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failure to establish chronic infection of the reproductive tract of the male horse with a south african asinine strain of equine arteritis virus (eav).eight sexually mature horse stallions were inoculated intranasally with a south african asinine strain of eav, a strain that was isolated from the semen of a donkey carrier. all horses developed fever, with maximum rectal temperatures of 38.9-39.9 degrees c recorded 3-6 d post challenge. six horses showed very mild clinical signs of equine viral arteritis and two were asymptomatic. the virus was recovered from the nasopharynxes of six horses 2-7 d after inoculation, and from buffy-coat samples o ...19979204499
orf1a-encoded replicase subunits are involved in the membrane association of the arterivirus replication complex.among the functions of the replicase of equine arteritis virus (eav; family arteriviridae, order nidovirales) are important viral enzyme activities such as proteases and the putative rna polymerase and rna helicase functions. the replicase is expressed in the form of two polyproteins (open reading frame 1a [orf1a] and orf1ab), which are processed into 12 nonstructural proteins by three viral proteases. in immunofluorescence assays, the majority of these cleavage products localized to the perinuc ...19989658116
equine viral arteritis in newborn foals: clinical, pathological, serological, microbiological and immunohistochemical observations.clinical, pathological, immunohistochemical, serological and microbiological findings are described for 2 geographically and temporally distinct equine arteritis virus (eav) epidemics in newborn foals. outbreak a occurred at a commercial standardbred breeding facility; outbreak b began in a group of research animals. clinical signs were severe and primarily referable to the respiratory tract. fever and leucopenia and/or thrombocytopenia were observed in foals surviving for more than 24 h. the mo ...19979234009
antigenic variants of j subgroup avian leukosis virus: sequence analysis reveals multiple changes in the env gene.hprs-103, the prototype of avian leukosis virus (alv) subgroup j, was isolated in 1989 from meat-type chickens from commercial flocks where it induces myelocytic myeloid leukosis (ml). the hprs-103 env gene differs considerably from other alv subgroups but shows high identity (75-97%) to env-like sequences of the different members of the eav family of endogenous avian retroviruses. recently, we have isolated several viruses related to hprs-103 from cases of ml. although these isolates showed pro ...19989568971
effect of heparin on hemagglutination by equine arteritis virus.heparin inhibited hemagglutination (ha) by equine arteritis virus (eav) as well as did ha by aujeszky's disease virus (adv), but failed to inhibit ha by parainfluenza virus type 3 (piv-3). the minimal concentration of heparin required to inhibit 8 ha u of eav was 0.1 u/ml. in addition, most eav hemagglutinin was retained by heparin acrylic beads, as was adv hemagglutinin, but was not piv-3 hemagglutinin. mouse erythrocytes failed to combine with the ha inhibitory factor of heparin. however, mous ...19989592716
genetic and amino acid analysis of the gl protein of canadian, american and european equine arteritis virus isolates.the genetic variation in equine arteritis virus (eav) gl protein encoding gene was investigated. nucleic and deduced amino acid sequences from 7 different eav isolates, including 4 eastern canadian field isolates, were compared with those of the bucyrus reference strain. nucleotide sequence identities between these isolates and the bucyrus reference strain ranged from 87.5% (vienna isolate) to 93.9% (11958 isolate). amino acid identities with the bucyrus reference strain varied from 90.2% (vienn ...19979008807
expression cloning and humoral immune response to the nucleocapsid and membrane proteins of equine arteritis virus.to provide a convenient and sensitive method for the detection of equine arteritis virus (eav)-specific serum antibodies, we developed an immunoblot assay employing the eav nucleocapsid (n) and membrane (m) proteins expressed in a procaryotic expression vector (pmal-c2) for the production of recombinant maltose-binding (mbp) fusion proteins (mbp-n and mbp-m). the antigenic reactivity of the recombinant fusion proteins and their xa factor cleavage eav products was confirmed by immunoblot using ho ...19979384283
hemagglutination with equine arteritis virus.equine arteritis virus (eav) grown on rk13 cell cultures was tested for hemagglutination (ha) with erythrocytes from a variety of species at 4 degrees c, room temperature and 37 degrees c. ha was observed at all temperatures with erythrocytes from mouse and chicken but not with those of cattle, horse, rabbit, guinea pig, mongolian gerbil, goose or chick embryo. chickens showed an individual variation in agglutinability of their erythrocytes, requiring selection of birds to obtain erythrocytes fo ...19979362047
nucleotide sequence and genetic analysis of the leader region of canadian, american and european equine arteritis virus isolates.the extreme 5' end, the entire leader sequence of the arvac vaccine strain, and 10 equine arteritis virus (eav) isolates, including the atcc bucyrus reference strain and 5 canadian field isolates, were determined and compared at the primary nucleotide and secondary structure levels. the leader sequence of eight eav isolates, including the bucyrus reference strain, and the leader sequence of the arvac vaccine strain was determined to be 206 nt in length (not including the putative 5' cap structur ...19989684053
molecular variation in the nucleoprotein gene (orf7) of the porcine reproductive and respiratory syndrome virus (prrsv).the nucleoprotein gene (orf7) of 15 european isolates of porcine reproductive and respiratory syndrome virus (prrsv) was sequenced and compared with corresponding sequences of other prrsv isolates (2 european and 13 american) and one isolate each of other arteriviruses (the lactate dehydrogenase elevating virus (ldv), the simian haemorrhagic fever virus (shfv) and the equine arteritis virus (eav)). their phylogenetic relationships were established using neighbour-joining and parsimony methods. f ...19989660067
the unique envelope gene of the subgroup j avian leukosis virus derives from ev/j proviruses, a novel family of avian endogenous viruses.a new subgroup of avian leukosis virus (alv), designated subgroup j, was identified recently. viruses of this subgroup do not cross-interfere with viruses of the avian a, b, c, d, and e subgroups, are not neutralized by antisera raised against the other virus subgroups, and have a broader host range than the a to e subgroups. sequence comparisons reveal that while the subgroup j envelope gene includes some regions that are related to those found in env genes of the a to e subgroups, the majority ...19989811756
development and application of polymerase chain reaction (pcr) tests for the detection of subgroup j avian leukosis virus.subgroup j avian leukosis virus (alv) is a recently identified avian retrovirus associated with myeloid leukosis in meat-type chickens. the env gene of the hprs-103 strain of alv, the prototype of this subgroup, differs considerably from that of other subgroups, but shows close homology to the env-like sequences of members of the eav family of endogenous retroviruses. polymerase chain reaction (pcr) tests using two sets of primers were developed for the specific detection of the members of this ...19989660074
serologic response of horses to the structural proteins of equine arteritis virus.equine arteritis virus (eav) is the causative agent of equine viral arteritis, an apparently emerging disease of equids. in this study, the antibody response of horses to the structural proteins of eav was evaluated using gradient-purified eav virions and baculovirus-expressed recombinant eav structural proteins (g(l), g(s), m, n) as antigens in a western immunoblotting assay. thirty-three sera from horses that previously had been naturally or experimentally infected with eav were evaluated, inc ...19989683071
screening of horse polyclonal antibodies with a random peptide library displayed on phage: identification of ligands used as antigens in an elisa test to detect the presence of antibodies to equine arteritis virus.a random hexapeptide fusion-phage library was screened to isolate phages that bind to antibodies present in horse sera positive for equine arteritis virus (eav). analysis of the peptide sequences displayed by isolated phages identified seven groups. 25% of the isolated phages used as antigens in an elisa test were specifically recognised by a pool of sera which was positive for eav in virus neutralisation test (vn). five of these, when used as antigen in elisa, detected greater than 50% of sera ...19989766888
sequence determination of the extreme 5' end of equine arteritis virus leader region.the extreme 5' end of the leader sequence of four equine arteritis virus (eav) strains was obtained by using rapid amplification of cdna end method (5' race), and sequenced. seventeen more nucleotides were added upstream of the 5' end of the eav published genomic sequence. a common feature among the analyzed eav isolates was the presence of an aug start codon within the added sequence and the appearance of an intraleader open reading frame (orf) of 111 nucleotides which was predicted to encode a ...19968883367
experimental exposure of pregnant mares to the asinine-94 strain of equine arteritis virus.clinical, virological and serological responses were evaluated in 10 pregnant mares after different challenge exposures to the asinine-94 strain of equine arteritis virus (eav). the outcome of maternal infection on the progeny was also investigated. mares were inoculated intranasally (n = 4), intramuscularly (n = 2), intravenously (n = 1), or contract-exposed (n = 3). all inoculated mares developed pyrexia, 5 showed mild clinical signs related to eav infection and 2 remained asymptomatic. viraem ...19979291073
enzyme-linked immunosorbent assay for serological survey of equine arteritis virus in racehorses.to examine antibodies against equine arteritis virus (eav), an enzyme-linked immunosorbent assay (elisa) using purified virus antigen was developed. the results of elisa were compared with those of serum neutralization (sn) tests. the elisa absorbance values and the sn titers in sera collected weekly from eav-infected horses showed a similar pattern. the elisa could detect antibody to eav in horses experimentally infected with not only a homologous virus strain, which was used as the elisa antig ...19989795910
venereal infection of mares by equine arteritis virus and use of killed vaccine against the infection.venereal infection with equine arteritis virus (eav) was established in each of seven mares by inoculation via the cervix with 20 ml of viral suspension (> or = 8 x 10(6) plaque-forming units; pfu), following treatment with prostaglandin and oestradiol. a dose of < or = 8 x 10(5) pfu produced infection in only five of eight mares. serum neutralizing antibody developed in mares manifesting clinical signs of equine viral arteritis (eva), and a weak antibody was detectable in one apparently healthy ...19979447481
equine viral arteritis risk from imported semen. 19989921633
genetic diversity of equine arteritis virus.equine arteritis viruses (eav) from europe and america were compared by phylogenetic analysis of 43 isolates obtained over four decades. an additional 22 virus sequences were retrieved from genbank. fragments of the glycoprotein g(l) and the replicase genes were amplified by rt-pcr, prior to sequencing and construction of phylogenetic trees. the trees revealed many distinctive lineages, consistent with prolonged diversification within geographically separated host populations. two large groups a ...199910092009
novel endogenous retroviral sequences in the chicken genome closely related to hprs-103 (subgroup j) avian leukosis virus.hprs-103, the prototype of avian leukosis virus (alv) subgroup j, is a recently identified retrovirus associated with myeloid leukosis in meat-type chickens. although this virus shows high sequence identity to other alv subgroups within the gag and pol genes, its env gene is highly diverged (with only about 40% sequence identity) from other alv subgroups. on the other hand, the sequence of the env gene of hprs-103 was 75% identical to that of e51, a member of the eav family of endogenous avian r ...19999934710
[detection of equine arteritis virus (eav) in stallions--a contribution to the improvement of eav diagnosis].serum samples from 72 stallions were examined for the occurrence of antibodies against equine arteritis virus, of which 41 animals (57%) were found to be positive. 32 of the seropositive stallions were then screened for persistent eav infection, before and after the breeding season. semen samples were investigated by rt-pcr followed by dot blot hybridization and nested pcr, and by virus isolation on cell cultures as well. the carrier state was virologically confirmed in 11 of 32 stallions (34%) ...199910028725
comparison of nucleic and amino acid sequences and phylogenetic analysis of open reading frames 3 and 4 of various equine arteritis virus isolates.the genetic variation in equine arteritis virus (eav) protein-encoding open reading frames (orfs) 3 and 4 genes was investigated. nucleic and deduced amino acid sequences from seven different eav isolates (one european, one american and five canadian isolates) and the arvac vaccine strain were compared with those of bucyrus reference strain. orf 3 nucleotide and amino acid sequence identities between these isolates (including the arvac vaccine strain) and the bucyrus reference strain ranged from ...19979428144
open reading frame 1a-encoded subunits of the arterivirus replicase induce endoplasmic reticulum-derived double-membrane vesicles which carry the viral replication complex.the replicase of equine arteritis virus (eav; family arteriviridae, order nidovirales) is expressed in the form of two polyproteins (the open reading frame 1a [orf1a] and orf1ab proteins). three viral proteases cleave these precursors into 12 nonstructural proteins, which direct both genome replication and subgenomic mrna transcription. immunofluorescence assays showed that most eav replicase subunits localize to membranes in the perinuclear region of the infected cell. using replicase-specific ...19999971782
proteolytic processing of the open reading frame 1b-encoded part of arterivirus replicase is mediated by nsp4 serine protease and is essential for virus replication.the open reading frame (orf) 1b-encoded part of the equine arteritis virus (eav) replicase is expressed by ribosomal frameshifting during genome translation, which results in the production of an orf1ab fusion protein (345 kda). four orf1b-encoded processing products, nsp9 (p80), nsp10 (p50), nsp11 (p26), and nsp12 (p12), have previously been identified in eav-infected cells (l. c. van dinten, a. l. m. wassenaar, a. e. gorbalenya, w. j. m. spaan, and e. j. snijder, j. virol. 70:6625-6633, 1996). ...19999971783
characterization of an equine arteritis virus replicase mutant defective in subgenomic mrna synthesis.equine arteritis virus (eav) is a positive-stranded rna virus that synthesizes a 5'- and 3'-coterminal nested set of six subgenomic mrnas. these mrnas all contain a common leader sequence which is derived from the 5' end of the genome. subgenomic mrna transcription and genome replication are directed by the viral replicase, which is expressed in the form of two polyproteins and subsequently processed into smaller nonstructural proteins (nsps). during the recent construction of an eav infectious ...199910364273
[veterinary recommendations for the handling of equine virus arteritis (eva) in practical breeding care].the equine virus arteritis (eva) consistently epidemically varying throughout the different breeds of the horse breeding countries is up to now only of lower significance by means of the typical clinical manifestation as well as an abortion causing factor. the susceptibility of the sexual mature stallions against the equine arteritis virus (eav) causes different infection response which may lead to some restrictions in their use in natural breeding especially in the artificial insemination. in a ...199910077817
control of equine viral arteritis. 199910097333
genetic divergence with emergence of novel phenotypic variants of equine arteritis virus during persistent infection of stallions.the persistently infected carrier stallion is the critical natural reservoir of equine arteritis virus (eav), as venereal infection of mares frequently occurs after breeding to such stallions. two thoroughbred stallions that were infected during the 1984 outbreak of equine viral arteritis in central kentucky subsequently became long-term eav carriers. eav genomes amplified from the semen of these two stallions were compared by sequence analysis of the six 3' open reading frames (orfs 2 through 7 ...199910196259
rapid and sensitive detection of equine arteritis virus in semen and tissue samples by reverse transcription-polymerase chain reaction, dot blot hybridisation and nested polymerase chain reaction.a reverse transcription-polymerase chain reaction (rt-pcr) assay using four different primer pairs for the detection of equine arteritis virus (eav) rna in semen and tissue samples was evaluated. a fragment encoding the leader sequence of the eav genome was most successfully amplified. the specificity and sensitivity of rt-pcr was assessed by virus isolation in cell culture, restriction analysis, dot blot hybridisation and nested pcr. to this end, 23 semen samples from seropositive stallions and ...199810358735
the reverse transcriptase activity in cell-free medium of chicken embryo fibroblast cultures is not associated with a replication-competent retrovirus.reverse transcriptase (rt) activity has previously been reported in concentrated medium of primary chicken embryo cell cultures using the traditional rt assay. recently, using the newly-developed and highly-sensitive product-enhanced reverse transcriptase (pert) assay, rt activity has been detected in live, attenuated vaccines grown in chicken cell substrates. furthermore, this activity has been associated with particles that contain rna related to an ancient, endogenous avian retrovirus family ...19989784139
identification of a novel structural protein of arteriviruses.arteriviruses are positive-stranded rna viruses with an efficiently organized, polycistronic genome. a short region between the replicase gene and open reading frame (orf) 2 of the equine arteritis virus (eav) genome was previously assumed to be untranslated. however, here we report that this segment of the eav genome contains the 5' part of a novel gene (orf 2a) which is conserved in all arteriviruses. the 3' part of eav orf 2a overlaps with the 5' part of the former orf 2 (now renamed orf 2b), ...199910400725
retrovirus-induced disease in poultry.three species of avian retrovirus cause disease in poultry: the avian leukosis/sarcoma virus (alsv), reticuloendotheliosis virus (rev), and lymphoproliferative disease virus (lpdv) of turkeys. the alsv can be classified as slowly transforming viruses, which lack a viral oncogene, and acutely transforming viruses, which possess a viral oncogene. slowly transforming viruses induce late onset leukoses of the b cell lymphoid, erythroid, and myeloid cell lineages, and other tumors, by viral promoter ...19989706091
equine viral arteritis. current status in finland.a serological study for antibodies against equine arteritis virus (eav) in finland was performed during 1996. all equine sera delivered to the virology unit at the national veterinary and food research institute were tested with a micro-neutralization test, using the arvac strain as antigen. the study also included imported horses to evaluate eav circulation in the countries of origin. nucleocapsid gene sequences of 2 finnish equine semen isolates were amplified with rt-pcr and sequenced. the ge ...19989782341
the arterivirus replicase. the road from rna to protein(s), and back again. 19989782270
sequence determination and genetic analysis of the leader region of various equine arteritis virus isolates.the entire leader sequence of ten equine arteritis virus (eav) isolates including the bucyrus reference strain was determined and analyzed at the primary nucleotide and secondary structure levels. the leader sequence of eight eav isolates was determined to be 206 nucleotides (nt) in length, whereas those of the 86ab-a1 and 86ny-a1 isolates were found to be 205 and 207 nt in length, respectively. the sequence identity of the leader sequences between the different isolates and the bucyrus referenc ...19989782362
equine arteritis virus derived from an infectious cdna clone is attenuated and genetically stable in infected stallions.virus derived from an infectious cdna clone of equine arteritis virus (eav030h) was intranasally inoculated into two stallions, neither of which subsequently developed clinical manifestations of equine viral arteritis (eva). virus was isolated from nasal swabs and mononuclear cells collected from both stallions </=14 days p.i. and from the semen of one stallion only at 7 days p.i. similarly, viral rna was detected by rt nested-pcr in nasal swabs and mononuclear cells for </=14 days p.i. and at 7 ...199910405372
arterivirus discontinuous mrna transcription is guided by base pairing between sense and antisense transcription-regulating sequences.to generate an extensive set of subgenomic (sg) mrnas, nidoviruses (arteriviruses and coronaviruses) use a mechanism of discontinuous transcription. during this process, mrnas are generated that represent the genomic 5' sequence, the so-called leader rna, fused at specific positions to different 3' regions of the genome. the fusion of the leader to the mrna bodies occurs at a short, conserved sequence element, the transcription-regulating sequence (trs), which precedes every transcription unit i ...199910518575
the open reading frame 3 of equine arteritis virus encodes an immunogenic glycosylated, integral membrane protein.open reading frame 3 (orf 3) of equine arteritis virus (eav) is predicted to encode a glycosylated membrane protein (gp3) that is uncharacterized. orf 3 of the american type culture collection strain of eav was in vitro transcribed and the encoded gp3 protein was in vitro translated with and without canine microsomal membranes. the gp3 protein was approximately 17 kda after in vitro translation without canine microsomal membranes whereas the glycosylated form, after translation with microsomal m ...199910544133
serological surveillance of equine viral arteritis in the united kingdom since the outbreak in 1993.serological analysis of blood samples submitted to the animal health trust showed that during 1995, 185 of 9203 unvaccinated horses (2.0 per cent) tested positive for antibodies to equine arteritis virus (eav), and that during 1996, 46 of 8851 unvaccinated horses (0.52 per cent) tested positive. during both years thoroughbreds were the predominant breed tested and only a small proportion of these (<0.3 per cent), consisting predominantly of imported mares, were seropositive. in contrast, among s ...199910576624
avian endogenous retrovirus eav-hp shares regions of identity with avian leukosis virus subgroup j and the avian retrotransposon art-ch.the existence of novel endogenous retrovirus elements in the chicken genome, designated eav-hp, with close sequence identity to the env gene of avian leukosis virus (alv) subgroup j has been reported (l. m. smith, a. a. toye, k. howes, n. bumstead, l. n. payne, and k. venugopal, j. gen. virol. 80:261-268, 1999). to resolve the genome structure of these retroviral elements, we have determined the complete sequence of two proviral clones of eav-hp from a line n chicken genomic dna yeast artificial ...200010627540
genetic variation and phylogenetic analysis of open reading frames 3 and 4 of various equine arteritis virus isolates.the genetic variation in equine arteritis virus (eav) nonstructural (ns) protein-encoding open reading frames (orf) 3 and 4 genes was investigated. nucleotide and deduced amino acid sequences from seven different eav isolates (one european, one american and five canadian isolates) and the arvac vaccine strain were compared with those of the bucyrus reference strain. orf 3 nucleotide and amino acid sequence identities amongst these isolates (including the arvac vaccine strain) and the bucyrus ref ...19989782363
detection of antibodies to equine arteritis virus by enzyme linked immunosorbant assays utilizing g(l), m and n proteins expressed from recombinant baculoviruses.indirect enzyme linked immunosorbant assays (elisas) utilizing the three major structural proteins (m, n, and g(l)) of equine arteritis virus (eav) expressed from recombinant baculoviruses were developed. a large panel of sera collected from uninfected horses, and from animals experimentally and naturally infected with eav or vaccinated with the modified live virus vaccine against equine viral arteritis, were used to characterize the humoral immune response of horses to the three major eav struc ...19989923747
isolation and characterization of an arterivirus defective interfering rna genome.equine arteritis virus (eav), the type member of the family arteriviridae, is a single-stranded rna virus with a positive-stranded genome of approximately 13 kb. eav uses a discontinuous transcription mechanism to produce a nested set of six subgenomic mrnas from which its structural genes are expressed. we have generated the first documented arterivirus defective interfering (di) rnas by serial undiluted passaging of a wild-type eav stock in bhk-21 cells. a cdna copy of the smallest di rna (5.6 ...200010708432
genetic manipulation of equine arteritis virus using full-length cdna clones: separation of overlapping genes and expression of a foreign epitope.equine arteritis virus (eav) is an enveloped, positive-stranded rna virus belonging to the family arteriviridae of the order nidovirales. the unsegmented, infectious genome of eav is 12,704 nt in length [exclusive of the poly(a) tail] and contains eight overlapping genes that are expressed from a 3'-coterminal nested set of seven leader-containing mrnas. to investigate the importance of the overlapping gene arrangement in the viral life-cycle and to facilitate the genetic manipulation of the vir ...200010772982
passive transfer, rate of decay, and protein specificity of antibodies against equine arteritis virus in horses from a standardbred herd with high seroprevalence.to determine rate of decay of passively acquired antibodies in standardbred foals on a farm with a high seroprevalence to equine arteritis virus (eav) and to determine whether vertical or horizontal transmission of the virus was responsible for infection on the farm.19989743724
evidence of avian leukosis virus subgroup e and endogenous avian virus in measles and mumps vaccines derived from chicken cells: investigation of transmission to vaccine recipients.reverse transcriptase (rt) activity has been detected recently in all chicken cell-derived measles and mumps vaccines. a study of a vaccine manufactured in europe indicated that the rt is associated with particles containing endogenous avian retrovirus (eav-0) rna and originates from the chicken embryonic fibroblasts (cef) used as a substrate for propagation of the vaccine. we investigated the origin of rt in measles and mumps vaccines from a u.s. manufacturer and confirm the presence of rt and ...199910364336
analysis of the effect of endogenous viral genes in the smyth line chicken model for autoimmune vitiligo.the smyth line (sl) chicken, an animal model for autoimmune human vitiligo, is characterized by a spontaneous posthatch pigment loss, determined to be the result of an autoimmune phenomenon. because endogenous virus (ev) genes have been reported to be associated with a number of autoimmune diseases of human and animal models, we designed this experiment to investigate the role of ev in the sl vitiligo by using the complete sequence of rous-associated virus-2 as a probe for ev. an f(2) resource p ...200010702426
detection of antibodies to equine arteritis virus by a monoclonal antibody-based blocking elisa.a potent elisa antigen was prepared from equine arteritis virus (eav) by differential centrifugation of eav-infected cell culture fluid, followed by solubilization of the preparation by triton x-100 treatment. using this antigen and a mouse monoclonal antibody against the g(l) protein of eav, a reliable blocking elisa (belisa) was developed for the detection of eav antibodies in equine sera. the belisa was evaluated using a total of 837 test serum samples. the relative sensitivity (n = 320) of t ...200010680655
assembly of the coronavirus envelope: homotypic interactions between the m proteins.the viral membrane proteins m and e are the minimal requirements for the budding of coronavirus particles. since the e protein occurs in particles only in trace amounts, the lateral interactions between the m proteins apparently generate the major driving force for envelope formation. by using coimmunoprecipitation and envelope incorporation assays, we provide extensive evidence for the existence of such m-m interactions. in addition, we determined which domains of the m protein are involved in ...200010799570
the predicted metal-binding region of the arterivirus helicase protein is involved in subgenomic mrna synthesis, genome replication, and virion biogenesis.equine arteritis virus (eav), the prototype arterivirus, is a positive-stranded rna virus that expresses its replicase in the form of two large polyproteins of 1,727 and 3,175 amino acids. the functional replicase subunits (nonstructural proteins), which drive eav genome replication and subgenomic mrna transcription, are generated by extensive proteolytic processing. subgenomic mrna transcription involves an unusual discontinuous step and generates the mrnas for structural protein expression. pr ...200010799597
efficient homologous rna recombination and requirement for an open reading frame during replication of equine arteritis virus defective interfering rnas.equine arteritis virus (eav), the prototype arterivirus, is an enveloped plus-strand rna virus with a genome of approximately 13 kb. based on similarities in genome organization and protein expression, the arteriviruses have recently been grouped together with the coronaviruses and toroviruses in the newly established order nidovirales. previously, we reported the construction of pedi, a full-length cdna copy of eav di-b, a natural defective interfering (di) rna of 5.6 kb (r. molenkamp et al., j ...200010982351
genetic stability of equine arteritis virus during horizontal and vertical transmission in an outbreak of equine viral arteritis.an imported carrier stallion (a) from europe was implicated in causing an extensive outbreak of equine viral arteritis (eva) on a warmblood breeding farm in pennsylvania, usa. strains of equine arteritis virus (eav) present in the semen of two carrier stallions (a and g) on the farm were compared to those in tissues of foals born during the outbreak, as well as viruses present in the semen of two other stallions that became persistently infected carriers of eav following infection during the out ...199910466790
avian leukosis virus subgroup j: a rapidly evolving group of oncogenic retroviruses.a strain of avian leukosis virus (alv) belonging to a new envelope subgroup j was isolated in the uk in 1988 from meat-type chickens. the disease caused by the members of this subgroup has since spread very rapidly worldwide and has become one of the major problems facing the broiler meat industry. molecular characterisation of hprs -103, the prototype of subgroup j, has shown that it has a structure of a typical alv with gag, pol and env genes. however the env gene was distinct from that of oth ...199910502478
expression of the two major envelope proteins of equine arteritis virus as a heterodimer is necessary for induction of neutralizing antibodies in mice immunized with recombinant venezuelan equine encephalitis virus replicon particles.rna replicon particles derived from a vaccine strain of venezuelan equine encephalitis virus (vee) were used as a vector for expression of the major envelope proteins (g(l) and m) of equine arteritis virus (eav), both individually and in heterodimer form (g(l)/m). open reading frame 5 (orf5) encodes the g(l) protein, which expresses the known neutralizing determinants of eav (u. b. r. balasuriya, j. f. patton, p. v. rossitto, p. j. timoney, w. h. mccollum, and n. j. maclachlan, virology 232:114- ...200011044106
equine viral arteritis in a gelding in the uk. 199910458581
biochemical characterization of the equine arteritis virus helicase suggests a close functional relationship between arterivirus and coronavirus helicases.the arterivirus equine arteritis virus nonstructural protein 10 (nsp10) has previously been predicted to contain a zn finger structure linked to a superfamily 1 (sf1) helicase domain. a recombinant form of nsp10, mbp-nsp10, was produced in escherichia coli as a fusion protein with the maltose-binding protein. the protein was partially purified by affinity chromatography and shown to have atpase activity that was strongly stimulated by poly(dt), poly(u), and poly(da) but not by poly(g). the prote ...200011000230
genetic manipulation of arterivirus alternative mrna leader-body junction sites reveals tight regulation of structural protein expression.to express its structural proteins, the arterivirus equine arteritis virus (eav) produces a nested set of six subgenomic (sg) rna species. these rna molecules are generated by a mechanism of discontinuous transcription, during which a common leader sequence, representing the 5' end of the genomic rna, is attached to the bodies of the sg rnas. the connection between the leader and body parts of an mrna is formed by a short, conserved sequence element termed the transcription-regulating sequence ( ...200011090163
no evidence of infectious retroviruses in measles virus vaccines produced in chicken embryo cell cultures.all vaccines that are prepared in chicken embryo fibroblasts (cefs) contain a low level of particle-associated reverse transcriptase (rt) activity, which is produced from the avian cell substrate. the rnas present in the particles have sequence homology to viral dnas belonging to the ancient endogenous avian virus (eav) family or to the avian sarcoma-leukosis virus (alv)-related subgroup e endogenous virus loci. although no replication-competent retrovirus has been associated with the rt activit ...200111158127
detection of equine arteritis virus in semen by reverse transcriptase polymerase chain reaction-elisa.the reverse transcriptase polymerase chain reaction (rt-pcr) assay was used to detect equine arteritis virus (eav) in the semen of 88 horses and 2 donkeys, with neutralising antibodies against eav, on the basis of the amplification of a 279 bp long fragment located in the viral polymerase gene. the rt-pcr assay revealed the virus at 4 tcid50/ml in cell culture and showed a greater sensitivity (54.4%) than cell culture isolation (33.3%). moreover, the two samples of donkey semen were found positi ...199910391506
in vitro detection of equine arteritis virus from seminal plasma for identification of carrier stallions.equine arteritis virus (eav) was readily isolated in rk-13 cell monolayers by plaque assay from seminal plasma of experimental carrier stallions when they contained high titers of virus regardless of the presence of non-viral cytotoxicity in the seminal plasma. the cytotoxicity interfered with virus isolation from seminal plasma which contained virus at titers less than 10 pfu/ml. however, it was possible to detect the virus in seminal plasma pretreated with peg (#6000). eav was consistently ide ...200010907693
a zinc finger-containing papain-like protease couples subgenomic mrna synthesis to genome translation in a positive-stranded rna virus.the genome expression of positive-stranded rna viruses starts with translation rather than transcription. for some viruses, the genome is the only viral mrna and expression is regulated primarily at the translational level and by limited proteolysis of polyproteins. other virus groups also generate subgenomic mrnas later in the reproductive cycle. for nidoviruses, subgenomic mrna synthesis (transcription) is discontinuous and yields a 5' and 3' coterminal nested set of mrnas. nidovirus transcrip ...200111172046
an orf-2a protein is not present at a significant level in virions of the arterivirus lactate dehydrogenase-elevating virus.mrna2 of the arteriviruses lactate dehydrogenase-elevating virus (ldv) and equine arteritis virus (eav) encodes two proteins that are read in different frames, an about 26 kda minor envelope glycoprotein and an about 8 kda protein that lacks n-glycosylation sites and a signal peptide, but possesses a central hydrophobic segment. recent studies have shown that both proteins of eav are translated from mrna 2 in eav infected bhk cells, that the 8 kda protein is membrane associated and that small am ...200111226573
diagnosis of equine arteritis virus infection in two horses by using monoclonal antibody immunoperoxidase histochemistry on skin biopsies.two 5-year-old grade male horses presented with epiphora, rhinorrhea, conjunctival and nasal mucosal hyperemia, and dorsal and thoracic macropapular rash. skin biopsies were collected from the affected areas, and serial sections were evaluated following hematoxylin and eosin and immunoperoxidase histochemistry staining by using a murine monoclonal antibody of the immunoglobulin g2a isotype recognizing the 30-kda membrane protein of equine arteritis virus (eav). in both horses, lesions consisted ...200011055877
phylogenetic characterization of a highly attenuated strain of equine arteritis virus from the semen of a persistently infected standardbred stallion.an avirulent, novel variant of equine arteritis virus (eav; ca95g) was isolated from the semen of a persistently infected standardbred stallion. the ca95g virus caused subclinical infection and seroconversion in susceptible horses, and virus was isolated only once from blood and nasal secretions collected from 6 experimentally infected horses. sequence analysis of genes encoding the known eav structural proteins shows that this highly attenuated strain of eav is genetically similar to virulent f ...199910365172
monoclonal antibodies directed against conserved epitopes on the nucleocapsid protein and the major envelope glycoprotein of equine arteritis virus.we recently developed a highly effective immunization procedure for the generation of monoclonal antibodies (mabs) directed against the porcine reproductive and respiratory syndrome virus (e. weiland, m. wieczorek-krohmer, d. kohl, k. k. conzelmann, and f. weiland, vet. microbiol. 66:171-186, 1999). the same method was used to produce a panel of 16 mabs specific for the equine arteritis virus (eav). ten mabs were directed against the eav nucleocapsid (n) protein, and five mabs recognized the maj ...200010834955
bluetongue and equine viral arteritis viruses as models of virus-induced fetal injury and abortion.a number of viruses have the capacity to cross the placenta and infect the fetus to cause, among other potential outcomes, developmental defects (teratogenesis), fetal death and abortion. bluetongue virus (btv) infection of fetal ruminants provides an excellent model for the study of virus-induced teratogenesis. this model has shown that only viruses modified by passage in cell culture, such as modified live virus vaccine strains, readily cross the ruminant placenta, and that the timing of fetal ...200010844231
induction of apoptosis by equine arteritis virus infection.equine arteritis virus (eav) is the etiological agent of equine viral arteritis, a contagious viral disease of equids. eav is the prototype virus of the arteriviruses, a group of small enveloped viruses with positive single-stranded rna genomes. because apoptosis or programmed cell death is believed to play an important role in the biogenesis of several cytopathogenic viruses, we examined whether eav was able to induce cell apoptosis in vitro. to do this, vero cells were infected with eav at a m ...200010872876
development and evaluation of elisa procedures to detect antibodies against the major envelope protein (g(l)) of equine arteritis virus.enzyme-linked immunosorbant assays (elisas) were developed for the detection of antibodies against the major envelope glycoprotein (g(l)) of equine arteritis virus (eav). a 6-histidine tagged recombinant protein expressing the complete g(l) ectodomain (g(l)-6his), a glutathione-s-transferase recombinant protein expressing amino acids 55-98 of g(l) (g(l)-gst) and an ovalbumin-conjugated synthetic peptide representing amino acids 81-106 of g(l) (g(l)-ova) were used as diagnostic antigens. an elisa ...200011064117
effects of chlorine, iodine, and quaternary ammonium compound disinfectants on several exotic disease viruses.the effects of three representative disinfectants, chlorine (sodium hypochlorite), iodine (potassium tetraglicine triiodide), and quaternary ammonium compound (didecyldimethylammonium chloride), on several exotic disease viruses were examined. the viruses used were four enveloped viruses (vesicular stomatitis virus, african swine fever virus, equine viral arteritis virus, and porcine reproductive and respiratory syndrome virus) and two non-enveloped viruses (swine vesicular disease virus (svdv) ...200010676896
recombinant equine arteritis virus as an expression vector.equine arteritis virus (eav) is the prototypic member of the family arteriviridae, which together with the corona- and toroviridae constitutes the order nidovirales. a common trait of these positive-stranded rna viruses is the 3'-coterminal nested set of six to eight leader-containing subgenomic mrnas which are generated by a discontinuous transcription mechanism and from which the viral open reading frames downstream of the polymerase gene are expressed. in this study, we investigated whether t ...200111384225
the arterivirus replicase is the only viral protein required for genome replication and subgenomic mrna transcription.equine arteritis virus (eav) (arteriviridae:) encodes several structural proteins. whether any of these also function in viral rna synthesis is unknown. for the related mouse hepatitis coronavirus (mhv), it has been suggested that the nucleocapsid protein (n) is involved in viral rna synthesis. as described for mhv, we established that the eav n protein colocalizes with the viral replication complex, suggesting a role in rna synthesis. using an infectious cdna clone, point mutations and deletion ...200010993938
equine immunity to viruses.the identification of some of the adaptive immune responses to infection with equine viruses has been the first step toward rational immunoprophylactic design. sufficient knowledge of infection-induced immunity and informed estimates of the requirements for long-term immunity for eiv have now been obtained. thus, the future for inactivated eiv vaccines is promising now that new adjuvants have been applied to induce cellular immunity and safe methods have been designed to stimulate virus-neutrali ...200010752138
intact eav-hp endogenous retrovirus in sonnerat's jungle fowl.the eav-hp group of chicken endogenous retrovirus elements was previously shown to be defective, with large deletions of the pol gene. in this report, we demonstrate that genomes of other gallus species also maintain eav-hp elements with similar deletions. the chicken eav-hp1 locus was detected in both red (gallus gallus gallus) and sonnerat's (gallus sonneratii) jungle fowl with identical integration sites, indicating that these elements had integrated before separation of the gallus species. f ...200111160706
segregation of eav-hp ancient endogenous retroviruses within the chicken population.avian leukosis virus subgroup j (alv-j), an exogenous avian retrovirus, is thought to have evolved by recombination with the highly identical env gene of the endogenous avian retrovirus eav-hp. embryonic expression of eav-hp env has been suggested to be associated with the induction of immunological tolerance, a feature observed in a significant proportion of meat-type chickens infected with alv-j. in support of this hypothesis, we demonstrate that eav-hp loci, some of which could be associated ...200111689680
construction of chimeric arteriviruses reveals that the ectodomain of the major glycoprotein is not the main determinant of equine arteritis virus tropism in cell culture.the recent development of arterivirus full-length cdna clones makes possible the construction of chimeric arteriviruses for fundamental and applied studies. using an equine arteritis virus (eav) infectious cdna clone, we have engineered chimeras in which the ectodomains of the two major envelope proteins, the glycoprotein gp(5) and the membrane protein m, were replaced by sequences from envelope proteins of related and unrelated rna viruses. using immunofluorescence microscopy, we monitored the ...200111601900
elisa and direct immunofluorescence test to detect equine arteritis virus (eav) using a monoclonal antibody directed to the eav-n protein.a monoclonal antibody (mab) directed against the equine arteritis virus (eav) nucleocapsid (n) protein was used for indirect enzyme-linked immunosorbent assays (elisas) using viral antigen from different sources. the same mab was labelled with fluorescein isothiocyanate for direct immunofluorescence tests (difts). the n-specific mab appeared to be suitable for the detection in both elisa and dift of different eav strains and field isolates from semen and tissue samples after passage in lines of ...200111254093
seroprevalence of antibodies against equine arteritis virus in horses residing in the united states and imported horses.to compare seroprevalence of antibodies against equine arteritis virus (eav) in horses residing in the united states with that of imported horses.200111601790
identification of equine herpesvirus 3 (equine coital exanthema virus), equine gammaherpesviruses 2 and 5, equine adenoviruses 1 and 2, equine arteritis virus and equine rhinitis a virus by polymerase chain reaction.to develop rapid (< 8 hour) tests using polymerase chain reaction (pcr) for the diagnosis of equine herpesvirus 3 (ehv3; equine coital exanthema virus), equine gammaherpesviruses 2 (ehv2) and ehv5, equine adenovirus 1 (eadv1), eadv2, equine arteritis virus (eav), equine rhinitis a virus (erav; formerly equine rhinovirus 1)200111712710
phylogenetic characterisation of the g(l) sequences of equine arteritis virus isolated from semen of asymptomatic stallions and fatal cases of equine viral arteritis in denmark.the study describes for the first time the phylogenetic relationship between equine arteritis virus (eav) isolated from asymptomatic virus-shedding stallions and fatal cases of equine viral arteritis (eva) in an european country. eav was isolated from three dead foals and an aborted foetus during three different outbreaks of eva. from these fatalities, the complete open reading frame 5, encoding the eav g(l) protein, was amplified by reverse transcription-polymerase chain reaction and subjected ...200111348770
testing for equine arteritis virus. 200312723636
evaluation of a prototype sub-unit vaccine against equine arteritis virus comprising the entire ectodomain of the virus large envelope glycoprotein (g(l)): induction of virus-neutralizing antibody and assessment of protection in ponies.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 ...200111562536
alphavirus replicon particles expressing the two major envelope proteins of equine arteritis virus induce high level protection against challenge with virulent virus in vaccinated horses.replicon particles derived from a vaccine strain of venezuelan equine encephalitis (vee) virus were used as vectors for expression in vivo of the major envelope proteins (g(l) and m) of equine arteritis virus (eav), both individually and in heterodimer form (g(l)/m). the immunogenicity of the different replicons was evaluated in horses, as was their ability to protectively immunize horses against intranasal and intrauterine challenge with a virulent strain of eav (eav ky84). horses immunized wit ...200211858869
equine viral arteritis.equine viral arteritis (eva) can cause prominent economic losses for the equine industry. the purpose of this review is to provide the pathologist some familiarity with the clinical history, lesions, pathogenesis, and diagnosis of eva. eva is caused by an arterivirus (equine arteritis virus, eav), and the vascular system is the principal but not unique viral target. eva has variable presentations, including interstitial pneumonia, panvasculitis with edema, thrombosis and hemorrhage, lymphoid nec ...200010896389
nuclear localization of non-structural protein 1 and nucleocapsid protein of equine arteritis virus.rna synthesis (genome replication and subgenomic mrna transcription) directed by equine arteritis virus (eav; family arteriviridae, order nidovirales) occurs on modified cytoplasmic membranes to which most viral replicase subunits localize. remarkably, a fraction of non-structural protein 1 (nsp1), a protein essential for transcription but dispensable for genome replication, is present in the host cell nucleus, in particular during the earlier stages of infection. expression of gfp-tagged fusion ...200211907328
characterization of endogenous avian leukosis viruses in chicken embryonic fibroblast substrates used in production of measles and mumps vaccines.previous findings of low levels of reverse transcriptase (rt) activity in chick cell-derived measles and mumps vaccines showed this activity to be associated with virus particles containing rna of both subgroup e endogenous avian leukosis viruses (alv-e) and endogenous avian viruses (eav). these particles originate from chicken embryonic fibroblast (cef) substrates used for propagating vaccine strains. to better characterize vaccine-associated alv-e, we examined the endogenous alv proviruses (ev ...200111264350
lack of evidence of endogenous avian leukosis virus and endogenous avian retrovirus transmission to measles, mumps, and rubella vaccine recipients.the identification of endogenous avian leukosis virus (alv) and endogenous avian retrovirus (eav) in chick cell-derived measles and mumps vaccines in current use has raised concern about transmission of these retroviruses to vaccine recipients. we used serologic and molecular methods to analyze specimens from 206 recipients of measles, mumps, and rubella (mmr) vaccine for evidence of infection with alv and eav. a western blot assay for detecting antibodies to endogenous alv was developed and val ...200111266296
non-structural proteins 2 and 3 interact to modify host cell membranes during the formation of the arterivirus replication complex.the replicase polyproteins of equine arteritis virus (eav; family arteriviridae, order nidovirales) are processed by three viral proteases to yield 12 non-structural proteins (nsps). the nsp2 and nsp3 cleavage products have previously been found to interact, a property that allows nsp2 to act as a co-factor in the processing of the downstream part of the polyprotein by the nsp4 protease. remarkably, upon infection of vero cells, but not of bhk-21 or rk-13 cells, eav nsp2 is now shown to be subje ...200111297673
characterization of an arterivirus defective interfering rna. replication and homologous recombination. 200111774517
detection of equine arteritis virus by real-time taqman reverse transcription-pcr assay.a one-tube real-time taqman reverse transcription-polymerase chain reaction (rt-pcr) assay was developed for the detection of equine arteritis virus (eav). the test was validated using the seminal plasma and nasal secretions of infected horses that were proven to contain eav by traditional virus isolation in rabbit kidney thirteen (rk-13) cells, as well as a variety of cell culture-propagated european and north american strains of eav. the primers and a fluorogenic taqman probe were designed to ...200211849680
functional properties of the predicted helicase of porcine reproductive and respiratory syndrome virus.porcine reproductive and respiratory syndrome virus (prrsv) is a member of the positive-strand rna virus family arteriviridae. although considerable research has focused on this important pathogen, little is known about the function of most prrsv proteins. to examine characteristics of putative nonstructural proteins (nsp) encoded in orf1b, which have been identified by nucleotide similarity to domains of equine arteritis virus, defined genomic regions were cloned and expressed in the prset expr ...200212127789
a replication-competent chimera of plant and animal viruses.human, animal, fungal, and plant viruses encode papain-like proteinases that function in polyprotein processing, rna synthesis, and virus-host interactions. to compare the functional profiles of diverse papain-like proteinases, we replaced a proteinase gene of the beet yellows virus (byv) with those derived from equine arteritis virus (eav), foot-and-mouth disease virus (fmdv), and the fungal virus chv1. we found that, although each of the foreign proteinases efficiently processed the viral poly ...200211886267
characterization of two new structural glycoproteins, gp(3) and gp(4), of equine arteritis virus.equine arteritis virus (eav) is an enveloped, positive-stranded rna virus belonging to the family arteriviridae of the order nidovirales. four envelope proteins have hitherto been identified in eav particles: the predominant membrane proteins m and g(l), the unglycosylated small envelope protein e, and the nonabundant membrane glycoprotein g(s). in this study, we established that the products of eav open reading frame 3 (orf3) and orf4 (designated gp(3) and gp(4), respectively) are also minor st ...200212368326
genetic variation of orfs 3 and 4 of equine arteritis virus. 200111774546
molecular epidemiology and evolution of equine arteritis virus. 200111774467
structure of arterivirus nsp4. the smallest chymotrypsin-like proteinase with an alpha/beta c-terminal extension and alternate conformations of the oxyanion hole.arteriviruses are enveloped, positive-stranded rna viruses and include pathogens of major economic concern to the swine- and horse-breeding industries. the arterivirus replicase gene encodes two large precursor polyproteins that are processed by the viral main proteinase nonstructural protein 4 (nsp4). the three-dimensional structure of the 21-kda nsp4 from the arterivirus prototype equine arteritis virus has been determined to 2.0 a resolution. nsp4 adopts the smallest known chymotrypsin-like f ...200212163505
identification and characterization of avian retroviruses in chicken embryo-derived yellow fever vaccines: investigation of transmission to vaccine recipients.all currently licensed yellow fever (yf) vaccines are propagated in chicken embryos. recent studies of chick cell-derived measles and mumps vaccines show evidence of two types of retrovirus particles, the endogenous avian retrovirus (eav) and the endogenous avian leukosis virus (alv-e), which originate from the chicken embryonic fibroblast substrates. in this study, we investigated substrate-derived avian retrovirus contamination in yf vaccines currently produced by three manufacturers (yf-vax [ ...200312502826
stable and long-lasting immune response in horses after dna vaccination against equine arteritis virus.equine arteritis virus (eav) is the causative agent of the equine viral arteritis. it is a small rna virus with a linear, non-segmented plus rna genome. eav is a member of the arteriviridae family that includes porcine reproductive and respiratory syndrome virus (prssv), simian haemorrhagic fever virus (shfv) and lactate dehydrogenase virus (ldv). the viral transmission is via respiratory and reproductive routes. clinical signs in horses vary, and severe infection can lead to abortions in pregna ...200212418451
the stability of the duplex between sense and antisense transcription-regulating sequences is a crucial factor in arterivirus subgenomic mrna synthesis.subgenomic mrnas of nidoviruses (arteriviruses and coronaviruses) are composed of a common leader sequence and a "body" part of variable size, which are derived from the 5'- and 3'-proximal part of the genome, respectively. leader-to-body joining has been proposed to occur during minus-strand rna synthesis and to involve transfer of the nascent rna strand from one site in the template to another. this discontinuous step in subgenomic rna synthesis is guided by short transcription-regulating sequ ...200312502834
heterodimerization of the two major envelope proteins is essential for arterivirus infectivity.the two major envelope proteins of arteriviruses, the membrane protein (m) and the major glycoprotein (gp(5)), associate into a disulfide-linked heterodimer that is incorporated into the virion and has been assumed to be a prerequisite for virus assembly. using an equine arteritis virus (eav) infectious cdna clone, we have analyzed the requirement for gp(5)-m heterodimerization and have identified the cys residues involved in the formation of the gp(5)-m disulfide bond. the single cys residue (c ...200312477814
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