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ABSTRACT: Canine coronavirus (CCoV) is an etiologic agent of diarrhea in dogs and is known to have spread worldwide. Mild disease or asymptomatic carriage are probably in many cases common outcomes of infection. To date, two different genotypes of CCoV are known, CCoV type I (CCoV-I) and CCoV type II (CCoV-II). CCoV type II is divided in two subtypes, CCoV-IIa (classical strains) and CCoV-IIb, with CCoV-IIb emerging as a result of a putative recombination between CCoV-IIa and transmissible gastroenteritis virus (TGEV). The aim of the present study was to investigate the presence of CCoV in Greece and to genetically analyze the circulating strains. Between December 2007 and December 2009, 206 fecal samples were collected from dogs with diarrhea from kennels, pet shops and veterinary clinics of different country regions. RT-PCR and real time RT-PCR assays were used for CCoV detection and characterization. CCoV was identified in 65.1% of the dogs presenting diarrhea, being more frequently detected in animals younger than 3months old and in animals housed in groups. In 47% of the positive samples more than one CCoV genotype/subtype were detected, with triple CCoV-I/CCoV-IIa/CCoV-IIb infections being identified for the first time. Molecular and phylogenetic analysis revealed that CCoV-I Greek strains share low genetic relatedness to each other and to the prototype CCoV-I strains in the 5' end of the S gene. Moreover, a divergent CCoV-IIa strain was identified. The circulation of highly variable CCoV-I and CCoV-IIb emerging strains, as well as the detection of the divergent strain, raise concerns on the importance of these new strains as primary pathogens of diarrhoeic syndromes diagnosed in dogs.
Infection, genetics and evolution: journal of molecular epidemiology and evolutionary genetics in infectious diseases 02/2013; · 3.22 Impact Factor
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Nicola Decaro,
Nathalie Cordonnier,
Zoltan Demeter,
Herman Egberink,
Gabriella Elia,
Aurélien Grellet,
Sophie Le Poder,
Viviana Mari,
Vito Martella, Vasileios Ntafis,
Marcela von Reitzenstein,
Peter J Rottier,
Miklos Rusvai,
Shelly Shields,
Eftychia Xylouri,
Zach Xu,
Canio Buonavoglia
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ABSTRACT: Highly virulent canine coronavirus (pantropic CCoV) strains belonging to subtype CCoV-IIa were recently identified in dogs. To assess the distribution of such strains in Europe, tissue samples were collected from 354 dogs that had died after displaying systemic disease in France (n = 92), Hungary (n = 75), Italy (n = 69), Greece (n = 87), The Netherlands (n = 27), Belgium (n = 4), and Bulgaria (n = 1). A total of 124 animals tested positive for CCoV, with 33 of them displaying the virus in extra-intestinal tissues. Twenty-four CCoV strains (19.35% of the CCoV-positive dogs) detected in internal organs were characterised as subtype IIa and consequently assumed to be pantropic CCoVs. Sequence and phylogenetic analyses of the 5' end of the spike-protein gene showed that pantropic CCoV strains are closely related to each other with the exception of two divergent French viruses that clustered with enteric strains.
Journal of clinical microbiology 10/2012; · 4.16 Impact Factor
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ABSTRACT: 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 characteristic of CB/05, indicates the need to identify new genetic markers for pantropic variants of CCoV, probably in the spike-protein gene region.
Archives of Virology 01/2012; 157(1):171-5. · 2.11 Impact Factor
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ABSTRACT: For first time in Greece equine influenza virus infection was confirmed, by isolation and molecular analysis, as the cause of clinical respiratory disease among unvaccinated horses during 2003 and 2007 outbreaks.
Equine influenza virus (EIV) H3N8 was isolated in MDCK cells from 30 nasal swabs from horses with acute respiratory disease, which were tested positive by Directigen Flu A. Isolation was confirmed by haemagglutination assay and RT-PCR assay of the M, HA and NA gene.
HA sequences of the Greek isolates appeared to be more closely related to viruses isolated in early 1990s in Europe. These results suggested that viruses with fewer changes than those on the main evolutionary lineage may continue to circulate. On the other hand, analysis of deduced NA amino acid sequences were more closely related to viruses isolated in outbreaks in Europe and Asia during 2003-2007. Phylogenetic analysis characterized the Greek isolates as a member of the Eurasian lineage by the haemagglutinin (HA) protein alignment, but appeared to be a member of the Florida sublineage clade 2 by the neuraminidase (NA) protein sequence suggesting that reassortment might be a possible explanation.
Our findings suggest that the Greek strains represent an example of "frozen evolution" and probably reassortment between genetically distinct co-circulated strains. Therefore expanding current equine influenza surveillance efforts is a necessity.
Virology Journal 01/2011; 8:350. · 2.34 Impact Factor
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ABSTRACT: The aim of the present study was to characterize Canine parvovirus 2 (CPV-2) variants currently circulating in Greece. Between March 2008 and March 2009, 167 fecal samples were collected from diarrheic dogs from different regions of Greece. Canine parvovirus 2 was detected by standard polymerase chain reaction, whereas minor groove binder probe assays were used to distinguish genetic variants and discriminate between vaccine and field strains. Of 84 CPV-2-positive samples, 81 CPV-2a, 1 CPV-2b, and 2 CPV-2c were detected. Vaccine strains were not detected in any sample. Sequence analysis of the VP2 gene of the 2 CPV-2c viruses revealed up to 100% amino acid identity with the CPV-2c strains previously detected in Europe. The results indicated that, unlike other European countries, CPV-2a remains the most common variant in Greece, and that the CPV-2c variant found in Europe is also present in Greece.
Journal of veterinary diagnostic investigation: official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 09/2010; 22(5):737-40. · 1.21 Impact Factor
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ABSTRACT: An outbreak of norovirus (NoV) infection was identified in a kennel. Sequence analysis of a short fragment in the polymerase complex indicated the clonal origin of the strains, which were similar to the prototype canine NoV strain GIV.2/Bari/170/07-4/ITA (94.7% nucleotide identity). The findings demonstrate that canine NoV circulates in dogs in Greece and that it can spread easily across a group of animals.
Journal of clinical microbiology 07/2010; 48(7):2605-8. · 4.16 Impact Factor
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ABSTRACT: 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 polymerase chain reaction in all 11 puppies, whereas 1 puppy presented dual infection with CCoV type II and canine parvovirus 2. Surprisingly, sequence analysis of the samples revealed higher similarity to the pantropic CCoV II strain CB/05 than to other reference strains, in the most variable region of the S gene.
Journal of veterinary diagnostic investigation: official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 03/2010; 22(2):320-3. · 1.21 Impact Factor
