Border Disease Virus Infection Research Articles

Detection and Genetic Characterization of Border Disease Virus (BDV) Isolated from a Persistently Infected Sheep in a Migratory Flock from Rajasthan State, Northwestern India.

Border disease virus (BDV) causes significant economic losses in sheep farming worldwide. In India, BDV has not yet been studied in sheep migrating for summer pasturing. This study aimed to determine the extent of BDV infection in migratory sheep and provide genetic characteristics of BDV. Blood and serum samples from 90 lambs of a migratory sheep flock (600) in Central India were collected and subjected to molecular detection, phylogenetic analysis and virus neutralization test (VNT). We detected BDV in two lambs through real-time RT-PCR, while 64.4% (58/90) of in-contact lambs had BDV neutralizing antibodies. One apparently healthy lamb was found to be persistently infected with BDV. Phylogenetic analysis of 5'-UTR and Npro genes and the concatenated datasets typed the BDV isolate from PI sheep as BDV-3 genotype. However, it showed a closer relationship with BDV-3 strains from China than the previously reported Indian BDV-3 strains. This is the first report on the detection of BDV persistently infected migratory sheep in India. Additionally, we provided evidence of genetic variability among BDV-3 strains in India. The findings improve our understanding of epidemiology and genetic characteristics of BDV in India and highlight the potential risks associated with the traditional practice of sheep migration for summer pasturing.

Pestiviruses in sheep and goats in Austria: Options for integration into the bovine viral diarrhea (BVDV) monitoring program

After the successful eradication of the bovine viral diarrhea virus (BVDV) in cattle in Austria, the risk of infections with the border disease virus (BDV) remains. Both viruses belong to the pestivirus genus. BDV infections lead to false-positive results in BVDV surveillance. This can be attributed to the contact to small ruminant populations. In particular, keeping cattle together with sheep or goats on a farm or alpine pasture are significant risk factors. Between 2015 and 2022, BDV type 3 was detected in 15 cattles in Austria. These animals were almost exclusively persistently infected calves. However, a positive antibody result for pestiviruses can lead to an extremely time-consuming and costly, and not always successful search for the source of the infection if no active virus excretor is found. This study documents how small ruminants can be integrated into pestivirus monitoring with a manageable amount of work and costs. 23 406 sheep and goat samples from two brucellosis surveillance programs in small ruminants were analyzed retrospectively. Blood samples were examined using pestivirus real-time pool RT-PCR (qPCR). Direct virus detection of BDV-3 was achieved in 40 sheep from five different federal states. Over the entire investigation period a further 37 detections of BDV-3 were found in cattle, sheep and goats outside of this study throughout Austria. This study accounts for 52 % of all border disease detections from 2015 to 2022. By including small ruminants in pestivirus monitoring, the disruptive factor BDV and the risk of its introduction into cattle herds can be significantly minimized in the future.

New Potential Role of European Mouflon (Ovis aries musimon) in the Epidemiology of Border Disease in the Pyrenees.

Since 2001, high-mortality outbreaks of border disease (BD) have negatively affected populations of Pyrenean chamois (Rupicapra pyrenaica pyrenaica). Studies in the affected areas determined that sympatric wild ruminants did not seem to have an epidemiologic role in the circulation of border disease virus (BDV). However, the recent increase in European mouflon (Ovis aries musimon) densities might enhance the risk of pathogen transmission among chamois and mouflons. We conducted a serologic and virologic investigation of BDV in European mouflon from the Spanish Pyrenees, with the aim of determining potential changes in the role of this species in BDV epidemiology. From 2018 to 2022, we detected antibodies against BDV in 31/185 (16.7%) animals but did not detect BDV RNA in any spleen sample (0/65). These results indicate that BDV infection is occurring in these mouflon populations to a greater extent than previously described, which could shift the current understanding of BD epidemiology in the Pyrenees and cause an unpredictable effect on both chamois and mouflon populations. Further studies on the molecular identification of BDV in mouflon and chamois are required to better understand the contribution of mouflon in the epidemiology of BD.

Circulation of Pestiviruses in Small Ruminants from Greece: First Molecular Identification of Border Disease Virus.

The incidence of small ruminant pestivirus infections in Greece remains unknown as they have not been diagnosed in the country since 1974 when the most recent Border Disease Virus (BDV) outbreak was reported. The objective of our study was to explore the possible occurrence of pestiviral infections among sheep and goat farms in Greece and to further determine the variants of major concern. Thus, serum samples were collected from 470 randomly selected animals belonging to 28 different flocks/herds. ELISA on p80 antibody revealed the existence of seropositive animals in four out of the 24 studied sheep flocks, whereas all the goats in the four studied herds were seronegative. Viral RNA and antigens were detected in two sheep out of the four seropositive flocks by RT-PCR and ELISA, respectively. Sequencing and phylogenetic analysis showed that the newly identified Greek variants were closely related to the strains of the BDV-4 genotype. One of the BDV-positive sheep demonstrated the diagnostic profile of a persistently infected (PI) animal, providing additional information regarding the source of the infection. This is the first molecular identification of BDV isolates in Greece. Our findings indicate that BDV infections are likely to remain undiagnosed, highlighting the need for further epidemiological studies and active surveillance programs to determine the prevalence and impact of BDV infections on a countrywide level.

Vaccination of Sheep with Bovine Viral Diarrhea Vaccines Does Not Protect against Fetal Infection after Challenge of Pregnant Ewes with Border Disease Virus.

Border Disease (BD) is a major sheep disease characterized by immunosuppression, congenital disorders, abortion, and birth of lambs persistently infected (PI) by Border Disease Virus (BDV). Control measures are based on the elimination of PI lambs, biosecurity, and frequent vaccination which aims to prevent fetal infection and birth of PI. As there are no vaccines against BDV, farmers use vaccines directed against the related Bovine Viral Diarrhea Virus (BVDV). To date, there is no published evidence of cross-effectiveness of BVDV vaccination against BDV infection in sheep. We tested three commonly used BVDV vaccines, at half the dose used in cattle, for their efficacy of protection against a BDV challenge of ewes at 52 days of gestation. Vaccination limits the duration of virus-induced leukopenia after challenge, suggesting partial protection in transient infection. Despite the presence of BDV neutralizing antibodies in vaccinated ewes on the day of the challenge, fetuses of vaccinated and unvaccinated sheep were, two months after, highly positive for BDV RNA loads and seronegative for antibodies. Therefore, BVDV vaccination at half dose was not sufficient to prevent ovine fetal infection by BDV in a severe challenge model and can only be reconsidered as a complementary mean in BD control.

Prevalence of border disease virus in sheep and goats in Mosul, Iraq

Globally, Border disease virus (BDV) has caused substantial economic losses among small ruminants (sheep and goats). This is the first molecular study carried out in Mosul city, Iraq. To determine the prevalence of Border Disease Virus and to examine problem of persistent infection (PI) using Reverse transcriptase polymerase chain reaction technique (RT-PCR) in female local breed of small ruminants. During the period between November 2018 to June 2019, 364 blood samples were collected from 264 local Awassi sheep and 100 local cross breed goats secure provided by private breeders. The animals were of ≥1.5 years old and the samples were obtained from various locations in Mosul city, with varying rearing methods and had not been vaccinated against BDV. This investigation indicated that the prevalence of BDV infection in sheep and goats were 15.9% (42/264) and 3% (3/100) respectively, whereas the occurrence of PI in sheep was 2.38% and in goats was 0%. Hence it was concluded that Border disease was circulating in small ruminants in Mosul city. This calls for a need to design programs to monitor and control the disease and eventually eradicate it is prevalence in Mosul city.

Pestivirus infection of sheep – testing times?

Bovine viral diarrhoea in cattle and border disease in sheep are caused by infection with the pestiviruses bovine viral diarrhoea virus (BVDV) and border disease virus (BDV), respectively, although either...

New insights on pestivirus infections in transhumant sheep and sympatric Pyrenean chamois (Rupicapra p. pyrenaica)

Border Disease Virus (BDV) causes health and economic impact on livestock and is also of importance in wildlife conservation as it causes high mortality outbreaks in Pyrenean chamois (Rupicapra pyrenaica pyrenaica). Pastoral practices are known as a main interspecies pathogen transmission. Hence, the presence of pestivirus in transhumant sheep flocks and sympatric chamois was assessed in areas with different epidemiological scenarios of chamois BDV infections. Moreover, the present study had also the goal to identify if inter-specific infections occurred and when they happened. Five sheep flocks grazing in two alpine areas in the Pyrenees with two different BDV epidemiological scenarios in chamois populations were studied during two transhumant seasons. Sheep were sampled before and after transhumance. Pyrenean chamois sera and spleen samples from both areas where also studied during the same period. Antibodies against BDV were assessed by means of ELISA and VNT. A qRT-PCR was used in order to detect the virus. Seroprevalence in sheep ranged between 0 and 91.1% at the flock level. Chamois were found to have high seroprevalences (52.9–77.7%) in both areas, and four new BDV isolates were sequenced. One sheep farm presented persistent BDV circulation and three showed low BDV circulation. The after-transhumance period was identified as the moment when viral transmission occured in the first farm, associated to BDV strains of domestic origin, according to VNT results. However, the BDV isolate was genetical closely related to previous BDV strains from chamois origin. In another farm, antibodies in two of the three positive sera were associated to infection with a chamois-like BDV strain. Altogether indicates that occasional viral transmission from chamois to sheep may occur.

Border Disease Virus Infection of Bovine Placentas.

Subsequent to a previous study of border disease virus (BDV) horizontal transmission from a persistently BDV-infected calf to 6 seronegative pregnant heifers, the heifers were slaughtered 60 days after exposure to the infected calf, and their fetuses and placentas were examined. Immunohistochemical examination of fetal organs and placenta showed positive labeling of moderate intensity for pestivirus antigen in 3 of 6 heifers. BDV infection in these 3 animals was confirmed by the detection of BDV RNA in different organs using reverse transcription quantitative polymerase chain reaction. In the placenta, the positive cells were visualized mostly on the fetal side. In those 3 heifers that harbored an infected fetus, the placental tissue in the placentome region showed a moderate to severe mononuclear and fibrosing placentitis and, in severe cases, necrotic areas. The inflammatory population was composed predominantly of T and B cells, a substantial number of macrophages, and, to a lesser extent, plasma cells. This is a novel report of placentitis in persistently BDV-infected fetuses from pregnant heifers that became acutely infected by cohousing with a calf persistently infected with BDV, which extends previous reports on bovine viral diarrhea virus-infected and BDV-infected cattle and sheep, respectively.

Spatial and Temporal Phylogeny of Border Disease Virus in Pyrenean Chamois (Rupicapra p. pyrenaica).

Border disease virus (BDV) affects a wide range of ruminants worldwide, mainly domestic sheep and goat. Since 2001 several outbreaks of disease associated to BDV infection have been described in Pyrenean chamois (Rupicapra pyrenaica pyrenaica) in Spain, France and Andorra. In order to reconstruct the most probable places of origin and pathways of dispersion of BDV among Pyrenean chamois, a phylogenetic analysis of 95 BDV 5’untranslated sequences has been performed on chamois and domestic ungulates, including novel sequences and retrieved from public databases, using a Bayesian Markov Chain Monte Carlo method. Discrete and continuous space phylogeography have been applied on chamois sequences dataset, using centroid positions and latitude and longitude coordinates of the animals, respectively.The estimated mean evolutionary rate of BDV sequences was 2.9×10−3 subs/site/year (95% HPD: 1.5–4.6×10−3). All the Pyrenean chamois isolates clustered in a unique highly significant clade, that originated from BDV-4a ovine clade. The introduction from sheep (dated back to the early 90s) generated a founder effect on the chamois population and the most probable place of origin of Pyrenean chamois BDV was estimated at coordinates 42.42 N and 1.9 E. The pathways of virus dispersion showed two main routes: the first started on the early 90s of the past century with a westward direction and the second arise in Central Pyrenees. The virus spread westward for more than 125 km and southward for about 50km and the estimated epidemic diffusion rate was about 13.1 km/year (95% HPD 5.2–21.4 km/year). The strong spatial structure, with strains from a single locality segregating together in homogeneous groups, and the significant pathways of viral dispersion among the areas, allowed to reconstruct both events of infection in a single area and of migrations, occurring between neighboring areas.

Influence of border disease virus (BDV) on serological surveillance within the bovine virus diarrhea (BVD) eradication program in Switzerland.

BackgroundIn 2008, a program to eradicate bovine virus diarrhea (BVD) in cattle in Switzerland was initiated. After targeted elimination of persistently infected animals that represent the main virus reservoir, the absence of BVD is surveilled serologically since 2012. In view of steadily decreasing pestivirus seroprevalence in the cattle population, the susceptibility for (re-) infection by border disease (BD) virus mainly from small ruminants increases. Due to serological cross-reactivity of pestiviruses, serological surveillance of BVD by ELISA does not distinguish between BVD and BD virus as source of infection.ResultsIn this work the cross-serum neutralisation test (SNT) procedure was adapted to the epidemiological situation in Switzerland by the use of three pestiviruses, i.e., strains representing the subgenotype BVDV-1a, BVDV-1h and BDSwiss-a, for adequate differentiation between BVDV and BDV. Thereby the BDV-seroprevalence in seropositive cattle in Switzerland was determined for the first time. Out of 1,555 seropositive blood samples taken from cattle in the frame of the surveillance program, a total of 104 samples (6.7%) reacted with significantly higher titers against BDV than BVDV. These samples originated from 65 farms and encompassed 15 different cantons with the highest BDV-seroprevalence found in Central Switzerland. On the base of epidemiological information collected by questionnaire in case- and control farms, common housing of cattle and sheep was identified as the most significant risk factor for BDV infection in cattle by logistic regression.ConclusionThis indicates that pestiviruses from sheep should be considered as a source of infection of domestic cattle and might well impede serological BVD surveillance.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-016-0932-0) contains supplementary material, which is available to authorized users.

Identification and molecular characterization of border disease virus (BDV) from sheep in India.

Pestiviruses isolated from sheep and goats in India thus far have been bovine viral diarrhoea virus 1 (BVDV-1) or BVDV-2. During routine genetic typing of pestiviruses in the years 2009-10, border disease virus (BDV) was detected in eight Indian sheep of a flock showing clinical signs of BD by real time RT-PCR. All the samples yielded positive virus isolates in cell culture but were found negative by a BVDV antigen ELISA. A representative BDV isolate was characterized at genetic and antigenic level. Phylogenetic analysis carried out in 5'-UTR, N(pro) and E2 regions of genome typed the Indian BDV isolate as BDV-3. A more detailed analysis in N(pro) and entire region coding structural proteins showed that the N(pro) (168), C (100 aa), E(rns) (227 aa), E1 (195 aa) and E2 (373 aa) proteins were of size characteristic for BDV reference strain X818. Antigenic differences were evident between the BDV-3 isolate and previously reported BDV-1, BDV-5 and BDV-7 strains. Although origin of BDV-3 in India is not clear, the results reflect probable introduction through trade in sheep between India and other countries or BDV-3 may be more widely distributed. Additionally, this study suggests that for diagnosis of BDV infection, the commercial BVDV Ag-ELISA should be used with caution. This is the first identification of BDV in sheep in India which highlights the need for continued pestivirus surveillance and assessing its impact on sheep and goat production.

Chinese border disease virus strain JSLS12-01 infects piglets and down-regulates the antibody responses of classical swine fever virus C strain vaccination

During 2012 and 2013, several border disease virus (BDV) strains were identified from Chinese goat and sheep herds. At the same time, pigs from the same areas were found to be seropositive to BDV by ELISA, without showing clinical signs (unpublished data). To examine the susceptibility of pigs to the Chinese BDV strains, BDV isolate JSLS12-01, isolated from naturally infected sheep, was used to infect pigs. Antibody responses, viremia, clinical signs and pathological changes of the infected animals were examined. It confirmed that the current BDV strain could infect the domestic pigs, the animals showed viremia during 4 to 14 days post infection (dpi) and sero-conversion from 14dpi; no clinical and pathological changes were observed. In addition, CSFV maternal antibody did not influence BDV infection. Subsequently, pigs were infected with the BDV isolate and vaccinated with Hog cholera lapinized virus (HCLV) 21 days later to determine the effect of BDV infection on antibody induction of CSFV vaccination. The specific CSFV antibody and neutralizing antibody titers of the BDV infected group remained negative after the primary vaccination. Even after the boost vaccination, they were still significantly lower than those of the uninfected groups (p<0.05). These results indicated that BDV infection could down-regulate the antibody responses of CSFV C-strain vaccination. It should be paid attention that BDV prevalence in pig herds and in live vaccines might hamper the vaccination of CSF.

Characterization of one sheep border disease virus in China.

BackgroundBorder disease virus (BDV) causes border disease (BD) affecting mainly sheep and goats worldwide. BDV in goat herds suffering diarrhea was recently reported in China, however, infection in sheep was undetermined. Here, BDV infections of sheep herds in Jiangsu, China were screened; a BDV strain was isolated and identified from the sheep flocks in China. The genomic characteristics and pathogenesis of this new isolate were studied.ResultsIn 2012, samples from 160 animals in 5 regions of Jiangsu province of China were screened for the presence of BDV genomic RNA and antibody by RT-PCR and ELISA, respectively. 44.4% of the sera were detected positively, and one slowly grown sheep was analyzed to be pestivirus RNA positive and antibody-negative. The sheep kept virus positive and antibody negative in the next 6 months of whole fattening period, and was defined as persistent infection (PI). The virus was isolated in MDBK cells without cytopathic effect (CPE) and named as JSLS12-01. Near-full-length genome sequenced was 12,227 nucleotides (nt). Phylogenetic analysis based on 5'-UTR and Npro fragments showed that the strain belonged to genotype 3, and shared varied homology with the other 3 BDV strains previously isolated from Chinese goats. The genome sequence of JSLS12-01 also had the highest homology with genotype BDV-3 (the strain Gifhorn). Experimental infections of sheep had mild clinical signs as depression and short-period mild fever (5 days). Viremia was detected in 1–7 days post-infection (dpi), and seroconversion began after 14 dpi.ConclusionsThis study reported the genomic and pathogenesis characterizations of one sheep BDV strain, which confirmed the occurrence of BDV infection in Chinese sheep. This sheep derived BDV strain was classified as BDV-3, together with the goat derived strains in China. These results might be helpful for further understanding of BDV infection in China and useful for prevention and control of BDV infections in the future.

First report on seroprevalence of bluetongue, border disease and peste des petits ruminants virus infections in sheep in Kyrgyzstan

In this study, the presence of antibodies against Bluetongue Virus (BTV), Border Disease Virus (BDV) and Peste des Petits Ruminants virus (PPRV), were evaluated for the first time in Kyrgyzstan. A total of 655 serum samples were collected from healthy sheep (Jaydara breed) from different regions of Kyrgyzstan (144 from Issik Gol region, 208 from Narin region, 189 from Talas region and 114 from Cuy region). Commercially available competitive ELISA kits were used to detect antibodies against tested samples. Seroprevalence was found to be 36.94%, 7.32% and 35.11% against BTV, BDV and PPRV infections, respectively. Significant differences in antibody prevalences were found among both BTV-BDV and BDV-PPRV groups (P< 0.001) only. This is the first report on seroprevalence of viral infections for BTV, BDV and PPRV in the Jaydara breed of sheep within Kyrgyzstan.

Molecular and serological investigation of border disease virus infection in sheep in the Kars District of Turkey

This study is a serological and virological examination of the border disease virus (BDV) in sheep at 1–5 years of age from private small scale production units of less than 20 sheep per unit, in the Kars District of Turkey. For this purpose, blood sera from 460 sheep were tested for antibodies against BDV using a commercial enzyme-linked immunosorbent assay (ELISA). Since BDV causes persistent infection, antigen-ELISA was also performed for this agent. Seropositivity rate was detected to be 74.57%. In addition, the BDV antigen was detected in one sample of seronegative sera (0.85%). Reverse transcription polymerase chain reaction (RT-PCR) technique was used to determine the presence of pestivirus nucleic acid by using 5’UTR primer pair. Pestivirus nucleic acid was found in 2 of 117 seronegative samples (1.71%) by RT-PCR. The results suggest that the infection was spreading in private small scale production units. Furthermore, recommendations for the control of BDV infection are presented. This study is the first molecular and serological study to determine viroprevalence and seroprevalence of BDV infection in sheep in the Kars District of Turkey.

Comparative Investigation of Border Disease Virus Infection in Sheep Flocks with Abortion Problems in Konya Province

The aim of the present study was to determine the serological and virological status of animals persistently infected (PI) with border disease virus (BDV), with focus on the prevalence of pestiviruses in sheep and lambs. This study is the first to our knowledge, to investigate the role of viral infections in abortions in the sheep population of Konya province. The prevalence of antibodies against BDV was as follows: sheep, 79%; lambs, 43.4%; and rams, 6%. Of 1000 leukocyte (sheep) samples examined with a direct ELISA, 11 (1.1%) (first sampling) were antibody-positive. Three sheep were determined to be PI and were sacrificed. Of 1011 sheep leukocyte samples, 14 were examined using a direct immunoperoxidase (IP) assay for BDV antigen, and 11 of 327 (3.36%) tissue samples were positive. The BDV genome was detected in 14 of 63 (22.2%) sheep leukocyte samples and 11 of 327 (3.36%) tissue samples by one-step RT-PCR. The BDV genome could not be detected in any vaginal swab samples. The sensitivity and specificity rates between direct ELISA and direct IP were respectively 100% and 98.8% and the values were, respectively, 78.57% and 98.6% between direct ELISA and one-step RT-PCR and 100% and 97.5% between one-step RT-PCR and direct IP. In conclusion it was determinate that BDV has an important role as an etiologic agent in abortion causes in sheep population in Konya.

Natural border disease virus infection in feedlot lambs

Border disease is caused by border disease virus (BDV, a pestivirus from the family Flaviviridae ) infection in sheep and goats (Vantsis and others 1976). BDV infection causes sizeable economic...

A survey on the status of the border disease virus infection in sheep with reproductive failure using cell culture and polymerase chain reaction (PCR) methods in Tabriz, Iran

The most important causal agent of border disease (BD), border disease virus (BDV), is a Pestivirus in the family Flaviviridae. In attention to the lack of molecular documented data on the prevalence of BDV in Iran, especially in East Azerbaijan Province, northwest of Iran, this research was designed to determine the prevalence of BDV in suspected ewes that have an antecedent of reproductive failure. Eighty-eight suspected ewes in the breed of Ghezel, Mogan, and mix (Ghezel × Mogan) in five various districts of East Azerbaijan Province (Azershahr, Lyqvan, Osku, Sofian, and Varzqan) were selected. Razi bovine kidney (RBK) cell lines were grown, and the buffy coat layer was separated and inoculated to the monolayer cell culture. All cell cultures were inspected for cytopathic effects after 5–7 days. BDV genomes were being checked using nested reverse transcription polymerase chain reaction method. Ten samples (10/88, 11.36 %) were positive for the genome of BDV. The prevalence of BDV infection was found to vary between different districts. The highest percentage of infected sheep was in animals at the 3–4.5-year-old range (15.38 %). There was no statistically significant difference in the prevalence among different age categories. The highest BDV infection rate was observed in full blood ewes in Mogan and Ghezel. Cytopathic effects of the virus in the culture on RBK cell line were not observed at all. This study proved that the infection of BDV is present in sheep in the East Azerbaijan, northwest of Iran, and cause some reproductive failure in the flocks. The survey is the first molecular evidence of the occurrence of the disease in the country.

Investigation of border disease and bovine virus diarrhoea in sheep from 76 mixed cattle and sheep farms in eastern Switzerland

The purpose of this study was to examine the occurrence of sheep persistently infected with Border disease virus (BDV) on 76 mixed cattle and sheep farms and whether seroconversion to BDV infection occurred in cattle of these farms. Seroprevalence of BDV and bovine viral disease virus (BVDV) infection in sheep was also investigated. Quantitative RT-PCR for pestivirus detection and an ELISA to detect pestivirus antibodies were used in 2'384 and 2'291 ovine blood samples, respectively. Another 27 seropositive sheep from ten flocks underwent serum neutralization testing to differentiate between BDV and BVDV antibodies. A BDV titre that was at least four times higher than the BVDV titre was interpreted as the result of BDV infection. Titres against BVDV were interpreted in an analogous fashion. All examined sheep were pestivirus-negative, 310 sheep were seropositive, 119 had an indeterminate titre and 1'862 were seronegative. The flock seroprevalence ranged from 0.0 to 73.9 %. Three of the 27 flocks that underwent serum neutralization testing were interpreted as BDV-infected because of 6 sheep with higher BDV titres, and 6 flocks were interpreted as BVDV-infected because of 14 sheep with higher BVDV titres.