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pcr assay for the detection and the identification of atypical canine coronavirus in dogs.comparative sequence analysis of the pcr products of the m gene and fragments of the pol1a and pol1b genes of canine coronavirus (ccov) have demonstrated that two separate clusters of ccov are present in dogs. this note describes a pcr assay to identify atypical ccov strains with nucleotide substitutions in the m gene. a total of 177 faecal samples from dogs ccov positive previously with the pcr assay were analysed. sixty-two of the 177 samples were amplified with the pcr described in the presen ...200212393151
identification of coronaviruses in dogs that segregate separately from the canine coronavirus genotype.the genetic diversity of 16 canine coronavirus (ccov) samples is described. samples were obtained from pups infected naturally living in different areas. sequence data were obtained from the m gene and pol1a and pol1b regions. the phylogenetic relationships among these sequences and sequences published previously were determined. the canine samples segregated in two separate clusters. samples of the first cluster were intermingled with reference strains of ccov genotype and therefore could be as ...200312505636
m gene evolution of canine coronavirus in naturally infected dogs.two stray pups (a and b), three and five months old, respectively, both naturally infected with canine coronavirus (ccov), were studied for 180 days. the virus was detected intermittently in the pups' faeces by pcr for periods of 156 and 146 days, respectively. sequence analysis of a fragment of the gene encoding the m protein revealed that the viruses detected at the onset of the infection were very similar to typical strains of ccov, whereas from 42 days after infection in pup a and 40 days af ...200212521247
recombinant m protein-based elisa test for detection of antibodies to canine coronavirus.the membrane (m) protein of canine coronavirus (ccov) was cloned and expressed in e. coli. the purified recombinant protein was then evaluated for its antigenicity and reliability in an enzyme-linked immunosorbent assay (elisa) for detection of ccov antibodies in dog sera. fifty serum samples, screened previously by whole virus elisa and western blotting, were tested. when the performance of the new test was compared with those of whole virus elisa and western blotting, an excellent correlation ...200312711056
efficacy of an inactivated canine coronavirus vaccine in pups.the efficacy of an inactivated ccov vaccine (duramune pc) was evaluated in four pups. two dogs were maintained non-vaccinated. ten days after the booster shot all the pups were challenged with a field ccov strain administered by oro-nasal route. the vaccinated pups did not display clinical signs and shed the challenge-virus for 11.25 days, evaluated by virus isolation, and 13.5 days, evaluated by pcr assay. the two non vaccinated pups displayed mild diarrhoea at day post-challenge 4 and shed the ...200312737196
genetic diversity of a canine coronavirus detected in pups with diarrhoea in italy.the sequence of the s gene of a field canine coronavirus (ccov), strain elmo/02, revealed low nucleotide (61%) and amino acid (54%) identity to reference ccov strains. the highest correlation (77% nt and 81.7% aa) was found with feline coronavirus type i. a pcr assay for the s gene of strain elmo/02 detected analogous ccovs of different geographic origin, all which exhibited at least 92-96% nucleotide identity to each other and to strain elmo/02. the evident genetic divergence between the refere ...200312757915
detection of a group 2 coronavirus in dogs with canine infectious respiratory disease.an investigation into the causes of canine infectious respiratory disease was carried out in a large rehoming kennel. tissue samples taken from the respiratory tract of diseased dogs were tested for the presence of coronaviruses using rt-pcr with conserved primers for the polymerase gene. sequence analysis of four positive samples showed the presence of a coronavirus with high similarity to both bovine and human coronavirus (strain oc43) in their polymerase and spike genes, whereas there was a l ...200312781709
small envelope protein e of sars: cloning, expression, purification, cd determination, and bioinformatics analysis.to obtain the pure sample of sars small envelope e protein (sars e protein), study its properties and analyze its possible functions.200312791175
fatal coronavirus infection in puppies following canine parvovirus 2b infection. 199912968743
molecular modelling of s1 and s2 subunits of sars coronavirus spike glycoprotein.the s1 and s2 subunits of the spike glycoprotein of the coronavirus which is responsible for the severe acute respiratory syndrome (sars) have been modelled, even though the corresponding amino acid sequences were not suitable for tertiary structure predictions with conventional homology and/or threading procedures. an indirect search for a protein structure to be used as a template for 3d modelling has been performed on the basis of the genomic organisation similarity generally exhibited by cor ...200314511651
fecal immunoglobulin a antibodies in dogs infected or vaccinated with canine coronavirus.fecal secretory immunoglobulin a (iga) antibodies in dogs infected or vaccinated with canine coronavirus (ccv) were evaluated by an enzyme-linked immunosorbent assay. the study was carried out with 32 fecal samples collected just before inoculation and at 28 days postinoculation. five groups were studied: naturally infected dogs, experimentally infected dogs, dogs inoculated with a modified live (ml) ccv vaccine by the intramuscular route, dogs inoculated with an ml ccv vaccine by the oronasal r ...200414715553
prevalence of feline coronavirus types i and ii in cats with histopathologically verified feline infectious peritonitis.feline coronaviruses (fcov) vary widely in virulence causing a spectrum of clinical manifestations reaching from subclinical course to fatal feline infectious peritonitis (fip). independent of virulence variations they are separated into two different types, type i, the original fcov, and type ii, which is closely related to canine coronavirus (ccv). the prevalence of fcov types in austrian cat populations without fip has been surveyed recently indicating that type i infections predominate. the ...200415019109
safety and efficacy of a modified-live canine coronavirus vaccine in dogs.the safety and the efficacy of a modified-live (ml) canine coronavirus (ccov) vaccine strain 257/98-3c was evaluated in 14 dogs seronegative and virus negative for ccov. for the safety test, four dogs were inoculated, two by intramuscular and two by oronasal route, with 10 times the vaccinal dose. during the observation period (28 days) all dogs did not display any local or systemic reaction. for the efficacy test, eight dogs were vaccinated by intramuscular (four dogs-group a) or by oronasal ro ...200415019110
cloning and expression of two fragments of the s gene of canine coronavirus type i.two fragments, s66 and s55, of the s glycoprotein of the newly identified canine coronavirus type i (ccov type i), were expressed in a procariotic system. the purified recombinant proteins of 350 and 366 amino acids in length, respectively, were employed to develop an enzyme-linked immunosorbent assay (elisa) for the detection of ccov type i antibodies in dog sera. four canine sera-positive for ccov type ii, four sera-positive for ccov type i and 10 negative control sera were examined. only the ...200415019260
two genotypes of canine coronavirus simultaneously detected in the fecal samples of dogs with diarrhea.sixty-nine fecal samples from diarrheic puppies were examined by reverse transcription-pcr assays for the m and the s genes of canine coronaviruses (ccovs). the isolates in 10 samples were recognized as ccov type i, and the isolates in 6 samples were recognized as ccov type ii, while isolates of both genotypes were simultaneously detected in 53 samples.200415071054
the pros and cons of vaccinating the dog population against canine coronavirus and one of the serovars of leptospira interrogans. 200315086088
quantitation of canine coronavirus rna in the faeces of dogs by taqman rt-pcr.a taqman fluorogenic reverse transcriptase-polymerase chain reaction (rt-pcr) assay was developed for the detection and quantitation of canine coronavirus (ccov) rna in the faeces of naturally or experimentally infected dogs. the ccov fluorogenic rt-pcr assay, which targeted the orf5 (m gene), was more sensitive than a conventional rt-pcr assay targeting the same gene, showing a detection limit of 10 copies of ccov standard rna, and was linear from 10 to 10(8) copies, allowing quantitation of sa ...200415158596
canine distemper and related diseases: report of a severe outbreak in a kennel.an outbreak of canine distemper in a kennel of german shepherds in the province of bari is reported. six 42-day-old pups developed typical signs of canine distemper (fever, conjunctivitis, respiratory distress and enteritis) and died within 7-10 days. neurological symptoms were observed only in one pup. four additional pups, which had shown no sign of illness, were separated and vaccinated, but two of these developed a severe, fatal nervous form 15 days later. post-mortem examination, carried ou ...200415164629
molecular characterization of a virulent canine coronavirus bgf strain.molecular characterisation of a canine coronavirus (ccov) isolate (bgf), associated with an outbreak of diarrhoea in puppies, showed 92.7% identity with attenuated insavc-1 strain. canine coronavirus bgf revealed a full length non-structural protein 3b (nsp 3b), associated with virulence in other coronaviruses, and a highly divergent region at the amino terminal domain of the membrane protein that may be implicated in avoiding the host immune reaction. this new canine coronavirus strain could he ...200415177889
coronavirus infection of spotted hyenas in the serengeti ecosystem.sera from 38 free-ranging spotted hyenas (crocuta crocuta) in the serengeti ecosystem, tanzania, were screened for exposure to coronavirus of antigenic group 1. an immunofluorescence assay indicated high levels of exposure to coronavirus among serengeti hyenas: 95% when considering sera with titer levels of > or = 1:10 and 74% when considering sera with titer levels of > or = 1:40. cubs had generally lower mean titer levels than adults. exposure among serengeti hyenas to coronavirus was also con ...200415288921
antibodies to selected pathogens in free-ranging terrestrial carnivores and marine mammals in canada.antibody titres to selected pathogens (canine adenovirus [cav-2], feline herpesvirus [fhv], phocine herpesvirus [phv-1], canine distemper virus, dolphin morbillivirus [dmv], phocine distemper virus [pdv], parainfluenza virus type 3 [pi3], rabies virus, dolphin rhabdovirus [drv], canine coronavirus, feline coronavirus, feline leukaemia virus, borrelia burgdorferi and toxoplasma gondii) were determined in whole blood or serum samples from selected free-ranging terrestrial carnivores and marine mam ...200415338705
canine coronavirus infection in turkish dog population.canine coronavirus (ccov) is one of the most important viral agents affecting the gastrointestinal system of dogs. in this study virological and serological investigations were performed to demonstrate the existence and prevalence of ccov infection in a turkish dog population. a total of 269 animals were subjected to the study. of 179 dogs tested for ccov antibodies, 112 (62.5%) were found to be positive by serum neutralization test, while 133 (74.3%) were positive by elisa. the highest prevalen ...200415525364
investigation into the causes of canine infectious respiratory disease: antibody responses to canine respiratory coronavirus and canine herpesvirus in two kennelled dog populations.two training centres for working dogs were monitored for one year to determine the presence of viruses and viral antibodies and their association with canine infectious respiratory disease (cird). tonsillar swabs and serum were obtained from dogs on entry into the kennels and in regular intervals thereafter. additional samples were collected during outbreaks of cird. the swabs were examined by virus culture and pcr for canine parainfluenza virus, canine adenovirus, canine herpesvirus (chv) and c ...200515841339
[two genotypes of canine coronavirus simultaneously detected in the fecal samples of healthy foxes and raccoon dogs].61 fecal samples from healthy foxes and 24 from healthy raccoon dogs were examined by rt-nested pcr assays for the presence and genotypic identification of canine coronaviruses (ccvs). 77.0% fox samples were recognized as ccv positive, 43 of which belonged to type ii and 29 to type i, as well as both genotypes were simultaneously detected in 25 samples. out of the total 24 fecal samples from raccoon dogs, 22 were ccv positive for type ii and 16 for type i. m gene fragments of 8 samples were sequ ...200515989282
epidemiologic evaluation of diarrhea in dogs in an animal shelter.to determine associations among infectious pathogens and diarrheal disease in dogs in an animal shelter and demonstrate the use of geographic information systems (giss) for tracking spatial distributions of diarrheal disease within shelters.200516008226
genotype-specific fluorogenic rt-pcr assays for the detection and quantitation of canine coronavirus type i and type ii rna in faecal samples of dogs.two genotype-specific fluorogenic rt-pcr assays were developed for the detection and quantitation of canine coronavirus (ccov) type i and type ii rna in the faeces of dogs with diarrhoea. both the fluorogenic assays showed high specificity, sensitivity and reproducibility, allowing a precise quantitation of ccov type i and type ii rna over a linear range of about eight orders of magnitude (from 10(1) to 10(8) copies of standard rna). comparison with genotype-specific gel-based rt-pcr assays reve ...200516024100
an update on aspects of viral gastrointestinal diseases of dogs and cats.viruses commonly cause gastrointestinal illnesses in dogs and cats that range in severity from mild diarrhoea to malignant neoplasia. perpetual evolution of viruses is reflected in changing disease patterns, so that familiar viruses are sometimes discovered to cause new or unexpected diseases. for example, canine parvovirus (cpv) has regained the ability to infect felids and cause a panleucopenia-like illness. feline panleucopenia virus (fpv) has been shown to cause fading in young kittens and h ...200316032339
[construction of recombinant canine adenovirus type 2 expressing canine coronavirus spike glycoprotein and its immunogenicity].in order to construct a recombinant canine adenovirus type 2 (cav-2) expressing the spike glycoprotein of canine coronavirus (ccv), the s1 gene fragment of ccv strain dxmv, encoding major antigenic region a, b, c and d of s protein, was amplified by rt-pcr and cloned into pvax1 vector. the complete s1 expression cassette was subcloned into the shuttle vector pvaxe3, then further cloned into the backbone vector ppoly2-cav2 containing complete genome of cav-2. to gain the recombinant canine adenov ...200516245877
detection of canine coronaviruses genotype i and ii in raised canidae animals in china.an rt-npcr assay was used for testing fecal samples of dogs, foxes, raccoon dogs and minks for the presence of canine coronavirus (ccv). the animals were raised in homes, dog schools or farms. seventy out of 81 healthy dog feces from three cities and 21 out of 48 diarrhea feces from pet dogs were positive for type ii ccv. from a total of 61 healthy fox feces, 43 were positive for type ii and 29 for type i ccv, out of which 25 were simultaneously positive for the two different genotypes. among 24 ...200616450706
the prevalence of a group 2 coronavirus in dogs in japan.canine coronavirus (ccov) has been reported to cause acute diarrhea mainly in young pups. ccov and feline coronavirus are classified as group 1 coronaviruses. however, it has recently been reported in the united kingdom that the group 2 coronavirus gene, which is more closely related to the bovine coronavirus (bcov) and human coronavirus strain oc43, has been detected in respiratory tract tissue samples from dogs with respiratory disease. in this study, we examined the prevalence of antibodies t ...200616462112
genetic evolution of canine coronavirus and recent advances in prophylaxis.since the first identification of the virus in 1971, the disease caused by canine coronavirus (ccov) has not been adequately investigated and the role that the virus plays in canine enteric illness has still not been well established. in the last decade, as a consequence of the relatively high mutation frequency of rna positive stranded viruses, ccov has evolved and a new genotype has been identified in the faeces of infected dogs. the several studies carried out by different researchers have fo ...200616472519
canine coronavirus-associated puppy mortality without evidence of concurrent canine parvovirus infection.this report presents 2 cases in which puppy fatalities were associated with canine coronavirus (ccv), but no evidence of concurrent canine parvovirus (cpv-2) disease was observed. case 1 involved a 7-week-old, male short-haired chihuahua, which had become lethargic 24 hours after purchase from a pet store. within 72 hours, the puppy began to vomit, had diarrhea, and was admitted to the veterinary clinic, where it was placed on iv fluids. the parvovirus cite test was negative. the puppy died with ...200516475526
molecular characterization of a novel coronavirus associated with epizootic catarrhal enteritis (ece) in ferrets.a novel coronavirus, designated as ferret enteric coronavirus (fecv), was identified in feces of domestic ferrets clinically diagnosed with epizootic catarrhal enteritis (ece). initially, partial sequences of the polymerase, spike, membrane protein, and nucleocapsid genes were generated using coronavirus consensus pcr assays. subsequently, the complete sequences of the nucleocapsid gene and the last two open reading frames at the 3' terminus of the fecv genome were obtained. phylogenetic analyse ...200616499943
serological prevalence of canine respiratory coronavirus.canine respiratory coronavirus (crcov) has recently been detected in dogs; it is a group 2 coronavirus showing similarity to bovine coronavirus (bcov) but is distinct from canine enteric coronavirus (cecov). crcov may play an important role in canine infectious respiratory disease (cird) either by predisposing to further and potentially more serious viral and bacterial infections or possibly as a primary pathogen. the prevalence of serum antibodies to crcov, in a population of dogs in the south ...200616551493
canine coronavirus highly pathogenic for dogs.canine coronavirus (ccov) is usually responsible for mild, self-limiting infections restricted to the enteric tract. we report an outbreak of fatal disease in puppies caused by a pathogenic variant of ccov that was isolated from organs with severe lesions.200616704791
survey of dogs in japan for group 2 canine coronavirus infection.specimens obtained from 96 dogs with respiratory and enteric clinical signs in japan were retrospectively examined for group 2 coronavirus by reverse transcription-pcr. two dogs were found to be positive. phylogenetic analysis of the spike gene indicated that they were most probably related to the canine respiratory coronavirus recently described in the united kingdom.200616825396
m gene analysis of atypical strains of feline and canine coronavirus circulating in an austrian animal shelter.coronavirus-positive samples of faeces collected in an austrian animal shelter from 12 cats and 10 dogs were analysed by reverse transcriptase-pcr with primers amplifying a segment of the m protein gene, and by sequence analysis. in addition, the samples were subjected to s gene typing, using primers that differentiated between feline coronavirus (fcov) types i and ii. a phylogenetic analysis of the m gene sequences revealed not only clearly segregating canine coronavirus (ccov) in the dogs, typ ...200616891424
sybr green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses.coronaviruses are etiologic agents of respiratory and enteric diseases in humans and in animals. in this study, a one-step real-time reverse transcription-polymerase chain reaction (rt-pcr) assay based on sybr green chemistry and degenerate primers was developed for the generic detection of coronaviruses. the primers, designed in the open reading frame 1b, enabled the detection of 32 animal coronaviruses including strains of canine coronavirus, feline coronavirus, transmissible gastroenteritis v ...200716941059
real-time rt-pcr detection of bovine viral diarrhoea virus in whole blood using an external rna reference.a novel two-step real-time rt-pcr assay using sybr green i was developed for the detection of acute bovine viral diarrhoea virus (bvdv) infection in whole blood from cattle. during infection animals experience a characteristic transient leucopenia and the number of cells per volume of blood changes over time; so quantitation of viral load by reference to a cellular housekeeping gene is not ideal as this may hide significant animal to animal variation. therefore, to facilitate comparison of diffe ...200617030066
isolation and sequence analysis of canine respiratory coronavirus.canine respiratory coronavirus (crcov) has frequently been detected in respiratory samples from dogs by rt-pcr. in this report the first successful isolation of crcov from a dog with respiratory disease is described. the isolate crcov-4182 was cultured in hrt-18 cells but failed to replicate in a number of other cell lines. the nucleotide sequence of the 3'-terminal portion of the crcov genome was determined including all open reading frames from the ns2 gene to the n gene. comparison with other ...200717092595
mutational analysis of aminopeptidase n, a receptor for several group 1 coronaviruses, identifies key determinants of viral host range.feline coronavirus (fcov), porcine transmissible gastroenteritis coronavirus (tgev), canine coronavirus (ccov), and human coronavirus hcov-229e, which belong to the group 1 coronavirus, use aminopeptidase n (apn) of their natural host and feline apn (fapn) as receptors. using mouse-feline apn chimeras, we identified three small, discontinuous regions, amino acids (aa) 288 to 290, aa 732 to 746 (called r1), and aa 764 to 788 (called r2) in fapn that determined the host ranges of these coronavirus ...200717093189
first detection of canine parvovirus type 2c in pups with haemorrhagic enteritis in spain.canine parvovirus type 2 (cpv-2), the aetiological agent of haemorrhagic enteritis in dogs, includes three antigenic variants, types 2a, 2b and 2c. cpv-2c has been detected initially in italy and subsequently in vietnam. we report the first identification of this novel antigenic variant in spain, where it caused an outbreak of fatal enteritis in basset hound pups in association with canine coronavirus type i and type ii. we suggest that this new antigenic variant of cpv-2 could spread throughout ...200617123424
infectious canine hepatitis: an "old" disease reemerging in italy.four outbreaks of infectious canine hepatitis (ich) occurring in italy between 2001 and 2006 are reported. three outbreaks were observed in animal shelters of southern italy, whereas a fourth outbreak involved two purebred pups imported from hungary few days before the onset of clinical symptoms. in all outbreaks canine adenovirus type 1 (cav-1) was identified by virus isolation and pcr. in three outbreaks, other canine viral pathogens were detected, including canine distemper virus, canine parv ...200717197003
serological and molecular evidence that canine respiratory coronavirus is circulating in italy.canine respiratory coronavirus (crcov) is a group ii coronavirus that was firstly identified in lung samples of dogs with canine infectious respiratory disease (cird) in uk in 2003. we report for the first time the identification of crcov in italy, together with serological evidence that the virus has been circulating in the italian dog population as from 2005. serological investigations on 216 dog sera, carried out by an elisa test using the strictly related bovine coronavirus (bcov) as antigen ...200717215093
canine coronavirus induces apoptosis in cultured cells.canine coronavirus (ccov) is widespread in dogs in several countries and causes mild enteric illness evolving to severe enteritis in young pups. in in vitro cultures canine coronaviruses generally induce extensive cell death, however nature of the events leading to cell death remains largely unknown. we analysed the induction of cytopathic effect by ccov in a canine fibrosarcoma cell line (a-72) in order to characterize the apoptotic effect in homologous cell system. following ccov infection a-7 ...200717254720
molecular characterisation of the virulent canine coronavirus cb/05 strain.this paper characterises a virulent strain (cb/05) of canine coronavirus (ccov) isolated from the internal organs of pups that had died of a systemic disease without evidence of other common canine pathogens. high viral rna titres were detected in the internal organs by a real-time rt-pcr assay specific for ccov type ii. sequence analysis of the 3' end (8.7kb) of the genomic rna of strain cb/05 revealed conserved structural as well as non-structural proteins, with the exception of a truncated fo ...200717275120
indomethacin has a potent antiviral activity against sars coronavirus.severe acute respiratory syndrome (sars) is a newly emerging, highly transmissible and fatal disease caused by a previously unknown coronavirus (sars-cov). existing in non-identified animal reservoirs, sars-cov continues to represent a threat to humans because there is no effective specific antiviral therapy for coronavirus infections.200617302372
exposure of free-ranging maned wolves (chrysocyon brachyurus) to infectious and parasitic disease agents in the noël kempff mercado national park, bolivia.maned wolves (chrysocyon brachyurus) are neotropic mammals, listed as a cites appendix ii species, with a distribution south of the amazon forest from bolivia, through northern argentina and paraguay and into eastern brazil and northern uruguay. primary threats to the survival of free-ranging maned wolves include habitat loss, road kills, and shooting by farmers. an additional threat to the conservation of maned wolves is the risk of morbidity and mortality due to infectious and parasitic diseas ...200517323558
feline aminopeptidase n is not a functional receptor for avian infectious bronchitis virus.coronaviruses are an important cause of infectious diseases in humans, including severe acute respiratory syndrome (sars), and have the continued potential for emergence from animal species. a major factor in the host range of a coronavirus is its receptor utilization on host cells. in many cases, coronavirus-receptor interactions are well understood. however, a notable exception is the receptor utilization by group 3 coronaviruses, including avian infectious bronchitis virus (ibv). feline amino ...200717324273
serological prevalence of canine respiratory coronavirus in southern italy and epidemiological relationship with canine enteric coronavirus.canine respiratory coronavirus (crcov) has been detected in dogs suffering from respiratory disease and is thought to be involved in canine infectious respiratory disease (cird) complex. canine enteric coronavirus (cecov) is a widespread pathogen of dogs, responsible for mild to severe diarrhea in pups. the purpose of this study was to establish the seroprevalence of crcov in italy and its relationship to cecov type ii seroprevalence. the age and year of sample collection from seropositive dogs ...200717402612
serosurvey of infectious disease agents of carnivores in captive red pandas (ailurus fulgens) in china.the future of the endangered red panda (ailurusfulgens) depends in part on the development of protective measures against infectious diseases. the present study is a first step toward improved understanding of infectious diseases in the species' home regions. serum samples obtained from 73 red pandas in 10 captive facilities in southwest, east, and northeast china from october to december 2004 were tested for antibodies against nine common infectious pathogens of carnivores. antibody titers agai ...200717469274
canine coronavirus inactivation with physical and chemical agents.canine coronavirus (ccov) is responsible for mild or moderate enteritis in puppies. the virus is highly contagious and avoiding contact with infected dogs and their excretions is the only way to ensure disease prevention. since no studies have yet focused on the sensitivity of ccov to chemical biocides the present investigation examined the efficiency of physical and chemical methods of viral inactivation. ccov infectivity was stable at +56 degrees c for up to 30 min, but tended to decrease rapi ...200817513145
characterization of divergent and atypical canine coronaviruses from sweden.field canine coronaviruses (ccvs) identified during a series of outbreaks of gastroenteritis in swedish dogs were subjected to genetic analysis involving the open reading frame 1b (orf1b) and the membrane (m) and spike (s) protein genes. four field viruses originating from the stockholm region presented identical sequences and segregated separately from other ccvs characterized so far and from got/05, the variant recovered in western sweden. a recombinant origin of the fifth virus identified in ...200717533554
systemic fatal type ii coronavirus infection in a dog: pathological findings and immunohistochemistry.a case of fatal systemic coronavirus infection is described in a 53-day-old pekinese dog. pathological findings and immunohistochemical identification using a monoclonal anti-canine coronavirus antibody are included. visceral lesions consisted of extensive fibrinopurulent bronchopneumonia, multiple renal cortical infarcts, severe coalescing centrilobular hepatic fatty change with minimal random hepatic necrosis, and multifocal splenic haemorrhage with lymphoid depletion. moderate chronic diffuse ...200817618660
aetiological role of viruses in puppies with diarrhoea. 200717675636
high-cell-passage canine coronavirus vaccine providing sterilising immunity.to evaluate the ability of a high-cell-passage canine coronavirus vaccine to immunise dogs against challenge with a field isolate of the virus.200717877547
experimental infection of dogs with a novel strain of canine coronavirus causing systemic disease and lymphopenia.a pantropic canine coronavirus (ccov) strain (cb/05) has been recently associated to a fatal outbreak of systemic disease in young dogs. we report the clinical, virological and serological findings in dogs experimentally infected with strain cb/05. the dogs, three 2.5-month-old and two 6-month-old pups, were successfully infected, shedding viral rna with their faeces for the entire observation period (21 days) and displaying systemic clinical signs resembling those observed during the course of ...200818031959
action of disinfectants on canine coronavirus replication in vitro.canine coronavirus (ccov) is responsible for enteric disease in pups. infected dogs generally have a rapid recovery, so the virus is highly contagious and the spread of infection is difficult to control. chemical disinfectants have been widely used in human disease-control programmes to prevent viral infectious diseases from spreading, but to date, there are no studies in the literature on the sensitivity of ccov to chemical biocides. the present study investigated the sensitivity of ccov to dis ...200718035977
comparison of the prevalence of enteric viruses in healthy dogs and those with acute haemorrhagic diarrhoea by electron microscopy.to evaluate prevalence of enteric viruses in healthy dogs and to compare it with prevalences in dogs with acute haemorrhagic diarrhoea.200818086157
serosurvey of ex situ giant pandas (ailuropoda melanoleuca) and red pandas (ailurus fulgens) in china with implications for species conservation.conservation strategies for the giant panda (ailuropoda melanoleuca) include the development of a self-sustaining ex situ population. this study examined the potential significance of infectious pathogens in giant pandas ex situ. serologic antibody titers against canine distemper virus (cdv), canine parvovirus (cpv), canine adenovirus (cav), canine coronavirus (ccv), canine herpesvirus, canine parainfluenza virus (cpiv), toxoplasma gondii, neospora caninum, and leptospira interrogans were measur ...200718229861
molecular characterization of the 9.36 kb c-terminal region of canine coronavirus 1-71 strain.a total of 9.36 kb nucleotides of the c-terminal one-third genome of the canine coronavirus (ccv) 1-71 strain, including all the structural protein genes and some non-structural protein (nsp) genes were cloned and sequenced. nucleotide and amino acid sequence alignment as well as phylogenetic analysis of all the structural orfs with reference coronavirus strains showed that ccv 1-71 was highly related to chinese ccv isolates. this indicates that these ccv stains may have the same ancestor. two l ...200818306032
detection of antigenic heterogeneity in feline coronavirus nucleocapsid in feline pyogranulomatous meningoencephalitis.a new monoclonal antibody (mab), ccv2-2, was compared with the widely used fipv3-70 mab, both directed against canine coronavirus (ccov), as a diagnostic and research tool. western blot showed that both anti-ccov mabs only reacted with a protein of 50 kd, a weight consistent with the feline coronavirus (fcov) viral nucleocapsid. a competitive inhibition enzyme-linked immunosorbent assay showed that the 2 recognized epitopes are distinct. preincubation of ccv2-2 mab with fcov antigen suppressed t ...200818424826
canine respiratory coronavirus: an emerging pathogen in the canine infectious respiratory disease complex.infectious respiratory disease in dogs is a constant challenge because of the involvement of several pathogens and environmental factors. canine respiratory coronavirus (crcov) is a new coronavirus of dogs, which is widespread in north america, japan, and several european countries. crcov has been associated with respiratory disease, particularly in kenneled dog populations. the virus is genetically and antigenically distinct from enteric canine coronavirus; therefore, specific tests are require ...200818501280
etiologic study of upper respiratory infections of household dogs.infectious tracheobronchitis (itb), also known as the kennel cough, is a respiratory syndrome of dogs and usually appears to be contagious among dogs housed in groups. etiologic agent of itb is multiple and sometimes complex. in the present study, 68 household dogs showing clinical signs of respiratory infection were examined, and 20 dogs (29.4%) were found to be positive for either of following agents. bordetella bronchiseptica (b.b.) was most frequently detected from nasal and oropharynx sites ...200818628596
an update on canine coronaviruses: viral evolution and pathobiology.the emergence of human severe acute respiratory syndrome incited renewed interest in animal coronaviruses (covs) as potential agents of direct and indirect zoonoses. the reinforced epidemiological surveillance on covs has led to the identification of new viruses, genotypes, pathotypes and host variants in animals and humans. in dogs, a cov associated with mild enteritis, canine coronavirus (ccov), has been known since 1970s. cov strains with different biological and genetic properties with respe ...200818635322
gain, preservation, and loss of a group 1a coronavirus accessory glycoprotein.coronaviruses are positive-strand rna viruses of extraordinary genetic complexity and diversity. in addition to a common set of genes for replicase and structural proteins, each coronavirus may carry multiple group-specific genes apparently acquired through relatively recent heterologous recombination events. here we describe an accessory gene, orf3, unique to canine coronavirus type i (ccov-i) and characterize its product, glycoprotein gp3. whereas orf3 is conserved in ccov-i, only remnants rem ...200818667517
prevalence of antibodies to canine respiratory coronavirus in some dog populations in japan. 200818806283
quantification of mrna encoding cytokines and chemokines and assessment of ciliary function in canine tracheal epithelium during infection with canine respiratory coronavirus (crcov).one of the first lines of defence against viral infection is the innate immune response and the induction of antiviral type i interferons (ifns). however some viruses, including the group 2 coronaviruses, have evolved mechanisms to overcome or circumvent the host antiviral response. canine respiratory coronavirus (crcov) has previously been shown to have a widespread international presence and has been implicated in outbreaks of canine infectious respiratory disease (cird). this study aimed to q ...200918977539
development of a quantitative real-time pcr for the detection of canine respiratory coronavirus.canine respiratory coronavirus (crcov) has been detected recently in dogs with canine infectious respiratory disease and is involved in the clinical disease complex. crcov is a group 2 coronavirus most closely related to bovine coronavirus and human coronavirus oc43. a real-time pcr assay was developed for the detection of crcov. the assay was validated against cell culture grown virus and shown to have a high level of sensitivity. a range of tissue samples were collected from dogs at a re-homin ...200919013196
recombinant canine coronaviruses related to transmissible gastroenteritis virus of swine are circulating in dogs.four canine coronavirus type ii (ccov-ii) strains were identified in the guts and internal organs of pups which had died of acute gastroenteritis. the ccov-ii strains were strictly related to porcine transmissible gastroenteritis virus (tgev) in the n-terminal domain of the spike protein, whereas in the other parts of the genome, a higher genetic relatedness to recent ccov-ii isolates was observed. experimental infection of dogs with a tgev-like isolate induced mild gastroenteritis without any s ...200919036814
molecular characterization of a canine respiratory coronavirus strain detected in italy.coronaviruses (covs) are positive-stranded, non-segmented rna viruses generally responsible for the emergence of respiratory and enteric disease in humans, companion animals and livestock. their aptitude to evolve by genetic recombination and/or point mutation is recognized, thus giving rise to new viral genotypes and mutants with different tissues or host tropism. in particular, a probable origin from the strictly related bovine coronavirus (bcov) or, alternatively, from a common ancestor has b ...200919162098
sequence analysis of divergent canine coronavirus strains present in a uk dog population.forty faecal samples were tested by rt-pcr using coronavirus consensus primers to determine faecal shedding of canine coronavirus (ccov) and canine respiratory coronavirus (crcov) in a dog population housed at a rescue centre. seven samples were positive for ccov while all samples were negative for crcov. sequence analysis of five ccov strains showed a high similarity with transmissible gastroenteritis virus (tgev) at the n-terminus of the spike protein. all strains contained an open reading fra ...200919162099
analysis of putative recombination hot sites in the s gene of canine coronaviruses.the s gene sequence of canine coronavirus strain 1-71 (ccov 1-71) was cloned, sequenced, and compared to those of other ccovs, transmissible gastroenteritis virus (tgev), and feline coronavirus (fcov). the sequence analysis showed that ccov 1-71 displayed a 98.8-99.8% identity with ccovs strains v1, k378, and gp. four putative recombination sites were found at the 5'-end of the s gene, namely at nt 53, 75, 425, 991. both sequences flanking each site were significantly different. three recombinat ...200919537912
prevalence of canine enteric coronavirus in a cross-sectional survey of dogs presenting at veterinary practices.in order to determine the prevalence of canine enteric coronavirus (cecov) in the general dog population, faecal samples were obtained in a cross-sectional study of 249 dogs presenting for any reason at veterinary practices randomly selected from across the uk. demographic and clinical data was obtained for each of the samples, including signalment, number of dogs in the household, reason for visiting the practice, and any recent history of diarrhoea. the samples were tested by rt-pcr for the pr ...201019647379
isolation and identification of a canine coronavirus strain from giant pandas (ailuropoda melanoleuca).two giant pandas (ailuropoda melanoleuca) died of unknown causes in a chinese zoo. the clinical disease profile suggested that the pandas may have suffered a viral infection. therefore, a series of detection including virus isolation, electron microscopy, cytobiological assay, serum neutralization and rt-pcr were used to identify the virus. it was determined that the isolated virus was a canine coronavirus (ccv), on the basis of coronavirus, neutralization by canine anti-ccv serum, and 84.3% to ...200919687628
bid cleavage, cytochrome c release and caspase activation in canine coronavirus-induced apoptosis.a previous study demonstrated that infection of a canine fibrosarcoma cell line (a-72 cells) by canine coronavirus (ccov) resulted in apoptosis (ruggieri et al., 2007). in this study, we investigated the cell death processes during infection and the underlying mechanisms. we found that ccov-ii triggers apoptosis in a-72 cells by activating initiator (caspase-8 and -9) and executioner (caspase-3 and -6) caspases. the proteolytic cleavage of poly(adp-ribose) polymerases (parps) confirmed the activ ...201019781871
the seroprevalence of canine respiratory coronavirus and canine influenza virus in dogs in new zealand.to determine whether canine respiratory coronavirus (crcov) and canine influenza virus (civ) are present in dogs in new zealand.200919802044
genetic analysis of canine group 2 coronavirus in korean dogs.three out of 109 mixed lung and tracheal tissue extracts originating from korean dogs tested positive for infection with canine respiratory coronavirus (crcov) of coronaviridae group 2. the three crcov-positive samples were found to be competent for viral propagation and isolation, using human rectal tumor cells (hrt-18), and the structure and nonstructure proteins encoded in the 3'-end of the crcov genome were sequenced. the small open reading frames situated between the spike and envelope gene ...201019819086
an elisa based on recombinant spike protein s for the detection of antibodies to transmissible gastroenteritis virus of swine-like canine coronaviruses.recombinant canine coronaviruses, resembling the transmissible gastroenteritis virus of swine (tgev) in a 5' fragment of the s glycoprotein, have been detected recently and showed to be present in canine populations. the 5' fragment of the s protein (s') of a tgev-like canine coronavirus (ccov), strain 174/06, was expressed in an escherichia coli cell-free system. the purified recombinant polypeptide was employed to develop an elisa test for the detection of tgev-like ccov-specific antibodies in ...201019878695
immunity after natural exposure to enteric canine coronavirus does not provide complete protection against infection with the new pantropic cb/05 strain.recently, an outbreak of fatal infection caused by a pantropic variant (strain cb/05) of canine coronavirus (ccov) has been reported. in this study, evidence is provided that immunity induced by natural exposure to enteric ccov is not fully protective against strain cb/05. twenty-two, 10-week-old beagles with a recent natural infection by enteric ccov were randomly distributed in two experimental groups of eight (groups a and b) and one control group of six (group c) dogs. dogs in groups a and b ...201019887130
serosurvey of selected viruses in captive giant pandas (ailuropoda melanoleuca) in china.serum samples from 92 giant pandas in three captive facilities were tested for antibodies against five viruses of carnivores. antibody titers against canine distemper virus (cdv) in two facilities in which giant pandas were vaccinated were variable. the canine adenovirus (cav-1) and canine parvovirus (cpv) titers in vaccinated group were both positive, but titers were not high and varied among individual except one vaccinated panda had extremely high cav-1 titer, indicating infection with the fi ...201019913371
western european epidemiological survey for parvovirus and coronavirus infections in dogs.an epidemiological survey for canine parvovirus (cpv) and canine coronavirus (ccov) infections was conducted in western europe. a total of 156 faecal samples were collected from dogs with diarrhoea in spain (n=47), italy (n=39), france (n=26), germany (n=21), the united kingdom (n=8), belgium (n=10), and the netherlands (n=5). using molecular assays for virus detection and characterisation, cpv and ccov were found to be widespread in european dog populations, either alone or in mixed infections. ...201119932978
comprehensive detection and identification of seven animal coronaviruses and human respiratory coronavirus 229e with a microarray hybridization assay.based on microarray hybridization, a diagnostic test for coronavirus infection was developed using eight coronavirus strains: canine coronavirus (ccov), feline infectious peritonitis virus (fipv), feline coronavirus (fcov), bovine coronavirus (bcov), porcine respiratory coronavirus (prcov), turkey enteritis coronavirus (tcov), transmissible gastroenteritis virus (tgev), and human respiratory coronavirus (hrcov). up to 104 cdna clones of eight viruses were obtained by reverse transcription pcr wi ...201019955814
effect of recombinant lactobacillus expressing canine gm-csf on immune function in dogs.many lactobacillus strains have been promoted as good probiotics for the prevention and treatment of diseases. we engineered recombinant lactobacillus casei, producing biologically active canine granulocyte macrophage colony stimulating factor (cgm-csf), and investigated its possibility as a good probiotic agent for dogs. expression of the cgm-csf protein in the recombinant lactobacillus was confirmed by sds-page and western blotting methods. for the in vivo study, 18 beagle puppies of 7 weeks o ...200919996694
recombinant canine coronaviruses in dogs, europe.coronaviruses of potential recombinant origin with porcine transmissible gastroenteritis virus (tgev), referred to as a new subtype (iib) of canine coronavirus (ccov), were recently identified in dogs in europe. to assess the distribution of the tgev-like ccov subtype, during 2001-2008 we tested fecal samples from dogs with gastroenteritis. of 1,172 samples, 493 (42.06%) were positive for ccov. ccov-ii was found in 218 samples, and ccov-i and ccov-ii genotypes were found in 182. approximately 20 ...201020031041
an outbreak of canine coronavirus in puppies in a greek kennel.canine coronavirus (ccov) is usually the cause of mild gastroenteritis in dogs and is known to have spread worldwide. however, to date, no ccov cases have been confirmed in greece. in the present work, the authors investigated an outbreak of enteritis in puppies from a greek kennel for the presence of ccov. dogs were presented with clinical signs of diarrhea, anorexia, weakness, depression, dehydration, and 1 death. canine coronavirus type ii was detected by reverse transcription nested polymera ...201020224103
a multiplex reverse transcription-nested polymerase chain reaction for detection and differentiation of wild-type and vaccine strains of canine distemper virus.a multiplex reverse transcription-nested polymerase chain reaction (rt-npcr) method was developed for the detection and differentiation of wild-type and vaccine strains of canine distemper virus (cdv). a pair of primers (p1 and p4) specific for cdv corresponding to the highly conserved region of the cdv genome were used as a common primer pair in the first-round pcr of the nested pcr. primers p2 specific for cdv wild-type strains, were used as the forward primer together with the common reverse ...201020433759
detection and genotyping of canine coronavirus rna in diarrheic dogs in japan.to clarify the prevalence of canine coronavirus (ccov) infection in japan, faecal samples from 109 dogs with diarrhoea were examined for ccov rna together with canine parvovirus type 2 (cpv-2) dna. the detection rates of ccov and cpv-2 for dogs aged less than 1 year were 66.3% and 43.8%, while those for dogs aged 1 year or older were 6.9% and 10.3%, respectively, which were significantly different (p<0.0001 and p=0.0003, respectively), indicating not cpv-2 but ccov is an important diarrhoea-caus ...201120557915
prolonged depletion of circulating cd4+ t lymphocytes and acute monocytosis after pantropic canine coronavirus infection in dogs.a hypervirulent strain (cb/05) of canine coronavirus was employed to infect oronasally 11-week-old pups. peripheral blood monocytes (cd14(+)), t lymphocytes (cd4(+) and cd8(+)) and b lymphocytes (cd21(+)) were studied by flow cytometry within 5 days post-infection (p.i.) and at later time points. infection with cb/05 resulted in a profound depletion of t cells and a slight loss of b cells in the first week p.i. in particular, while the cd8(+) and the b lymphocytes returned to baseline levels by ...201020558216
epidemiology of canine parvovirus and coronavirus in dogs presented with severe diarrhoea to pdsa petaid hospitals.canine parvovirus (cpv) and canine enteric coronavirus (cecov) are often cited as causes of diarrhoea in dogs. this study aimed to determine the prevalence of cpv and cecov in dogs presenting with severe diarrhoea to pdsa petaid hospitals throughout the uk. a total of 355 samples were collected from the pdsa between 2006 and 2008. all samples were tested for cpv using a long range pcr and for cecov using rt-pcr. the prevalence of cpv was 58 per cent (95 per cent confidence interval [ci] 52 to 63 ...201020693502
detection and genetic characterization of canine parvoviruses and coronaviruses in southern ireland.canine parvovirus (cpv) and canine coronavirus (ccov) are considered the main pathogens responsible for acute gastroenteritis in dogs. from a collection of 250 samples, seven cpv strains and three ccov strains were identified in symptomatic irish dogs. samples were screened for the viruses using polymerase chain reaction (pcr) and typed via dna sequence analysis. three cpv strains were characterized as cpv-2a, while four others were characterized as cpv-2b. to date, cpv-2c remains unreported in ...201021107617
immunogenicity and protective efficacy in dogs of an mf59™-adjuvanted vaccine against recombinant canine/porcine coronavirus.recently, canine coronavirus (ccov) strains with putative recombinant origin with porcine transmissible gastroenteritis virus (tgev) were shown to be widespread in europe. in this study, a killed vaccine against tgev-like ccov strains, included in the new subtype ccov-iib, was developed through inactivation with betapropiolactone and emulsification with mf59™ adjuvant. safety, immunogenicity and efficacy of the developed vaccine were evaluated in vivo. five 10-week-old beagle pups were administe ...201121272607
basic science track. entry and release of canine coronavirus from polarized epithelial cells.virus entry into and release from epithelial cells are polarized as a result of the distribution of the viral receptors. in order to establish the polarity of entry and release of ccov from epithelial cells, the interactions of the virus with a72 and crfk cells grown on permeable supports was evaluated, and the amount of infective virus in the apical and in the basolateral media was determined and compared. infection of a72 cells after different times post seeding demonstrated that ccov grow aft ...201121344143
isolation, tissue distribution and molecular characterization of two recombinant canine coronavirus strains.canine coronavirus (ccov) is an enveloped rna virus, responsible for gastrointestinal infection in dogs. to date, two different ccov genotypes have been recognized, ccov type i and ccov type ii. recently, ccov type ii strains of potential recombinant origin with transmissible gastroenteritis virus (tgev) were detected and characterized as a new subtype (ccov-iib) of canine coronavirus, in order to be differentiated from the "classical" ccov type ii strains (ccov-iia). in the present study, two c ...201121481551
Canine coronavirus: not only an enteric pathogen.This article reviews the currently available literature on pantropic canine coronavirus (CCoV), providing a meaningful update on the virologic, epidemiologic, clinical, diagnostic, and prophylactic aspects of the infections caused by this emerging pathogen of dogs. It also describes pantropic CCoV-induced disease reproduced under experimental conditions.201122041207
involvement of foxo transcription factors, trail-fasl/fas, and sirtuin proteins family in canine coronavirus type ii-induced apoptosis.n our previous study, we have shown that canine coronavirus type ii (ccov-ii) activates both extrinsic and intrinsic apoptotic pathway in a canine fibrosarcoma cell line (a-72 cells). herein we investigated the role of sirtuin and forkhead box o (foxo) families in this experimental model using nortern blot and western blot analysis. our results demonstrated that mitochondrial sirt3 and sirt4 protein expression increased from 12 and 24 h post infection (p.i.) onwards, respectively, whereas the nu ...201122087287
molecular characterization of a canine coronavirus na/09 strain detected in a dog's organs.in the present study, the detection of a pantropic canine coronavirus (ccov) strain in a dog with lethal diarrhoea is reported. rt-pcr and real-time rt-pcr assays were used for the detection, characterization and quantitation of ccov. sequence and phylogenetic analysis of the ccov na/09 revealed a high degree of sequence identity with the pantropic strain cb/05, indicating the presence of cb/05-like pantropic strains in greece. the absence of the 38-nucleotide deletion in orf3b, which is charact ...201122002680
development of a combined canine distemper virus specific rt-pcr protocol for the differentiation of infected and vaccinated animals (diva) and genetic characterization of the hemagglutinin gene of seven chinese strains demonstrated in dogs.a combined reverse-transcription polymerase chain reaction (rt-pcr) method was developed for the detection and differentiation of wild-type and vaccine strains of the canine distemper virus (cdv). a pair of primers (p1/p2) was used to detect both cdv wild-type strains and vaccines. another pair (p3/p4) was used to detect only cdv wild-type strains. a 335bp fragment was amplified from the genomic rna of the vaccine and wild-type strains. a 555bp fragment was amplified specifically from the genomi ...201222108430
coronavirus genomics and bioinformatics analysis.the drastic increase in the number of coronaviruses discovered and coronavirus genomes being sequenced have given us an unprecedented opportunity to perform genomics and bioinformatics analysis on this family of viruses. coronaviruses possess the largest genomes (26.4 to 31.7 kb) among all known rna viruses, with g + c contents varying from 32% to 43%. variable numbers of small orfs are present between the various conserved genes (orf1ab, spike, envelope, membrane and nucleocapsid) and downstrea ...201021994708
contact with domestic dogs increases pathogen exposure in endangered african wild dogs (lycaon pictus).infectious diseases have contributed to the decline and local extinction of several wildlife species, including african wild dogs (lycaon pictus). mitigating such disease threats is challenging, partly because uncertainty about disease dynamics makes it difficult to identify the best management approaches. serious impacts on susceptible populations most frequently occur when generalist pathogens are maintained within populations of abundant (often domestic) "reservoir" hosts, and spill over into ...201222238695
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