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1.  Feline fecal virome reveals novel and prevalent enteric viruses 
Veterinary microbiology  2014;171(0):102-111.
Humans keep more than 80 million cats worldwide, ensuring frequent contacts with their viruses. Despite such interactions the enteric virome of cats remains poorly understood. We analyzed a fecal sample from a single healthy cat from Portugal using viral metagenomics and detected five eukaryotic viral genomes. These viruses included a novel picornavirus (proposed genus “Sakobuvirus”) and bocavirus (feline bocavirus 2), a variant of feline astrovirus 2 and sequence fragments of a highly divergent feline rotavirus and picobirnavirus. Feline sakobuvirus A represents the prototype species of a proposed new genus in the Picornaviridae family, distantly related to human salivirus and kobuvirus. Feline astroviruses (mamastrovirus 2) are the closest relatives of the classic human astroviruses (mamastrovirus 1), suggestive of past cross-species transmission. Presence of these viruses by PCR among Portuguese cats was detected in 13% (rotavirus), 7% (astrovirus), 6% (bocavirus), 4% (sakobuvirus), and 4% (picobirnavirus) of 55 feline fecal samples. Co-infections were frequent with 40% (4/10) of cats shedding more than one of these viruses. Our study provides an initial unbiased description of the feline fecal virome indicating a high level of asymptomatic infections. Availability of the genome sequences of these viruses will facilitate future tropism and disease association studies.
PMCID: PMC4080910  PMID: 24793097
metagenomics; virome; enteric virus; Felis catus; sakobuvirus
2.  Fecal virome analysis of three carnivores reveals a novel nodavirus and multiple gemycircularviruses 
Virology Journal  2015;12:79.
More knowledge about viral populations in wild animals is needed in order to better understand and assess the risk of zoonotic diseases. In this study we performed viral metagenomic analysis of fecal samples from three healthy carnivores: a badger (Meles meles), a mongoose (Herpestes ichneumon) and an otter (Lutra lutra) from Portugal.
We detected the presence of novel highly divergent viruses in the fecal material of the carnivores analyzed, such as five gemycircularviruses. Four of these gemycircularviruses were found in the mongoose and one in the badger. In addition we also identified an RNA-dependent RNA polymerase gene from a putative novel member of the Nodaviridae family in the fecal material of the otter.
Together these results underline that many novel viruses are yet to be discovered and that fecal associated viruses are not always related to disease. Our study expands the knowledge of viral species present in the gut, although the interpretation of the true host species of such novel viruses needs to be reviewed with great caution.
PMCID: PMC4459443  PMID: 25986582
Virome; Gemycircularvirus; Metagenomics; viral discovery
3.  Presence of Antibodies against Genogroup VI Norovirus in Humans 
Virology Journal  2013;10:176.
Noroviruses are important enteric pathogens in humans and animals. Recently, we reported a novel canine norovirus (CaNoV) in dogs with diarrhea belonging to a new genogroup (GVI). No data are available on exposure of humans to this virus.
Sera from 373 small animal veterinarians and 120 age-matched population controls were tested for IgG antibodies to CaNoV by a recombinant virus like particle based enzyme-linked immunosorbent assay.
Antibodies to CaNoV were found in 22.3% of the veterinarians and 5.8% of the control group (p < 0.001). Mean corrected OD450 values for CaNoV antibodies were significantly higher in small animal veterinarians compared to the control group.
These findings suggest that CaNoV may infect humans and small animal veterinarians are at an increased risk for exposure to this virus. Additional studies are needed to assess if this virus is able to cause disease in humans.
PMCID: PMC3680240  PMID: 23735311
4.  Serosurvey of veterinary conference participants for evidence of zoonotic exposure to canine norovirus – study protocol 
Virology Journal  2012;9:250.
Noroviruses have emerged as the leading cause of outbreaks and sporadic cases of acute gastroenteritis in humans worldwide. Person-to-person contact and consumption of contaminated food are considered the most important ways of transmission of noroviruses however zoonotic transmission has been suggested. Recently, noroviruses have been found in dogs which, unlike bovine and swine noroviruses, may present a higher risk of zoonotic transfer, given to the often close contacts between humans and pet dogs in many societies across the world. The present paper describes a seroepidemiologic study aiming to provide information on the exposure level of humans to canine norovirus.
A case–control study was designed to address the potential exposure to canine norovirus based on the presence of antibodies against canine norovirus. Sera from veterinarians (a population repeatedly in close contact with dogs) will be collected in an annual Veterinary Sciences Congress in Portugal. In addition, sera from general population will be obtained and used as controls for comparative purposes. All sera will be tested for the presence of canine norovirus antibodies using a virus-like particle-based enzyme immune assay. Risk factors for canine norovirus antibodies presence in veterinarians will be investigated through the delivery of an anonymized questionnaire to the participants.
The present study aims to identify seropositive individuals to canine norovirus and to assess risk profiles among veterinary professionals with occupational exposure to dogs. To our knowledge this is the first study providing information on the potential zoonotic risk of canine norovirus, thus allowing the development of preventive measures and ascertaining potential risks for Public Health resulting from contact to dogs.
PMCID: PMC3495783  PMID: 23110789
Canine norovirus; Occupational exposure; Zoonosis; Veterinarians; Public Health; Risk factors
5.  Molecular epidemiology of canine norovirus in dogs from Portugal, 2007–2011 
Canine noroviruses (NoVs) have been recently described in south European countries and associated with outbreaks of diarrhea in kennels. Unlike human NoV which are known as an important cause of acute gastroenteritis, little is known about the role of canine NoV as pathogens in dogs as well as its epidemiological features.
From 2007–2011, 256 stool samples were collected from dogs across Portugal and tested by RT-PCR for canine NoV. Viral fecal shedding was found to be 23% (60/256). All sequences contained the GLPSG amino acid motif characteristic of the RNA-dependent RNA-polymerase gene of NoVs and had a high nucleotide identity (range 98%–100%) to the canine NoV first described in Portugal. The highest shedding rate was detected during the winter months.
This study shows that canine NoV infection is endemic in the dog population of Portugal. Peak shedding was detected in the winter months, a well-known epidemiologic feature of human NoV infections.
PMCID: PMC3410785  PMID: 22776749
Canine norovirus; Dog; Fecal shedding; Winter; Seasonality
6.  Novel Norovirus in Dogs with Diarrhea 
Emerging Infectious Diseases  2010;16(6):980-982.
To identify the prevalence and genetic variability of noroviruses in dogs, we tested fecal samples by using reverse transcription–PCR. We found canine norovirus in 40% and 9% of dogs with and without diarrhea, respectively. The virus was genetically unrelated to other noroviruses and constitutes a tentative new genogroup.
PMCID: PMC3086253  PMID: 20507751
Norovirus; canine; genogroup; dogs; viruses; zoonoses; dispatch

Results 1-6 (6)