Bluetongue Virus Serotype Research Articles

Immunological and Pathogenic Differences of Two Experimental Bluetongue Virus Serotype Infections Evaluated in Two Disparate Host Species.

Bluetongue virus (BTV) is a prevalent midge-borne pathogen that infects ruminant species worldwide. BTV infections range from asymptomatic to lethal, with mechanisms that determine the severity of infection remaining largely undefined. Although it is relatively poorly understood, the immune response to BTV infection is thought to be critical for both the propagation of disease as well as the resolution of infection. To bridge this gap in knowledge, we infected cohorts of sheep and muntjac deer with two serotypes of BTV (BTV10 and BTV17) for longitudinal analysis (30 days). Interestingly, species-specific differences were observed. Circulating virus was detected early and remained detectable for the duration of the sheep study, while infections in muntjac showed faltering detection of BTV10 at 3 weeks post infection. The magnitude of the immune response was subdued in the muntjac when compared to the sheep cohorts, though similar responses were observed. We also assessed midge viral uptake and the ability to replicate BTV. Midges successfully fed on both species, yet those that fed on sheep resulted in more efficient BTV transmission. Our findings demonstrate that differences in BTV infections, immune responses, and vector competence across host species and serotypes will impact global BTV emergence and strategies for mitigation.

Identification and characterization of two atypical strains of bluetongue virus in sheep, Tunisia

Bluetongue virus (BTV) is the causative agent of the bluetongue disease (BT), an infectious disease of domestic and wild ruminants that is primarily transmitted by Culicoides biting midges. In recent years, several novel BTV serotypes (often referred to as “atypical” BTVs) have been documented. These strains are usually asymptomatic in animals and seem to be unable to replicate efficiently in the arthropod vector.Here we report the detection of two putative atypical BTV strains in the Governorate of Gafsa, in the southwest region of Tunisia. Specifically, we recognised the recurrence of an atypical BTV strain (BTV-Y TUN2022) and a novel BTV-W TUN2022.

Impact of VP2 structure on antigenicity: comparison of BTV1 and the highly virulent BTV8 serotype.

Bluetongue virus (BTV) is an agriculturally and economically significant insect-borne virus that causes serious illness and death in sheep and other domestic and wild ruminants in large areas of the world. Numerous BTV serotypes exist, and distant serotypes exhibit unique neutralizing antibody profiles, which target the outermost capsid protein VP2. The predominant serotype-specific nature of the antibody response to VP2 is a barrier to the development of broad-spectrum prophylactic BTV vaccine candidates. Although VP2 is the main serotype determinant of BTV, the structural basis of serotype specificity has not been investigated. In this study, we utilized the recently available atomic structure of VP2 with a modeled tip domain to carry out in silico structural comparisons between distant serotypes BTV1 and BTV8. These analyses identified structural differences in the tip domain, positioned at the apex of VP2, and informed the design of mutant VP2 constructs. Dissection of tip domain antigenicity demonstrated that the region of structural difference between BTV1 and highly virulent BTV8 was a target of BTV neutralizing antibodies and that mutation of this region resulted in a loss of neutralizing antibody recognition. This study has for the first time provided insights into the structural differences, which underpin the serotype-specific neutralizing antibody response to BTV.IMPORTANCEThe immune system can protect against virus infection by producing antibodies, which bind and inhibit the virus from infecting the susceptible host. These antibodies are termed neutralizing antibodies and generally target the viral receptor binding protein, such as the VP2 of bluetongue virus (BTV). This pressure from the immune system can drive mutation of the viral protein resulting in escape from antibody-mediated neutralization and the evolution of serotypes, as is the case for BTV. Understanding the structural differences, which underpin the different BTV serotypes, could help guide the design of a BTV vaccine that targets multiple serotypes. In this study, we have mapped the VP2 structural differences between distant serotypes, to a region targeted by neutralizing antibodies, and have demonstrated for the first time how VP2 structure is the fundamental basis of serotype specificity.

Bluetongue virus serotype 3 in ruminants in the Netherlands: Clinical signs, seroprevalence and pathological findings.

The bluetongue virus serotype 3 (BTV-3) outbreak in the Netherlands in 2023 caused severe clinical signs in ruminants. The clinical and pathological signs in ruminants and their spread during the outbreak in 2023 are described. Data from the Dutch monitoring and surveillance system were available to describe clinical signs and pathological findings related to BTV-3 in sheep, cattle and goats. During the outbreak, 13 farms (five sheep, five cattle and three dairy goats) were closely monitored. In 2023, BTV-3 infections were confirmed by real-time polymerase chain reaction in sheep flocks (n = 1807), cattle herds (n = 1864), goat herds (n = 62), alpaca and/or llama herds (n = 15) and one dog. Sheep exhibited the most severe clinical signs and had the highest mortality. In other animal species, a large variation in both occurrence and severity of clinical signs was observed. Only 13 farms were closely monitored. The clinical signs observed in affected animals during the 2023 BTV-3 outbreak seem to be more severe than those observed during the BTV-8 outbreak between 2006 and 2008. It seems likely that BTV-3 will overwinter, similar to BTV-8. Therefore, the availability of an effective and safe vaccine is crucial to limit the future impact of BTV-3.

Emergence of Bluetongue Virus Serotype 3, the Netherlands, September 2023.

Since 1998, notifiable bluetongue virus (BTV) serotypes 1-4, 6, 8, 9, 11, and 16 have been reported in Europe. In August 2006, a bluetongue (BT) outbreak caused by BTV serotype 8 began in northwestern Europe. The Netherlands was declared BT-free in February 2012, and annual monitoring continued. On September 3, 2023, typical BT clinical manifestations in sheep were notified to the Netherlands Food and Product Safety Consumer Authority. On September 6, we confirmed BTV infection through laboratory diagnosis; notifications of clinical signs in cattle were also reported. We determined the virus was serotype 3 by whole-genome sequencing. Retrospective analysis did not reveal BTV circulation earlier than September. The virus source and introduction route into the Netherlands remains unknown. Continuous monitoring and molecular diagnostic testing of livestock will be needed to determine virus spread, and new prevention strategies will be required to prevent BTV circulation within the Netherlands and Europe.

The impact of the bluetongue serotype 3 outbreak on sheep and goat mortality in the Netherlands in 2023

In September 2023, bluetongue virus serotype 3 (BTV-3) emerged in the Netherlands, infecting over five thousand livestock farms. In sheep, high morbidity and mortality rates were reported that were unlike previously described bluetongue outbreaks. This study aimed to quantify the impact of BTV-3 in the small ruminant population in the Netherlands in 2023. Sheep and goat movement census data and BTV-3 notification data were available from 2020 until the end of 2023. Data were aggregated to farm and week level and mortality indicators were calculated for lambs (<1 year) and adult animals (≥1 year). Population averaged GEE models with a Negative-binomial distribution and a log-link function correcting for repeated measures per farm in time were used to quantify the association between BTV-3 and mortality. In 2023, 2994 sheep farmers and 89 goat farmers notified clinical signs of BTV-3 to the NVWA. During this BTV-3 outbreak period, an additional 55,000 sheep died compared to the same period in 2020–2022. At flock level a high variety in mortality was observed, with a clear increase in mortality in both flocks that were not notified but that were located in infected areas and in flocks of which the farmer notified clinical signs. During the BTV-3 outbreak period, mortality in infected areas increased 4.2 (95 % CI: 4.0–4.3) times in sheep lambs (<1 year) and 4.6 (95 % CI: 4.4–4.8) times in sheep (≥1 year) compared to BTV-3 free areas. Flocks with a confirmed BTV-3 infection that were notified in September showed a 12.8 (95 % CI: 11.4–14.3) times higher mortality in lambs and a 15.1 (95 % CI: 13.7–16.6) times higher mortality in sheep compared to flocks in BTV-3 areas. In flocks of which the farmer notified clinical signs after September, mortality was 4.6 (95 % CI: 4.2–5.0) and 5.6 (95 % CI: 5.1–6.0) times higher in lambs and sheep compared BTV-3 areas respectively. In goats, around 4000 additional deaths were recorded during the BTV-3 outbreak period. In farms that were notified, mortality of goats (≥1 year) was 1.8 (95 % CI: 1.2–2.8) times higher compared to BTV-3 free areas. Since May 2024, multiple BTV-3 vaccines are available in the Netherlands. In June 2024, the first new infections of BTV-3 were confirmed in Dutch sheep flocks. Hopes are that with the possibility to vaccinate, the spread and impact of BTV-3 in the Netherlands will rapidly decline and that losses as observed in 2023 will no longer be seen.

Sero-epidemiology of bluetongue in ruminants raised on private holdings in North-Western Pakistan.

This study investigated the sero-epidemiology of bluetongue in ruminants in North-Western Pakistan. A total of 3,173 serum samples were collected from small (n = 1,651) and large (n = 1,522) ruminants being reared by farmers in 14 districts. Antibodies to bluetongue virus (BTV) were detected usingcompetitive ELISA. The overall prevalence of BTV antibodies was 65%. A significant association (P < 0.05) between the prevalence of BTV antibodies and the risk factors including sex, species, age, area, husbandry practices and breed was shown by univariate analysis. In multivariate analysis, the seroprevalence was 6.5 (95% CL = 3.7-11.4), 5.9 (95% CL = 3.8-9.4) and 2.4 (95% CL = 1.5-3.7) timeshigher in buffaloes, cattle and goats than sheep, respectively. The seroprevalence was 1.4 (95% CL= 1.1-1.7) times higher in local breeds than in cross/exotic breeds. The seroprevalence was 1.6 (95% CL = 1.1 to 2.3) times higher in sedentary animals than in nomadic animals. The seroprevalence was significantly associated with age. Further work is required to determine the BTV serotypes prevalent in the study area for effective control of the disease.

Quantitative risk assessment for the introduction of bluetongue virus into mainland Europe by long-distance wind dispersal of Culicoides spp.: A case study from Sardinia.

Europe faces regular introductions and reintroductions of bluetongue virus (BTV) serotypes, most recently exemplified by the incursion of serotype 3 in the Netherlands. Although the long-distance wind dispersal of the disease vector, Culicoides spp., is recognized as a virus introduction pathway, it remains understudied in risk assessments. A Quantitative Risk Assessment framework was developed to estimate the risk of BTV-3 incursion into mainland Europe from Sardinia, where the virus has been present since 2018. We used an atmospheric transport model (HYbrid Single-Particle Lagrangian Integrated Trajectory) to infer the probability of airborne dispersion of the insect vector. Epidemiological disease parameters quantified the virus prevalence in vector population in Sardinia and its potential first transmission after introduction in a new area. When assuming a 24h maximal flight duration, the risk of BTV introduction from Sardinia is limited to the Mediterranean Basin, mainly affecting the southwestern area of the Italian Peninsula, Sicily, Malta, and Corsica. The risk extends to the northern and central parts of Italy, Balearic archipelago, and mainland France and Spain, mostly when maximal flight duration is longer than 24h. Additional knowledge on vector flight conditions and Obsoletus complex-specific parameters could improve the robustness of the model. Providing both spatial and temporal insights into BTV introduction risks, our framework is a key tool to guide global surveillance and preparedness against epizootics.

Bluetongue Virus Serotype 3 and Schmallenberg Virus in Culicoides Biting Midges, Western Germany, 2023.

In October 2023, bluetongue virus serotype 3 (BTV-3) emerged in Germany, where Schmallenberg virus is enzootic. We detected BTV-3 in 1 pool of Culicoides biting midges collected at the time ruminant infections were reported. Schmallenberg virus was found in many vector pools. Vector trapping and analysis could elucidate viral spread.

Immuno-informatics study identifies conserved T cell epitopes in non-structural proteins of Bluetongue virus serotypes: formulation of a computationally optimized next-generation broad-spectrum multi-epitope vaccine.

Bluetongue (BT) poses a significant threat to the livestock industry, affecting various animal species and resulting in substantial economic losses. The existence of numerous BT virus (BTV) serotypes has hindered control efforts, highlighting the need for broad-spectrum vaccines. In this study, we evaluated the conserved amino acid sequences within key non-structural (NS) proteins of BTV and identified numerous highly conserved murine- and bovine-specific MHC class I-restricted (MHC-I) CD8+ and MHC-II-restricted CD4+ epitopes. We then screened these conserved epitopes for antigenicity, allergenicity, toxicity, and solubility. Using these epitopes, we developed in silico-based broad-spectrum multiepitope vaccines with Toll-like receptor (TLR-4) agonists. The predicted proinflammatory cytokine response was assessed in silico using the C-IMMSIM server. Structural modeling and refinement were achieved using Robetta and GalaxyWEB servers. Finally, we assessed the stability of the docking complexes through extensive 100-nanosecond molecular dynamics simulations before considering the vaccines for codon optimization and in silico cloning. We found many epitopes that meet these criteria within NS1 and NS2 proteins and developed in silico broad-spectrum vaccines. The immune simulation studies revealed that these vaccines induce high levels of IFN-γ and IL-2 in the vaccinated groups. Protein-protein docking analysis demonstrated promising epitopes with strong binding affinities to TLR-4. The docked complexes were stable, with minimal Root Mean Square Deviation and Root Mean Square Fluctuation values. Finally, the in silico-cloned plasmids have high % of GC content with > 0.8 codon adaptation index, suggesting they are suitable for expressing the protein vaccines in prokaryotic system. These next-generation vaccine designs are promising and warrant further investigation in wet lab experiments to assess their immunogenicity, safety, and efficacy for practical application in livestock. Our findings offer a robust framework for developing a comprehensive, broad-spectrum vaccine, potentially revolutionizing BT control and prevention strategies in the livestock industry.

Treatment considerations for bluetongue virus serotype‐3 cases in sheep

Background: Bluetongue virus (BTV) serotype‐3 (BTV‐3) has been circulating in the Netherlands since September 2023 and was first reported in Kent in the UK in November 2023, picked up via the annual bluetongue surveillance programme. It is a highly pathogenic strain in sheep with limited preventive measures once virus is circulating in the biting midge population. At the time of publication there is no BTV‐3 vaccine available and no cross‐protection from other BTV vaccine strains. There are no specific antiviral medicines available, and therapy focuses on nursing care and anti‐inflammatory medicines to reduce clinical signs, although early euthanasia decisions need to be considered on the grounds of animal welfare.Aim of the article: This article takes the experiences of vets from the Netherlands and describes what practitioners might be able to do to treat clinical cases of BTV‐3, with thanks to Schapendokter (2023) for sharing their practice protocols.

Spatial Transmission Characteristics of the Bluetongue Virus Serotype 3 Epidemic in The Netherlands, 2023.

A devastating bluetongue (BT) epidemic caused by bluetongue virus serotype 3 (BTV-3) has spread throughout most of the Netherlands within two months since the first infection was officially confirmed in the beginning of September 2023. The epidemic comes with unusually strong suffering of infected cattle through severe lameness, often resulting in mortality or euthanisation for welfare reasons. In total, tens of thousands of sheep have died or had to be euthanised. By October 2023, more than 2200 locations with ruminant livestock were officially identified to be infected with BTV-3, and additionally, ruminants from 1300 locations were showing BTV-associated clinical symptoms (but not laboratory-confirmed BT). Here, we report on the spatial spread and dynamics of this BT epidemic. More specifically, we characterized the distance-dependent intensity of the between-holding transmission by estimating the spatial transmission kernel and by comparing it to transmission kernels estimated earlier for BTV-8 transmission in Northwestern Europe in 2006 and 2007. The 2023 BTV-3 kernel parameters are in line with those of the transmission kernel estimated previously for the between-holding spread of BTV-8 in Europe in 2007. The 2023 BTV-3 transmission kernel has a long-distance spatial range (across tens of kilometres), evidencing that in addition to short-distance dispersal of infected midges, other transmission routes such as livestock transports probably played an important role.

Evaluating Temperature Effects on Bluetongue Virus Serotype 10 and 17 Coinfection in Culicoides sonorensis.

Bluetongue virus (BTV) is a segmented, double-stranded RNA virus transmitted by Culicoides midges that infects ruminants. As global temperatures increase and geographical ranges of midges expand, there is increased potential for BTV outbreaks from incursions of novel serotypes into endemic regions. However, an understanding of the effect of temperature on reassortment is lacking. The objectives of this study were to compare how temperature affected Culicoides survival, virogenesis, and reassortment in Culicoides sonorensis coinfected with two BTV serotypes. Midges were fed blood meals containing BTV-10, BTV-17, or BTV serotype 10 and 17 and maintained at 20 °C, 25 °C, or 30 °C. Midge survival was assessed, and pools of midges were collected every other day to evaluate virogenesis of BTV via qRT-PCR. Additional pools of coinfected midges were collected for BTV plaque isolation. The genotypes of plaques were determined using next-generation sequencing. Warmer temperatures impacted traits related to vector competence in offsetting ways: BTV replicated faster in midges at warmer temperatures, but midges did not survive as long. Overall, plaques with BTV-17 genotype dominated, but BTV-10 was detected in some plaques, suggesting parental strain fitness may play a role in reassortment outcomes. Temperature adds an important dimension to host-pathogen interactions with implications for transmission and evolution.

Assessing Reassortment between Bluetongue Virus Serotypes 10 and 17 at Different Coinfection Ratios in Culicoides sonorenesis.

Bluetongue virus (BTV) is a segmented, double-stranded RNA orbivirus listed by the World Organization for Animal Health and transmitted by Culicoides biting midges. Segmented viruses can reassort, which facilitates rapid and important genotypic changes. Our study evaluated reassortment in Culicoides sonorensis midges coinfected with different ratios of BTV-10 and BTV-17. Midges were fed blood containing BTV-10, BTV-17, or a combination of both serotypes at 90:10, 75:25, 50:50, 25:75, or 10:90 ratios. Midges were collected every other day and tested for infection using pan BTV and cox1 (housekeeping gene) qRT-PCR. A curve was fit to the ∆Ct values (pan BTV Ct-cox1 Ct) for each experimental group. On day 10, the midges were processed for BTV plaque isolation. Genotypes of the plaques were determined by next-generation sequencing. Pairwise comparison of ∆Ct curves demonstrated no differences in viral RNA levels between coinfected treatment groups. Plaque genotyping indicated that most plaques fully aligned with one of the parental strains; however, reassortants were detected, and in the 75:25 pool, most plaques were reassortant. Reassortant prevalence may be maximized upon the occurrence of reassortant genotypes that can outcompete the parental genotypes. BTV reassortment and resulting biological consequences are important elements to understanding orbivirus emergence and evolution.

Seven-year prospective study on yearly incidence of Orbivirus infection of captive white-tailed deer and potential Culicoides vectors.

Bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) are arthropod-borne viruses that are transmitted by biting midges in the genus Culicoides (Diptera: Ceratopogonidae) and can cause hemorrhagic disease in certain ruminants. The objectives of this study were to measure the incidence of BTV and EHDV infections in captive white-tailed deer herd as well as tissues and corresponding presence of Culicoides midges at a location near Clinton, LA. During a 7-yr study with yearly outbreaks of hemorrhagic disease in the deer herd, 15 species of Culicoides were captured using Centers for Disease Control (CDC) black light traps. Reverse transcriptase quantitative polymerase chain reaction (PCR) was performed to screen for BTV and EHDV in pools of midges and tissues of deer. From 2012 to 2018, 1,711 pools of midges representing 24,859 specimens were tested, and specimens from 5 of the 15 collected species (Culicoides debilipalpis, Culicoides stellifer, Culicoides venustus, Culicoides haematopotus, and Culicoides crepuscularis) were found to be PCR positive for BTV and EHDV. Most of the BTV-positive pools of biting midges were from specimens of C. debilipalpis and C. stellifer, and most of the EHDV-positive pools were from specimens of C. venustus and C. stellifer. During the 7-yr period, 112 white-tailed deer that died at the study location were PCR positive for BTV or EHDV: detected BTV serotypes were 10 and 12 and EHDV serotypes were 1, 2, and 6. There was a significant increase in BTV/EHDV antibody prevalence in white-tailed deer during the study; antibody-positive rates increased from 15% to 78% in the deer herd of approximately 100 animals.

Detection, Characterization and Sequencing of BTV Serotypes Circulating in Cuba in 2022.

In Cuba, despite a high sero-prevalence of bluetongue virus (BTV), circulating serotypes remain unknown. The aim of this study was to identify circulating BTV serotypes in farms throughout the western region of Cuba. Blood samples were collected from 200 young cattle and sheep between May and July 2022 for virological analyses (PCR, viral isolation and virus neutralization) and genome sequencing. The results confirmed viral circulation, with viro-prevalence of 25% for BTV. The virus was isolated from 18 blood samples and twelve BTV serotypes were identified by sequencing RT-PCR products targeting the segment 2 of the BTV genome (BTV-1, 2, 3, 6, 10, 12, 13, 17, 18, 19, 22 and 24). Finally, the full genome sequences of 17 Cuban BTV isolates were recovered using a Sequence Independent Single Primer Amplification (SISPA) approach combined to MinION Oxford Nanopore sequencing technology. All together, these results highlight the co-circulation of a wide diversity of BTV serotypes in a quite restricted area and emphasize the need for entomological and livestock surveillance, particularly in light of recent changes in the global distribution and nature of BTV infections.

The First Report on the Complete Sequence Characterization of Bluetongue Virus Serotype 3 in the Republic of Korea.

The bluetongue virus (BTV) is a significant animal pathogen with economic implications in the ruminant industry. Despite global reports on BTV detection and epidemiologic investigations, limited studies have focused on the virus in the ROK. In this study, BTV epidemiological research was conducted on blood samples from cattle and goat farms across nine regions during 2013-2014. The results showed that 3.33% of bovine blood samples (194/5824) and 0.19% of goat blood samples (2/1075) tested positive for BTV antibodies using ELISA. In Jeju-do, BTV RNA amplification occurred in 51 of 422 samples (12.1%) using real-time reverse transcription (RT-qPCR). The isolation of one sample revealed it as serotype 3, as indicated by the sequence of segments 2 (Seg-2) and 6 (Seg-6), associated with the eastern BTV topotype. However, based on Seg-1, -3, -4, -5, -7, -8, -9, and -10 analyses, the BTV-3/JJBB35 strain is more closely related to distinct BTV strains. These findings imply BTV circulation and that the Korean-isolated BTV might originate from Asian BTV strains due to multiple reassortment events. This study provides foundational data for ongoing BTV monitoring and disease-control policies in the ROK.

A novel bluetongue virus serotype 2 strain isolated from a farmed Florida white-tailed deer (Odocoileus virginianus) arose from reassortment of gene segments derived from co-circulating serotypes in the Southeastern USA.

Bluetongue disease is a reportable animal disease that affects wild and farmed ruminants, including white-tailed deer (WTD). This report documents the clinical findings, ancillary diagnostics, and genomic characterization of a novel reassortant bluetongue virus serotype 2 (BTV-2) strain isolated from a dead Florida farmed WTD in 2022. Our analyses support that this BTV-2 strain likely stemmed from the acquisition of genome segments from co-circulating BTV strains in Florida and Louisiana. In addition, our analyses also indicate that genetically uncharacterized BTV strains may be circulating in the Southeastern USA; however, the identity and reassortant status of these BTV strains cannot be determined based on the VP2 and VP5 genome sequences. Hence, continued surveillance based on complete genome characterization is needed to understand the genetic diversity of BTV strains in this region and the potential threat they may pose to the health of deer and other ruminants.

Seroepidemiological Survey on Bluetongue Virus (BTV) among Cattle, Sheep, and Goats in Gadarif State, Eastern Sudan.

Bluetongue (BT) is an arthropod-borne viral disease that primarily affects ruminants in tropical and temperate regions. In the present study, a cross-sectional survey was conducted to define the seroprevalence of Bluetongue virus and to identify the possible risk factors correlated with BTV seropositivity among cattle, sheep, and goats during the period 2015-2016 in Gadarif State. A total of 420 cattle, 877 sheep, and 641 goat serum samples were collected randomly from 12 localities. Information about age, sex, breed, area ecology, and location was obtained for each sample. Bluetongue seroprevalence was estimated using competitive enzyme-linked immunosorbent assay (cELISA). The overall seroprevalence of BTV was 92.9% (390/420), 76.4% (670/877), and 85.3% (547/641) among cattle, sheep, and goats, respectively. Multivariate analysis followed univariate analysis showed that there was a significant difference (p < 0.05) between location, area ecology and age groups of cattle, sheep, and goats, and seropositivity to BTV. In addition, a significant association (p < 0.05) was observed between sex and seropositivity to BTV in sheep. In conclusion, BTV antibodies are highly prevalent in Gadarif State and susceptible livestock are at risk of exposition with BTV. Consequently, these animals have protection against specific BTV serotypes.

Co-expression of VP2, NS1 and NS2-Nt proteins by an MVA viral vector induces complete protection against bluetongue virus.

Bluetongue (BT), caused by bluetongue virus (BTV), is an important arthropod-borne livestock disease listed by the World Organization for Animal Health. Live-attenuated and inactivated vaccines have permitted to control BT but they do not simultaneously protect against the myriad of BTV serotypes. Recently, we identified the highly conserved BTV nonstructural protein NS1 and the N-terminal region of NS2 as antigens capable of conferring multiserotype protection against BTV. Here, we designed Modified Vaccinia Ankara (MVA) viral vectors that expressed BTV-4 proteins VP2 or VP7 along with NS1 and NS2-Nt as well as MVAs that expressed proteins VP2, VP7 or NS1 and NS2-Nt. Immunization of IFNAR(-/-) mice with two doses of MVA-NS1-2A-NS2-Nt protected mice from BTV-4M infection by the induction of an antigen-specific T cell immune response. Despite rMVA expressing VP7 alone were not protective in the IFNAR(-/-) mouse model, inclusion of VP7 in the vaccine formulation amplified the cell-mediated response induced by NS1 and NS2-Nt. Expression of VP2 elicited protective non-cross-reactive neutralizing antibodies (nAbs) in immunized animals and improved the protection observed in the MVA-NS1-2A-NS2-Nt immunized mice when these three BTV antigens were co-expressed. Moreover, vaccines candidates co-expressing VP2 or VP7 along with NS1 and NS2-Nt provided multiserotype protection. We assessed protective efficacy of both vaccine candidates in sheep against virulent challenge with BTV-4M. Immunization with MVA-VP7-NS1-2A-NS2-Nt partially dumped viral replication and clinical disease whereas administration of MVA-VP2-NS1-2A-NS2-Nt promoted a complete protection, preventing viraemia and the pathology produced by BTV infection.