302 European bison ( Bison bonasus) from 12 different sites in Poland were tested for the presence of SBV antibodies. In addition, 86 sera were collected between 2013 and 2016 from three species of cervids for testing for SBV antibodies. After the first detection of the virus in Poland in October 2012, the proportion of SBV-seropositive European bison reached 81% (95% confidence interval [CI]: 77.1-85.8%), whereas in cervids seroprevalence was 34% (95% CI: 23.5-43.9%). There was an increase in seroprevalence in European bison from 2012 to 2014. Biting midges ( Culicoides spp.), the primary vectors of SBV, were monitored entomologically for the identification of the biting midge populations and virologically for SBV infections in the Bialowieza Forest region, which contains the world's largest European bison population. We detected SBV by PCR in 3% of Culicoides pools from 2015. In addition, seven fetal brain samples from European bison or cervids were tested and were negative for SBV RNA. Our results indicate a high seroprevalence with reduced transmission of SBV in subsequent years in the European bison populations and lower seroprevalence in cervids.

Were Polish Wild Boars Exposed to Schmallenberg Virus?

INTRODUCTION: A novel to Europe Schmallenberg virus (SBV) causes clinical disease manifested by reproduction disorders in farm ruminants. In free-living ruminants, SBV antibodies as well as the virus were detected. Recent studies also revealed SBV antibodies in wild boars. The study investigates SBV antibodies occurring in wild boars in Poland at the peak of recent virus epidemics in the country. MATERIAL AND METHODS: Samples collected from 203 wild boars culled during the 2012/2013 and 2013/2014 hunting season were serologically tested using multi-species cELISA. Attempted neutralisation tests failed due to poor serum quality. RT-PCR was implemented in seropositive and doubtful animals. RESULTS: Two samples collected from wild boar in the winter of 2013 gave a positive result in ELISA, while another two from the 2012/2013 hunting season were doubtful. No SBV RNA was detected in spleen and liver tissues. CONCLUSION: Low SBV seroprevalence in wild boars, despite high incidence of SBV infections occurring simultaneously in wild ruminants, suggests that boars are unlikely to be a significant reservoir of the virus in the sylvatic environment in Poland.

Cross-sectional study of Schmallenberg virus seroprevalence in wild ruminants in Poland at the end of the vector season of 2013.

BACKGROUND: In view of recurrent Schmallenberg virus (SBV) infections all over Europe between 2011 and 2013, a lively scientific debate over the importance of the sylvatic transmission cycle of the virus has emerged. The study presents results of serosurvey which included wild ruminants representing species of red deer (Cervus elaphus), roe deer (Capreolus capreolus), European bison (Bison bonasus), fallow deer (Dama dama), mouflon (Ovis orientalis musimon) hunted or immobilized at 34 different locations of Poland in the autumn/winter 2013. RESULTS: Out of 580 sera, 145 (25%) were considered positive for SBV antibodies. The overall SBV seroprevalence calculated using district probability weights was estimated at 27.7% (95% CI: 24.0-31.4). The seroprevalences at the district level varied between 0 and 80.0% (95% CI: 24.5-135.0%) with the mean within-district prevalence of 24.0% (95% CI: 16.5-31.4). Significantly higher seroprevalence was observed in animals from the Eastern provinces (36.6%) compared to the Western provinces (22.8%). SBV infection impact varied significantly between different species (higher SBV seroprevalence in larger species such as European bison), population type (free-ranging; captive), age, body weight, percent of the district forest area, part of Poland, and the densities of wild and domestic ruminants at the district and province level. Using statistical multivariable logistic model, population type, age, part of Poland and domestic ruminant density were identified as the main risk factors for SBV infection in wild ruminants in Poland. CONCLUSIONS: SBV seroprevalence in wild ruminants, similarly to the epizootic situation in domestic ruminants in the country, varied significantly between districts and provinces. Association between SBV seropositivity, species, animal body weight and age group expressed by a higher prevalence in larger ruminants may be explained by more frequent exposure to midge-vector bites of the latter, however it might also be related to the different species susceptibility to SBV infection. The positive effect of higher domestic ruminant density on the risk of SBV infection in wildlife and lower SBV seroprevalences in the latter suggested that the sylvatic cycle of SBV transmission is an effect of the pathogen spillover from the domestic animals.