bovine torovirus: sequencing of the structural genes and expression of the nucleocapsid protein of breda virus.breda virus (brv), a member of the genus torovirus, is an established etiological agent of diarrhea of cattle, which is found as two separate serotypes, brv-1 and brv-2. in this study, a 7.5 kb fragment of the brv-1 genome that bracketed the genes for the structural proteins of brv was amplified by long rt-pcr and the amplicon purified and sequenced directly. sequence analysis revealed the presence of four open reading frames (orf) corresponding to the peplomer (s), envelope (m), and nucleocapsi ...19989879765
torovirus detection in faecal specimens of calves and pigs in hungary: short communication.bovine torovirus is an established aetiological agent of disease in cattle, while porcine torovirus has only been isolated from healthy animals. evidence for the presence of torovirus has been described in several european countries and also in the united states. a survey was performed to detect toroviruses in hungary by means of sampling ten swine and nine bovine herds. rectal swabs and faecal specimens were collected from diarrhoeic calves and from weaned piglets. the samples were tested by th ...200212237970
association of enteric shedding of bovine torovirus (breda virus) and other enteropathogens with diarrhea in veal determine the prevalence, fecal shedding pattern, and association of bovine torovirus (botv) with diarrhea in veal calves at time of arrival and periodically throughout the first 35 days after their arrival on a veal farm.200312693541
phylogenetic and evolutionary relationships among torovirus field variants: evidence for multiple intertypic recombination events.toroviruses (family coronaviridae, order nidovirales) are enveloped, positive-stranded rna viruses that have been implicated in enteric disease in cattle and possibly in humans. despite their potential veterinary and clinical relevance, little is known about torovirus epidemiology and molecular genetics. here, we present the first study into the diversity among toroviruses currently present in european swine and cattle herds. comparative sequence analysis was performed focusing on the genes for ...200312915570
detection of bovine torovirus in neonatal calf diarrhoea in lower austria and styria (austria).faeces of 230 calves with and without diarrhoea collected during the winter period 2004/2005 in 100 austrian farms (styria and lower austria) were examined for viral, bacterial and parasitic enteropathogens. torovirus-specific nucleic acid confirmed by reverse transcriptase-polymerase chain reaction was found in 12 of 230 calves (5.2%). ten of these calves were clinically ill, several of them showing signs of dehydration and abnormal faecal consistency at the time of sampling. computer assisted ...200616629982
bovine torovirus (breda virus) revisited.bovine torovirus (botv) is a pleomorphic virus with a spike-bearing envelope and a linear, non-segmented, positive-sense single-stranded rna genome. this kidney-shaped virus is associated with diarrhea in calves and apparently has a worldwide distribution. this review provides details of the history and taxonomy of botv since its discovery in 1979. information about virion morphology and architecture, antigenic and biological properties, viral genome, protein composition, thermal and chemical st ...200415984322
detection of bovine torovirus and other enteric pathogens in feces from diarrhea cases in cattle.the objectives of this study were to determine the prevalence of bovine torovirus (botv) in bovine fecal samples from diarrhea cases submitted to the ohio animal disease diagnostic laboratory (addl) and to assess if a relationship exists between botv and the other enteric pathogens detected. from november 1999 to may 2001, 259 specimens from 53 calves (< or = 6 months old), 27 young adults (52 years), 125 adults (> or = 2 years), and 54 animals of unknown age were examined by an antigen-capture ...200312735342
comparison of elisa and rt-pcr versus immune electron microscopy for detection of bovine torovirus (breda virus) in calf fecal specimens.bovine torovirus (botv) is an uncultivable enteric pathogen of cattle. its failure to grow in vitro limits epidemiological studies, characterization of the virus, and development of diagnostic techniques. the objectives of this study were to develop and standardize an antigen-capture enzyme-linked immunosorbent assay (elisa) and a reverse transcriptase-polymerase chain reaction (rt-pcr) assay for the detection of botv in fecal specimens. these assays were compared with immunoelectron microscopy ...200312661719
enteric and nasal shedding of bovine torovirus (breda virus) in feedlot assess fecal and nasal shedding patterns of bovine torovirus (botv) in cattle at time of arrival and periodically throughout the first 21 days after arrival at a feedlot.200211911568
the novel hemagglutinin-esterase genes of human torovirus and breda virus.human torovirus (htv) and breda virus (brv), members of the genus torovirus in the family coronaviridae, are established infectious agents of humans and cattle, respectively. the hemagglutinin-esterase (he) gene of breda virus serotype 2 (brv-2) has been identified and the nucleotide sequence for brv serotype 1 (brv-1) genome which contains the open reading frames for the viral structural proteins has been reported revealing the presence of a 1. 25 kb gene whose nucleotide sequence is identical ...199910518710
detection of bovine torovirus in fecal specimens of calves with diarrhea from ontario farms.breda virus (brv), a member of the genus torovirus, is an established etiological agent of disease in cattle. brv isolates have been detected in the stools of neonatal calves with diarrhea in both iowa and ohio and in several areas of europe. however, this virus has been reported only once in canada. therefore, a study was performed to determine the extent to which bovine torovirus is present in calves with diarrhea from farms in southern ontario. a total of 118 fecal samples from symptomatic ca ...19989574689
identification and characterization of a porcine torovirus.a porcine torovirus (potv) was identified and characterized; it is a novel member of the genus torovirus (family coronaviridae, order nidovirales), closely related to but clearly distinct from the already recognized equine torovirus (etv) and bovine torovirus (botv) representatives. immunoelectron microscopy of feces from piglets revealed elongated, 120- by 55-nm particles which were recognized by a torovirus-specific antiserum. amplification by reverse transcriptase (rt) pcr with primers design ...19989557628
hemagglutinin-esterase, a novel structural protein of torovirus.we have characterized the 3'-most 3 kb of the genome of bovine torovirus (botv) strain breda. a novel 1.2-kb gene, located between the genes for the membrane and nucleocapsid proteins, was identified. this gene, the 3'-most 0.5 kb of which is also present in the genome of the equine torovirus isolate berne virus (bev), codes for a class i membrane protein displaying 30% sequence identity with the hemagglutinin-esterases (hes) of coronaviruses and influenza c viruses. heterologous expression of t ...19979188596
enzyme-linked immunosorbent assay reactivity of torovirus-like particles in fecal specimens from humans with diarrhea.toroviruses are recognized enteric pathogens of cattle and horses; in humans, similar pleomorphic particles have been described, but doubt has been raised concerning their identity as viruses. we screened fecal samples from humans with diarrhea for the presence of torovirus-like particles (tvlps) by electron microscopy and subsequently used an enzyme-linked immunosorbent assay (elisa) with bovine torovirus reference reagents to test for the presence of torovirus antigens. to add another selectio ...19938253975
cdna probes for the diagnosis of bovine torovirus (breda virus) infection.a genomic cdna library of rna from breda virus (brv), a bovine torovirus, was prepared. the nucleotide sequence of the 3' end of the genome was found to be highly conserved (93% identical) between brv and berne virus, the torovirus prototype. cross-hybridization experiments were performed to select berne virus cdna clones for use as probes in a dot hybridization assay; the objective was to detect heterologous torovirus rna in fecal material. a rapid rna extraction method was employed to make the ...19912037666
case-control study of microbiological etiology associated with calf diarrhea.calf diarrhea is a major economic burden for the us cattle industry. a variety of infectious agents are implicated in calf diarrhea and co-infection of multiple pathogens is not uncommon in diarrheic calves. a case-control study was conducted to assess infectious etiologies associated with calf diarrhea in midwest cattle farms. a total of 199 and 245 fecal samples were obtained from diarrheic and healthy calves, respectively, from 165 cattle farms. samples were tested by a panel of multiplex pcr ...201323886509
detection and molecular characterisation of bovine corona and toroviruses from croatian cattle.bovine coronavirus (bcov) together with bovine torovirus (btov), both members of the coronaviridae family, order nidovirales are the most common viral enteric pathogens. although studied separately, their joint occurrence and the molecular diversity in cattle in croatia have not been investigated.201526268320
development of a novel detection system for microbes from bovine diarrhea by real-time pcr.diarrhea in cattle is one of the most economically costly disorders, decreasing milk production and weight gain. in the present study, we established a novel simultaneous detection system using taqman real-time pcr designed as a system for detection of microbes from bovine diarrhea using real-time pcr (referred to as dembo-pcr). dembo-pcr simultaneously detects a total of 19 diarrhea-causing pathogens, including viruses, bacteria and protozoa. specific primer-probe sets were newly designed for 7 ...201626616156
molecular detection of bovine coronavirus in a diarrhea outbreak in pasture-feeding nellore steers in southern brazil.worldwide diarrhea outbreaks in cattle herds are more frequently detected in calves being that diarrhea outbreaks in adult cattle are not common. winter dysentery (wd) is a bovine coronavirus (bcov) enteric infection that is more reported in northern hemisphere. seasonal outbreaks of wd in adult cattle occur mainly in dairy cows. wd has not been described in beef cattle herds of tropical countries. this study describes the molecular detection of bcov in a diarrhea outbreak in beef cattle steers ...201626712361
Characterization of epidemic diarrhea outbreaks associated with bovine torovirus in adult cows.Bovine torovirus (BToV) is recognized as an enteric pathogen of calves, but its etiological role in diarrhea and epidemiological characterization in adult cows remain unclear. In 2007-2008, three outbreaks of epidemic diarrhea occurred in adult cows at three dairy farms in Niigata Prefecture, Japan. BToV was the only enteric pathogen detected in these outbreaks, as determined by electron microscopy, reverse transcription-PCR, bacteria and parasite tests of fecal samples, and antibody tests with ...201122167249
Design and validation of consensus-degenerate hybrid oligonucleotide primers for broad and sensitive detection of corona- and toroviruses.The ssRNA+ family Coronaviridae includes two subfamilies prototyped by coronaviruses and toroviruses that cause respiratory and enteric infections. To facilitate the identification of new distantly related members of the family Coronaviridae, we have developed a molecular assay with broad specificity. The consensus-degenerated hybrid oligonucleotide primer (CODEHOP) strategy was modified to design primers targeting the most conserved motifs in the RNA-dependent RNA polymerase locus. They were ev ...201121864579
genetic and antigenic characterization of newly isolated bovine toroviruses from japanese cattle.torovirus, a member of the coronaviridae family, is a gastrointestinal infectious agent that has been identified in humans, cattle, pigs, and equines. toroviruses, except equine torovirus, are difficult to propagate in cell culture; indeed, to date, only the aichi/2004 strain of bovine torovirus (btov) has been isolated among the human, bovine, and porcine toroviruses. in the present study, four cytopathogenic btovs were isolated from diarrheal feces of the cattle using the hrt-18 cell line, and ...201020220164
structural basis for ligand and substrate recognition by torovirus hemagglutinin esterases.hemagglutinin esterases (hes), closely related envelope glycoproteins in influenza c and corona- and toroviruses, mediate reversible attachment to o-acetylated sialic acids (sias). they do so by acting both as lectins and as receptor-destroying enzymes, functions exerted by separate protein domains. he divergence was accompanied by changes in quaternary structure and in receptor and substrate specificity. the selective forces underlying he diversity and the molecular basis for sia specificity ar ...200919721004
detection and characterization of bovine torovirus from the respiratory tract in japanese cattle.bovine torovirus (btov), a member of the coronaviridae family, is a causative agent of diarrhea in cattle, but it may also possess tropism for the respiratory tract. however, no surveys concerning with the relation between respiratory symptoms and the detection of btov have been conducted in wide range. among 311 nasal samples, btov gene products were detected in seven samples (rbtov-1 to -7) derived only from calves with respiratory symptoms, suggesting that btov may be a predisposing factor an ...200919128894
molecular epidemiology of bovine toroviruses circulating in south korea.the prevalence of the bovine torovirus (btov) and its genetic characterization have been reported in north america, europe and japan. therefore, this study examined the prevalence and genetic diversity of the btov in a total of 645 diarrheic fecal samples from 629 korean native beef calf herds using rt-pcr and nested pcr with the primer pairs specific to a part of the btov membrane (m) gene. overall, 19 (2.9%) out of 645 diarrheic samples from 19 herds (6.9%) tested positive for btovs by either ...200817719729
first isolation of cytopathogenic bovine torovirus in cell culture from a calf with diarrhea.a cytopathogenic virus (designated the aichi/2004 strain) was isolated in a human rectal adenocarcinoma cell line (hrt-18) from the ileum contents of a calf with diarrhea. oval and elongated particles, approximately 100 to 170 nm in diameter, with club-shaped projections were seen in the infected culture supernatant, and torovirus-like (tubular and torus nucleocapsid) structures were seen in the infected cells by electron microscopy. an antiserum against bovine torovirus (btov) reacted with the ...200717567770
detection of bovine torovirus in fecal specimens of calves with diarrhea in japan.the aim of this study was to determine the prevalence of bovine torovirus (botv) in bovine fecal samples and to determine whether a relationship exists between botv and diarrhea in japan. ninety-nine diarrheic and 114 normal fecal samples from calves in hokkaido prefecture and 38 diarrheic fecal samples from calves in 10 other prefectures were examined by reverse transcription (rt)-pcr with primers designed in the spike (s) gene for the presence of botv. the specimens were also examined for the ...200717551218
molecular epidemiology of bovine noroviruses in south korea.since the prevalence of bovine norovirus (bnov) and their genetic diversity have only been reported in the usa, england, germany and the netherlands, this study examined the prevalence and genetic diversity of bnovs in diarrheic calves in south korea using 645 diarrheic fecal specimens from calves by rt-pcr and nested pcr assays. overall, 9.3% of the diarrheic fecal samples tested positive for bnovs by either rt-pcr or nested pcr, of which 5.9% samples also tested positive for other enteric path ...200717466472
epidemiological analysis of bovine torovirus in japan.bovine torovirus (btov), a member of the coronaviridae family, is an established gastrointestinal infectious agent in cattle. no epidemiological research on btov has been reported from japan. in this study, we performed a survey to detect btov in japan in 2004 and 2005 using 231 fecal samples (167 from diarrheic cattle and 64 from asymptomatic cattle) that were analyzed by nested reverse transcription (rt) pcr using primers located in the consensus sequences of the reported btov nucleocapsid (n) ...200717320234
the complete sequence of the bovine torovirus genome.viruses in the family coronaviridae have elicited new interest, with the outbreaks caused by sars-hcov in 2003 and the recent discovery of a new human coronavirus, hcov-nl63. the genus torovirus, within the family coronaviridae, is less well characterized, in part because toroviruses cannot yet be grown in cell culture (except for the berne virus). in this study, we determined the sequence of the complete genome of breda-1 (botv-1), a bovine torovirus. this is the first complete torovirus genome ...200616137782
development and application of one-step multiplex reverse transcription pcr for simultaneous detection of five diarrheal viruses in adult cattle.a one-step multiplex reverse transcription (rt)-pcr method was developed for the simultaneous detection of five viruses causing diarrhea in adult cattle: bovine group a rotavirus (gar), bovine group b rotavirus (gbr), bovine group c rotavirus (gcr), bovine coronavirus (bcv), and bovine torovirus (btov). the detection limit of the one-step multiplex rt-pcr for gar, gcr, bcv, and btov was 10(2), 10(0), 10(1), and 10(2) tcid(50)/ml, respectively, and that for gbr was 10(6) copies/ml. the one-step m ...201222407445
whole genome analysis of japanese bovine toroviruses reveals natural recombination between porcine and bovine toroviruses.bovine toroviruses (btovs), belong to the subfamily toroviridae within the family coronaviridae, and are pathogens, causing enteric disease in cattle. in japan, btovs are distributed throughout the country and cause gastrointestinal infection of calves and cows. in the present study, complete genome sequences of two japanese btovs and partial genome sequences of two japanese btovs and one porcine torovirus (ptov) from distant regions in japan were determined and genetic analyses were performed. ...201626708248
detection of bovine torovirus in fecal specimens from calves with diarrhea in turkey.bovine torovirus (btov), a member of the family coronaviridae, is an established gastrointestinal infectious agent in cattle. in this study, we performed a survey to detect btov in turkey between 2009 and 2011 using 235 fecal samples from neonatal calves with diarrhea that were analyzed by the nested reverse transcription (rt) pcr method using primers located in the consensus sequences of the btov membrane (m) gene. the btov m gene was detected in 4.7 % (11/235) of the samples using the nested r ...201424420162
evolution and homologous recombination of the hemagglutinin-esterase gene sequences from porcine torovirus.the objective of the present study was to gain new insights into the evolution, homologous recombination, and selection pressures imposed on the porcine torovirus (ptov), by examining the changes in the hemagglutinin-esterase (he) gene. the most recent common ancestor of ptov was estimated to have emerged 62 years ago based upon he gene sequence data obtained from ptov isolates originating from spain, south korea, netherlands, hungary, and italy and using the he gene of bovine torovirus isolates ...201323749172
first detection and molecular diversity of brazilian bovine torovirus (btov) strains from young and adult cattle.bovine torovirus (btov) is an established enteric pathogen of cattle, but its occurrence in brazilian cattle had not been reported until now. this article describes a survey on btov in brazil carried out on 80 fecal samples from diarrheic young and adult cattle, using a nested-rt-pcr targeting the nucleocapsid (n) gene. btov was detected in 6.25% (5/80) of stool samples from three different geographic regions. sequences analysis showed that brazilian btovs have a high degree of identity with eur ...201323648077
hemagglutination mediated by the spike protein of cell-adapted bovine torovirus.bovine torovirus (btov)-aichi, recently isolated in cultured cells, showed hemagglutination (ha) activity, although the virus has a truncated hemagglutinin-esterase (he) protein, judging from its gene structure, indicating the existence of another viral protein with ha activity. we examined whether the spike (s) protein possesses ha activity. a btov antiserum used in this study, reactive to s but not to he, inhibited ha activity. furthermore, cells infected with btov and those expressing s showe ...201323420207
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