Equine Virus Research Articles

Short communication: Retrospective analysis of obligatory testing results for Equine virus arteritis reveals a decrease of its seroprevalence in stallions used for artificial insemination

Equine viral arteritis (EVA) can induce a persistent carrier state in stallions which then shed the virus via semen. About 30 years ago, obligatory EVA testing of stallions used for artificial insemination (AI) was implemented in the European Union. Information on the efficacy of these regulations on the prevalence of EVA in stallions are not yet available. Therefore, we retrospectively analyzed results of serological and virus antigen testing for EVA in sires of different age and breed referred to Vetmeduni Vienna for semen preservation or veterinary diagnostic procedures between 2001 and 2021. For analysis, stallions were grouped by age (1–5, 6–8, 9–12, >12 years) and breed. The EVA antibody titer was determined by serum neutralization test and semen was analyzed for EVA virus by PCR and/or virus isolation test. Of 308 stallions tested, 14.9% (n = 46) were EVA seropositive and in 12 stallions EVA virus was detected in semen (26% of seropositive stallions). The incidence of seropositive stallions decreased over time (P < 0.05, χ2 test). Differences in the seroprevalence of EVA antibodies existed among stallion age groups (P < 0.01, Fisher’s test) with the highest percentage of seropositive stallions being > 12 years old (43.5%). The EVA antibody titer increased with age (P < 0.01, Kruskal-Wallis test), potentially reflecting repeated virus challenge. In conclusion, analysis of monitoring results revealed a decrease of EVA seroprevalence and virus shedding in a European sire population. As monitoring for EVA was the only measure implemented Europe-wide, testing might be a major contributor to this development.

Detection of Selected Equine Respiratory Pathogens in Stall Samples Collected at a Multi-Week Equestrian Show during the Winter Months.

The aim of this study was to use environmental sampling to determine the frequency of detection of selected equine respiratory viruses and bacteria in horses attending a multi-week equestrian show during the winter months. At four time points during showing, environmental sponge samples were collected from all stalls on the property and tested for the presence of equine herpesvirus-1 (EHV-1), EHV-2, EHV-4, equine influenza virus (EIV), equine rhinitis B virus (ERBV), Streptococcus equi ss. equi (S. equi), and S. equi ss. zooepidemicus (S. zooepidemicus) using real-time PCR (PCR). Environmental sponges were collected from all 53 barns by using one sponge for up to 10 stalls. Further, 2/53 barns were randomly selected for individual stall sampling in order to compare the results between individual and pooled stall samples. A total of 333/948 (35.13%, 95% CI 32.09-38.26%) pooled environmental stall sponges tested PCR-positive for at least one of the selected respiratory pathogens. Streptococcus zooepidemicus was the most commonly detected pathogen in pooled samples (28.69%, 95% CI 25.83-31.69%), followed by EHV-2 (14.45%, 95% CI 12.27-16.85%), EHV-4 (1.37%, 95% CI 0.73-2.33%), and a very small percentage of pooled stall sponges tested PCR-positive for EHV-1, ERBV, EIV, and S. equi. In individual samples, 171/464 (36.85%, 95% CI 32.45-41.42%) environmental stall sponges tested PCR-positive for at least one of the selected pathogens, following a similar frequency of pathogen detection as pooled samples. The detection frequency of true respiratory pathogens from environmental samples was higher during the winter months compared to previous studies performed during spring and summer, and this testing highlights that such pathogens circulate with greater frequency during the colder months of the year. The strategy of monitoring environmental stall samples for respiratory pathogens circumvents the often labor-intensive collection of respiratory secretions from healthy horses and allows for a more efficient assessment of pathogen buildup over time. However, environmental stall testing for respiratory pathogens should not replace proper biosecurity protocols, but it should instead be considered as an additional tool to monitor the silent circulation of respiratory pathogens in at-risk horses.

Interactions of Equine Viruses with the Host Kinase Machinery and Implications for One Health and Human Disease.

Zoonotic pathogens that are vector-transmitted have and continue to contribute to several emerging infections globally. In recent years, spillover events of such zoonotic pathogens have increased in frequency as a result of direct contact with livestock, wildlife, and urbanization, forcing animals from their natural habitats. Equines serve as reservoir hosts for vector-transmitted zoonotic viruses that are also capable of infecting humans and causing disease. From a One Health perspective, equine viruses, therefore, pose major concerns for periodic outbreaks globally. Several equine viruses have spread out of their indigenous regions, such as West Nile virus (WNV) and equine encephalitis viruses (EEVs), making them of paramount concern to public health. Viruses have evolved many mechanisms to support the establishment of productive infection and to avoid host defense mechanisms, including promoting or decreasing inflammatory responses and regulating host machinery for protein synthesis. Viral interactions with the host enzymatic machinery, specifically kinases, can support the viral infectious process and downplay innate immune mechanisms, cumulatively leading to a more severe course of the disease. In this review, we will focus on how select equine viruses interact with host kinases to support viral multiplication.

Epidemiological aspects of some equine viral diseases

Although different equine viruses’ outbreaks have been recorded. However, the most important ones in are the African horse sickness virus (AHSV), equine influenza virus (EIV), equine viral arteritis (EVA), Equine infectious anaemia virus (EIAV), and equine herpes viruses (EHV). To combat these diseases, it is imperative to understand their epidemiological aspects. So, the current review aims to highlight some epidemiological aspects including; causative agents, clinical forms, history, prevalence and geographical distribution, source of infection, and methods of transmission. The AHSV mainly causes pulmonary, and cardiac forms with high morbidity and mortality rates in Africa. The EIV is found all over the world and results in respiratory signs. The EVA has low morbidity and mortality rates and is mainly found in the Americas and Europe, its significance is due to the reproductive problems as abortion in mares and subfertility in stallions. The EIAV has low morbidity and mortality rates and causes long time course disease mainly of fever, and chronic anaemia or death. The EHVs are the current most important pathogens due to their endemicity all over the world and their high morbidity. It causes respiratory, abortion, neonatal, and sometimes neurological manifestations. Aerosols and body excretions are the main sources of infection with EIV, EVA, and EHV. Venereal EVA transmission occurs through natural breeding or artificial insemination with the semen of infected or carrier stallions. The spreading of arboviruses is greatly affected by the vector activity like the AHSV which transmitted by the the Culicoide. imicola biting midges, and the EIAV by family Tabanidae. In general, it is recommended to take all epidemiological measures, including vaccinations and vector control, to limit the spread of such diseases and reduce economic losses.

Seroprevalence of Infectious Respiratory Agents in Thoroughbred Race Horses at the Seoul Race Park, Republic of Korea

Infectious respiratory disease is one of the most frequent causes of lost days in training and reduced performance of Thoroughbred racehorses. Equine influenza virus (EIV), Equine herpesvirus-1 (EHV-1) and -4 (EHV-4), equine rhinitis virus A (ERAV) and B (ERBV), and Streptococcus equi subspecies equi (S. equi) are important infectious agents of the respiratory tract of horses. The purpose of this study was to determine the seroprevalence of EHV-1, EHV-4, ERAV, ERBV, and S. equi and to measure EIV antibody levels of Thoroughbred racehorses at Seoul Race Park (SRP), Republic of Korea. All horses had previously been vaccinated against EIV and S. equi, but not against any of the other pathogens that were tested. A total of 94 serum samples, which were collected from race participants at the SRP were tested using the single radial haemolysis (SRH) test for EIV (H3N8), the complement-fixation (CF) test for EHV-1, EHV-4, ERAV, ERBV and an indirect enzyme-linked immunosorbent assay (iELISA) for S. equi. Serum samples from seventy eight out of 94 horses (83%) generated zones of over 85 mm2 in the SRH test, which classified them as clinically protected against EIV (H3N8). The most sero-prevalent agent detected was EHV-4 (30.9%, 29/94), followed by EHV-1 (9.6%, 9/94), S. equi (2.1%, 2/94), ERAV (1.1%, 1/94) and ERBV (1.1%, 1/94). All horses showed no visual clinical signs. The present study showed that the seroprevalence of infectious respiratory agents was relatively low and provides evidence of low risk of respiratory infectious agents in Thoroughbred race horses at SRP.

Equine Hepacivirus: A Systematic Review and a Meta-Analysis of Serological and Biomolecular Prevalence and a Phylogenetic Update.

Simple SummaryThis is a comprehensive review containing the most up-to-date information on Equine Hepacivirus, one of the recently discovered hepatic equine viruses, together with an analysis of serological and biomolecular presence presented in apreviously published papers, and an update on its genetic relationship within the species and with similar species. Extensive description of the EqHV features is included, and results are presented with several tables and figures, providing a valuable reference guide for further studies.Viral hepatitis has recently assumed relevance for equine veterinary medicine since a variety of new viruses have been discovered. Equine Hepacivirus (EqHV) is an RNA virus belonging to the Flaviviridae family that can cause subclinical hepatitis in horses, occasionally evolving into a chronic disease. EqHV, to date, is considered the closest known relative of human HCV. EqHV has been reported worldwide therefore assessing its features is relevant, considering both the wide use of blood products and transfusions in veterinary therapies and its similitude to HCV. The present review resumes the actual knowledge on EqHV epidemiology, risk factors and immunology, together with potential diagnostics and good practices for prevention. Moreover, adhering to PRISMA guidelines for systematic reviews a meta-analysis of serological and biomolecular prevalence and an updated phylogenetic description is presented as a benchmark for further studies.

Complete Coding Genome Sequence of an Influenza A/H3N8 Equine Virus Isolated in Kazakhstan in 2007.

ABSTRACTHere, we reported the complete coding sequence of the influenza A/equine/Otar/3/2007 (H3N8) equine virus, first isolated in Kazakhstan in 2007. The hemagglutinin (HA) sequences of the Kazakhstan isolates appeared to be closely related to viruses isolated in early 2000 in Asia. Phylogenetic analysis characterized the Kazakhstan isolates as a member of the Florida sublineage clade 2 by the HA protein sequence.

Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus.

Equine infectious anaemia (EIA) is controlled by the identification of seropositive animals. The official diagnostic method is the agar gel immunodiffusion (AGID) test, which detects antibodies against a viral core protein (p26). Although AGID is inexpensive and specific, the report of results takes considerable time and the test has low analytical sensitivity. To validate our in-house indirect ELISAgp90/45 , following the World Organization of Animal Health (OIE) criteria. Test validation. Synthetic peptides gp90 and gp45 were used as antigens in ELISAgp90/45 . Tests used for validation, calibration and linear working operating range, analytical and diagnostic sensitivity and specificity, repeatability and reproducibility were assessed by comparing them with the AGID test and using 1844 equine sera grouped into five different panels. We were able to replace the National References Sera with our Internal Reference Sera. ELISAgp90/45 had acceptable repeatability and reproducibility. Analytical sensitivity of the ELISAgp90/45 was 800 times greater than that of AGID test for positive sera and 400 times greater for weak positive sera. ELISAgp90/45 also showed optimal analytical specificity, since no cross-reactivity was detected with antibodies against other equine viruses. One sample was positive by AGID test and negative by ELISAgp90/45. ELISAgp90/45 was performed using 243 EIA positive and 878 negative equid sera, and showed a diagnostic sensitivity of 99.59% [CI 97.73%-99.99%] and a diagnostic specificity of 90.32% [CI 88.17%-92.19%], compared to AGID test; thus, it was demonstrated to be a robust test. Samples were derived from naturally infected equid populations showing heterogeneous clinical states: therefore, their status was uncertain and some horses were sampled more than once. The AGID test may not be the most useful gold standard. ELISAgp90/45 is a useful tool for the diagnosis of EIAV infection and meets validation requirements established by the OIE.

Development of a high‐throughput screening assay to identify equine virus arteritis replication inhibitors

Development of a high‐throughput screening assay to identify equine virus arteritis replication inhibitors

Madariaga Virus in Colombia, Detection of Alphavirus in Colombian Caribbean

Abstracts Alphaviruses are agents of neurotropic arboviral infections, several complications include encephalitis, meningoencephalitis and hemorrhagic fever. In Colombia, Venezuelan Encephalitis Equine virus (VEEV) is an endemic virus detected in Caribbean and Santander region, however, scarce information has evidenced about Eastern Equine Encephalitis virus complex (EEEV sensu estricto and Madariaga virus), with reports in Antioquia and Pacific region. We detected Madariaga virus infecting mosquito species (Culex erraticus, Culex quinquefasciatus, Psorophora ferox, Mansonia titillans and Culex spp.), in the rural locality of “Alto Cotoca” - Lorica, belongs to Northern – Cordoba (Colombia). The phylogenetic analysis allows verified to Lineage III for Madariaga virus.

Serological Evidence of Common Equine Viral Infections in a Semi-Isolated, Unvaccinated Population of Hucul Horses.

Simple SummaryThe aim of the following research was an analysis of the occurrence of common equine viral infections in a Hucul herd, based on serological studies. This study provides epidemiological data concerning animals representing a semi-isolated herd, devoid of any specific prophylaxis. The obtained results provide a source of information on a primitive breed of horses, as well as giving an insight into viruses (among them, arboviruses) circulating within a local ecosystem.Huculs (Equus caballus) are an old breed of primitive mountain horses, originating from the Carpathian Mountains. To the best of our knowledge, data concerning the epidemiology of viral infections observed within this breed are sparse. The objective of this study was to estimate the serological status of a semi-isolated, unvaccinated Hucul herd, with respect to both common equine viral infections and horse-infecting arboviruses, the presence of which was previously reported in Poland. Twenty horses of the Hucul breed, living in a remote area in Poland, were studied in 2018 from March to May. Using nasal secretion swabs as a specimen source, isolation attempts were negative regarding ERAV, EHV-1, EAV, and EIV. According to the virus neutralisation method, in the sera obtained from the animals, antibodies against the following viruses were detected: EHV-1 in 12 horses (60%; with titres from 1:8 to 1:64), EIV A/H7N7 in 13 (65%; titres from 1:20 to 1:80), EIV A /H3N8 in 12 (60%; titres from 1:20 to 1:80), USUV in 5 (25%; titres from 1:10 to 1:80), and ERAV in 1 (5%; titre 1:32). Antibodies against EAV, EIAV, and WNV were not present in the tested sera. The detected presence of specific antibodies associated with five out of the eight equine viruses investigated indicates that the Hucul herd, due to its partial separation and lack of specific prophylaxis, could serve as a sentinel animal group for the detection of equine viruses/arboviruses present within the local ecosystem. The detection of common equine viral infections within the herd provides additional epidemiological data concerning the breed.

Isothermal Nucleic Acid Amplification Technologies for the Detection of Equine Viral Pathogens

Simple SummaryEquine viral diseases remain a prominent concern for human and equine health globally. Many of these viruses are of primary biosecurity concern to countries that import equines where these viruses are not present. In addition, several equine viruses are zoonotic, which can have a significant impact on human health. Current diagnostic techniques are both time consuming and laboratory-based. The ability to accurately detect diseases will lead to better management, treatment strategies, and health outcomes. This review outlines the current modern isothermal techniques for diagnostics, such as loop-mediated isothermal amplification and insulated isothermal polymerase chain reaction, and their application as point-of-care diagnostics for the equine industry.The global equine industry provides significant economic contributions worldwide, producing approximately USD $300 billion annually. However, with the continuous national and international movement and importation of horses, there is an ongoing threat of a viral outbreak causing large epidemics and subsequent significant economic losses. Additionally, horses serve as a host for several zoonotic diseases that could cause significant human health problems. The ability to rapidly diagnose equine viral diseases early could lead to better management, treatment, and biosecurity strategies. Current serological and molecular methods cannot be field-deployable and are not suitable for resource-poor laboratories due to the requirement of expensive equipment and trained personnel. Recently, isothermal nucleic acid amplification technologies, such as loop-mediated isothermal amplification (LAMP) and insulated isothermal polymerase chain reaction (iiPCR), have been developed to be utilized in-field, and provide rapid results within an hour. We will review current isothermal diagnostic techniques available to diagnose equine viruses of biosecurity and zoonotic concern and provide insight into their potential for in-field deployment.

Equine respiratory viruses, airway inflammation and performance in thoroughbred racehorses

Equine asthma is a common cause of poor performance in racehorses but it is unclear if respiratory viruses contribute to its etiology. The objective of the study was to determine if respiratory viruses were associated with clinical signs and bronchoalveolar lavage fluid (BALF) cytology in Thoroughbred racehorses. Equine herpesviruses (EHV-1, 2, 4, 5) and equine rhinitis A and B viruses (ERBV, ERAV) genomes were quantified by qPCR in nasopharyngeal, tracheal, and BALF samples collected after racing. The relationships between virus detection and load and clinical signs, performance, BALF cytology, and environmental exposures were examined with generalized linear mixed models. Ninety-two samples were collected from 31 horses. EHV-1 and ERAV were not found; EHV-4 was detected in only one sample. EHV-2, EHV-5 and ERBV were more likely to be detected in upper airway samples than in BALF (P < 0.0001). Neither respiratory virus detection nor load was associated with clinical signs or performance. Nasopharyngeal detection and load of ERBV and tracheal detection and load of EHV-5 were associated with increased proportions of neutrophils in BALF (P < 0.003). However, nasopharyngeal detection and load of EHV-5 was not (P = 0.11). Nasopharyngeal detection and load of EHV-2 were associated with decreased BALF mast cell proportions. Respirable dust exposures were significantly higher in horses with detection of ERBV when compared to horses with no detectable ERBV (P < 0.001). Our results suggest that ERBV, EHV-2 and EHV-5 are commonly present in upper airways of healthy racehorses; however, the role they play in the etiology of equine asthma remains unclear.

Serological evidence of Eastern equine encephalitis circulation in equids in Pará state, Brazil.

Serum samples from 89 equids were analyzed (75 horses, 9 donkeys, and 5 mules) from the municipality of Viseu, Pará state, Brazil. Samples were collected in November 2014 and August 2015. The antibody prevalence against the following alphaviruses was estimated: Eastern equine encephalitis virus, Western equine encephalitis virus, Mucambo virus, and Mayaro virus. Seroprevalence was determined by the hemagglutination inhibition (HI) technique. Sera that exhibited HI antibodies with heterotypic reactions for the analyzed viruses were subjected to the 90% plaque reduction neutralization test (PRNT90). The HI prevalence of monotypic reactions to EEEV was 7.9%, and that of WEEV was 1.1%, as confirmed by PRNT90. Viral isolation attempts were negative for all tested blood samples. Our results suggest the circulation of equine encephalitis complex viruses. Future studies should evaluate the possible involvement of arthropod hosts and residents in the viral transmission in the study area.

EQUINE ZOONOTIC DISEASES: A REVIEW

Zoonotic diseases are those diseases that are transferred from animals to humans. Several equine zoonotic diseases affected humans from mild to severe such as vesicular stomatitis, brucellosis, methicillin-resistant staphylococcus aureus, equine Hendra virus, dermatophytosis, leptospirosis, anthrax, giardiasis, rabies, arboviral encephalitis, acute diarrhea, salmonellosis, clostridium difficile, and cryptosporidiosis. These serious diseases did not eliminate due to many reasons. Personal hygiene, protective clothing, recognition of zoonotic agents and identification of potential fomites can reduce zoonotic diseases or prevent the transfer of zoonotic diseases.

First identification and genomic characterization of equine hepacivirus sub-type 3 strain in China.

Equine Hepacivirus (EqHV) is a newly discovered equine virus that is classified under the Hepacivirus genus of the Flaviviridae family. There are three sub-types of EqHV worldwide namely; sub-types 1-3. The majority of EqHV sub-type 1 strains were found in China. While different sub-types have been found in Japan and USA, therefore, to investigate whether the other sub-types of EqHV strains were present in China, a total of 60 horse serum samples were collected and screened for EqHV RNA through RT-PCR. The results revealed that 19 serum samples were RNA-positive (19/60) and the EqHV detection rate was 31.67%. One EqHV strain named GD23 was obtained and its near-complete genome sequence was acquired. Analysis of nucleotide p-distance with reference to the entire polyprotein gene revealed that GD23 was classified into sub-type 3. In addition, the phylogenetic analysis demonstrated that GD23 was clustered together with EqHV strains of sub-type 3 in other countries. The present study is the first to identify an EqHV sub-type 3 strain in China.

Molecular and genomic characterization of a novel equine molluscum contagiosum-like virus.

Cases of pox-like lesions in horses and donkeys have been associated with poxviruses belonging to different genera of the family Poxviridae. These include the orthopoxviruses vaccinia virus (VACV), horsepoxvirus (HPXV) and cowpoxvirus (CPXV), as well as a potentially novel parapoxvirus and molluscum contagiosum virus (MOCV). However, with the exception of VACV, HPXV and CPXV, the genomic characterization of the causative agents remains largely elusive with only single short genome fragments available. Here we present the first full-length genome sequence of an equine molluscum contagiosum-like virus (EMCLV) directly determined from skin biopsies of a horse with generalized papular dermatitis. Histopathological analysis of the lesions revealed severe epidermal hyperplasia with numerous eosinophilic inclusion bodies within keratinocytes. Virions were detected in the lesions in embedded tissue by transmission electron microscopy. The genome sequence determined by next- and third-generation sequencing comprises 166 843 nt with inverted terminal repeats (ITRs) of 3473 nt. Overall, 20 of the predicted 159 ORFs have no equivalents in other poxviruses. Intriguingly, two of these ORFs were identified to encode homologues of mammalian proteins involved in immune signalling pathways, namely secreted and transmembrane protein 1 (SECTM1) and insulin growth factor-like family receptor 1 (IGFLR1), that were not described in any virus family so far. Phylogenetic analysis with all relevant representatives of the Poxviridae suggests that EMCLV should be nominated as a new species within the genus Molluscipoxvirus.

Detection of the epidemic of the H3N8 subtype of the equine influenza virus in large-scale donkey farms

ABSTRACT To monitor the occurrence of equine influenza in large-scale donkey farms in Liaocheng City, Shandong Province, serological investigation and sequence analysis of HA/M protein gene of equine influenza virus (EIV) were carried out. Samples (n = 65) of the lung and nasal swab were collected in six different large-scale donkey farms and detected with RT-PCR for HA and M protein gene. The homology and evolution of HA and M genes were analysed with known sequences. Antibody titres of serum samples (n = 120, unvaccinated) level was determined by the HI test. The average seropositive rate was 32.5% (39/120) with great diversity among different populations. The positive rate of EIV HA/M protein gene was 21.5% (14/65) by RT-PCR. The equine influenza H3N8 virus was confirmed by gene sequencing, and the homology of the sequence was 99.77% with isolates from Northeast China (equine/heilongjiang/1/2010), consistent with the input of donkeys. This suggested that EIV has become an important threat to large-scale donkey farms in Liaocheng and threats from the input area must be vigilant.

Smartphone-based multiplex 30-minute nucleic acid test of live virus from nasal swab extract.

Rapid, sensitive and specific detection and reporting of infectious pathogens is important for patient management and epidemic surveillance. We demonstrated a point-of-care system integrated with a smartphone for detecting live virus from nasal swab media, using a panel of equine respiratory infectious diseases as a model system for corresponding human diseases such as COVID-19. Specific nucleic acid sequences of five pathogens were amplified by loop-mediated isothermal amplification on a microfluidic chip and detected at the end of reactions by the smartphone. Pathogen-spiked horse nasal swab samples were correctly diagnosed using our system, with a limit of detection comparable to that of the traditional lab-based test, polymerase chain reaction, with results achieved in ∼30 minutes.

Association between inflammatory airway disease of horses and exposure to respiratory viruses: a case control study

Background: Inflammatory airway disease (IAD) in horses, similar to asthma in humans, is a common cause of chronic poor respiratory health and exercise intolerance due to airway inflammation and exaggerated airway constrictive responses. Human rhinovirus is an important trigger for the development of asthma; a similar role for viral respiratory disease in equine IAD has not been established yet.&#x0D; Methods: In a case–control study, horses with IAD (n = 24) were compared to control animals from comparable stabling environments (n = 14). Horses were classified using pulmonary function testing and bronchoalveolar lavage. PCR for equine rhinitis virus A and B (ERAV, ERBV), influenza virus (EIV), and herpesviruses 2, 4, and 5 (EHV-2, EHV-4, EHV-5) was performed on nasal swab, buffy coat from whole blood, and cells from BAL fluid (BALF), and serology were performed. Categorical variables were compared between IAD and control using Fisher’s exact test; continuous variables were compared with an independent t-test. For all analyses, a value of P &lt;0.05 was considered significant.&#x0D; Results: There was a significant association between diagnosis of IAD and history of cough (P = 0.001) and exercise intolerance (P = 0.003) but not between nasal discharge and IAD. Horses with IAD were significantly more likely to have a positive titer to ERAV (68 %) vs. control horses (32 %). Horses with IAD had higher log-transformed titers to ERAV than did controls (2.28 ± 0.18 v.1.50 ± 0.25, P = 0.038). There was a significant association between nasal shedding (positive PCR) of EHV-2 and diagnosis of IAD (P = 0.002).&#x0D; Conclusions: IAD remains a persistent problem in the equine population and has strong similarities to the human disease, asthma, for which viral infection is an important trigger. The association between viral respiratory infection and development or exacerbation of IAD in this study suggests that viral infection may contribute to IAD susceptibility; there is, therefore, merit in further investigation into the relationship between respiratory virus exposure and development of IAD.