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epizotiology and phylogeny of equine arteritis virus in hucul horses.the aim of the study was to determine the situation of equine arteritis virus (eav) infections in hucul horses. a total of 176 horses (154 mares and 22 stallions) from the biggest hucul horse stud in poland were tested. antibodies against eav were detected in 97 (55.1%) horses. the eav seroprevalence among mares was 53.2% while in stallions - 68.2%. the percentage of positive mares increased with their age, thus amongst the mares of less than 2 years of age the percentage was 32.5%, while in the ...201020956062
resonance assignment of nsp7α from arterivirus.the (1)h, (15)n and (13)c resonance assignment of nsp7α, a non-structural protein of unknown function from the equine arteritis virus, is reported.201120703834
the prrsv replicase: exploring the multifunctionality of an intriguing set of nonstructural proteins.our knowledge about the structure and function of the nonstructural proteins (nsps) encoded by the arterivirus replicase gene has advanced in recent years. the continued characterization of the nsps of the arterivirus prototype equine arteritis virus has not only corroborated several important functional predictions, but also revealed various novel features of arteriviral replication. for porcine reproductive and respiratory syndrome virus (prrsv), based on bioinformatics predictions and experim ...201020696193
extended phylogeny of the equine arteritis virus sequences including south american sequences.to perform genetic analysis of the orf5 of equine arteritis virus (eav) may provide new insights into the genetic evolution and origin of the argentinean eav sequences.201120689314
equine viral arteritis: further characterization of the carrier state in stallions.further characterization of the carrier state in stallions infected with equine arteritis virus revealed that there is considerable variation in the frequency of its occurrence among breeds. the frequency ranged from 12.5% (holsteiner stallions) to 72.7% (dutch warmblood stallions), with a mean occurrence of 40.8% in the seropositive stallions (n=561) examined. more than 70% of the virus shedders were standardbred stallions. the carrier state was not confirmed in any of the stallions that had be ...200020681110
autophagy protein atg5 interacts transiently with the hepatitis c virus rna polymerase (ns5b) early during infection.autophagy is an important cellular process by which atg5 initiates the formation of double membrane vesicles (dmvs). upon infection, dmvs have been shown to harbor the replicase complex of positive-strand rna viruses such as mhv, poliovirus, and equine arteritis virus. recently, it has been shown that autophagy proteins are proviral factors that favor initiation of hepatitis c virus (hcv) infection. here, we identified atg5 as an interacting protein for the hcv ns5b. atg5/ns5b interaction was co ...201020580051
evaluation of neutralization patterns of the five unique argentine equine arteritis virus field strains reported.equine viral arteritis (eva) is a contagious viral disease that frequently causes mild or subclinical infections in adult horses. only one eav serotype has been described. however, there are differences in antigenicity, pathogenicity and neutralization characteristics of virus field strains. the interaction of two viral proteins, gp5 and m, is critical for infectivity and amino acid changes in the gp5 sequences have an effect on the neutralizing phenotype, regardless the effects of other viral p ...201020461287
molecular epidemiology and genetic characterization of equine arteritis virus isolates associated with the 2006-2007 multi-state disease occurrence in the usa.in 2006-2007, equine viral arteritis (eva) was confirmed for the first time in quarter horses in multiple states in the usa. the entire genome of an equine arteritis virus (eav) isolate from the index premises in new mexico was 12 731 nt in length and possessed a previously unrecorded unique 15 nt insertion in the nsp2-coding region in orf1a and a 12 nt insertion in orf3. sequence analysis of additional isolates made during this disease occurrence revealed that all isolates from new mexico, utah ...201020444993
complex interactions between the major and minor envelope proteins of equine arteritis virus determine its tropism for equine cd3+ t lymphocytes and cd14+ monocytes.extensive cell culture passage of the virulent bucyrus (vb) strain of equine arteritis virus (eav) to produce the modified live virus (mlv) vaccine strain has altered its tropism for equine cd3(+) t lymphocytes and cd14(+) monocytes. the vb strain primarily infects cd14(+) monocytes and a small subpopulation of cd3(+) t lymphocytes (predominantly cd4(+) t lymphocytes), as determined by dual-color flow cytometry. in contrast, the mlv vaccine strain has a significantly reduced ability to infect cd ...201020219931
curing of hela cells persistently infected with equine arteritis virus by a peptide-conjugated morpholino oligomer.a significant consequence of equine arteritis virus (eav) infection of horses is persistence of the virus in a variable percentage of infected stallions. we recently established an in vitro model of eav persistence in cell culture for the purpose of furthering our understanding of eav biology in general and viral persistence in the stallion in particular. in this study we investigated whether persistently infected hela cells could be cured of eav infection by treatment with an antisense peptide- ...201020206215
arterivirus nsp1 modulates the accumulation of minus-strand templates to control the relative abundance of viral mrnas.the gene expression of plus-strand rna viruses with a polycistronic genome depends on translation and replication of the genomic mrna, as well as synthesis of subgenomic (sg) mrnas. arteriviruses and coronaviruses, distantly related members of the nidovirus order, employ a unique mechanism of discontinuous minus-strand rna synthesis to generate subgenome-length templates for the synthesis of a nested set of sg mrnas. non-structural protein 1 (nsp1) of the arterivirus equine arteritis virus (eav) ...201020174607
antiviral effect of recombinant equine interferon-gamma on several equine viruses.recombinant equine interferon-gamma (reifn-gamma) was prepared using a baculovirus expression system and its antiviral activity was investigated using several equine viruses. the reifn-gamma suppressed the replication of all equine viruses used in the present experiment in horse cell cultures, but did not affect the growth of host cells at concentrations of less than 1000 u/ml. a strong antiviral effect was observed, especially against rna viruses. equine picornavirus, equine rhinovirus and equi ...201020004981
a virus-induced epizootic hemorrhagic disease of the virginia white-tailed deer (odocoileus virginianus).a circumscribed natural outbreak of a highly fatal disease of deer, which we have designated epizootic hemorrhagic disease (ehd), has been studied. the disease has proven readily transmissible in deer but not in other experimental or domestic animals tested, nor in embryonating eggs or deer kidney cell cultures. the causative agent is a virus which is readily filterable and is capable of storage, either frozen or in glycerol, for relatively long periods of time. it produces a solid immunity in t ...196019867168
myristoylation of the arterivirus e protein: the fatty acid modification is not essential for membrane association but contributes significantly to virus infectivity.the envelope of equine arteritis virus (eav) contains two glycoprotein complexes (gp2b/gp3/gp4 and gp5/m) and the small, non-glycosylated e protein. as e is essential for the production of infectious progeny but dispensable for assembly and release of virus-like particles, it probably mediates virus entry into cells, putatively in concert with the gp2b/gp3/gp4 complex. the e protein contains a central hydrophobic domain and a conserved potential site for n-terminal myristoylation, a hydrophobic ...200919656967
structure and cleavage specificity of the chymotrypsin-like serine protease (3clsp/nsp4) of porcine reproductive and respiratory syndrome virus (prrsv).biogenesis and replication of the porcine reproductive and respiratory syndrome virus (prrsv) include the crucial step of replicative polyprotein processing by self-encoded proteases. whole genome bioinformatics analysis suggests that nonstructural protein 4 (nsp4) is a 3c-like serine protease (3clsp), responsible for most of the nonstructural protein processing. the gene encoding this protease was cloned and expressed in escherichia coli in order to confirm this prediction. the purified protein ...200919646449
the lactate dehydrogenase-elevating virus capsid protein is a nuclear-cytoplasmic protein.arteriviruses replicate in the cytoplasm and do not require the nucleus function for virus multiplication in vitro. however, nucleocapsid (n) protein of two arteriviruses, porcine reproductive respiratory syndrome virus and equine arteritis virus, has been observed to localize in the nucleus and nucleolus of virus-infected and n-gene-transfected cells in addition to their normal cytoplasmic distribution. in the present study, the n protein of lactate dehydrogenase-elevating virus (ldv) of mice w ...200919517211
testing for antibodies to equine arteritis virus. 200919346547
novel approach for detection of enteric viruses to enable syndrome surveillance of acute viral gastroenteritis.acute gastroenteritis is one of the most common diseases worldwide, with viruses, particularly noroviruses, being the leading cause in developed countries. in the netherlands, systematic surveillance of gastroenteritis outbreaks of suspected viral etiology was established by the national institute for public health and the environment in 1994. since 2002, the total number of outbreaks reported has been increasing, and with that comes the need for sensitive assays that can be performed quickly. i ...200919339472
[use of multiplex pcr method with real-time detection for differential diagnosis of respiratory viral infections].multiplex real-time polymerase chain reaction test-system with fluorescent detection (rt-pcr) for simultaneous identification of main agents of acute respiratory viral infections: influenza a (iav) and b viruses (ibv), parainfluenza viruses types 1, 2, 3, 4 (piv 1 - 4), adenoviruses (adv), respiratory syncitial virus (rsv), rhinoviruses (rv) and enteroviruses (ev), in presence internal positive control (ipc) represented by vaccine strain of rubella virus ra 27/3. using multiplex rt-pcr method, r ...200919338239
biochemical characterization of arterivirus nonstructural protein 11 reveals the nidovirus-wide conservation of a replicative endoribonuclease.nidoviruses (arteriviruses, coronaviruses, and roniviruses) are a phylogenetically compact but diverse group of positive-strand rna viruses that includes important human and animal pathogens. nidovirus rna synthesis is mediated by a cytoplasmic membrane-associated replication/transcription complex that includes up to 16 viral nonstructural proteins (nsps), which carry common enzymatic activities, like the viral rna polymerase, but also unusual and poorly understood rna-processing functions. of t ...200919297500
[examination of biocides for their effectiveness against animal viruses according to european union standards with emphasis on the selection of a suitable test virus].because of the changes to be expected in the methods for testing disinfectants deemed to be used in the veterinary field, candidate viral species were evaluated for their suitability as test virus. considered viral species included different non-enveloped viruses [bovine enterovirus type 1 (ecbo (enteric cytopathogenic bovine orphan) virus), mammalian reovirus type 1, feline calici virus (fcv), and bovine parvovirus (bpv)], as well as enveloped viruses, as equine arteritisvirus (eav), bovine her ...200919226936
[molecular biological progression of equine arteritis virus]. 200819035332
rna interference protects horse cells in vitro from infection with equine arteritis virus.equine arteritis virus (eav) belongs to the arteriviridae and causes viral arteritis in horses. in an attempt to develop novel and save therapies against the infection it was tested whether eav is susceptible to rna interference (rnai) in an equine in vitro system. horse cells were transfected with chemically synthesized small interfering rna oligonucleotides (sirnas) and challenged with eav. application of these sirnas led to a significant protection of the cells, and virus titers decreased dra ...200919007819
equine arteritis virus: a new isolate from the presumable first carrier stallion in argentina and its genetic relationships among the four reported unique argentinean strains.equine arteritis virus (eav) was isolated from a testicle of the presumable first stallion infected with eav in argentina. this virus isolate (named lt-lp-arg) was confirmed by gp5-specific pcr and indirect immunofluorescence assays. the pcr product was sequenced, and the phylogenetic analysis revealed that the lt-lp-arg strain of eav forms a monophyletic group, together with other strains previously isolated in our laboratory (lp02 group). however, all argentinean eav strains belong to a polyph ...200818937029
identification of an additional neutralization determinant of equine arteritis virus.we recently established an in vitro model of equine arteritis virus (eav) persistence in hela cells. the objective of this study was to determine whether viral variants with novel neutralization phenotypes emerged during persistent eav infection of hela cells, as occurs during viral persistence in carrier stallions. viruses recovered from persistently infected hela cells had different neutralization phenotypes than the virus in the original inoculum, as determined by neutralization assays using ...200818851997
amino acid substitutions in the structural or nonstructural proteins of a vaccine strain of equine arteritis virus are associated with its attenuation.comparative sequence analysis of a series of strains of equine arteritis virus (eav) of defined virulence for horses, ranging from the horse-adapted virulent bucyrus (vb) strain to a fully attenuated vaccine strain derived from it, identified 13 amino acid substitutions associated with attenuation. these include 4 substitutions in the replicase proteins and 9 in the structural proteins. using reverse genetic techniques, these amino acid substitutions were introduced into a virulent infectious cd ...200818619638
persistent equine arteritis virus infection in hela cells.the horse-adapted virulent bucyrus (vb) strain of equine arteritis virus (eav) established persistent infection in high-passage-number human cervix cells (hela-h cells; passages 170 to 221) but not in low-passage-number human cervix cells (hela-l cells; passages 95 to 115) or in several other cell lines that were evaluated. however, virus recovered from the 80th passage of the persistently infected hela-h cells (hela-h-eavp80) readily established persistent infection in hela-l cells. comparative ...200818579588
equine arteritis virus is delivered to an acidic compartment of host cells via clathrin-dependent endocytosis.equine arteritis virus (eav) is an enveloped, positive-stranded rna virus belonging to the family arteriviridae. infection by eav requires the release of the viral genome by fusion with the respective target membrane of the host cell. we have investigated the entry pathway of eav into baby hamster kidney cells (bhk). infection of cells assessed by the plaque reduction assay was strongly inhibited by substances which interfere with clathrin-dependent endocytosis and by lysosomotropic compounds. f ...200818570963
analysis of the eiav rev-responsive element (rre) reveals a conserved rna motif required for high affinity rev binding in both hiv-1 and eiav.a cis-acting rna regulatory element, the rev-responsive element (rre), has essential roles in replication of lentiviruses, including human immunodeficiency virus (hiv-1) and equine infection anemia virus (eiav). the rre binds the viral trans-acting regulatory protein, rev, to mediate nucleocytoplasmic transport of incompletely spliced mrnas encoding viral structural genes and genomic rna. because of its potential as a clinical target, rre-rev interactions have been well studied in hiv-1; however ...200818523581
the in vitro rna synthesizing activity of the isolated arterivirus replication/transcription complex is dependent on a host factor.the cytoplasmic replication of positive-stranded rna viruses is associated with characteristic, virus-induced membrane structures that are derived from host cell organelles. we used the prototype arterivirus, equine arteritis virus (eav), to gain insight into the structure and function of the replication/transcription complex (rtc) of nidoviruses. rtcs were isolated from eav-infected cells, and their activity was studied using a newly developed in vitro assay for viral rna synthesis, which repro ...200818411274
comparison of two real-time reverse transcription polymerase chain reaction assays for the detection of equine arteritis virus nucleic acid in equine semen and tissue culture fluid.two previously developed taqman fluorogenic probe-based 1-tube real-time reverse transcription polymerase chain reaction (real-time rt-pcr) assays (t1 and t2) were compared and validated for the detection of equine arteritis virus (eav) nucleic acid in equine semen and tissue culture fluid (tcf). the specificity and sensitivity of these 2 molecular-based assays were compared to traditional virus isolation (vi) in cell culture. the t1 real-time rt-pcr had a higher sensitivity (93.4%) than the t2 ...200818319426
formation of the arterivirus replication/transcription complex: a key role for nonstructural protein 3 in the remodeling of intracellular membranes.the replication/transcription complex of the arterivirus equine arteritis virus (eav) is associated with paired membranes and/or double-membrane vesicles (dmvs) that are thought to originate from the endoplasmic reticulum. previously, coexpression of two putative transmembrane nonstructural proteins (nsp2 and nsp3) was found to suffice to induce these remarkable membrane structures, which are typical of arterivirus infection. here, site-directed mutagenesis was used to investigate the role of ns ...200818305048
the efficacy of a commercial elisa as an alternative to virus neutralisation test for the detection of antibodies to eav.infection with equine arteritis virus is a notifiable disease with sporadic occurrence in the uk. as stallions may harbour the virus after infection, horses are screened for exposure by serological testing prior to breeding. the virus neutralisation test is considered the 'gold standard' serological screening test, but it is time-consuming and labour intensive; consequently there is a move towards more rapid screening methodology. in this study, a commercially available eva antibody elisa is ass ...200818267889
molecular analysis of endogenous avian leukosis/sarcoma virus genomes in korean chicken embryos.since the status of endogenous avian leucosis/sarcoma virus (alsv) infections in korean broiler chickens is unclear, this study examined embryonated eggs obtained from broiler farms and korean native chicken breeds in korea using pcr with the primer sets specific for endogenous alsvs. the pcr assays detected the genomes of eav, ev, ev/j and art-ch belonging to the endogenous alsv from all embryos tested. phylogenetically, the korean eav genomes were more closely related to the prototype eav-0 th ...200818250567
characterization of equine arteritis virus particles and demonstration of their hemolytic activity.equine arteritis virus (eav), a member of the newly established family arteriviridae, is a small, positive-stranded rna virus. it carries two protein complexes in its envelope, gp5/m and the recently described gp2b/gp3/gp4 complex. we report here on several basic features of eav replication in cell culture and on the protein composition of virus particles. we have also characterized gp2b, gp3, and gp4 expressed using a baculovirus system in insect cells. finally, we provide evidence that eav pos ...200818219439
analysis of orfs 2b, 3, 4, and partial orf5 of sequential isolates of equine arteritis virus shows genetic variation following experimental infection of horses.samples from horses experimentally infected with the "large plaque variant (lp3a+)" of equine arteritis virus were analysed. these included 182 nasal swabs collected from day 1 to 14 post-infection (p.i.), and 21 virus isolates obtained from white blood cells of animals that showed a prolonged viraemia between days 30 to 72 p.i. in order to determine the genetic stability of the virus and particularly to characterise the genetic variants found during the prolonged viraemia, partial sequences of ...200818191505
development of a fluorescent-microsphere immunoassay for detection of antibodies specific to equine arteritis virus and comparison with the virus neutralization test.the development and validation of a microsphere immunoassay (mia) to detect equine antibodies to the major structural proteins of equine arteritis virus (eav) are described. the assay development process was based on the cloning and expression of genes for full-length individual major structural proteins (gp5 amino acids 1 to 255 [gp5(1-255)], m(1-162), and n(1-110)), as well as partial sequences of these structural proteins (gp5(1-116), gp5(75-112), gp5(55-98), m(88-162), and n(1-69)) that cons ...200818032597
testing for antibodies to equine arteritis virus. 200717965374
molecular epizootiology of equine arteritis virus isolates from poland.phylogenetic analysis was performed on the sequences of 44 polish isolates of equine arteritis virus that were isolated from the semen of stallions from national and private studs, collected during 2001--2005. these sequences were also compared with 41 reference strains previously described and commonly used in phylogenesis. on the basis of the nucleotide sequence analysis of the orf5 gene, encoding the glycoprotein gp5, it was demonstrated that the polish eav isolates belonged to two subgroups ...200817964086
[european biologists versus rome and berlin]. 200717892151
comparison of different molecular methods for assessment of equine arteritis virus (eav) infection: a novel one-step mgb real-time rt-pcr assay, pcr-elisa and classical rt-pcr for detection of highly diverse sequences of slovenian eav variants.in the present study, a new one-step real-time reverse transcription-polymerase chain reaction (rt-pcr) strategy with minor-groove-binder (mgb) technology for the detection of eav from 40 semen samples of slovenian carrier stallions was tested. a novel mgb probe (eavmgbpr) and a reverse primer (eav-r) based on the multiple sequence alignment of 49 different eav strain sequences of the highly conserved orf7 (nucleocapsid gene) were designed. the performance of the assay was compared with differen ...200717854913
detection of antibodies to equine arteritis virus in horse sera using recombinant chimaeric n/g(l) protein. 200717827478
genetic variation and phylogenetic analysis of 22 french isolates of equine arteritis virus.genetic variation and phylogenetic relationships among 22 french isolates of equine arteritis virus (eav) obtained over four breeding seasons (2001-2004) were determined by sequencing open reading frames (orfs) 2a-7. the orfs 2a-7 of 22 isolates differed from the prototype virulent bucyrus strain of eav by between 14 (99.5% identity) and 328 (88.7% identity) nucleotides, and differed from each other by between 0 (100% identity) and 346 (88.1% identity) nucleotides, confirming genetic diversity a ...200717680321
arterivirus subgenomic mrna synthesis and virion biogenesis depend on the multifunctional nsp1 autoprotease.many groups of plus-stranded rna viruses produce additional, subgenomic mrnas to regulate the expression of part of their genome. arteriviruses and coronaviruses (order nidovirales) are unique among plus-stranded rna viruses for using a mechanism of discontinuous rna synthesis to produce a nested set of 5'- and 3'-coterminal subgenomic mrnas, which serve to express the viral structural protein genes. the discontinuous step presumably occurs during minus-strand synthesis and joins noncontiguous s ...200717626105
the equine arteritis virus induces apoptosis via caspase-8 and mitochondria-dependent caspase-9 activation.we have previously showed that equine arteritis virus (eav), an arterivirus, induces apoptosis in vitro. to determine the caspase activation pathways involved in eav-induced apoptosis, target cells were treated with peptide inhibitors of apoptosis z-vad-fmk (pan-caspase inhibitor), z-ietd-fmk (caspase-8-specific inhibitor) or z-lehd-fmk (caspase-9-specific inhibitor) 4 h prior to infection with the eav t1329 canadian isolate. significant inhibition of apoptosis was obtained with all peptide inhi ...200717583760
an rna pseudoknot in the 3' end of the arterivirus genome has a critical role in regulating viral rna synthesis.in the life cycle of plus-strand rna viruses, the genome initially serves as the template for both translation of the viral replicase gene and synthesis of minus-strand rna and is ultimately packaged into progeny virions. these various processes must be properly balanced to ensure efficient viral proliferation. to achieve this, higher-order rna structures near the termini of a variety of rna virus genomes are thought to play a key role in regulating the specificity and efficiency of viral rna sy ...200717581985
antiviral activity of carbohydrate-binding agents against nidovirales in cell culture.coronaviruses are important human and animal pathogens, the relevance of which increased due to the emergence of new human coronaviruses like sars-cov, hku1 and nl63. together with toroviruses, arteriviruses, and roniviruses the coronaviruses belong to the order nidovirales. so far antivirals are hardly available to combat infections with viruses of this order. therefore, various antiviral strategies to counter nidoviral infections are under evaluation. lectins, which bind to n-linked oligosacch ...200717560666
de novo initiation of rna synthesis by the arterivirus rna-dependent rna polymerase.all plus-strand rna viruses encode an rna-dependent rna polymerase (rdrp) that functions as the catalytic subunit of the viral replication/transcription complex, directing viral rna synthesis in concert with other viral proteins and, sometimes, host proteins. rna synthesis essentially can be initiated by two different mechanisms, de novo initiation and primer-dependent initiation. most viral rdrps have been identified solely on the basis of comparative sequence analysis, and for many viruses the ...200717537850
structure of the equine arteritis virus nucleocapsid protein reveals a dimer-dimer arrangement.equine arteritis virus (eav) is an enveloped positive-sense rna virus belonging to the arteriviridae family, which also includes the porcine pathogen prrsv and is genetically and structurally related to the coronaviruses. eav is an important equine pathogen that has caused significant economic losses to the horse-breeding industry and has been difficult to control. the eav virion consists of a genome-containing nucleocapsid core made of nucleocapsid (n) protein surrounded by a lipid envelope con ...200717452783
ontario. equine arteritis virus isolated from a standardbred foal with pneumonia. 198817423174
equine viral arteritis.equine viral arteritis is reviewed with specific reference to clinical features, etiology, transmission, diagnosis, epidemiology, and current methods for the control of this disease. there is evidence of variation in pathogenicity among strains of equine arteritis virus. virus transmission occurs primarily by the respiratory and venereal routes during the acute phase of the infection. the long-term carrier stallion appears to play a major epidemiological role in dissemination and perpetuation of ...198717422919
an infectious recombinant equine arteritis virus expressing green fluorescent protein from its replicase gene.thus far, systems developed for heterologous gene expression from the genomes of nidoviruses (arteriviruses and coronaviruses) have relied mainly on the translation of foreign genes from subgenomic mrnas, whose synthesis is a key feature of the nidovirus life cycle. in general, such expression vectors often suffered from relatively low and unpredictable expression levels, as well as genome instability. in an attempt to circumvent these disadvantages, the possibility to express a foreign gene [en ...200717374763
development and characterization of an infectious cdna clone of the virulent bucyrus strain of equine arteritis virus.strains of equine arteritis virus (eav) differ in the severity of the disease that they induce in horses. infectious cdna clones are potentially useful for identification of genetic determinants of eav virulence; to date, two clones have been derived from a cell culture-adapted variant of the original (bucyrus) isolate of eav, and it has previously been shown that recombinant virus derived from one of these (reav030) is attenuated in horses. a complete cdna copy of the genome of the virulent buc ...200717325365
genetic typing of equine arteritis virus isolates from argentina.we report the nucleotide sequence and genetic diversity of four equine arteritis virus (eav) orf 5 and 6 from argentina isolates, obtained from asymptomatic virus-shedding stallions. nucleic acid recovered from the isolates were amplified by rt-pcr and sequenced. nucleotide and deduced amino acid sequences from the argentine isolates were compared with 17 sequences available from the genbank. phylogenetic analysis revealed that the argentine isolates grouped together in a definite cluster near e ...200717294142
experiences with new generation vaccines against equine viral arteritis, west nile disease and african horse sickness.viral diseases constitute an ever growing threat to the horse industry worldwide because of the rapid movement of large numbers of horses for competition and breeding. a number of different types of vaccines are available for protective immunization of horses against viral diseases. traditional inactivated and live-attenuated (modified live virus, mlv) virus vaccines remain popular and efficacious but recombinant vaccines are increasingly being developed and used, in part because of the perceive ...200717267078
proteolytic maturation of replicase polyprotein pp1a by the nsp4 main proteinase is essential for equine arteritis virus replication and includes internal cleavage of nsp7.the positive-stranded rna genome of the arterivirus equine arteritis virus (order nidovirales) encodes the partially overlapping replicase polyproteins pp1a (1727 aa) and pp1ab (3175 aa). previously, three viral proteinases were reported to cleave these large polyproteins into 12 non-structural proteins (nsps). the chymotrypsin-like viral main proteinase residing in nsp4 is responsible for eight of these cleavages. processing of the c-terminal half of pp1a (the nsp3-8 region) was postulated to o ...200617098961
equine viral arteritis.eva is an important if uncommon disease of horses. potential economic losses attributable to eva include direct losses from abortion, pneumonia in neonates, and febrile disease in performance horses. indirect losses are those associated with national and international trade/animal movement regulations, particularly those pertaining to persistently infected carrier stallions and their semen. however, eav infection and eva are readily prevented through serological and virological screening of hors ...200617037573
[prokaryotic expression of the major antigenic domain of equine arteritis virus gl protein and the establishment of putative indirect elisa assay].according to the antigenic analysis of equine arteritis virus (eav) gl protein, one pair of primers were designed, with which the gene fragment coding the high antigenic domain of eav gl protein was amplified from the eav genome. the cloned gene was digested with bamh i and xho i and then inserted into pet-32a and resulted pet-gl1. the pet-gl1 was transformed into the host cell bl21(de3) and the expression was optimized including cultivation temperature and concentration of iptg. the aim protein ...200616933616
application of polymerase chain reaction and virus isolation techniques for the detection of viruses in aborted and newborn foals.the occurrence of two important pathogens, equine herpesvirus 1 (ehv1) and equine arteritis virus (eav) causing abortions, perinatal foal mortality and respiratory disease, was investigated by polymerase chain reaction (pcr) and virus isolation to demonstrate the presence of abortigenic viruses in samples from 248 horse fetuses in hungary. we found 26 ehv1- and 4 eav-positive aborted or prematurely born foals from 16 and 4 outbreaks, respectively, proving that despite the widely applied vaccinat ...200616841764
highly diverse type of equine arteritis virus (eav) from the semen of a south african donkey: short communication.equine arteritis virus (eav) was detected by rt-nested pcr in semen samples from a naturally infected south african donkey. sequence analysis of the amplified orf5 fragment revealed only 60 to 70% nucleotide identity to a panel of eav reference sequences. the unique donkey eav sequence was also found to be stable during passage in horses. the sequence data reported in this study indicate that the south african donkey variant might represent a new genotype of eav. the distinct genetic properties ...200616841763
the intraleader aug nucleotide sequence context is important for equine arteritis virus replication.the 5(-terminal leader sequence of the equine arteritis virus (eav) genome contains an open reading frame (orf) with an aug codon in a suboptimal context for initiation of protein synthesis. to investigate the significance of this intraleader orf (ilo), an expression plasmid was generated carrying a dna copy of the subgenomic mrna7 behind a t7 promoter. capped rna transcribed from this construct was shown to direct, in an in vitro translation system, the synthesis of leader peptide as well as n ...200616791420
rna signals in the 3' terminus of the genome of equine arteritis virus are required for viral rna synthesis.rna virus genomes contain cis-acting sequences and structural elements involved in virus replication. both full-length and subgenomic negative-strand rna synthesis are initiated at the 3' terminus of the positive-strand genomic rna of equine arteritis virus (eav). to investigate the molecular mechanism of eav rna synthesis, the rna secondary structure of the 3'-proximal region of the genome was analysed by chemical and enzymic probing. based on the rna secondary structure model derived from this ...200616760399
equine arteritis virus.equine arteritis virus (eav) is a small, enveloped, positive-stranded rna virus, in the family arteriviridae , w.h.ich can infect both horses and donkeys. while the majority of eav infections are asymptomatic, acutely infected animals may develop a wide range of clinical signs, including pyrexia, limb and ventral edema, depression, rhinitis, and conjunctivitis. the virus may cause abortion and has caused mortality in neonates. after natural eav infection, most horses develop a solid, long-term i ...199716728076
[case report: equine arteritis virus (eav) as the cause of abortion in alpacas?].here we report a case of a late abortion of a primiparous alpaca where genome fragments of the equine viral arteritis virus (eav) could be detected in fetal tissues using reverse transcription polymerase chain reaction (rt-pcr). all five alpacas of the herd had virus neutralizing antibodies against eav. eav thus must be regarded as a potential agent for abortions in alpacas. possible routes of introduction of the virus are discussed.200616716054
effective removal of equine arteritis virus from stallion semen.a method of removing equine arteritis virus (eav) from equine semen used for artificial insemination is urgently needed. recent medical studies suggest that a double semen processing technique of density gradient centrifugation followed by a 'swim-up' can provide virus-free sperm preparations for assisted reproduction.200616706276
extended phylogeny of equine arteritis virus: division into new subgroups.to determine a conclusive phylogeny, equine arteritis viruses from italy, austria, hungary, sweden, south africa and other parts of the world were analysed by reverse-transcription polymerase chain reaction amplification and direct sequencing. the nucleotide sequences corresponding to the variable part of the large glycoprotein gp5, specified by open reading frame 5, were compared and added to a previously published phylogenetic tree in which a clear division between 'european' and 'american' ty ...200616626399
equine viral arteritis. 200616582004
mutagenesis analysis of the nsp4 main proteinase reveals determinants of arterivirus replicase polyprotein autoprocessing.nonstructural protein 4 (nsp4; 204 amino acids) is the chymotrypsin-like serine main proteinase of the arterivirus equine arteritis virus (order nidovirales), which controls the maturation of the replicase complex. nsp4 includes a unique c-terminal domain (ctd) connected to the catalytic two-beta-barrel structure by the poorly conserved residues 155 and 156. this dipeptide might be part of a hinge region (hr) that facilitates interdomain movements and thereby regulates (in time and space) autopr ...200616537610
expression, purification, and in vitro activity of an arterivirus main proteinase.to allow the biochemical and structural characterization of the chymotrypsin-like "main proteinase" (non-structural protein 4; nsp4) of the arterivirus prototype equine arteritis virus (eav), we developed protocols for the large-scale production of recombinant nsp4 in escherichia coli. the nsp4 proteinase was expressed either fused to maltose binding protein or carrying a c-terminal hexahistidine tag. following purification, the nsp4 moiety of mbp-nsp4 was successfully used for structural studie ...200616527369
site-directed mutagenesis of the nidovirus replicative endoribonuclease nendou exerts pleiotropic effects on the arterivirus life cycle.the highly conserved nendou replicative domain of nidoviruses (arteriviruses, coronaviruses, and roniviruses) belongs to a small protein family whose cellular branch is prototyped by xendou, a xenopus laevis endoribonuclease involved in nucleolar rna processing. recently, sequence-specific in vitro endoribonuclease activity was demonstrated for the nendou-containing nonstructural protein (nsp) 15 of several coronaviruses. to investigate the biological role of this novel enzymatic activity, we ha ...200616439522
recovery of swedish equine arteritis viruses from semen by cell culture isolation and rna transfection.recovery of infectious equine arteritis virus (eav) from the semen of persistently infected swedish stallions was attempted by classical cell culture isolation and by transfection of extracted total rna. whereas virus from semen samples stored for several months at -20 degrees c or from extended semen could only be recovered by transfection of extracted rna, isolation in cell culture was achieved readily with fresh, unextended semen stored at -70 degrees c or directly used after sampling. in par ...200616297456
testing for antibodies to equine arteritis virus. 200516244241
antiviral activity of morpholino oligomers designed to block various aspects of equine arteritis virus amplification in cell culture.the antiviral efficacy of ten antisense phosphorodiamidate morpholino oligomers (pmos) directed against equine arteritis virus (eav), a nidovirus belonging to the family arteriviridae, was evaluated in mammalian (vero-e6) cells. peptide-conjugated pmos (p-pmos) supplied in cell culture medium at micromolar concentrations were efficiently taken up by vero-e6 cells and were minimally cytotoxic. the p-pmos were designed to base pair to rna sequences involved in different aspects of eav amplificatio ...200516227231
testing for antibodies to equine arteritis virus. 200516170008
the 2b protein as a minor structural component of prrsv.porcine reproductive and respiratory syndrome virus (prrsv) orf2 contains an internal orf that codes for a small non-glycosylated protein known as 2b. previous work had identified the presence of a 10kda 2b protein in virus-infected cells and the induction of an anti-2b response in prrsv-infected pigs, as well as a possible association of 2b with the virion (, virology 287:183-191). in this study, we utilized two experimental approaches, including the use of a 2b peptide-specific monoclonal anti ...200516095746
modulating reproductive activity in stallions: a review.situations in which suppression or stimulation of reproductive activity in stallions has been attempted, or is desired, include resolution of the equine arteritis virus 'shedding' state, induction of testicular descent in inguinal cryptorchids, and the improvement of sperm production capacity and/or semen quality in sub-fertile stallions. however, the most common reason for wanting to modulate reproductive activity in a stallion is to alter the expression of sexual behaviour. in the case of inta ...200516061332
equine respiratory viruses in foals in new zealand.to identify the respiratory viruses that are present among foals in new zealand and to establish the age at which foals first become infected with these viruses.200216032260
viruses associated with outbreaks of equine respiratory disease in new zealand.to identify viruses associated with respiratory disease in young horses in new zealand.200216032259
study on the epidemiology of equine arteritis virus infection with different diagnostic techniques by investigating 96 cases of equine abortion in hungary.the occurrence of equine arteritis virus (eav) induced equine abortions was studied with different laboratory methods during a 3-year period. tissue samples from 96 aborted equine foetuses or newborn foals were collected from 57 farms located in different parts of hungary. virus isolation, polymerase chain reaction (pcr), immunohistochemistry and serology were used for the detection of eav infection. the overall seroprevalence of eav infection in mares was 65%. eav induced abortion was diagnosed ...200515925460
a novel subgroup among genotypes of equine arteritis virus: genetic comparison of 40 strains.the authors determined partial nucleic sequences of the variable regions of open-reading frame (orf5) from 151 nucleotide to 668 nucleotide and deduced amino acid sequences of 518 nucleotide respectively of 20 equine arteritis virus (eav) isolates. about 19 hungarian and one austrian eav strains were subjected to sequence analysis, the further data of 20 eav strains: six north american and 14 european were obtained from the genbank. comparative sequence analysis of the hungarian eav strains indi ...200515876222
discontinuous subgenomic rna synthesis in arteriviruses is guided by an rna hairpin structure located in the genomic leader region.nidoviruses produce an extensive 3'-coterminal nested set of subgenomic (sg) mrnas, which are used to express structural proteins and sometimes accessory proteins. in arteriviruses and coronaviruses, these mrnas contain a common 5' leader sequence, derived from the genomic 5' end. the joining of the leader sequence to different segments derived from the 3'-proximal part of the genome (mrna bodies) presumably involves a unique mechanism of discontinuous minus-strand rna synthesis in which base pa ...200515858015
antibodies against equine herpesviruses and equine arteritis virus in burchell's zebras (equus burchelli ) from the serengeti ecosystem.a total of 51 sera from a migratory population of burchell's zebras (equus burchelli) were collected in the serengeti national park (tanzania) between 1999 and 2001 to assess levels of exposure to equine herpesvirus types 1, 2, 4, 9 (ehv-1, -2, -4, -9), ehv-1 zebra isolate t965, and equine arteritis virus (eav). using virus-specific neutralizing antibody tests, seroprevalence was high for ehv-9 (60% of 45), moderate for eav (24% of 51), and lower for the ehv-1-related zebra isolate (17% of 41), ...200515827213
equine viral arteritis control scheme: a brief review with emphasis on laboratory aspects of the scheme in new zealand.to review laboratory aspects of the equine viral arteritis (eva) control scheme in new zealand between 1989 and 2002.200415768101
binding of cellular proteins to the leader rna of equine arteritis virus.the genome of equine arteritis virus (eav) produces a 3' coterminal-nested set of six subgenomic (sg) viral rnas during virus replication cycle, and each set possesses a common leader sequence of 206 nucleotides (nt) in length derived from the 5' end of the viral genome. given the presence of the leader region within both genomic and sg mrnas, it is likely to contain cis-acting signals that may interact with cellular or viral proteins for rna synthesis. gel mobility shift assays indicated that p ...200515744570
diagnostic application of immunoperoxidase monolayer assay using monoclonal antibodies produced against equine arteritis virus 14-kda nucleocapsid protein.two monoclonal antibodies against the bucyrus strain of equine arteritis virus (eav) were produced, and according to immunoperoxidase reaction following western blot of electrophoresed eav structural proteins, they recognized the nucleocapsid (n) protein antigen (14-kda protein). besides reacting with the blotted polypeptide, the antibodies of the two clones (designated 1h1 and 4g6) selected from 576 have shown high affinity and specificity to intracellular virus antigen as well. both antibodies ...200415684664
equine viral arteritis in a stallion. 200515658568
a complex zinc finger controls the enzymatic activities of nidovirus helicases.nidoviruses (coronaviridae, arteriviridae, and roniviridae) encode a nonstructural protein, called nsp10 in arteriviruses and nsp13 in coronaviruses, that is comprised of a c-terminal superfamily 1 helicase domain and an n-terminal, putative zinc-binding domain (zbd). previously, mutations in the equine arteritis virus (eav) nsp10 zbd were shown to block arterivirus reproduction by disrupting rna synthesis and possibly virion biogenesis. here, we characterized the atpase and helicase activities ...200515613297
testing for equine arteritis virus. 200415573796
rescue of disabled infectious single-cycle (disc) equine arteritis virus by using complementing cell lines that express minor structural glycoproteins.equine arteritis virus (eav) contains seven structural proteins that are all required to produce infectious progeny. alphavirus-based expression vectors have been generated for each of these proteins to explore the possibilities for their constitutive expression in cell lines. this approach was successful for minor glycoproteins gp(2b), gp(3) and gp(4) and for the e protein. subsequently, it was demonstrated that cell lines expressing these proteins could rescue eav mutants that were disabled in ...200415557244
structural protein requirements in equine arteritis virus assembly.equine arteritis virus (eav) is an enveloped, positive-stranded rna virus belonging to the family arteriviridae of the order nidovirales. eav particles contain seven structural proteins: the nucleocapsid protein n, the unglycosylated envelope proteins m and e, and the n-glycosylated membrane proteins gp(2b) (previously named g(s)), gp(3), gp(4), and gp(5) (previously named g(l)). proteins n, m, and gp(5) are major virion components, e occurs in virus particles in intermediate amounts, and gp(4), ...200415542653
the immune response to equine arteritis virus: potential lessons for other arteriviruses.the members of the family arteriviridae, genus arterivirus, include equine arteritis virus (eav), porcine reproductive and respiratory syndrome virus (prrsv), lactate dehydrogenase-elevating virus (ldv) of mice, and simian hemorrhagic fever virus (shfv). prrsv is the newest member of the family (first isolated in north america and europe in the early 1990s), whereas the other three viruses were recognized earlier (eav in 1953, ldv in 1960, and shfv in 1964). although arterivirus infections are s ...200415507299
assessing the roles of endogenous retrovirus eav-hp in avian leukosis virus subgroup j emergence and tolerance.avian leukosis virus (alv) subgroup j is thought to have emerged through a recombination event between an unknown exogenous alv and the endogenous retrovirus elements designated eav-hp. all eav-hp elements identified to date in the chicken genome show large deletions, including that of the entire pol gene. here we report the identification of four segregating chicken eav-hp proviruses with complete pol genes, one of which shows exceptionally high sequence identity and a close phylogenetic relati ...200415367619
evidence that use of an inactivated equine herpesvirus vaccine induces serum cytotoxicity affecting the equine arteritis virus neutralisation test.several laboratories worldwide have recently experienced problems related to serum cytotoxicity with the equine arteritis virus (eav) neutralisation test (vn) when using office international des epizooties (oie) reference laboratory prescribed rabbit kidney (rk-13) indicator cells. cytotoxicity can be mistaken for viral cytopathic effect and has led to increasing difficulties in test interpretation, consequently causing disruption to both equine breeding and disease surveillance. results from ex ...200415364465
regulation of relative abundance of arterivirus subgenomic mrnas.the subgenomic (sg) mrnas of arteriviruses (order nidovirales) form a 5'- and 3'-coterminal nested set with the viral genome. their 5' common leader sequence is derived from the genomic 5'-proximal region. fusion of sg rna leader and "body" segments involves a discontinuous transcription step. presumably during minus-strand synthesis, the nascent rna strand is transferred from one site in the genomic template to another, a process guided by conserved transcription-regulating sequences (trss) at ...200415254182
phage display of the equine arteritis virus nsp1 zf domain and examination of its metal interactions.a putative zinc finger (zf) domain in the equine arteritis virus (eav) nsp1 protein was described recently to be required for viral transcription. the nsp1 zf (50 aa) was expressed on the surface of m13ke giii phage, fused to the n terminus of the phage piii protein. to evaluate the functionality of the zf domain, a binding assay was developed, based on the use of immobilized ni(2+) ions (ni-nta). phages displaying zf bound significantly better to ni-nta than did phages displaying negative-contr ...200415158598
exploring the binding mechanism of the main proteinase in sars-associated coronavirus and its implication to anti-sars drug design.the main proteinase of sars-associated coronavirus (sars-cov) plays an important role in viral transcription and replication, and is an attractive target for anti-sars drug development. the important thing is to understand its binding mechanism with possible ligands. in this study, we investigated possible noncanonical interactions, potential inhibitors, and binding pockets in the main proteinase of sars-cov based on its recently determined crystal structure. these findings provide a wide clue t ...200415080921
characterization of the neutralization determinants of equine arteritis virus using recombinant chimeric viruses and site-specific mutagenesis of an infectious cdna clone.we have used an infectious cdna clone of equine arteritis virus (eav) and reverse genetics technology to further characterize the neutralization determinants in the gp5 envelope glycoprotein of the virus. we generated a panel of 20 recombinant viruses, including 10 chimeric viruses that each contained the orf5 (which encodes gp5) of different laboratory, field, and vaccine strains of eav, a chimeric virus containing the n-terminal ectodomain of gp5 of a european strain of porcine reproductive an ...200415051384
the first isolation of equine arteritis virus in argentina.this paper describes the first isolation of equine arteritis virus (eav) in argentina. the virus was isolated from the semen of an imported seropositive stallion held in isolation at a breeding farm in tandil in the buenos aires province. in addition, viral nucleic acid was detected in seminal plasma using the reverse-transcription polymerase chain reaction. the isolated virus was propagated in cell cultures and confirmed as eav by indirect immunofluorescence and virus neutralisation, using a se ...200315005559
secondary structure and function of the 5'-proximal region of the equine arteritis virus rna genome.nidoviruses produce an extensive 3'-coterminal nested set of subgenomic mrnas, which are used to express their structural proteins. in addition, arterivirus and coronavirus mrnas contain a common 5' leader sequence, derived from the genomic 5' end. the joining of this leader sequence to different segments (mrna bodies) from the genomic 3'-proximal region presumably involves a unique mechanism of discontinuous minus-strand rna synthesis. key elements in this process are the so-called transcriptio ...200414970388
genetic characterization of equine arteritis virus during persistent infection of stallions.equine arteritis virus (eav) causes a persistent infection of the reproductive tract of carrier stallions. the authors determined the complete genome sequences of viruses (cw96 and cw01) that were present 5 years apart in the semen of a carrier stallion (cw). the cw96 and cw01 viruses respectively had only 85.6 % and 85.7 % nucleotide identity to the published sequence of eav (eav030). the cw96 and cw01 viruses had two 1 nt insertions and a single 1 nt deletion in the leader sequence, and a 3 nt ...200414769895
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