Virulent African horse sickness virus serotype 4 interferes with the innate immune response in horse peripheral blood mononuclear cells in vitro.

African horse sickness (AHS) is caused by African horse sickness virus (AHSV), a double stranded RNA (dsRNA) virus of the genus Orbivirus, family Reoviridae. For the development of new generation AHS vaccines or antiviral treatments, it is crucial to understand the host immune response against the virus and the immune evasion strategies the virus employs. To achieve this, the current study used transcriptome analysis of RNA sequences to characterize and compare the innate immune responses activated during the attenuated AHSV serotype 4 (attAHSV4) (in vivo) and the virulent AHSV4 (virAHSV4) (in vitro) primary and secondary immune responses in horse peripheral blood mononuclear cells (PBMC) after 24 h. The pro-inflammatory cytokine and chemokine responses were negatively regulated by anti-inflammatory cytokines, whereas the parallel type I and type III IFN responses were maintained downstream of nucleic acid sensing pattern recognition receptor (PRR) signalling pathways during the attAHSV4 primary and secondary immune responses. It appeared that after translation, virAHSV4 proteins were able to interfere with the C-terminal IRF association domain (IAD)-type 1 (IAD1) containing IRFs, which inhibited the expression of type I and type III IFNs downstream of PRR signalling during the virAHSV4 primary and secondary immune responses. Viral interference resulted in an impaired innate immune response that was not able to eliminate virAHSV4-infected PBMC and gave rise to prolonged expression of pro-inflammatory cytokines and chemokines during the virAHSV4 induced primary immune response. Indicating that virAHSV4 interference with the innate immune response may give rise to an excessive inflammatory response that causes immunopathology, which could be a major contributing factor to the pathogenesis of AHS in a naïve horse. Viral interference was overcome by the fast kinetics and increased effector responses of innate immune cells due to trained innate immunity and memory T cells and B cells during the virAHSV4 secondary immune response.

Sero-surveillance of emerging viral diseases in camels and cattle in Nouakchott, Mauritania: an abattoir study.

This study reports the monitoring of several emerging viral pathogens in Mauritania, which was carried out by the analysis of bovine and camel samples taken at the slaughterhouse of Nouakchott. Blood and serum were collected by random sampling from 159 camels and 118 cattle in March 2013 at the large animals abattoir in Nouakchott. Serological tests for Rift Valley Fever (RVF), Peste des Petits Ruminants (PPR), West Nile disease (WND), epizootic haemorrhagic disease (EHD) and African horse sickness (AHS) were carried out using commercial ELISA kits. The samples, which resulted positives for PPR, WND and AHS, were tested with the confirmatory virus neutralization test (VNT). According to ELISA results, serological prevalence of RVF was 45% (95% CI 52.3-37.7) in camels and 16% (95% CI 22.6-9.4) in cattle. The difference between the observed prevalences in camels and in cattle was significant (p value ≤ 0.01). PPR was absent in camels and had 12% prevalence (95% CI, 17.86-6.14) in cattle. Furthermore, camels showed 92% (95% CI, 96.1-87.9) prevalence of WNV, 73% (95% CI, 82.3-63.64) of EHD and 3% (95% CI, 5.6-0.4) of AHS. This data are of relevance since provided useful feedbacks on the circulation of the pathogens in field. Moreover, this survey provided new information on the susceptibility of camels to several emerging pathogens and on the possible use of this species as sentinel animal.

Inhibition of Orbivirus Replication by Aurintricarboxylic Acid.

Bluetongue virus (BTV) and African horse sickness virus (AHSV) are vector-borne viruses belonging to the Orbivirus genus, which are transmitted between hosts primarily by biting midges of the genus Culicoides. With recent BTV and AHSV outbreaks causing epidemics and important economy losses, there is a pressing need for efficacious drugs to treat and control the spread of these infections. The polyanionic aromatic compound aurintricarboxylic acid (ATA) has been shown to have a broad-spectrum antiviral activity. Here, we evaluated ATA as a potential antiviral compound against Orbivirus infections in both mammalian and insect cells. Notably, ATA was able to prevent the replication of BTV and AHSV in both cell types in a time- and concentration-dependent manner. In addition, we evaluated the effect of ATA in vivo using a mouse model of infection. ATA did not protect mice against a lethal challenge with BTV or AHSV, most probably due to the in vivo effect of ATA on immune system regulation. Overall, these results demonstrate that ATA has inhibitory activity against Orbivirus replication in vitro, but further in vivo analysis will be required before considering it as a potential therapy for future clinical evaluation.

Assessing the potential of plains zebra to maintain African horse sickness in the Western Cape Province, South Africa.

African horse sickness (AHS) is a disease of equids that results in a non-tariff barrier to the trade of live equids from affected countries. AHS is endemic in South Africa except for a controlled area in the Western Cape Province (WCP) where sporadic outbreaks have occurred in the past 2 decades. There is potential that the presence of zebra populations, thought to be the natural reservoir hosts for AHS, in the WCP could maintain AHS virus circulation in the area and act as a year-round source of infection for horses. However, it remains unclear whether the epidemiology or the ecological conditions present in the WCP would enable persistent circulation of AHS in the local zebra populations. Here we developed a hybrid deterministic-stochastic vector-host compartmental model of AHS transmission in plains zebra (Equus quagga), where host populations are age- and sex-structured and for which population and AHS transmission dynamics are modulated by rainfall and temperature conditions. Using this model, we showed that populations of plains zebra present in the WCP are not sufficiently large for AHS introduction events to become endemic and that coastal populations of zebra need to be >2500 individuals for AHS to persist >2 years, even if zebras are infectious for more than 50 days. AHS cannot become endemic in the coastal population of the WCP unless the zebra population involves at least 50,000 individuals. Finally, inland populations of plains zebra in the WCP may represent a risk for AHS to persist but would require populations of at least 500 zebras or show unrealistic duration of infectiousness for AHS introduction events to become endemic. Our results provide evidence that the risk of AHS persistence from a single introduction event in a given plains zebra population in the WCP is extremely low and it is unlikely to represent a long-term source of infection for local horses.

Modeling the global distribution of Culicoides imicola: an Ensemble approach.

Culicoides imicola is a midge species serving as vector for a number of viral diseases of livestock, including Bluetongue, and African Horse Sickness. C. imicola is also known to transmit Schmallenberg virus experimentally. Environmental and demographic factors may impose rapid changes on the global distribution of C. imicola and aid introduction into new areas. The aim of this study is to predict the global distribution of C. imicola using an ensemble modeling approach by combining climatic, livestock distribution and land cover covariates, together with a comprehensive global dataset of geo-positioned occurrence points for C. imicola. Thirty individual models were generated by 'biomod2', with 21 models scoring a true skill statistic (TSS) >0.8. These 21 models incorporated weighted runs from eight of ten algorithms and were used to create a final ensemble model. The ensemble model performed very well (TSS = 0.898 and ROC = 0.991) and indicated high environmental suitability for C. imicola in the tropics and subtropics. The habitat suitability for C. imicola spans from South Africa to southern Europe and from southern USA to southern China. The distribution of C. imicola is mainly constrained by climatic factors. In the ensemble model, mean annual minimum temperature had the highest overall contribution (42.9%), followed by mean annual maximum temperature (21.1%), solar radiation (13.6%), annual precipitation (11%), livestock distribution (6.2%), vapor pressure (3.4%), wind speed (0.8%), and land cover (0.1%). The present study provides the most up-to-date predictive maps of the potential distributions of C. imicola and should be of great value for decision making at global and regional scales.

Evidence of Intragenic Recombination in African Horse Sickness Virus.

Intragenic recombination has been described in various RNA viruses as a mechanism to increase genetic diversity, resulting in increased virulence, expanded host range, or adaptability to a changing environment. Orbiviruses are no exception to this, with intragenic recombination previously detected in the type species, bluetongue virus (BTV). African horse sickness virus (AHSV) is a double-stranded RNA virus belonging to the Oribivirus genus in the family Reoviridae. Genetic recombination through reassortment has been described in AHSV, but not through homologous intragenic recombination. The influence of the latter on the evolution of AHSV was investigated by analyzing the complete genomes of more than 100 viruses to identify evidence of recombination. Segment-1, segment-6, segment-7, and segment-10 showed evidence of intragenic recombination, yet only one (Segment-10) of these events was manifested in subsequent lineages. The other three hybrid segments were as a result of recombination between field isolates and the vaccine derived live attenuated viruses (ALVs).

Evaluating African horse sickness virus in horses and field-caught Culicoides biting midges on the East Rand, Gauteng Province, South Africa.

A prospective study was undertaken during 2013 and 2014, to determine the prevalence of African horse sickness virus (AHSV) in Culicoides midges and the incidence of infection caused by the virus in 28 resident horses on two equine establishments on the East Rand, Gauteng Province, South Africa. Field caught Culicoides midges together with whole blood samples from participating horses were collected every two weeks at each establishment. Culicoides midges and blood samples were tested for the presence of AHSV RNA by real-time quantitative reverse transcription polymerase chain reaction. Nine immunised horses became infected with AHSV during the study period, although infections were subclinical. African horse sickness virus was also identified from a field-collected midge pool. The observations recapitulate previously published data in another setting, where further investigation is warranted to determine what role subclinical infection plays in the diseases epidemiology.

Targeted mutational analysis to unravel the complexity of African horse sickness virus NS3 function in mammalian cells.

The African horse sickness virus non-structural protein 3 (NS3) is involved in the final stages of infection. To gain insight into the function of different NS3 domains, we generated reverse genetics-derived mutants, each expressing a modified version of the protein. A functional comparison of these mutants to the wild-type virus in mammalian cells indicated the variable contribution of the different domains to the cytopathic effect and in ensuring effective virus trafficking and release. The transmembrane domains were determined as essential mediators of NS3 localisation, as the abnormal processing of these mutant proteins resulted in their nuclear localisation and interaction with NS1. NS3 cytoplasmic domain disruptions resulted in increased cytosolic virus particle accumulation and abnormal virion tethering to plasma membranes. Other aspects of infection were also affected, such as VIB formation and distribution of the outer capsid proteins. Overall, these results illustrate the intricate role of NS3 in the infection cycle.

Using a new serotype-specific Polymerase Chain Reaction (PCR) and sequencing to differentiate between field and vaccine-derived African Horse Sickness viruses submitted in 2016/2017.

The outer capsid viral protein 2 (VP2) of African horse sickness virus, encoded by the most variable genome segment 2 (Seg-2), is the primary target for AHSV-specific neutralising antibodies and thus determines the virus serotype. Full length segment 2 sequences from more than 100 AHSVs isolated over the last 80 years were compared and single nucleotide polymorphisms (SNPs) identified between the reference strains and recent field viruses. Regions unique to each individual serotype were identified and primers designed to differentially amplify each of the nine serotypes. The sequences of resulting amplicons contained a significant amount of SNPs to discriminate between field viruses and reference strains or live attenuated viruses. The new serotype specific RT-PCR were subsequently used to determine the prevalence of different AHSV serotypes associated with samples submitted to the Agricultural Research Council - Onderstepoort Veterinary Research Institute during the 2016 / 2017 season. Subsequent sequencing of the PCR products were used to determine if the infections were caused by field or vaccine-derived strains. The serotypes of 70 AHSV positive diagnostic samples submitted to the ARC-OVR were determined. Serotypes 2 and 6 were the most prevalent, while Serotype 1 was the only serotype where sequences identical to the ALV or reference strains were detected in field samples. Based on this study, the incidence of vaccine-derived AHS infections submitted from southern Africa were low. This serotype-specific RT-PCR and sequencing assay could assist with the surveillance and control of equines movement nationally and internationally. It could also provide valuable scientific guidance on the policies and guidelines regulating vaccination and trade of equines in South Africa.

A field investigation of an African horse sickness outbreak in the controlled area of South Africa in 2016.

An outbreak of African horse sickness (AHS) caused by AHS virus type 1 occurred within the South African AHS surveillance zone during April and May 2016. The index case was detected by a private veterinarian through passive surveillance. There were 21 cases in total, which is relatively low compared to case totals during prior AHS outbreaks in the same region (and of the same AHS virus type) in 2004, 2011 and 2014. The affected proportion of horses on affected properties was 0.07 (95% CI 0.04, 0.11). Weather conditions were conducive to high midge activity immediately prior to the outbreak but midge numbers decreased rapidly with the advent of winter. The outbreak was localized, with 18 of the 21 cases occurring within 8 km of the index property and the three remaining cases on two properties within 21 km of the index property, with direction of spread consistent with wind-borne dispersion of infected midges. Control measures included implementation of a containment zone with movement restrictions on equids. The outbreak was attributed to a reversion to virulence of a live attenuated vaccine used extensively in South Africa. Outbreaks in the AHS control zones have a major detrimental impact on the direct export of horses from South Africa, notably to the European Union.

Sero-epidemioloical survey on African horse sickness virus among horses in Khartoum State, Central Sudan.

BACKGROUND: African horse sickness virus (AHSV) is an infectious non contagious insect-transmitted double-stranded (ds) RNA orbivirus of the family Reoviridae. AHSV causes an often fatal hemorrhagic infection with high mortality among selected breeds of Arabian horses. This study was conducted to avail some information with regard to the prevalence and associated risk factors of AHSV among ecotype breeds of horses in central Sudan. METHODS: Sera were collected from 320 horses, which were selected randomly from four localities and employed in the study. A competitive enzyme-linked immunosorbent assay (cELISA) was used to screen sampled sera for AHSV-specific immunoglobulin G (Ig G) antibodies. RESULTS: Seropositivity to AHSV Ig G was detected in 275 out of the 320 horse sera, thus accounting for a prevalence rate of 85.9%. Potential risk factors to AHSV infection were reported to be associated with horse breed (OR = 5.0, CI = 0.07-2.104, p-value = 0.039) and activity of the horse (OR = 3.21, CI = 0.72-1.48, p- value = 0.008). CONCLUSIONS: The high prevalence of AHSV in Khartoum State of Central Sudan necessitates the need for continuous surveillance for AHSV infection to prevent a possible disease outbreak in this region of the African continent.

Spatial distribution modelling of Culicoides (Diptera: Ceratopogonidae) biting midges, potential vectors of African horse sickness and bluetongue viruses in Senegal.

BACKGROUND: In Senegal, the last epidemic of African horse sickness (AHS) occurred in 2007. The western part of the country (the Niayes area) concentrates modern farms with exotic horses of high value and was highly affected during the 2007 outbreak that has started in the area. Several studies were initiated in the Niayes area in order to better characterize Culicoides diversity, ecology and the impact of environmental and climatic data on dynamics of proven and suspected vectors. The aims of this study are to better understand the spatial distribution and diversity of Culicoides in Senegal and to map their abundance throughout the country. METHODS: Culicoides data were obtained through a nationwide trapping campaign organized in 2012. Two successive collection nights were carried out in 96 sites in 12 (of 14) regions of Senegal at the end of the rainy season (between September and October) using OVI (Onderstepoort Veterinary Institute) light traps. Three different modeling approaches were compared: the first consists in a spatial interpolation by ordinary kriging of Culicoides abundance data. The two others consist in analyzing the relation between Culicoides abundance and environmental and climatic data to model abundance and investigate the environmental suitability; and were carried out by implementing generalized linear models and random forest models. RESULTS: A total of 1,373,929 specimens of the genus Culicoides belonging to at least 32 different species were collected in 96 sites during the survey. According to the RF (random forest) models which provided better estimates of abundances than Generalized Linear Models (GLM) models, environmental and climatic variables that influence species abundance were identified. Culicoides imicola, C. enderleini and C. miombo were mostly driven by average rainfall and minimum and maximum normalized difference vegetation index. Abundance of C. oxystoma was mostly determined by average rainfall and day temperature. Culicoides bolitinos had a particular trend; the environmental and climatic variables above had a lesser impact on its abundance. RF model prediction maps for the first four species showed high abundance in southern Senegal and in the groundnut basin area, whereas C. bolitinos was present in southern Senegal, but in much lower abundance. CONCLUSIONS: Environmental and climatic variables of importance that influence the spatial distribution of species abundance were identified. It is now crucial to evaluate the vector competence of major species and then combine the vector densities with densities of horses to quantify the risk of transmission of AHS virus across the country.

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 safe and efficacious vaccines, while DIVA (Differentiating Infected from Vaccinated) vaccines would accelerate control of AHS. Previously, we have shown that highly virulent AHSV with an in-frame deletion of 77 amino acids (aa) in NS3/NS3a is completely safe, does not cause viremia and shows protective capacity. This deletion mutant is a promising DISA (Disabled Infectious Single Animal) vaccine platform, since exchange of serotype specific virus proteins has been shown for all nine serotypes. Here, we show that a prototype NS3 competitive ELISA is DIVA compliant to AHS DISA vaccine platforms. Epitope mapping of NS3/NS3a shows that more research is needed to evaluate this prototype serological DIVA assay regarding sensitivity and specificity, in particular for AHSVs expressing antigenically different NS3/NS3a proteins. Further, an experimental panAHSV PCR test targeting genome segment 10 is developed that detects reference AHSV strains, whereas AHS DISA vaccine platforms were not detected. This DIVA PCR test completely guarantees genetic DIVA based on in silico and in vitro validation, although test validation regarding diagnostic sensitivity and specificity has not been performed yet. In conclusion, the prototype NS3 cELISA and the PCR test described here enable serological and genetic DIVA accompanying AHS DISA vaccine platforms.

A correlation between capsid protein VP2 and the plaque morphology of African horse sickness virus in cell culture.

The attenuated live virus vaccine that is used in South Africa to protect against African horse sickness infection was developed more than 50 years ago. With the selection of the vaccine strains by cell culture passage, a correlation between the size of plaques formed in monolayer Vero cultures and attenuation of virus virulence in horses was found. The large plaque phenotype was used as an indication of cell culture adaptation and strongly correlated with attenuation of virulence in horses. There was never any investigation into the genetic causes of either the variation in plaque size, or the loss of virulence. An understanding of the underlying mechanisms of attenuation would benefit the production of a safer AHSV vaccine. To this end, the genomes of different strains of two African horse sickness isolates, producing varying plaque sizes, were compared and the differences between them identified. This comparison suggested that proteins VP2, VP3, VP5 and NS3 were most likely involved in the determination of the plaque phenotype. Comparison between genome sequences (obtained from GenBank) of low and high passage strains from two additional serotypes indicated that VP2 was the only protein with amino acid substitutions in all four serotypes. The amino acid substitutions all occurred within the same hydrophilic area, resulting in increased hydrophilicity of VP2 in the large plaque strains.

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 related bluetongue virus. AHS outbreaks cause huge economic losses in developing countries. In the developed world, outbreaks will result in losses in the equestrian industry and will have an enormous emotional impact on owners of pet horses. Live-attenuated vaccine viruses (LAVs) have been developed, however, safety of these LAVs are questionable due to residual virulence, reversion to virulence, and risk on virulent variants by reassortment between LAVs or with field AHSV. Research aims vaccines with improved profiles. Reverse genetics has recently being developed for AHSV and has opened endless possibilities including development of AHS vaccine candidates, such as Disabled Infectious Single Animal (DISA) vaccine. Here, virulent AHSV5 was recovered and its high virulence was confirmed by experimental infection of ponies. 'Synthetically derived' virulent AHSV5 with an in-frame deletion of 77 amino acids codons in genome segment 10 encoding NS3/NS3a protein resulted in similar in vitro characteristics as published NS3/NS3a knockout mutants of LAV strain AHSV4LP. In contrast to its highly virulent ancestor virus, this deletion AHSV5 mutant (DISA5) was completely safe for ponies. Two vaccinations with DISA5 as well as two vaccinations with DISA vaccine based on LAV strain AHSV4LP showed protection against lethal homologous AHSV. More research is needed to further improve efficacy, to explore the AHS DISA vaccine platform for all nine serotypes, and to study the vaccine profile in more detail.

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 reaction (RT-PCR) (n = 31) and enzyme-linked immunosorbent assay (ELISA) (n = 18) to determine the presence of AHSV and/or antibodies against AHSV. The presence of antibodies against AHSV was demonstrated in all 18 samples assayed, and AHSV double stranded RNA was detected in 26% of the animals. This is evidence that E. z. hartmannae can become infected with AHSV.

B-cell epitopes of African horse sickness virus serotype 4 recognised by immune horse sera.

Identifying antigenic proteins and mapping their epitopes is important for the development of diagnostic reagents and recombinant vaccines. B-cell epitopes of African horse sickness virus (AHSV) have previously been mapped on VP2, VP5, VP7 and NS1, using mouse, rabbit and chicken monoclonal antibodies. A comprehensive study of the humoral immune response of five vaccinated horses to AHSV-4 antigenic peptides was undertaken. A fragmented-genome phage display library expressing a repertoire of AHSV-4 peptides spanning the entire genome was constructed. The library was affinity selected for binders on immobilised polyclonal immunoglobulin G (IgG) isolated from horse sera collected pre- and post-immunisation with an attenuated AHSV-4 monovalent vaccine. The DNA inserts of binding phages were sequenced with Illumina high-throughput sequencing. The data were normalised using preimmune IgG-selected sequences. More sequences mapped to the genes coding for NS3, VP6 and VP5 than to the other genes. However, VP2 and VP5 each had more antigenic regions than each of the other proteins. This study identified a number of epitopes to which the horse's humoral immune system responds during immunisation with AHSV-4.

African Horse Sickness Virus: History, Transmission, and Current Status.

African horse sickness virus (AHSV) is a lethal arbovirus of equids that is transmitted between hosts primarily by biting midges of the genus Culicoides (Diptera: Ceratopogonidae). AHSV affects draft, thoroughbred, and companion horses and donkeys in Africa, Asia, and Europe. In this review, we examine the impact of AHSV critically and discuss entomological studies that have been conducted to improve understanding of its epidemiology and control. The transmission of AHSV remains a major research focus and we critically review studies that have implicated both Culicoides and other blood-feeding arthropods in this process. We explore AHSV both as an epidemic pathogen and within its endemic range as a barrier to development, an area of interest that has been underrepresented in studies of the virus to date. By discussing AHSV transmission in the African republics of South Africa and Senegal, we provide a more balanced view of the virus as a threat to equids in a diverse range of settings, thus leading to a discussion of key areas in which our knowledge of transmission could be improved. The use of entomological data to detect, predict and control AHSV is also examined, including reference to existing studies carried out during unprecedented outbreaks of bluetongue virus in Europe, an arbovirus of wild and domestic ruminants also transmitted by Culicoides.

Molecular and serological surveillance of African horse sickness virus in eastern and central Saudi Arabia.

African horse sickness virus (AHSV) is one of the most devastating viral diseases of the family Equidae. Infection with AHSV threatens not only the Saudi equine industry but also the equine industry worldwide. This is due to the high morbidity and mortality rates among the infected population of up to 100%. The World Organisation for Animal Health (OIE) lists AHSV among its notifiable diseases; this requires Member Countries to monitor the situation with regard to AHSV very carefully in order to avoid the spread of the virus. The OIE also suggests the systematic monitoring of AHSV in the equine population at regular intervals. The main aim of the current study is to perform molecular and serological surveillance on different horse populations in eastern and central regions of Saudi Arabia. To achieve this aim, the authors collected 361 serum samples, 103 whole blood samples and 323 swabs from Al-Hasa, Dammam, Al-Jubail, Al-Qateef, Riyadh and Al-Qassim. Commercial enzyme-linked immunosorbent assay (ELISA) kits were used to detect AHSV antibodies and commercial real-time reverse transcriptase-polymerase chain reaction (RT-PCR) kits were used to detect AHSV nucleic acids in blood and swabs. The results of this study demonstrate the absence of anti-AHSV antibodies in the sera of tested animals. Furthermore, no viral nucleic acids were detected in the collected blood and swab samples, as evaluated by real-time AHSV-RT-PCR. Moreover, all tested samples collected during 2014-2016 were negative for AHSV. This confirms that the horse populations studied in the eastern and central regions of Saudi Arabia during 2014-2016 were AHSV free.

Le virus de la peste équine est responsable d'une des maladies virales les plus dévastatrices affectant les membres de la famille des Equidae. Les infections par le virus de la peste équine sont une menace pour le secteur équin saoudien et plus largement pour celui du monde entier. La gravité de cette menace est due aux taux de morbidité et de mortalité extrêmement élevés dans les populations atteintes, pouvant atteindre 100 %. L'infection par le virus de la peste équine fait partie des maladies à déclaration obligatoire de l'Organisation mondiale de la santé animale (OIE) ; de ce fait, les Pays membres doivent suivre la situation sanitaire de leur cheptel au regard du virus de la peste équine afin d'éviter sa propagation. L'OIE recommande également de réaliser un dépistage systématique et régulier du virus de la peste équine dans la population équine. Les auteurs présentent les résultats d'une étude basée sur la surveillance moléculaire et sérologique de plusieurs populations de chevaux dans les régions orientale et centrale de l'Arabie saoudite. Pour les besoins de cette étude, les auteurs ont prélevé 323 échantillons de sérum, 103 échantillons de sang entier et 323 écouvillons de chevaux provenant des localités d'Al-Hasa, Dammam, Al-Jubail, Al-Qatif, Riyad et Al-Qasim. Une épreuve immuno-enzymatique (ELISA) sous forme de kits du commerce a été utilisée pour détecter la présence d'anticorps dirigés contre le virus de la peste équine ; la présence dans le sang et les écouvillons d'acides nucléiques spécifiques du virus de la peste équine a été détectée au moyen d'une amplification en chaîne par polymérase couplée à une transcription inverse (RT­PCR) du commerce. Les résultats de cette étude ont montré l'absence d'anticorps dirigés contre le virus de la peste équine dans le sérum des animaux testés. De même, la RT­PCR en temps réel n'a pas détecté d'acides nucléiques spécifiques du virus de la peste équine dans les prélèvements de sang ni les écouvillons analysés. En outre, tous les échantillons collectés entre 2014 et 2016 et soumis à un test ont donné des résultats négatifs pour le virus de la peste équine. Ces résultats confirment que les populations de chevaux étudiées entre 2014 et 2016 dans les régions orientale et centrale de l'Arabie saoudite étaient indemnes de peste équine.

El virus de la peste equina provoca una de las enfermedades víricas más devastadoras que afectan a la familia de los équidos. La infección por este virus amenaza al sector equino no solo de Arabia Saudí, sino del mundo entero, dado que en las poblaciones infectadas las tasas de morbilidad y mortalidad pueden llegar al 100%. La Organización Mundial de Sanidad Animal (OIE) tiene incluida esta infección en su lista de enfermedades de declaración obligatoria, lo que obliga a sus Países Miembros a seguir muy de cerca la situación sanitaria al respecto para evitar que el virus se disemine. La OIE también sugiere hacer periódicamente controles sistemáticos de la presencia del virus en la población equina. Los autores describen un estudio encaminado básicamente a realizar operaciones de vigilancia molecular y serológica de diferentes poblaciones de caballos de las regiones oriental y central de Arabia Saudí. Para ello, los autores obtuvieron 361 muestras de suero, 103 muestras de sangre entera y 323 hisopados en las áreas de Al Hasa, Dammam, Jubail, Qatif, Riad y Casim. Para detectar anticuerpos contra el virus de la peste equina utilizaron un estuche comercial de ensayo inmunoenzimático (ELISA) y para detectar la presencia de ácidos nucleicos del virus en muestras sanguíneas e hisopados un estuche comercial de reacción en cadena de la polimerasa con retrotranscriptasa (RT­PCR) en tiempo real. Los resultados del estudio demuestran la ausencia de anticuerpos contra el virus en el suero de los animales analizados. La técnica de RT­PCR en tiempo real tampoco deparó indicio alguno de la presencia de ácido nucleico vírico en las muestras de sangre e hisopados. Además, todas las muestras analizadas obtenidas entre 2014 y 2016 resultaron negativas para el virus, lo que confirma que las poblaciones equinas estudiadas durante ese periodo en las regiones central y oriental de Arabia Saudí estaban libres del virus de la peste equina.

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of African Horse Sickness Virus.

African horse sickness (AHS) is a disease of equids caused by African Horse Sickness Virus (AHSV) and is transmitted by Culicoides midges. AHS is endemic in sub-Saharan Africa, but during the past century, outbreaks of significant economic importance and elevated mortality have been recorded in Northern African countries, the Iberian and Arabian Peninsula, the Middle East and the Indian subcontinent. Effective control combines the application of early warning systems, accurate laboratory diagnosis and reporting, animal movement restrictions, suitable vaccination and surveillance programs, and the coordination of all these measures by efficient veterinary services. Conventional reverse-transcriptase (RT) PCR (RT-PCR) and real-time RT-PCR (rRT-PCR) assays have improved the sensitivity and rapidity of diagnosing AHS, resulting in the adoption of these methods as recommended tests by the World Organisation for Animal Health (OIE). However, currently these assays are only performed within laboratory settings; therefore, the development of field diagnostics for AHS would improve the fast implementation of control policies. Loop-mediated isothermal amplification (LAMP) is an isothermal, autocycling, strand-displacement nucleic acid amplification technique which can be performed in the field. LAMP assays are attractive molecular assays because they are simple to use, rapid, portable and have sensitivity and specificity within the range of rRT-PCR. This study describes the development of a novel RT-LAMP assay for the detection of AHSV. The AHSV RT-LAMP assay has an analytical sensitivity of 96.1% when considering an rRT-PCR cut-off value of CT  > 36, or 91.3% when no rRT-PCR cut-off is applied. Diagnostic sensitivity and specificity were 100%. This assay provides for a rapid and low cost AHS diagnostic for use in the field.