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Topic:Virology

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.
A prospective longitudinal study of naturally infected horses to evaluate the performance characteristics of rapid diagnostic tests for equine influenza virus.
Veterinary microbiology    November 6, 2011   Volume 156, Issue 3-4 246-255 doi: 10.1016/j.vetmic.2011.10.031
Read AJ, Arzey KE, Finlaison DS, Gu X, Davis RJ, Ritchie L, Kirkland PD.An outbreak of equine influenza (EI) occurred in Australia in 2007. During the laboratory support for this outbreak, real-time reverse transcriptase polymerase chain reaction (qRT-PCR) assays and a blocking enzyme linked immunosorbent assay (bELISA) were used as testing methods to detect infection with the virus. The qRT-PCR and bELISA tests had not been used for EI diagnosis before, so it was not known how soon after infection these tests would yield positive results, or for how long these results would remain positive. To answer these questions, nasal swabs and blood samples were collected d...
Functional characterization of EUL47 in productive replication, morphogenesis and infectivity of equine herpesvirus 1.
Virus research    October 26, 2011   Volume 163, Issue 1 310-319 doi: 10.1016/j.virusres.2011.10.021
Yu MH, Kasem S, Yoshizaki N, Pagamjav O, Yamaguchi T, Ohya K, Fukushi H.EUL47 is a major component of the tegument of equine herpesvirus 1 (EHV-1). To determine its function, we used Red/ET cloning to delete its gene (gene 13) from EHV-1 strain Ab4p inserted into a bacterial artificial chromosome (BAC), yielding Ab4pattBΔ13. We also examined the reverted virus (Ab4pattB13R). Ab4pattBΔ13 replicated in rabbit kidney (RK)-13 cells, indicating that ORF13 is dispensable for virus replication in cell culture. Its intracellular and extracellular titers were about 10- and 100-fold lower than those of the revertant and parent strains, respectively. In addition, the plaqu...
Isolation and phylogenetic grouping of equine encephalosis virus in Israel.
Emerging infectious diseases    October 18, 2011   Volume 17, Issue 10 1883-1886 doi: 10.3201/eid1710.110350
Aharonson-Raz K, Steinman A, Bumbarov V, Maan S, Maan NS, Nomikou K, Batten C, Potgieter C, Gottlieb Y, Mertens P, Klement E.During 2008-2009 in Israel, equine encephalosis virus (EEV) caused febrile outbreaks in horses. Phylogenetic analysis of segment 10 of the virus strains showed that they form a new cluster; analysis of segment 2 showed ≈92% sequence identity to EEV-3, the reference isolate. Thus, the source of this emerging EEV remains uncertain.
Pathology in practice. EHV-1 infection.
Journal of the American Veterinary Medical Association    October 18, 2011   Volume 239, Issue 9 1193-1195 doi: 10.2460/javma.239.9.1193
Cushing TL, Schlafer DH.No abstract available
Gene expression analysis in the thalamus and cerebrum of horses experimentally infected with West Nile virus.
PloS one    October 4, 2011   Volume 6, Issue 10 e24371 doi: 10.1371/journal.pone.0024371
Bourgeois MA, Denslow ND, Seino KS, Barber DS, Long MT.Gene expression associated with West Nile virus (WNV) infection was profiled in the central nervous system of horses. Pyrosequencing and library annotation was performed on pooled RNA from the CNS and lymphoid tissues on horses experimentally infected with WNV (vaccinated and naïve) and non-exposed controls. These sequences were used to create a custom microarray enriched for neurological and immunological sequences to quantitate gene expression in the thalamus and cerebrum of three experimentally infected groups of horses (naïve/WNV exposed, vaccinated/WNV exposed, and normal).From the sequ...
Outbreak of equine herpesvirus myeloencephalopathy in France: a clinical and molecular investigation.
Transboundary and emerging diseases    October 4, 2011   Volume 59, Issue 3 256-263 doi: 10.1111/j.1865-1682.2011.01263.x
Pronost S, Legrand L, Pitel PH, Wegge B, Lissens J, Freymuth F, Richard E, Fortier G.Equid herpesvirus 1 (EHV-1)-associated myeloencephalopathy (EHM) is a disease affecting the central nervous system of horses. Despite the constantly increasing interest about this syndrome, epidemiological data are limited especially when related to the description of large outbreaks. The aim of this article is to describe clinical, virological and molecular data obtained throughout a severe outbreak of EHM, with emphasis on laboratory diagnostic methods. The epidemic disease concerned a riding school in France where 7/66 horses aged 12-22 years developed signs of neurological disease in July ...
In vitro characterization of EHV-4 gG-deleted mutant.
Virus genes    September 29, 2011   Volume 44, Issue 1 109-111 doi: 10.1007/s11262-011-0677-6
Azab W, El-Sheikh A, Abdel-Gawad A.Equine herpesvirus 4 (EHV-4) is an important pathogen that causes respiratory tract disease in horse populations worldwide. Glycoprotein G (gG) homologs have been identified in several alphaherpesviruses as minor non-essential membrane-anchored glycoproteins. In this study, EHV-4 gG deletion mutant has been generated by using bacterial artificial chromosome technology to investigate the role of gG in viral pathogenesis. Our findings reported here revealed no significant difference between parental EHV-4 and gG-negative strain in their replication cycle in cell culture. Furthermore, virus titer...
Single amino acid residue in the A2 domain of major histocompatibility complex class I is involved in the efficiency of equine herpesvirus-1 entry.
The Journal of biological chemistry    September 26, 2011   Volume 286, Issue 45 39370-39378 doi: 10.1074/jbc.M111.251751
Sasaki M, Kim E, Igarashi M, Ito K, Hasebe R, Fukushi H, Sawa H, Kimura T.Equine herpesvirus-1 (EHV-1), an α-herpesvirus of the family Herpesviridae, causes respiratory disease, abortion, and encephalomyelitis in horses. EHV-1 utilizes equine MHC class I molecules as entry receptors. However, hamster MHC class I molecules on EHV-1-susceptible CHO-K1 cells play no role in EHV-1 entry. To identify the MHC class I molecule region that is responsible for EHV-1 entry, domain exchange and site-directed mutagenesis experiments were performed, in which parts of the extracellular region of hamster MHC class I (clone C5) were replaced with corresponding sequences from equine...
Development and evaluation of a reverse transcription loop-mediated isothermal amplification assay for H3N8 equine influenza virus.
Journal of virological methods    August 30, 2011   Volume 178, Issue 1-2 239-242 doi: 10.1016/j.jviromet.2011.07.015
Nemoto M, Yamanaka T, Bannai H, Tsujimura K, Kondo T, Matsumura T.Reverse transcription loop-mediated isothermal amplification (RT-LAMP) was applied to the detection of equine influenza virus (EIV). Because equine influenza is caused currently by EIV of the H3H8 subtype, the RT-LAMP primer set was designed to target the hemagglutinin gene of this subtype. The detection limit of the RT-LAMP assay was a virus dilution of 10(-5); which was 10(3) times more sensitive than the Espline Influenza A&B-N test and 10 times more sensitive than a reverse transcription polymerase chain reaction (RT-PCR) assay. The specificity of the RT-LAMP assay was examined by usin...
Mapping B-cell epitopes in equine rhinitis B viruses and identification of a neutralising site in the VP1 C-terminus.
Veterinary microbiology    August 30, 2011   Volume 155, Issue 2-4 128-136 doi: 10.1016/j.vetmic.2011.08.022
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...
Genetic characterization of equine adenovirus type 1.
Veterinary microbiology    August 22, 2011   Volume 155, Issue 1 33-37 doi: 10.1016/j.vetmic.2011.08.014
Cavanagh HM, Mahony TJ, Vanniasinkam T.Two known serotypes of equine adenovirus (EAdV), equine adenovirus type 1 (EAdV-1) and equine adenovirus type 2 (EAdV-2) have been isolated from horses. EAdV-1 is predominantly associated with upper respiratory tract infections while EAdV-2 appears to be associated with gastrointestinal infections in horses. In this report the EAdV-1 genome has been sequenced for the first time. The EAdV-1 genome encoded genes are characteristic of the Mastadenovirus genus such as protein V and IX. Unexpectedly, phylogenetic reconstructions also revealed a close relationship between EAdV-1 and two recently cha...
Factors affecting recombinant Western equine encephalitis virus glycoprotein production in the baculovirus system.
Protein expression and purification    August 16, 2011   Volume 80, Issue 2 274-282 doi: 10.1016/j.pep.2011.08.002
Toth AM, Geisler C, Aumiller JJ, Jarvis DL.In an effort to produce processed, soluble Western equine encephalitis virus (WEEV) glycoproteins for subunit therapeutic vaccine studies, we isolated twelve recombinant baculoviruses designed to express four different WEEV glycoprotein constructs under the transcriptional control of three temporally distinct baculovirus promoters. The WEEV glycoprotein constructs encoded full-length E1, the E1 ectodomain, an E26KE1 polyprotein precursor, and an artificial, secretable E2E1 chimera. The three different promoters induced gene expression during the immediate early (ie1), late (p6.9), and very lat...
Evaluation of two magnetic-bead-based viral nucleic acid purification kits and three real-time reverse transcription-PCR reagent systems in two TaqMan assays for equine arteritis virus detection in semen.
Journal of clinical microbiology    August 10, 2011   Volume 49, Issue 10 3694-3696 doi: 10.1128/JCM.01187-11
Miszczak F, Shuck KM, Lu Z, Go YY, Zhang J, Sells S, Vabret A, Pronost S, Fortier G, Timoney PJ, Balasuriya UB.This study showed that under specifically defined conditions with respect to nucleic acid extraction method and testing reagents, a previously described real-time reverse transcription-PCR (rRT-PCR) assay (T1 assay) provides sensitivity equal to or higher than that of virus isolation for the detection of equine arteritis virus in semen.
Prevalence of equine herpesvirus type 1 strains of neuropathogenic genotype in a major breeding area of Japan.
The Journal of veterinary medical science    August 9, 2011   Volume 73, Issue 12 1663-1667 doi: 10.1292/jvms.11-0140
Tsujimura K, Oyama T, Katayama Y, Muranaka M, Bannai H, Nemoto M, Yamanaka T, Kondo T, Kato M, Matsumura T.A single non-synonymous nucleotide substitution of guanine (G) for adenine (A) at position 2254 in the viral DNA polymerase gene (encoded by open reading frame [ORF] 30) of equine herpesvirus type 1 (EHV-1) has been significantly associated with neuropathogenic potential in strains of this virus. To estimate the prevalence of EHV-1 strains with the neuropathogenic genotype (ORF30 G(2254)) in the Hidaka district--a major horse breeding area in Japan--we analyzed the ORF30 genomic region in cases of EHV-1 infection in this area during the years 2001-2010. Of the 113 cases analyzed, 3 (2.7%) were...
Identification of functional domains of the IR2 protein of equine herpesvirus 1 required for inhibition of viral gene expression and replication.
Virology    July 26, 2011   Volume 417, Issue 2 430-442 doi: 10.1016/j.virol.2011.06.023
Kim SK, Kim S, Dai G, Zhang Y, Ahn BC, O'Callaghan DJ.The equine herpesvirus 1 (EHV-1) negative regulatory IR2 protein (IR2P), an early 1,165-amino acid (aa) truncated form of the 1487-aa immediate-early protein (IEP), lacks the trans-activation domain essential for IEP activation functions but retains domains for binding DNA, TFIIB, and TBP and the nuclear localization signal. IR2P mutants of the N-terminal region which lack either DNA-binding activity or TFIIB-binding activity were unable to down-regulate EHV-1 promoters. In EHV-1-infected cells expressing full-length IR2P, transcription and protein expression of viral regulatory IE, early EICP...
Phylogenetic and molecular characterization of equine H3N8 influenza viruses from Greece (2003 and 2007): evidence for reassortment between evolutionary lineages.
Virology journal    July 14, 2011   Volume 8 350 doi: 10.1186/1743-422X-8-350
Bountouri M, Fragkiadaki E, Ntafis V, Kanellos T, Xylouri E.For first time in Greece equine influenza virus infection was confirmed, by isolation and molecular analysis, as the cause of clinical respiratory disease among unvaccinated horses during 2003 and 2007 outbreaks. Methods: Equine influenza virus (EIV) H3N8 was isolated in MDCK cells from 30 nasal swabs from horses with acute respiratory disease, which were tested positive by Directigen Flu A. Isolation was confirmed by haemagglutination assay and RT-PCR assay of the M, HA and NA gene. Results: HA sequences of the Greek isolates appeared to be more closely related to viruses isolated in early 19...
The pathogenic and vaccine strains of equine infectious anemia virus differentially induce cytokine and chemokine expression and apoptosis in macrophages.
Virus research    July 14, 2011   Volume 160, Issue 1-2 274-282 doi: 10.1016/j.virusres.2011.06.028
Lin YZ, Cao XZ, Li L, Li L, Jiang CG, Wang XF, Ma J, Zhou JH.The attenuated equine infectious anemia virus (EIAV) vaccine was the first attenuated lentivirus vaccine to be used in a large-scale application and has been used to successfully control the spread of equine infectious anemia (EIA) in China. To better understand the potential role of cytokines in the pathogenesis of EIAV infection and resulting immune response, we used branched DNA technology to compare the mRNA expression levels of 12 cytokines and chemokines, including IL-1α, IL-1β, IL-4, IL-10, TNF-α, IFN-γ, IP-10, IL-8, MIP-1α, MIP-1β, MCP-1, and MCP-2, in equine monocyte-derived mac...
Decreased infectivity of a neutralization-resistant equine infectious anemia virus variant can be overcome by efficient cell-to-cell spread.
Journal of virology    July 13, 2011   Volume 85, Issue 19 10421-10424 doi: 10.1128/JVI.05349-11
Wu W, Blythe DC, Loyd H, Mealey RH, Tallmadge RL, Dorman KS, Carpenter S.Two variants of equine infectious anemia virus (EIAV) that differed in sensitivity to broadly neutralizing antibody were tested in direct competition assays. No differences were observed in the growth curves and relative fitness scores of EIAVs of principal neutralizing domain variants of groups 1 (EIAV(PND-1)) and 5 (EIAV(PND-5)), respectively; however, the neutralization-resistant EIAV(PND-5) variant was less infectious in single-round replication assays. Infectious center assays indicated similar rates of cell-to-cell spread, which was approximately 1,000-fold more efficient than cell-free ...
Neutralising antibodies for West Nile virus in horses from Brazilian Pantanal.
Memorias do Instituto Oswaldo Cruz    July 9, 2011   Volume 106, Issue 4 467-474 doi: 10.1590/s0074-02762011000400014
Pauvolid-Corrêa A, Morales MA, Levis S, Figueiredo LT, Couto-Lima D, Campos Z, Nogueira MF, da Silva EE, Nogueira RM, Schatzmayr HG.Despite evidence of West Nile virus (WNV) activity in Colombia, Venezuela and Argentina, this virus has not been reported in most South American countries. In February 2009, we commenced an investigation for WNV in mosquitoes, horses and caimans from the Pantanal, Central-West Brazil. The sera of 168 horses and 30 caimans were initially tested using a flaviviruses-specific epitope-blocking enzyme-linked immunosorbent assay (blocking ELISA) for the detection of flavivirus-reactive antibodies. The seropositive samples were further tested using a plaque-reduction neutralisation test (PRNT90) for ...
Real-time RT-PCR for detection of equine influenza and its evaluation using samples from horses infected with A/equine/Sydney/2007 (H3N8).
Australian veterinary journal    July 8, 2011   Volume 89 Suppl 1 37-38 doi: 10.1111/j.1751-0813.2011.00739.x
Foord AJ, Selleck P, Colling A, Klippel J, Middleton D, Heine HG.No abstract available
Application of high-throughput systems for the rapid detection of DNA and RNA viruses during the Australian equine influenza outbreak.
Australian veterinary journal    July 8, 2011   Volume 89 Suppl 1 38-39 doi: 10.1111/j.1751-0813.2011.00744.x
Kirkland PD, Davis RJ, Gu X, Frost M.No abstract available
Application of real-time PCR and ELISA assays for equine influenza virus to determine the duration of viral RNA shedding and onset of antibody response in naturally infected horses.
Australian veterinary journal    July 8, 2011   Volume 89 Suppl 1 42-43 doi: 10.1111/j.1751-0813.2011.00740.x
Read AJ, Finlaison DS, Gu X, Davis RJ, Arzey KE, Kirkland PD.During the equine influenza (EI) outbreak, two assays were used in parallel to diagnose the disease, to demonstrate freedom from infection in disease control zones and ultimately to demonstrate that EI virus had been eliminated from the Australian horse population. A longitudinal study of a population of naturally infected horses was established to determine the performance characteristics of these assays.
Use of a blocking ELISA for antibodies to equine influenza virus as a test to distinguish between naturally infected and vaccinated horses: proof of concept studies.
Australian veterinary journal    July 8, 2011   Volume 89 Suppl 1 45-46 doi: 10.1111/j.1751-0813.2011.00743.x
Kirkland PD, Delbridge G.An important consideration in the selection of a vaccine during the Australian equine influenza (EI) outbreak in 2007 was the ability to differentiate between infected and vaccinated animals (DIVA). A blocking enzyme-linked immunosorbent assay (bELISA) targeted for the nucleoprotein of influenza A viruses was developed to differentiate between naturally infected horses and horses vaccinated with the ProteqFlu® vaccine, which only induces a response to viral haemagglutinin. This bELISA assay met the DIVA requirements and was used extensively during the EI control and eradication programs and '...
Isolation and characterisation of an H3N8 equine influenza virus in Australia, 2007.
Australian veterinary journal    July 8, 2011   Volume 89 Suppl 1 35-37 doi: 10.1111/j.1751-0813.2011.00738.x
Watson J, Halpin K, Selleck P, Axell A, Bruce K, Hansson E, Hammond J, Daniels P, Jeggo M.Before 2007, equine influenza had never been diagnosed in Australia. On 22 August 2007, infection was confirmed in horses at Eastern Creek Animal Quarantine Station near Sydney. The virus subsequently isolated (A/equine/Sydney/2888-8/2007) was confirmed by sequence analysis of the haemagglutinin (HA) gene as an H3 virus of the variant American Florida lineage that is now referred to as Clade 1. The HA sequence of the virus was identical to that of a virus isolated from a contemporaneous outbreak in Japan and showed high homology to viruses circulating in North America.
Clinical outcomes and virology of equine influenza in a naïve population and in horses infected soon after receiving one dose of vaccine.
Australian veterinary journal    July 8, 2011   Volume 89 Suppl 1 139-142 doi: 10.1111/j.1751-0813.2011.00768.x
Kannegieter NJ, Frogley A, Crispe E, Kirkland PD.As part of the control measures of the equine influenza (EI) outbreak, in addition to the strategic use of vaccination to provide buffer zones around infected populations, approval was obtained to vaccinate Thoroughbred racing horses. We review the clinical expression of the disease and virus excretion in a population of racehorses that were exposed to EI approximately 7 days after administration of a single dose of the canarypox-vectored recombinant compared with a similar unvaccinated population of horses at a nearby racetrack. Although this study was undertaken opportunistically and under t...
Role of the diagnostic laboratories during the 2007 equine influenza outbreak in Australia.
Australian veterinary journal    July 8, 2011   Volume 89 Suppl 1 29-32 doi: 10.1111/j.1751-0813.2011.00736.x
Kirkland PD.During the 2007 equine influenza (EI) outbreak in Australia, diagnostic laboratories and the use of appropriate tests played a pivotal role in the response to the crisis. This role began with the detection of EI virus in New South Wales (NSW) on the evening of 24 August 2007 and culminated in providing the final 'proof of freedom' from EI in March 2008. The tests that were used during the EI response were able to provide results quickly, and with high sensitivity and specificity. This section of the supplement describes the roles and functions of the Australian laboratories; tests used and the...
Validation of an influenza virus A 5’Taq nuclease assay for the detection of equine influenza virus A RNA in nasal swab samples.
Australian veterinary journal    July 8, 2011   Volume 89 Suppl 1 39-42 doi: 10.1111/j.1751-0813.2011.00747.x
Oakey J, Hawkesford T, Smith C, Hewitson G, Tolosa X, Wright L, Moody N, Rodwell B, Corney B, Waltisbuhl D.Describe the in-house validation of a previously reported influenza virus type A 5'Taq nuclease assay for detecting equine influenza virus A RNA in nasal swab material. Methods: The validation compares the 5'Taq nuclease assay with a gel-based reverse transcription nested polymerase chain reaction (PCR) previously reported by the Irish Equine Centre for detection of H3N8 and H7N7 equine influenza viruses. This test was chosen because it targets a different region of the viral genome to the real-time test, so it is not merely a repeat of the same test in a different format. Moreover, nested PCR...
Responding to the equine influenza outbreak: challenges from a laboratory perspective.
Australian veterinary journal    July 8, 2011   Volume 89 Suppl 1 32-35 doi: 10.1111/j.1751-0813.2011.00737.x
Brown L, Townsend W, Waltisbuhl D.The unique challenges that laboratories in Queensland and New South Wales faced during the response to the 2007 equine influenza outbreak and how these were managed are described.
A recombinant Hendra virus G glycoprotein-based subunit vaccine protects ferrets from lethal Hendra virus challenge.
Vaccine    July 1, 2011   Volume 29, Issue 34 5623-5630 doi: 10.1016/j.vaccine.2011.06.015
Pallister J, Middleton D, Wang LF, Klein R, Haining J, Robinson R, Yamada M, White J, Payne J, Feng YR, Chan YP, Broder CC.The henipaviruses, Hendra virus (HeV) and Nipah virus (NiV), are two deadly zoonotic viruses for which no vaccines or therapeutics have yet been approved for human or livestock use. In 14 outbreaks since 1994 HeV has been responsible for multiple fatalities in horses and humans, with all known human infections resulting from close contact with infected horses. A vaccine that prevents virus shedding in infected horses could interrupt the chain of transmission to humans and therefore prevent HeV disease in both. Here we characterise HeV infection in a ferret model and show that it closely mirror...
Equine torovirus (BEV) induces caspase-mediated apoptosis in infected cells.
PloS one    June 15, 2011   Volume 6, Issue 6 e20972 doi: 10.1371/journal.pone.0020972
Maestre AM, Garzón A, Rodríguez D.Toroviruses are gastroenteritis causing agents that infect different animal species and humans. To date, very little is known about how toroviruses cause disease. Here, we describe for the first time that the prototype member of this genus, the equine torovirus Berne virus (BEV), induces apoptosis in infected cells at late times postinfection. Observation of BEV infected cells by electron microscopy revealed that by 24 hours postinfection some cells exhibited morphological characteristics of apoptotic cells. Based on this finding, we analyzed several apoptotic markers, and observed protein syn...
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