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development and evaluation of a sybr green real-time rt-pcr assay for evaluation of cytokine gene expression in horse.cytokine secretion is one of the main mechanisms by which the immune system is regulated in response to pathogens. therefore, the measurement of cytokine expression is fundamental to characterizing the immune response to infections. real-time quantitative reverse transcriptase-polymerase chain reaction (rt-qpcr) is widely used to measure cytokine mrna levels, but assay conditions should be properly evaluated before analyzing important equine infections through relative quantification of gene exp ...201323103121
african horse sickness outbreaks caused by multiple virus types in ethiopia.african horse sickness (ahs) is associated with high morbidity and mortality in equids, especially horses. a retrospective analysis was carried out concerning 737 ahs outbreaks that occurred during 2007-2010 in ethiopia. a total of ten outbreaks were investigated in the study period. all four forms of the disease (pulmonary, cardiac, horse sickness fever and the combined form) were observed, with the cardiac form being the most prevalent. multiple african horse sickness virus serotypes (ahsv-2, ...201423083078
utilization of cellulose microcapillary tubes as a model system for culturing and viral infection of mammalian cells.cryofixation by high-pressure freezing (hpf) and freeze substitution (fs) gives excellent preservation of intracellular membranous structures, ideal for ultrastructural investigations of virus infected cells. conventional sample preparation methods of tissue cultured cells can however disrupt the association between neighboring cells or of viruses with the plasma membrane, which impacts upon the effectiveness whereby virus release from cells can be studied. we established a system for virus infe ...201222865476
bluetongue virus genetic and phenotypic diversity: towards identifying the molecular determinants that influence virulence and transmission potential.bluetongue virus (btv) is the prototype member of the orbivirus genus in the family reoviridae and is the aetiological agent of the arthropod transmitted disease bluetongue (bt) that affects both ruminant and camelid species. the disease is of significant global importance due to its economic impact and effect on animal welfare. bluetongue virus, a dsrna virus, evolves through a process of quasispecies evolution that is driven by genetic drift and shift as well as intragenic recombination. quasi ...201222835527
an african horse sickness virus serotype 4 recombinant canarypox virus vaccine elicits specific cell-mediated immune responses in horses.a recombinant canarypox virus vectored vaccine co-expressing synthetic genes encoding outer capsid proteins, vp2 and vp5, of african horse sickness virus (ahsv) serotype 4 (alvac(®)-ahsv4) has been demonstrated to fully protect horses against homologous challenge with virulent field virus. guthrie et al. (2009) detected weak and variable titres of neutralizing antibody (ranging from <10 to 40) 8 weeks after vaccination leading us to hypothesize that there could be a participation of cell mediate ...201222763149
african horse sickness in naturally infected, immunised horses.to determine whether subclinical cases, together with clinical cases, of african horse sickness (ahs) occur in immunised horses in field conditions, whole blood samples were collected and rectal temperatures recorded weekly from 50 nooitgedacht ponies resident in open camps at the faculty of veterinary science, university of pretoria, onderstepoort, during 2008-2010. the samples were tested for the presence of african horse sickness virus (ahsv) rna by a recently developed real-time rt-pcr. it w ...201322612775
sustained high-throughput polymerase chain reaction diagnostics during the european epidemic of bluetongue virus serotype 8.a real-time reverse transcription polymerase chain reaction assay (pcr test) based on genome segment 10 of bluetongue virus (btv) was developed. the pcr test consists of robotized viral rna isolation from blood samples and an all-in-one method including initial denaturation of genomic double-stranded rna, reverse transcription polymerase chain reaction (rt-pcr), and real-time detection and analysis. reference strains of the 24 recognized btv serotypes, isolates from different years, and geograph ...201222529113
virus-specific cd8⁺ t-cells detected in pbmc from horses vaccinated against african horse sickness virus.african horsesickness (ahs) is an infectious but noncontagious viral disease affecting all species of equidae. the recall immune response of ahsv naïve horses immunised with an attenuated african horsesickness virus serotype 4 (ahsv4) was characterised using immune assays including elispot, real-time pcr (qpcr) and flow cytometry. the recall immune response detected in pbmc isolated from three inoculated horses showed an upregulation of circulating b lymphocytes that correlated with elevated il- ...201222365333
risk of introducing african horse sickness virus into the netherlands by international equine movements.african horse sickness (ahs) is a vector-borne viral disease of equines that is transmitted by culicoides spp. and can have severe consequences for the horse industry in affected territories. a study was performed to assess the risk of introducing ahs virus (ahsv) into the netherlands (p_ahs) by international equine movements. the goal of this study was to provide more insight into (a) the regions and equine species that contribute most to this risk, (b) the seasonal variation in this risk, and ...201222341773
immunogenicity of plant-produced african horse sickness virus-like particles: implications for a novel vaccine.african horse sickness (ahs) is a debilitating and often fatal viral disease affecting horses in much of africa, caused by the dsrna orbivirus african horse sickness virus (ahsv). vaccination remains the single most effective weapon in combatting ahs, as there is no treatment for the disease apart from good animal husbandry. however, the only commercially available vaccine is a live-attenuated version of the virus (lav). the threat of outbreaks of the disease outside its endemic region and the f ...201728650085
application of an embryonated chicken egg model to assess the vector competence of australian culicoides midges for bluetongue viruses.culicoides biting midges (diptera: ceratopogonidae) are vectors of a number of globally important arboviruses that affect livestock, including bluetongue virus (btv), african horse sickness virus and the recently emerged schmallenberg virus. in this study, a model using embryonated chicken eggs (eces) was utilized to undertake vector competence studies of australian culicoides spp. for 13 laboratory-adapted or wild-type virus strains of btv. a total of 7393 culicoides brevitarsis were reared fro ...201728429824
evidence of african horse sickness virus infection of equus zebra hartmannae in the south-western khomas region, namibia.equine mortalities suspected to be due to african horse sickness (ahs) were reported from the arid khomas region, namibia, in 2008. the area was previously considered a localized ahs-free area. hartmann's mountain zebra (equus zebra hartmannae), a potential but unconfirmed reservoir host of african horse sickness virus (ahsv), occurs in the region. between 2009 and 2010 serum, blood and tissue samples from 31 culled e. z. hartmannae were analysed by reverse transcription-polymerase chain reactio ...201728406577
phosphoproteomic analysis reveals the importance of kinase regulation during orbivirus infection.bluetongue virus (btv) causes infections in wild and domesticated ruminants with high morbidity and mortality and is responsible for significant economic losses in both developing and developed countries. btv serves as a model for the study of other members of the orbivirus genus. previously, the importance of casein kinase 2 for btv replication was demonstrated. to identify intracellular signalling pathways and novel host-cell kinases involved during btv infection, the phosphoproteome of btv in ...201728851738
seroprevalence and risk factor analysis for exposure to equine encephalosis virus in israel, palestine and jordan.equine encephalosis virus (eev) is an orbivirus transmitted by culicoides species. most infected horses show mild clinical signs and mortality is usually very low. eev is closely related and similarly transmitted to other, more pathogenic and economically important, orbiviruses such as african horse sickness virus (ahsv), bluetongue virus (btv) and epizootic haemorrhagic disease viruses (ehdv), and may serve as an indicator for possible transmission of the latter. israel has been reported to be ...201728713576
protective efficacy of multivalent replication-abortive vaccine strains in horses against african horse sickness virus challenge.african horse sickness virus (ahsv) is an orbivirus, a member of the reoviridae family. nine different serotypes have been described so far. ahsv is vectored by culicoides spp. to equids, causing high mortality, particularly in horses, with considerable economic impacts. for development of a safe attenuated vaccine, we previously established an efficient reverse genetics (rg) system to generate entry competent replication-abortive (ecra) virus strains, for all nine serotypes and demonstrated the ...201728625521
isolation of tibet orbivirus from culicoides and associated infections in livestock in yunnan, china.culicoides-borne orbiviruses, such as bluetongue virus (btv) and african horse sickness virus (ahsv), are important pathogens that cause animal epidemic diseases leading to significant loss of domestic animals. this study was conducted to identify culicoides-borne arboviruses and to investigate the associated infections in local livestock in yunnan, china.201728595631
the sero-prevalence and sero-incidence of african horse sickness and equine encephalosis in selected horse and donkey populations in zimbabwe.sentinel herds and samples submitted by private equine practitioners were used to determine the sero-prevalence and sero-incidence of african horse sickness virus (ahsv) and equine encephalosis virus (eev) in horse and donkey populations in the highveld region of zimbabwe. the sero-prevalence and sero-incidence of antibodies against these viruses were determined using the competitive enzyme-linked immunosorbent assay (elisa) for the detection of serum antibodies. in donkeys, the median sero-prev ...201728582979
vaccination with recombinant modified vaccinia ankara (mva) viruses expressing single african horse sickness virus vp2 antigens induced cross-reactive virus neutralising antibodies (vnab) in horses when administered in combination.african horse sickness is a lethal viral disease of equids transmitted by biting midges of the genus culicoides. the disease is endemic to sub-saharan africa but outbreaks of high mortality and economic impact have occurred in the past in non-endemic regions of africa, asia and southern europe. vaccination is critical for the control of this disease but only live attenuated vaccines are currently available. however, there are bio-safety concerns over the use of this type of vaccines, especially ...201728438410
african horse sickness virus (ahsv) with a deletion of 77 amino acids in ns3/ns3a protein is not virulent and a safe promising ahs disabled infectious single animal (disa) vaccine platform.african horse sickness virus (ahsv) is a virus species in the genus orbivirus of the family reoviridae. currently, nine serotypes have been defined showing limited cross neutralization. ahsv is transmitted by species of culicoides biting midges and causes african horse sickness (ahs) in equids with a mortality up to 95% in naïve domestic horses. ahs has become a serious threat for countries outside africa, since endemic culicoides species in moderate climates are competent vectors of closely rel ...201829525278
culicoides species composition and molecular identification of host blood meals at two zoos in the uk.culicoides biting midges are biological vectors of arboviruses including bluetongue virus (btv), schmallenberg virus (sbv) and african horse sickness virus (ahsv). zoos are home to a wide range of 'at risk' exotic and native species of animals. these animals have a high value both in monetary terms, conservation significance and breeding potential. to understand the risk these viruses pose to zoo animals, it is necessary to characterise the culicoides fauna at zoos and determine which potential ...202032178710
diagnostic diva tests accompanying the disabled infectious single animal (disa) vaccine platform for african horse sickness.african horse sickness virus (ahsv) (orbivirus genus, reoviridae family) causes high mortality in naïve domestic horses with enormous economic and socio-emotional impact. there are nine ahsv serotypes showing limited cross neutralization. ahsv is transmitted by competent species of culicoides biting midges. ahs is a serious threat beyond the african continent as endemic culicoides species in moderate climates transmit the closely related prototype bluetongue virus. there is a desperate need for ...201829759377
safety and efficacy of inactivated african horse sickness (ahs) vaccine formulated with different adjuvants.african horse sickness virus (ahsv) is a virus species in the genus orbivirus of the family reoviridae causing african horse sickness (ahs) in equids with a mortality of about 95% in naïve horses. ahs causes serious losses in developing countries where horses play a central role in draft power and transportation. there are nine ahsv serotypes inducing no or low cross-neutralizing antibodies. ahsv is spread by biting culicoides midges. ahs is endemic in sub-saharan africa, and a serious threat ou ...202032921506
safety and immunogenicity of plant-produced african horse sickness virus-like particles in horses.african horse sickness (ahs) is caused by multiple serotypes of the dsrna ahsv and is a major scourge of domestic equids in africa. while there are well established commercial live attenuated vaccines produced in south africa, risks associated with these have encouraged attempts to develop new and safer recombinant vaccines. previously, we reported on the immunogenicity of a plant-produced ahs serotype 5 virus-like particle (vlp) vaccine, which stimulated high titres of ahs serotype 5-specific n ...201830309390
humoral and cell-mediated immune responses to plant-produced african horse sickness virus vp7 quasi-crystals.african horse sickness (ahs) is a devastating viral disease affecting equines and has resulted in many disastrous epizootics. to date, no successful therapeutic treatment exists for ahs, and commercially used live-attenuated vaccines have various undesirable side effects. previous studies have shown that mice inoculated with insoluble african horse sickness virus (ahsv) vp7 crystals are protected from live challenge with a lethal dose of ahsv. this study investigates the humoral and cell-mediate ...202133421520
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