Virology in horses encompasses the study of viruses that affect equine species, including their biology, transmission, and impact on horse health. This field investigates viral pathogens that can lead to a range of diseases, from respiratory infections to neurological disorders. Common viruses affecting horses include equine influenza virus, equine herpesvirus, and West Nile virus. Understanding these viruses involves examining their genetic makeup, modes of transmission, and interactions with the equine immune system. This page compiles peer-reviewed research studies and scholarly articles that explore the epidemiology, pathogenesis, and control measures of viral infections in horses.
Telford EA, Watson MS, Aird HC, Perry J, Davison AJ.The complete DNA sequence of equine herpesvirus 2 (EHV-2) strain 86/67 was determined. The genome is 184,427 bp in size and has a base composition of 57.5% G + C. Unusually for a herpesvirus, about a third of the sequence distributed in several large blocks appears not to encode proteins. The 79 open reading frames that were identified as probably polypeptide-coding are predicted to encode 77 distinct proteins. Amino acid sequence comparisons confirmed that EHV-2 is a gamma-herpesvirus that is genetically collinear with herpesvirus saimiri (HVS; a gamma 2-herpesvirus) and Epstein-Barr virus (E...
Rico-Hesse R, Weaver SC, de Siger J, Medina G, Salas RA.One of the most important questions in arbovirology concerns the origin of epidemic Venezuelan equine encephalitis (VEE) viruses; these viruses caused periodic, extensive epidemics/epizootics in the Americas from 1938-1973 (reaching the United States in 1971) but had recently been presumed extinct. We have documented the 1992 emergence of a new epidemic/epizootic VEE virus in Venezuela. Phylogenetic analysis of strains isolated during two outbreaks indicated that the new epidemic/epizootic virus(es) evolved recently from an enzootic VEE virus in northern South America. These results suggest co...
Schlocker N, Gerber-Bretscher R, von Fellenberg R.In a search of viral agents in pulmonary macrophages of horses with chronic pulmonary disease, equine herpesvirus 2 was found to be unique. In 8 of 9 horses with chronic pulmonary disease, antigens of equine herpesvirus 2 were detected by indirect immunofluorescence staining of scattered foamy macrophages immediately after harvesting by bronchoalveolar lavage and fractionation on metrizamide gradients. In a healthy horse, antigens were not found. After 1 week of cultivation of bronchoalveolar lavage cells from a second group of 9 horses with chronic pulmonary disease, viral antigens were detec...
Sato K, Tokuhisa S, Inaba Y.The growth of group A human, bovine, equine and porcine rotaviruses were enhanced by pretreatment of virus with pancreatin, trypsin, protease, alkaline phosphatase or pepsin and incorporation of these enzymes in maintenance medium. In contrast, alpha-amylase or lipase inhibited the growth of equine and porcine rotaviruses. The other enzymes, adenosine deaminase, lactase, lysozyme, ribonuclease or triose-phosphate isomerase gave little or no change in the growth of all four rotaviruses.
Morley PS, Hanson LK, Bogdan JR, Townsend HG, Appleton JA, Haines DM.Antibodies specific for equine influenza viruses are usually quantified using single radial hemolysis (SRH), hemagglutination inhibition (HI) or virus neutralization (VN). Neutralizing antibodies are thought to provide optimum protection to challenged animals. The purpose of this study was to determine the extent to which SRH and HI assays detect antibodies which neutralize equine influenza viruses. Acute and convalescent sera from 41 horses were analyzed using VN, SRH, and HI assays. These horses were present in a population of Thoroughbred racehorses during an epidemic of upper respiratory t...
Lichtenstein DL, Rushlow KE, Cook RF, Raabe ML, Swardson CJ, Kociba GJ, Issel CJ, Montelaro RC.As an important enzyme in DNA synthesis, dUTPase is present in a wide variety of organisms and viruses and has been identified as a component of the equine infectious anemia virus (EIAV) pol gene. The role of EIAV dUTPase, designated DU, in virus replication in vitro and in vivo was investigated with a recently described infectious molecular clone of EIAV. A deletion mutant that was deficient in dUTPase activity was constructed, and its replication kinetics was examined in fetal equine kidney (FEK) cells and primary equine bone marrow macrophage (EBMM) cells. In FEK cells, which are permissive...
Zientara S, Sailleau C, Moulay S, Wade-Evans A, Cruciere C.The development of a coupled reverse transcriptase-polymerase chain reaction assay (RT-PCR) is described for the detection of African horse sickness virus (AHSV) double-stranded RNA. Genome segments 7 and 10 were chosen as target templates for primers selected for use in the RT-PCR. Using these AHSV-specific primers all 9 serotypes were detectable. The sensitivity and specificity of the RT-PCR results were compared to those obtained by competition ELISA.
Zhao Y, Holden VR, Smith RH, O'Callaghan DJ.The UL3 protein of equine herpesvirus 1 (EHV-1) KyA strain is a homolog of the ICP27 alpha regulatory protein of herpes simplex virus type 1 (HSV-1) and the ORF 4 protein of varicella-zoster virus. To characterize the regulatory function of the UL3 gene product, a UL3 gene expression vector (pSVUL3) and a vector expressing a truncated version of the UL3 gene (pSVUL3P) were generated. These effector plasmids, in combination with an EHV-1 immediate-early (IE) gene expression vector (pSVIE) and chimeric EHV-1 promoter-chloramphenicol acetyltransferase (CAT) reporter constructs, were used in trans...
Murray K, Selleck P, Hooper P, Hyatt A, Gould A, Gleeson L, Westbury H, Hiley L, Selvey L, Rodwell B.A morbillivirus has been isolated and added to an increasing list of emerging viral diseases. This virus caused an outbreak of fatal respiratory disease in horses and humans. Genetic analyses show it to be only distantly related to the classic morbilliviruses rinderpest, measles, and canine distemper. When seen by electron microscopy, viruses had 10- and 18-nanometer surface projections that gave them a "double-fringed" appearance. The virus induced syncytia that developed in the endothelium of blood vessels, particularly the lungs.
Batt RM, Embaye H, van de Waal S, Burgess D, Edwards GB, Hart CA.We used organ culture of jejunal mucosal explants obtained from ponies aged between 2 and 12 months to study enterocyte damage by group A strains of equine rotavirus. Electron microscopy of jejunal explants maintained for < or = 48 h in the presence of organ culture medium alone showed that enterocytes were structurally intact and had a densely packed brush border and overlying mucus. Similarly, examination of explants maintained in the presence of rotavirus for 48 h revealed no apparent ultrastructural abnormalities. However, obvious replication and assembly of virus in enterocytes had occ...
Flowers CC, Flowers SP, Jennings SR, O'Callaghan DJ.Previous studies (C. C. Flowers and D. J. O'Callaghan, 1992, Virology 190, 307-315) employed peptide-specific antibodies to identify the product of the glycoprotein D (gD) gene of equine herpesvirus 1 strain Kentucky A (KyA). gD polypeptides of 55 and 58 kDa were detected in EHV-1-infected L-M cells, and the 58-kDa protein was observed in the membrane fraction of EHV-1 virions. In this report, the kinetics of synthesis and processing of gD polypeptides are described. One-hour pulse-labeling of EHV-1-infected L-M cells revealed that gD proteins are first detected at 6 hr after infection and tha...
Baba SS, Fagbami AH, Ojeh CK, Olaleye OD, Omilabu SA.Retrospective and prospective serological surveys to determine the prevalence of Wesslsbron (WSL) virus infections in animal populations were carried out in different vegetational zones in Nigeria. Sera from 1,492 animals comprising 292 camels, 81 horses, 4 donkeys, 320 cattle, 235 sheep, 260 goats, 114 pigs, 101 dogs and 85 domestic fowls were assayed by haemagglutination-inhibition (HI) test for presence of antibodies to WSL virus and other flavivirus antigens: Yellow Fever (YF), Potiskum (POT), Banzi (BAN), Uganda S (UGS) and West Nile (WN) viruses. Four hundred and eighty one (32%) of the ...
Southgate CD, Green MR.Lentivirus Tat proteins comprise a novel class of RNA-binding transcriptional activators that are essential for viral replication. In this study, we performed a series of protein fusion experiments to delineate the minimal protein domains and promoter elements required for Tat action. We show that a 15-amino-acid region of equine infectious anemia virus (EIAV) Tat protein, when fused to the GAL4 or LexA DNA binding domain, can activate transcription in appropriate promoter contexts. In the natural human immunodeficiency virus type 1 long terminal repeat, activation by Tat is dependent on multi...
Ellis JA, Bogdan JR, Kanara EW, Morley PS, Haines DM.The ability of monovalent and bivalent equine herpesvirus (EHV) vaccines to stimulate cellular and antibody responses to EHV-1 and EHV-4 was compared in healthy horses. Comparison of data from lymphocyte blastogenesis tests in which live viruses were used as antigens and that were conducted prior to vaccination and after 2 vaccinations revealed that horses given modified-live EHV-1 had significant increases in proliferative responses to EHV-1 (P = 0.03) and EHV-4 (P = 0.04). Responses to EHV-1 and EHV-4 in horses given the inactivated-virus bivalent vaccine were less; however, significant diff...
McCann SH, Mumford JA, Binns MM.A search for variable restriction sites has been carried out for equine herpesvirus-1 (EHV-1) in an attempt to develop markers which can be used to group epidemiologically related viruses into groups, and to learn more about the dynamics of EHV-1 disease. Crude viral DNA extracts of EHV-1, prepared by Hirt extraction, were digested with AluI, HaeIII, or RsaI, and Southern blotted following electrophoresis. DNA fingerprints, produced by probing the Southern blots with the EHV-1 EcoR1-I fragment, separated 56 isolates into 16 groups. The variable sites within the EcoR1-I fragment were mapped app...
Morley PS, Bogdan JR, Townsend HG, Haines DM.The Directigen Flu A assay (Becton Dickinson, Microbiology Systems, Mississauga, Ontario, Canada) is a commercially available immunoassay designed for rapid in vitro recognition of influenza A nucleoprotein. The purpose of this study was to evaluate this assay for detection of influenza virus in nasal secretions of naturally infected horses. The assay was shown to react with representative strains of influenza virus which cause disease in horses and did not react with nasal secretions from uninfected horses kept in isolation. Between 33% and 45% of nasal secretions specimens obtained from clin...
Cook RF, Berger SL, Rushlow KE, McManus JM, Cook SJ, Harrold S, Raabe ML, Montelaro RC, Issel CJ.Serial passage of the prototype (PR) cell-adapted Wyoming strain of equine infectious anemia virus (EIAV) in fetal donkey dermal (FDD) rather than fetal horse (designated fetal equine kidney [FEK]) cell cultures resulted in the generation of a variant virus strain which produced accelerated cytopathic effects in FDD cells and was 100- to 1,000-fold more sensitive to neutralizing antibodies than its parent. This neutralization-sensitive variant was designated the FDD strain. Although there were differences in glycosylation between the PR and FDD strains, passage of the FDD virus in FEK cells di...
Blunden AS, Smith KC, Binns MM, Zhang L, Gower SM, Mumford JA.Equid herpesvirus 4 (EHV-4) infection was diagnosed as the cause of interstitial pneumonia in a 6-week-old conventionally reared Welsh pony foal, by cocultivation and immunolabelling with specific monoclonal antibodies, EHV-4 specific amplification of viral DNA, and immunohistological examination of infected tissues. The case was novel in that replication of the EHV-4 isolate in endothelial cells and in the synovial epithelium was a feature. Restriction digests of this isolate were compared with those of seven respiratory and one abortigenic EHV-4 isolate, and no differences in restriction pat...
McCollum WH, Timoney PJ, Tengelsen LA.The clinical, virological and serological responses of seven female donkeys (Equus asinus) to inoculation with the KY-84 strain of equine arteritis virus (EAV), a strain that causes moderate to severe clinical signs in horses, was investigated. In the donkeys, the only clinical signs observed were fever (mainly 3-9 days after inoculation), mild depression in four animals, and a slight nasal or ocular discharge in three. All of the donkeys became infected with EAV as shown by recovery of the virus for periods of up to 14 days from the nasopharynx and buffy coat and, in three out of four donkeys...
McCartan CG, Russell MM, Wood JL, Mumford JA.An outbreak of equine herpesvirus-1 (EHV-1) occurred on a large stud farm with 133 mares, 54 foals and four stallions, and at least 85 mares, 22 foals and three stallions were infected. Clinical disease was observed in 16 mares, two stallions and 13 foals and the predominant clinical signs were scrotal oedema, ataxia and loss of libido in the stallions, ataxia and recumbency in the mares and uveitis and nasal discharge in the foals, although pneumonia and colic with intussusception were also recorded at autopsy. Neurological disease was more common in the mares nursing foals (12 of 38 infected...
Singh G.A/eq-1 and A/eq-2 influenza viruses were isolated simultaneously from an epidemic of equine influenza in north India. Evidently, both types of equine influenza viruses circulated in the equine population at the same time.
Ficorilli N, Studdert MJ, Crabb BS.The nucleotide sequence of the glycoprotein G (gG) homologue of asinine herpesvirus 3 (AHV3), a respiratory alphaherpesvirus of donkeys, was determined. The AHV3 gG gene consists of 1233 base pairs (bp) and codes for a predicted protein of 411 amino acids. This is identical in size to the equine herpesvirus 1 (EHV1) gG gene and 6 amino acids longer than the equine herpesvirus 4 (EHV4) gG gene. The predicted amino acid sequence of AHV3 gG has characteristics of a class 1 membrane protein. The amino acid sequence of AHV3 gG shows 92% and 60% identity to EHV1 gG and EHV4 gG respectively. Two regi...
Sekiguchi K, Sugita S, Fukunaga Y, Kondo T, Wada R, Kamada M, Yamaguchi S.A polymerase chain reaction (PCR) based assay capable of detecting and differentiating seven strains of equine arteritis virus (EAV) from around the world was developed. The primers for the PCR were chosen from the ORF6 gene encoding the unglycosylated membrane protein (M). Viral RNA from cell culture fluids infected with each of the seven EAV strains and RNA from the live vaccine, Arvac, was detected by PCR using four sets of primers. The sensitivity of detection was increased from 100 to 1,000 times by performing nested PCR enabling the detection of RNA at a level of 0.5-5 PFU. Differentiati...
Allen G, Yeargan M, Costa LR, Cross R.An experimental system that permits sensitive and reproducible detection of equine herpesvirus 1 (EHV-1)-specific cytotoxic T-lymphocyte (CTL) activity in the horse was developed. Peripheral blood mononuclear cells (PBMC) collected from immune horses were restimulated in vitro by culture with live EHV-1. Cytotoxic activity against virus-infected, pokeweed mitogen-stimulated lymphoblast targets was assessed in a 4-h 51Cr release assay. The optimal conditions for in vitro stimulation of equine memory CTLs and for preparation of EHV-1-infected target cells expressing viral antigens were systemati...
Reubel GH, Crabb BS, Studdert MJ.Nested polymerase chain reaction (PCR) assays were developed for the detection of equine herpesvirus 2 (EHV2) and equine herpesvirus 5 (EHV5) using the nucleotide sequences from the glycoprotein B (gB) gene of EHV2 and the thymidine kinase (TK) gene of EHV5. The simultaneous use of EHV2 specific and EHV5 specific primers in one nested amplification assay (multiplex PCR) enabled a rapid, specific and sensitive diagnosis for each virus. PCR was found to be 10(3) times more sensitive than virus isolation by cell culture for EHV2 and 10(6) for EHV5. In separate PCR assays, the routine detection li...
Dolby CA, Hannant D, Mumford JA.Five yearling ponies were vaccinated with inactivated Equid herpesvirus type 1 (EHV-1) in Freund's complete adjuvant as a double emulsion and revaccinated 6 weeks later with EHV-1 in Freund's incomplete adjuvant. These ponies and three age-matched controls were challenged intra-nasally after a further 6 weeks with homologous live virus and monitored clinically, biologically and serologically. After challenge, clinical signs were mild in both groups. No cell-associated viraemias were detected in vaccinated ponies. Vaccination induced high levels of complement-fixing (CF) and virus-neutralizing ...
Binz T, Lebelt J, Niemann H, Hagenau K.By reverse transcriptase/PCR amplification and subsequent sequence determination of the p24 gene, the relatedness of Borna disease virus (BDV) in various naturally infected animal species was determined. These results are indicative of a common ancestral virus pool and a remarkably low species barrier of BDV. Comparison of 11 sequences to that of tissue culture adapted virus revealed that the homology among all isolates was at least 96.2% at the nucleotide level, and 97% at the amino acid level. Viral sequences from sheep, donkey and horse were found to be not more distantly related to each ot...
Clark GG, Crabbs CL, Bailey CL, Calisher CH, Craig GB.An arbovirus survey was conducted during August 1985 at White Sands Missile Range in southcentral New Mexico following a suspected arboviral disease epizootic among feral horses. A total of 20,566 mosquitoes (18,505 females and 2,061 males) and 8,900 biting gnats were collected and assayed for virus. Female mosquitoes were principally Aedes campestris (54.8%), Aedes dorsalis (30.4%) and Culex tarsalis (13.2%). Arboviruses were not isolated from biting gnats, but mosquitoes yielded a total of 37 viral isolates, including western equine encephalitis (WEE) (18), California serogroup (15), Cache V...
Castillo-Olivares J, de Vries AAF, Raamsman MJB, Rottier PJM, Lakhani K, Westcott D, Tearle JP, Wood JLN, Mumford JA, Hannant D, Davis-Poynter NJ.An Escherichia coli-expressed recombinant protein (6hisG(L)ecto) comprising the entire ectodomain (aa 18-122) of equine arteritis virus (EAV) glycoprotein G(L), the immunodominant viral antigen, induced higher neutralizing antibody titres than other G(L)-derived polypeptides when compared in an immunization study in ponies. The potential of the recombinant G(L) ectodomain to act as a sub-unit vaccine against EAV was evaluated further in three groups of four ponies vaccinated with doses of 35, 70 or 140 microg of protein. All vaccinated animals developed a virus-neutralizing antibody (VNAb) res...
Cymerys J, Słońska A, Brzezicka J, Tucholska A, Chmielewska A, Rola J, Malik P, Bańbura MW.Equine herpesvirus type 1 (EHV-1) causes respiratory infections, abortion and neurological disorders in horses. Molecular epidemiology studies have demonstrated that a single-point mutation in DNA polymerase gene, resulting in an amino acid variation (N752/D752), is significantly associated with the neuropathogenic potential of EHV-1 strains. The aim of the study was to elucidate if there are any differences between neuropathogenic (EHV-1 26) and non-neuropathogenic (Jan-E and Rac-H) EHV-1 strains in their ability to infect neuronal cells. For the tested EHV-1 strains, cytopathic effect (CPE) ...
Silva Serra AC, Júnior EC, Cruz JF, Lobo PS, Júnior ET, Bandeira RS, Bezerra DA, Mascarenhas JD, Santos Guerra SF, Soares LS. To perform a molecular analysis of rotavirus A (RVA) G3P[6] strains detected in 2012 and 2017 in the Amazon region of Brazil. Eighteen RVA G3P[6] strains were collected from children aged under 10 years hospitalized with acute gastroenteritis, and partial sequencing of each segment genome was performed using Sanger sequencing. Phylogenetic analysis showed that all G3P[6] strains had a DS-1-like genotype constellation. Two strains had the highest nucleotide identities with equine-like G3P[6]/G3P[8] genotypes. Several amino acid alterations in VP4 and VP7 neutralizing epitopes of equine-l...
Wilks CR, Coggins L.Twenty-two ponies were examined for serum-neutralizing (SN) antibody to equine herpesvirus type 1 and for in vitro lymphocyte transformation in the presence of viral antigen. Six ponies had undetectable levels of neutralizing antibody (titer less than 1:2) and had lymphocytes which did not respond in culture with viral antigen (stimulation index less than 2.0). Four ponies which had SN antibody to equine herpesvirus type 1 did not manifest lymphocyte transformation in vitro. The 12 remaining seropositive ponies had lymphocyte transformation with viral antigen in vitro (stimulation indexes from...
Galosi CM, Vila Roza MV, Oliva GA, Pecoraro MR, Echeverría MG, Corva S, Etcheverrigaray ME.Equine herpesvirus 1 (EHV-1) is the causative agent of abortion, perinatal foal mortality, neurological and acute respiratory diseases in horses. Conventional laboratory diagnosis involving viral isolation from aborted foetuses is laborious and lengthy and requires processing of samples within 24 h of collection, which is problematic for samples that come from long distances. The aim of this study was to develop a polymerase chain reaction (PCR) assay useful in Argentina to detect DNA sequences of EHV-1 in different tissues from aborted equine foetuses with variable quality of preservation and...
Lai AC, Lin YP, Powell DG, Shortridge KF, Webster RG, Daly J, Chambers TM.An outbreak of influenza occurred among thoroughbred racehorses in Hong Kong in November-December 1992, with morbidity of 37%. All horses involved had been vaccinated against equine-1 and equine-2 influenza viruses but not against the virus responsible for the 1989 equine influenza outbreak in northern China (influenza A/equine/Jilin/89, subtype H3N8). Therefore the source and nature of the virus causing the Hong Kong outbreak was investigated. Virus isolated from a horse infected during the outbreak was used for genetic analysis. All the viral gene segments were similar to those of equine-2 (...
Rice NR, Coggins L.In the endogenous reverse transcriptase reaction, equine infectious anemia virus is able to synthesize complementary DNA (cDNA) of 8,000 nucleotides in high yield. After 2 h in 50 muM dNTP, about 2.8 mug of cDNA per mg of protein is produced, almost 30% of which is long cDNA. The system thus compares favorably with the other two well-characterized endogenous reaction systems, Moloney murine leukemia virus and avian sarcoma virus. Elongation rates of 100 to 150 nucleotides per min have been observed; these rates are comparable to those seen with purified avian myeloblastosis virus reverse trans...
Langsjoen RM, Key A, Shariatzadeh N, Jackson CR, Mahmood F, Arkun K, Alexandrescu S, Solomon IH, Piantadosi A.Eastern equine encephalitis virus (EEEV) is a relatively little-studied alphavirus that can cause devastating viral encephalitis, potentially leading to severe neurological sequelae or death. Although case numbers have historically been low, outbreaks have been increasing in frequency and scale since the 2000 s. It is critical to investigate EEEV evolutionary patterns, especially within human hosts, to understand patterns of emergence, host adaptation, and within-host evolution. To this end, we obtained formalin-fixed paraffin-embedded tissue blocks from discrete brain regions from five contem...
Lee DH, Lee EB, Seo JP, Ko EJ.Despite vaccination, equine influenza virus (EIV) and equine herpesvirus (EHV) infections still cause highly contagious respiratory diseases in horses. Recently, concurrent vaccination with EIV and EHV was suggested as a new approach; however, there have been no reports of concurrent vaccination with recombinant canarypox EIV and inactivated EHV vaccines. In this study, we aimed to compare the EIV-specific immune responses induced by concurrent administrations of a recombinant canarypox EIV vaccine and an inactivated bivalent EHV vaccine with those induced by a single recombinant canarypox EIV...
Studdert MJ, Fitzpatrick DR, Horner GW, Westbury HA, Gleeson LJ.Representative strains of EHV isolated from an aborted foetus and from a horse with rhinopneumonitis in New Zealand had restriction endonuclease DNA fingerprints typical of those usually associated with these syndromes elsewhere and now designated EHV1 and 4 respectively. EHV1 was isolated from the brain and spinal cord of a 4-year-old gelding that died of myeloencephalitis. A mare on the same farm, at about the same time as the gelding developed myeloencephalitis, aborted and EHV1 was isolated from the tissues of the aborted foetus. Restriction endonuclease DNA fingerprints of the viruses iso...
Yin X, Lu G, Guo W, Qi T, Ma J, Zhu C, Zhao S, Pan J, Xiang W.An outbreak of equine influenza was observed in the Asian wild horse population in Xinjiang Province, China, in 2007. Nasal swabs were collected from wild horses and inoculated into 9-10-day SPF embryonated eggs. The complete genome of the isolate was sequenced. A comparison of the amino acid sequence revealed that the isolate was an equine influenza virus strain, which we named A/equine/Xinjiang/4/2007. Each gene of the virus was found to have greater than 99 % homology to equine influenza virus strains of the Florida-2 sublineage, which were circulating simultaneously in China, and a lesser ...
Shen DT, Crawford TB, Gorham JR, McGuire TC.Twelve chemicals and commercial disinfectants were tested for inactivation of equine infectious anemia virus. In the presence of 10% bovine serum, all chemicals inactivated 4 log10 (based on 0.1 ml) of the virus within 5 minutes at 23 C. A reduction of at least 4 log10 was observed when the virus was exposed for 1 minute to substituted phenolic disinfectants (3 commercial preparations and sodium orthophenylphenate), halogen derivatives (iodophor and sodium hypochlorite), chlorhexidine, and 70% ethanol. Sodium hydroxide (5%), 2% formalin, and 2% glutaraldehyde were slower to inactivate the viru...
Hassanien RT, Hussein HA, Abdelmegeed HK, Abdelwahed DA, Khattab OM, Ali MH, Habashi AR, Ibraheem EM, Shahein MA, Abohatab EM.Due to climatic changes, arthropod-borne viruses have become a global health concern. In Egypt, West Nile virus (WNV) was initially detected in humans in 1950 and then in 1951, 1954, 1968, and 1989. Although WNV infection has been recorded in numerous Middle Eastern countries, its prevalence among the equine population in Egypt is unknown. This study aimed to investigate the current situation of vector-borne WNV in Egypt, estimate its seroprevalence, and assess the associated risk factors. Unassigned: We screened 1100 sera samples and nasal swabs from the same equids, 156 mosquito pools, and 3...
Cheevers WP, Fatemi-Nainie S, Anderson LW.A retrovirus is spontaneously released into the culture medium of the equine sarcoid-derived MC-1 cell line. The MC-1 virus did not exhibit in vitro transforming activity or replication when tested on equine fibroblasts or a variety of other mammalian cell cultures. Complementary DNA, synthesized using detergent-activated MC-1 virus RNA-dependent DNA polymerase, detected homologous sequences in the DNA of an established equine dermal cell line and in the DNA of primary equine dermal fibroblasts. Iododeoxyuridine or azacytidine induced a replication-deficient endogenous retrovirus in the normal...
Dale B, Brown R, Miller J, White RT, Air GM, Cordell B.Equine influenza is caused by two serotypes of type A influenza virus, EIV-A1 and EIV-A2. The complete nucleotide sequence of the neuraminidase (NA) genes of both the A1 (N7 subtype) and A2 (N8 subtype) serotype has been determined following cloning of full-length viral NA cDNAs into pBR322. Analysis of the deduced amino acid sequences reveals that the N7 and N8 genes share expected extensive homologies with the previously sequenced N1, N2, and N9 NA subtypes. These homologies include conservation of basic NA gene and protein structure, cysteine residues, potential glycosylation sites, and res...
Madrigal-Valencia TL, Saavedra-Montañez M, Pérez-Torres A, Hernández J, Segalés J, Hernández YD, Candanosa-Aranda IE, Pérez-Guiot A....Ovine gammaherpesvirus 2 (OvHV-2), a member of the genus Macavirus, causes sheep-associated malignant catarrhal fever (SA-MCF), a fatal lymphoproliferative disease affecting a wide variety of ungulates in addition to horses. This study described an outbreak of SA-MCF in Mexico and the identification of the OvHV-2 virus in primary rabbit testis cultures through the generation of intranuclear inclusion bodies, syncytia, immunofluorescence (IF), immunocytochemistry (ICC), immunohistochemistry (IHC), endpoint polymerase chain reaction (PCR), and partial sequencing of the ORF75 gene. The animals in...
Keane DP, Little PB, Wilkie BN, Artsob H, Thorsen J.A survey was conducted by testing 115 paired equine serum and cerebrospinal fluid samples by hemagglutination-inhibition for antibodies to Powassan and snowshoe hare viruses, and by virus neutralization for antibodies to equine herpesvirus type 1. Twenty-five samples were from horses with spontaneous neurological disease and the remainder from horses euthanized because of various nonneurological disorders. All sera and cerebrospinal fluids were negative for antibodies to Powassan virus. Fifty-one sera (44.3%) and 15 cerebrospinal fluids (13.0%) had antibodies to snowshoe hare virus. Ninety-eig...
Horsington JJ, Gilkerson JR, Hartley CA.Erbovirus is a genus of the family Picornaviridae and equine rhinitis B virus (ERBV) is the sole species. Erboviruses infect horses causing acute respiratory disease and sub-clinical and persistent infections. Despite the high seroprevalence and worldwide distribution of these viruses, the pathogenesis and antigenic structure of the three ERBV serotypes (ERBV1, 2 and 3) is poorly understood. To characterise linear epitopes on ERBV structural proteins, a set of fusion proteins were expressed in Escherichia coli. These proteins were tested in Western blot and ELISA and reactive proteins were als...
Hannant D, O'Neill T, Ostlund EN, Kydd JH, Hopkin PJ, Mumford JA.A paresis isolate of equid herpesvirus 1 (EHV1, Ab4/8) and a plaque-purified virus derived from it (EHV1, Ab4/13), induced long-term suppression of both mitogenic and antigen-specific lymphocyte proliferations in adult outbred ponies. Peripheral blood mononuclear cells (PBMC) taken from a pony after EHV1 infection suppressed the in vitro function of normal cells but serum did not. This showed that the observed immune suppression was associated with circulating PBMC and/or their products rather than circulating soluble factors such as antigen or immune complexes. The results suggested that prod...
Palfi V, Christensen LS.Twenty-five strains of equine herpesvirus 1 (EHV-1) and one strain of equine herpesvirus 4 (EHV-4) isolated from material from various clinical cases in Denmark, together with reference EHV-1 and EHV-4 strains, were compared by restriction fragment pattern (RFP) analysis and inoculation of baby mice. The RFP analyses revealed that all EHV-1 strains belonged to genome type Ip. Four fetal isolates exhibited genomic characteristics that have been suggested as specific markers of the attenuated strain Rac H, widely used as a live vaccine. As the use of five vaccines against EHV-1 and EHV-4 has nev...
Kitai Y, Shirafuji H, Kanehira K, Kamio T, Kondo T, Konishi E.West Nile virus (WNV) and Japanese encephalitis (JE) virus are distributed separately in the world with some exceptions. There is a concern that WNV may invade into Asia where JE virus exists. On and after such invasion, any differential diagnosis could be complicated by serological crossreactivities. We previously demonstrated experimentally using horses infected with WNV that preimmunization with inactivated JE vaccine considerably affected the ability of neutralization tests and immunoglobulin M (IgM) antibody-capture enzyme-linked immunosorbent assay (ELISA) to diagnose WNV infection. Here...
Rizvi SM, Slater JD, Wolfinger U, Borchers K, Field HJ, Slade AJ.The distribution of equine herpesvirus 2 (EHV-2) DNA within neurological and lymphoid tissues from 12 EHV-2 seropositive Welsh mountain ponies was determined by PCR. The lymphoid sites sampled in this study were almost universally PCR positive, thus confirming the existing virus co-cultivation data which suggest that the lymph nodes draining the respiratory tract are the main reservoirs of EHV-2 DNA. In addition, EHV-2 DNA was also detected, albeit with lower frequency, within both the peripheral and central nervous systems (PNS and CNS) of these animals. Of the CNS sites sampled 11% were PCR-...
Sharav T, Konnai S, Ochirkhuu N, Ts EO, Mekata H, Sakoda Y, Umemura T, Murata S, Chultemdorj T, Ohashi K.The genetic characterization and actual prevalence of EIAV in Mongolian horse in the disease endemic region is currently unknown. Here, 11 of 776 horse serum samples from four Mongolian provinces tested positive on agar gel immunodiffusion test. Genomic DNA extracted from all seropositive samples was subjected to nested PCR assay. Among these, three samples tested positive with nested PCR assay and were identified by sequencing analysis based on long termination repeat and tat gene of the virus. Two of the three sequences were identical, with 94.0% identity with the third. These two independen...
Oladunni FS, Sarkar S, Reedy S, Balasuriya UBR, Horohov DW, Chambers TM.Equine herpesvirus-1 (EHV-1) infection is an important and highly prevalent disease in equine populations worldwide. Previously we have demonstrated that a neuropathogenic strain of EHV-1, T953, suppresses the host cell's antiviral type-I interferon (IFN) response in vitro. Whether or not this is unique to EHV-1 strains possessing the neuropathogenic genotype has been undetermined. Here, we examined whether there is any direct relationship between neuropathogenic genotype and the induced IFN-β response in equine endothelial cells (EECs) infected with 10 different strains of EHV-1. The extent ...
Meister TL, Arroyo LG, Shanahan R, Papapetrou MA, Reinecke B, Brüggemann Y, Todt D, Stang A, Hazlett M, Baird JD, Steinmann E.Theiler's disease (TD) is a (sub-)acute hepatitis in adult horses and one of the most common causes of acute hepatic failure. Recent findings indicate that equine parvovirus hepatitis (EqPV-H) likely causes TD and that its transmission occurs via iatrogenic and/or natural routes. Following the death of an EqPV-H positive mare with TD, close-contact mares and foals in the same paddock were monitored to evaluate if there was any evidence of EqPV-H. For this purpose, the serum of close contact horses was examined 6 and 42 days after the mare's death for the presence of EqPV-H DNA and changes in l...
Horsington JJ, Gilkerson JR, Hartley CA.Equine rhinitis B virus (ERBV) is the single species in the genus Erbovirus, family Picornaviridae. Equine rhinitis B viruses exist in three serotypes and are associated with respiratory disease in horses. Members of the species vary in stability at acid pH. To date there has been discordance in genotype, serotype and acid stability phenotype groupings. To identify capsid regions associated with acid stability, two viruses were serially treated at pH 3.3 to isolate acid-stable mutants. An acid-stable mutant of the prototype acid-labile serotype 1 virus contained a single amino acid change in t...
Steinbach F, Mauel S, Beier I.Interferons (IFNs) are important mediators of the immune system. Their antiviral activity is an integral part of the innate immune defence, but all IFNs have immune regulatory functions also. Besides rec.eq.IFN-beta detailed descriptions on other rec.IFNs were lacking and none of the proteins was available. To compare the equine IFNs and allow detailed studies on proteins and bioactivity, we performed the expression cloning of rec.eq.IFN-alpha, -beta and -gamma. To achieve maximal expression, a bacterial expression system was chosen. Additionally, rec.eq.IFN-beta and -gamma were expressed in m...
van der Meulen KM, Nauwynck HJ, Pensaert MB.In the present study, equine herpesvirus-1 (EHV-1)-infected cells were identified in ionomycin/phorbol dibutyrate (IONO/PDB)-stimulated peripheral blood mononuclear cells (PBMC) and the mechanism by which stimulation increases the percentage of infected cells was examined. In the population of viral antigen-positive PBMC, 38.4+/-4.5% were CD5(+) T-lymphocytes (18.1+/-3.2% CD4(+) 13.6+/-1.8% CD8(+)), 18.1+/-5.4% were B-lymphocytes, 8.5+/-3.9% were monocytes and 35% remained unidentified. The role of the cell cycle in the increased susceptibility to EHV-1 upon stimulation was examined by stimula...
du Plessis DH, van Wyngaardt W, Bremer CW.African horsesickness virus (AHSV), an important disease of equines is caused by an orbivirus. Because of the need to contain the spread of the disease, it is often essential to make a rapid diagnosis. For this purpose, an ELISA capable of detecting viral antigen in animal tissue and in cell culture fluid was developed. Immobilised F(ab')2 fragments prepared by digestion of AHSV-specific IgG with pepsin were used to trap virus from tissue homogenates or cell culture supernatant. After addition of intact IgG as detecting antibody, Staphylococcus aureus protein A labelled with horseradish peroxi...
