Canine parvovirus type 2 (CPV-2) serological and molecular patterns in dogs with viral gastroenteritis from southern Brazil.

Canine Parvovirus type 2 (CPV-2) is a highly contagious virus that can cause severe systemic disease with gastroenteric symptoms in dogs, particularly in young puppies. Originating from the feline parvovirus in the late 1970s, it swiftly propagated globally, instigating a pandemic in dogs. Despite vaccination advancements, CPV-2 remains a substantial challenge for veterinary professionals and pet owners. This study aimed to contribute knowledge about the current situation of CPV-2 among dogs in southern Brazil. In this study, the sera of 125 dogs (mostly with gastroenteritis symptoms) were screened for antibodies against CPV-2 and their faeces for the virus itself. The results showed that 40% (50/125) of dogs were infected with CPV-2. Most animals (65.5%) had previously been exposed to CPV-2 (with serotitres equal or above 1:40), and only 37.6% had protective antibody titres equal or above 1:80. The findings have also demonstrated that vaccination against CPV-2 significantly reduced the risk of infection, with positive cases decreasing from 56.9% (unvaccinated) to 2.0% (fully vaccinated). Furthermore, the prevalence of CPV-2 decreased as dogs aged, with younger dogs and those with an incomplete or non-existent vaccination history at the highest risk of infection. In conclusion, this study provides valuable insight into the prevalence and risk factors associated with CPV-2 infection in dogs in southern Brazil, thereby providing valuable knowledge for the improvement of veterinary care and pet health.

Recovery of complete genomes of canine parvovirus from clinical samples.

Canine parvovirus (CPV) is a highly pathogenic virus that affects dogs, especially puppies. CPV is believed to have evolved from feline panleukopenia virus (FPV), eventually giving rise to three antigenic types, CPV-2a, 2b, and 2c. CPV-2 is recognized for its resilience in contaminated environments, ease of transmission among dogs, and pathogenicity for puppies. Despite the relevance of the virus, complete genome sequences of CPV available at GenBank, to date, are scarce. In the current study, we have developed a methodology to allow the recovery of complete CPV-2 genomes directly from clinical samples. For this, seven fecal samples from Gurupi, Tocantins, North Brazil, were collected from puppies with clinical signals of viral enteritis, and submitted to viral DNA isolation and amplification. Two multiplex PCR strategies were designed including primers targeting fragments of 400 base pairs (bp) and 1,000 bp along the complete genome. Sequencing was performed with the Nanopore® technology and results obtained with the two approaches were compared. Genome assembly revealed that the 400 bp amplicons generated larger numbers of reads, allowing a more reliable coverage of the whole genome than those attained with primers targeting the larger (1000 bp) amplicons. Nevertheless, both enrichment methodologies were efficient in amplification and sequencing. Viral genome sequences were of high quality and allowed more precise typing and subtyping of viral genomes compared to the commonly employed strategy relying solely on the analysis of the VP2 region, which is limited in scope. The CPV-2 genomes recovered in this study belong to the CPV2a and CPV-2c subtypes, closely related to isolates from the neighboring Amazonian region. In conclusion, the technique reported here may contribute to increase the number of full CPV genomes available, which is essential for understanding the genetic mechanisms underlying the evolution and spread of CPV-2.

Molecular characterization of carnivore protoparvovirus strains circulating in cats in Turkey / Caracterização molecular de cepas de protoparvovírus carnívoros circulantes em gatos na Turquia

Cats are susceptible to feline panleukopenia virus (FPV) and canine parvovirus type 2 (CPV-2). Therefore, coinfection and superinfection with multiple parvovirus strains may occur, resulting in high heterogeneity and recombination. Considering the importance of cats as a potential source of genetic diversity for parvoviruses, we investigated the frequency of parvovirus infection in cats using their blood and fecal samples and performed molecular characterization of parvovirus strains circulating in cat populations. Accordingly, the fecal and blood samples of 60 cats with gastroenteritis symptoms were collected from Turkey's Burdur, Isparta, and Izmit provinces. Of these 15 fecal samples tested as parvovirus-positive by PCR, 14 were confirmed to have been infected with true FPV strains by sequencing analysis. Through the phylogeny analysis, those were located in the FPV cluster, closely related to CPV-2, and one was discriminated in the CPV-2b cluster. Additionally, sequence analysis of the VP2 gene of CPV and FPV revealed that the FPV strains detected in Turkey and the vaccine strains were highly related to each other, with a nucleotide identity of 97.7- 100%. Furthermore, 13 variable positions were detected in VP2 of the field and reference FPV strains. Three synonymous mutations were determined in the VP2 gene. Some amino acid mutations in the VP2 protein-affected sites were considered responsible for the virus's biological and antigenic properties. The partial sequence analysis of the VP2 gene revealed that four FPV strains detected in Turkey have a single nucleotide change from T to G at the amino acid position 384 between the nucleotides 3939-3941, which was reported for the first time. Therefore, these four isolates formed a different branch in the phylogenetic tree. The results suggest that both FPV and CPV-2b strains are circulating in domestic cats in Turkey and cats should be considered as potential sources of new parvovirus variants for cats, dogs and other animals.

Os gatos são suscetíveis ao vírus da panleucopenia felina (FPV) e ao parvovírus canino tipo 2 (CPV-2). Portanto, coinfecção e superinfecção com múltiplas cepas de parvovírus podem ocorrer, resultando em alta heterogeneidade e recombinação. Considerando a importância dos gatos como uma fonte potencial de diversidade genética para parvovírus, investigamos a frequência da infecção por parvovírus em gatos usando suas amostras de sangue e fezes e realizamos a caracterização molecular de cepas de parvovírus circulantes nas populações de gatos. Amostras fecais e de sangue de 60 gatos com sinais de gastroenterite foram coletadas nas províncias de Burdur, Isparta e Izmit, na Turquia. Destas, 15 amostras fecais testaram positivas para parvovírus por PCR e 14 foram confirmadas como infectadas com cepas verdadeiras de FPV por análise de sequenciamento. Através da análise filogenética, aqueles foram localizados no agrupamento FPV que está intimamente relacionado com o CPV-2, e um foi discriminado no agrupamento CPV-2b. Além disso, a análise da sequência do gene VP2 de CPV e FPV revelou que as cepas de FPV detectadas na Turquia e as cepas vacinais eram altamente relacionadas entre si, com uma identidade de nucleotídeos de 97,7-100%. Além disso, 13 posições variáveis foram detectadas em VP2 das cepas de campo e FPV de referência. Três mutações sinônimas foram determinadas no gene VP2. Algumas mutações de aminoácidos nos locais afetados pela proteína VP2 foram consideradas responsáveis pelas propriedades biológicas e antigênicas do vírus. A análise da sequência parcial do gene VP2 revelou que quatro cepas de FPV detectadas na Turquia têm uma única mudança de nucleotídeo de T para G na posição do aminoácido 384 entre os nucleotídeos 3939-3941, o que foi relatado pela primeira vez. Portanto, esses quatro isolados formaram um ramo diferente na árvore filogenética. Os resultados sugerem que ambas as cepas FPV e CPV-2b estão circulando em gatos domésticos na Turquia e os gatos devem ser considerados como fontes potenciais de novas variantes de parvovírus para gatos, cães e outros animais.

Carnivore protoparvovirus 1 in Peruvian dogs: Temporal/geographical and evolutionary dynamics of virus.

Canine parvovirus (CPV) has been recognized all around the world as the causal agent of a contagious and highly mortal disease in domestic dogs. In Peru, the infection is endemic and unvaccinated animals and puppies are the most at risk. In order to analyze viral diversity and determine the evolutionary genetic relationships and transmission dynamic of Peruvian CPV-2, were collected during the period of 2016-2017 rectal swabs from puppies with parvovirosis compatible symptoms. Viral DNA was amplified by PCR using primers that flanked the ends of the viral genome and sequenced by Illumina Miseq platform. Twenty-six genomic sequences (NSP1-VP1) of CPV from several districts in Lima Metropolitan area were obtained. The VP2 gene analysis demonstrated the presence of the New CPV-2a, New CPV-2b and 2c variants. The phylodynamic analysis of the viral genomes determined that all Peruvian sequences were clustered into a big clade named South American clade that emerged from the west region of Europe (Italy). The Time to the Most Recent Common Ancestor (TMRCA) of the South American clade was dated to 1993. Peruvian sequences were distributed into three subclades, and the 92% of these sequences were related to Ecuadorian CPV-2. The results suggests that three independent introduction events of virus from other countries could have occurred, in two of these events, CPV-2 from Ecuador were introduced in Peru in 2003 and 2009, and another introduction event, in 2000, from Europe. Overall, these results indicate a viral genetic relationship between Peruvian with Ecuadorian and European virus, and the circulation of several viral subpopulations in Lima Metropolitan.

Molecular analysis of the full-length VP2 gene of Brazilian strains of canine parvovirus 2 shows genetic and structural variability between wild and vaccine strains.

Canine parvovirus 2 (CPV-2), a highly contagious virus, affects dogs worldwide. Infected animals present severe and acute gastroenteritis which may culminate in death. CPV-2 VP2 protein is responsible for important biological functions related to virus-host interactions. Herein we obtained VP2 full-length gene sequences from Brazilian dogs with bloody diarrhea (n=15) and vaccine strains (n=7) produced by seven different laboratories and marketed in Brazil. All wild sequences and one vaccine strain were classified as CPV-2b and six vaccines were the classic CVP-2. Mutations in VP2 protein from vaccine and wild strains obtained in Brazil and worldwide were analyzed (n=906). Amino acid sequences from vaccine strains remarkably diverge from each other, even that classic CPV-2. Phylogenetic analysis based on VP2 gene and conducted with sequences displaying mutations in epitope regions previously described shows that vaccine strains are distantly related from the wide range of wild CPV-2. The impact of amino acid mutations over VP2 protein structure shows that vaccine and wild strains obtained in this study diverge in loop 3, an epitope region that plays a role in the CPV-2 host range. This is the first analysis of CPV-2 VP2 from commercial vaccine strains in Brazil and wild ones from Minas Gerais State, Brazil, and the first detailed attempt to vaccinal VP2 molecular and structural analyses.

Molecular Investigation of Canine Parvovirus-2 (CPV-2) Outbreak in Nevis Island: Analysis of the Nearly Complete Genomes of CPV-2 Strains from the Caribbean Region.

To date, there is a dearth of information on canine parvovirus-2 (CPV-2) from the Caribbean region. During August-October 2020, the veterinary clinic on the Caribbean island of Nevis reported 64 household dogs with CPV-2-like clinical signs (hemorrhagic/non-hemorrhagic diarrhea and vomiting), of which 27 animals died. Rectal swabs/fecal samples were obtained from 43 dogs. A total of 39 of the 43 dogs tested positive for CPV-2 antigen and/or DNA, while 4 samples, negative for CPV-2 antigen, were not available for PCR. Among the 21 untested dogs, 15 had CPV-2 positive littermates. Analysis of the complete VP2 sequences of 32 strains identified new CPV-2a (CPV-2a with Ser297Ala in VP2) as the predominant CPV-2 on Nevis Island. Two nonsynonymous mutations, one rare (Asp373Asn) and the other uncommon (Ala262Thr), were observed in a few VP2 sequences. It was intriguing that new CPV-2a was associated with an outbreak of gastroenteritis on Nevis while found at low frequencies in sporadic cases of diarrhea on the neighboring island of St. Kitts. The nearly complete CPV-2 genomes (4 CPV-2 strains from St. Kitts and Nevis (SKN)) were reported for the first time from the Caribbean region. Eleven substitutions were found among the SKN genomes, which included nine synonymous substitutions, five of which have been rarely reported, and the two nonsynonymous substitutions. Phylogenetically, the SKN CPV-2 sequences formed a distinct cluster, with CPV-2b/USA/1998 strains constituting the nearest cluster. Our findings suggested that new CPV-2a is endemic in the region, with the potential to cause severe outbreaks, warranting further studies across the Caribbean Islands. Analysis of the SKN CPV-2 genomes corroborated the hypothesis that recurrent parallel evolution and reversion might play important roles in the evolution of CPV-2.

First detection of canine parvovirus 2b DNA in a crab-eating fox pup (Cerdocyon thous, Linnaeus, 1766).

The crab-eating fox (Cerdocyon thous) is a small wild mammal present in all Brazilian biomes and in some countries of South America. This study aimed to verify the involvement of viral infectious agents in the death of a wild crab-eating fox pup (Cerdocyon thous) in Brazil. The Center for Medicine and Research of Wild Animals of the Universidade Estadual Paulista received a free-living crab-eating fox aged approximately 21 days and apparently healthy. After 13 days, the animal presented anorexia, diarrhea, fever, prostration, and neurological signs progressing to death with an inconclusive diagnosis. In a retrospective study, tissue fragments stored at - 80 °C were used to identify nucleic acids from major canine viruses, such as canine parvovirus-2 (CPV-2), canine adenovirus A types 1 and 2, canid alphaherpesvirus 1, and canine distemper virus. The amplified product with the expected length for CPV-2 was obtained from the heart fragment. After performing nucleotide (nt) sequencing of the amplicon, it was possible to demonstrate that the crab-eating fox strain exhibited high (99.8%) nt identity with the CPV-2b prototype (CPV-39 strain). Additionally, deduced amino acid (aa) sequence analysis showed the GAT codon for the aa Asp (D) at position 426 of the CPV-2 viral protein VP2, which characterizes the subtype 2b. To the best of the authors' knowledge, this report describes the first detection of CPV-2b DNA in tissue fragments from a crab-eating fox.

Canine papillomavirus type 16 associated to squamous cell carcinoma in a dog: virological and pathological findings.

Papillomaviruses (PVs) are circular double-stranded DNA virus belonging to Papillomaviridae family. During the infection cycle, PVs translate proteins that can influence cell growth and differentiation, leading to epidermal hyperplasia and papillomas (warts) or malignant neoplasms. Canis familiaris papillomaviruses (CPVs) have been associated with different lesions, such as oral and cutaneous papillomatosis, pigmented plaques, and squamous cell carcinomas (SCCs). Here, we report a clinical case of a mixed bred female dog with pigmented plaques induced by CPV16 (Chipapillomavirus 2) that progressed to in situ and invasive SCCs. Gross and histological findings were characterized, and the lesions were mainly observed in ventral abdominal region and medial face of the limbs. In situ hybridization (ISH) revealed strong nuclear hybridization signals in the neoplastic epithelial cells, as well as in the keratinocytes and koilocytes of the pigmented viral plaques. The full genome of the CPV16 recovered directly from the lesions was characterized, and the phylogenetic relationships were determined. The identification of oncoprotein genes (E5, E6, and E7) by high throughput sequencing (HTS) and their expected domains are suggestive of the malignant transformation by CPV16.

First Molecular Identification of Canine Parvovirus Type 2 (CPV2) in Chile Reveals High Occurrence of CPV2c Antigenic Variant.

Canine parvovirus type 2 (CPV2) is one of the most important intestinal pathogens in dogs and puppies. CPV2 has been evolved into three genetic and antigenic variants (2a, 2b, and 2c), which are distributed worldwide. We reported the first study of genetic diversity of CPV2 in Chile. Sixty-five samples were collected from puppies presenting with severe gastroenteritis and different vaccination statuses. PCR, restriction fragment length polymorphism (RFLP), and partial sequencing of the coding region of the structural viral protein VP2 was performed. Thirty of a total of 65 samples tested positive by PCR out of which 19 were further classified as CPV2c and one as CPV2a using RFLP and Sanger sequencing. The phylogeny was in concordance with the RFLP analysis. This is the first report of the genetic characterization of CPV2 in Chile and reveals a high occurrence of CPV2c.

Canine parvovirus 2b in fecal samples of asymptomatic free-living South American coatis (Nasua nasua, Linnaeus, 1766).

Canine parvovirus type 2 (CPV-2) is classified into three subtypes (CPV-2a, CPV-2b, and CPV-2c) and is the main cause of enteritis and myocarditis in young domestic and wild animals. This study aimed to evaluate the presence of CPV-2 in the feces of asymptomatic free-living coatis from Garden Forest Reserve, Palmital city, SP, Brazil. Fecal samples from 21 coatis (both sexes, different ages, and different aspects of feces) were collected in August 2014 and March 2015. The nucleic acid extracted was submitted to a polymerase chain reaction (PCR) assay to amplify a fragment of the VP2 gene of CPV-2. Eight (38%) fecal samples were positive in the PCR assay and were confirmed by sequencing. The 7 nucleotide (nt) sequences analyzed showed 100% nt identity with the prototype strain of CPV-2b (CPV-39 strain). The analysis of the deduced amino acid (aa) sequence revealed the presence of the GAT codon (aa D-Asp) at position 426 of the VP2 viral protein (subtype 2b). This study describes for the first time the identification of CPV-2b in asymptomatic free-living coatis (Nasua nasua) and suggests that coatis are susceptible to Carnivore protoparvovirus 1 infection and are important as a reservoir and an asymptomatic carrier to other wild and domestic animal species.

Epidemiological, clinical and pathological features of canine parvovirus 2c infection in dogs from southern Brazil / Aspectos epidemiológicos, clínicos e patológicos da infecção pelo parvovirus canino 2c em cães do Sul do Brasil

Canine parvovirus type 2c (CPV-2c) emerged in Europe in the early 2000's and rapidly spread out worldwide. Clinical and molecular data have demonstrated its circulation in Brazilian dogs, yet detailed descriptions of cases are still lacking. This article describes the epidemiological, clinical and pathological features of 24 cases of CPV-2c-associated disease in dogs submitted to veterinary clinics and laboratory diagnosis in southern Brazil (2014-2016). Most affected dogs presented signs/lesions suggestive of parvovirus enteritis: diarrhea, vomiting, hyperemia and hemorrhage of the serous membrane of the small intestine, diffuse segmental granulation, atrophy of the villi, necrosis and fusion of crypts, squamous metaplasia and epithelial syncytia. A number of cases presented features divergent from the classical presentations, including a wide variation in the color of feces (reddish and/or yellowish, light-brownish, orange-brown and brownish), involvement of adults (4/24) and vaccinated dogs (12/24), extensive involvement of the small intestine (8/20) and the presence of pulmonary edema (7/24) and convulsions (3/24). Feces and intestinal fragments submitted to PCR for the CPV-2 VP2 gene and to virus isolation in cell culture yielded positive results in 100% and 58.3% (14/24) of the cases, respectively. Nucleotide sequencing revealed a high nucleotide identity in VP2 (99.4 to 100%) and a consistent mutation at amino acid 426 (asparagine to glutamic acid), considered a signature of CPV-2c. These results confirm the involvement of CPV-2c in the described cases and demonstrate the importance of CPV-2c infection among Brazilian dogs, calling attention of veterinarians to correctly diagnose the disease, mainly considering the frequent atypical presentations.(AU)

O parvovírus canino tipo 2c (CPV-2c) surgiu na Europa no início do ano 2000 e rapidamente se espalhou pelas populações de cães ao redor do mundo. Dados clínicos e moleculares demonstraram a sua circulação em cães brasileiros, porém descrições detalhadas desses casos ainda são escassas. Este artigo descreve os aspectos epidemiológicos, clínicos e patológicos de 24 casos de doença gastroentérica associada com a infecção pelo CPV-2c em cães atendidos em clínicas veterinárias e submetidos ao diagnóstico laboratorial no Sul do Brasil (2014-2016). A maioria dos cães afetados apresentaram sinais e/ou lesões sugestivas de enterite por parvovírus: diarreia, vômitos, hiperemia e hemorragia na membrana serosa do intestino delgado, granulação segmentar difusa, atrofia das vilosidades, necrose e fusão de criptas, metaplasia escamosa e sincícios epiteliais. Alguns casos apresentaram características divergentes das apresentações clássicas, incluindo uma grande variação na cor das fezes (avermelhada e/ou amarelada, marrom-claro, marrom-alaranjada ou amarronzada), a participação dos adultos (4/24) e cães vacinados (12/24), um amplo envolvimento do intestino delgado (8/20), a presença de edema pulmonar (7/24) e convulsões (3/24). As fezes e fragmentos intestinais foram submetidos ao teste de PCR para o gene VP2 do CPV-2, e ao isolamento do vírus em cultura de células produziram resultados positivos em 100% e 58,3% (14/24) dos casos, respectivamente. O sequenciamento dos nucleótidos revelou uma alta identidade de nucleótidos na VP2 (99,4-100%) e uma mutação no aminoácido 426 (asparagina para ácido glutâmico), considerada uma assinatura de CPV-2c. Estes resultados confirmam o envolvimento do CPV-2c nos casos descritos e demonstra a importância da infecção pelo CPV-2c entre os cães do Brasil, chamando a atenção de veterinários para diagnosticar corretamente a doença, principalmente considerando-se as apresentações atípicas frequentes.(AU)

Epidemiological, clinical and pathological features of canine parvovirus 2c infection in dogs from southern Brazil / Aspectos epidemiológicos, clínicos e patológicos da infecção pelo parvovirus canino 2c em cães do Sul do Brasil

Canine parvovirus type 2c (CPV-2c) emerged in Europe in the early 2000's and rapidly spread out worldwide. Clinical and molecular data have demonstrated its circulation in Brazilian dogs, yet detailed descriptions of cases are still lacking. This article describes the epidemiological, clinical and pathological features of 24 cases of CPV-2c-associated disease in dogs submitted to veterinary clinics and laboratory diagnosis in southern Brazil (2014-2016). Most affected dogs presented signs/lesions suggestive of parvovirus enteritis: diarrhea, vomiting, hyperemia and hemorrhage of the serous membrane of the small intestine, diffuse segmental granulation, atrophy of the villi, necrosis and fusion of crypts, squamous metaplasia and epithelial syncytia. A number of cases presented features divergent from the classical presentations, including a wide variation in the color of feces (reddish and/or yellowish, light-brownish, orange-brown and brownish), involvement of adults (4/24) and vaccinated dogs (12/24), extensive involvement of the small intestine (8/20) and the presence of pulmonary edema (7/24) and convulsions (3/24). Feces and intestinal fragments submitted to PCR for the CPV-2 VP2 gene and to virus isolation in cell culture yielded positive results in 100% and 58.3% (14/24) of the cases, respectively. Nucleotide sequencing revealed a high nucleotide identity in VP2 (99.4 to 100%) and a consistent mutation at amino acid 426 (asparagine to glutamic acid), considered a signature of CPV-2c. These results confirm the involvement of CPV-2c in the described cases and demonstrate the importance of CPV-2c infection among Brazilian dogs, calling attention of veterinarians to correctly diagnose the disease, mainly considering the frequent atypical presentations.(AU)

O parvovírus canino tipo 2c (CPV-2c) surgiu na Europa no início do ano 2000 e rapidamente se espalhou pelas populações de cães ao redor do mundo. Dados clínicos e moleculares demonstraram a sua circulação em cães brasileiros, porém descrições detalhadas desses casos ainda são escassas. Este artigo descreve os aspectos epidemiológicos, clínicos e patológicos de 24 casos de doença gastroentérica associada com a infecção pelo CPV-2c em cães atendidos em clínicas veterinárias e submetidos ao diagnóstico laboratorial no Sul do Brasil (2014-2016). A maioria dos cães afetados apresentaram sinais e/ou lesões sugestivas de enterite por parvovírus: diarreia, vômitos, hiperemia e hemorragia na membrana serosa do intestino delgado, granulação segmentar difusa, atrofia das vilosidades, necrose e fusão de criptas, metaplasia escamosa e sincícios epiteliais. Alguns casos apresentaram características divergentes das apresentações clássicas, incluindo uma grande variação na cor das fezes (avermelhada e/ou amarelada, marrom-claro, marrom-alaranjada ou amarronzada), a participação dos adultos (4/24) e cães vacinados (12/24), um amplo envolvimento do intestino delgado (8/20), a presença de edema pulmonar (7/24) e convulsões (3/24). As fezes e fragmentos intestinais foram submetidos ao teste de PCR para o gene VP2 do CPV-2, e ao isolamento do vírus em cultura de células produziram resultados positivos em 100% e 58,3% (14/24) dos casos, respectivamente. O sequenciamento dos nucleótidos revelou uma alta identidade de nucleótidos na VP2 (99,4-100%) e uma mutação no aminoácido 426 (asparagina para ácido glutâmico), considerada uma assinatura de CPV-2c. Estes resultados confirmam o envolvimento do CPV-2c nos casos descritos e demonstra a importância da infecção pelo CPV-2c entre os cães do Brasil, chamando a atenção de veterinários para diagnosticar corretamente a doença, principalmente considerando-se as apresentações atípicas frequentes.(AU)

Epidemiological, clinical and pathological features of canine parvovirus 2c infection in dogs from southern Brazil

ABSTRACT: Canine parvovirus type 2c (CPV-2c) emerged in Europe in the early 2000s and rapidly spread out worldwide. Clinical and molecular data have demonstrated its circulation in Brazilian dogs, yet detailed descriptions of cases are still lacking. This article describes the epidemiological, clinical and pathological features of 24 cases of CPV-2c-associated disease in dogs submitted to veterinary clinics and laboratory diagnosis in southern Brazil (2014-2016). Most affected dogs presented signs/lesions suggestive of parvovirus enteritis: diarrhea, vomiting, hyperemia and hemorrhage of the serous membrane of the small intestine, diffuse segmental granulation, atrophy of the villi, necrosis and fusion of crypts, squamous metaplasia and epithelial syncytia. A number of cases presented features divergent from the classical presentations, including a wide variation in the color of feces (reddish and/or yellowish, light-brownish, orange-brown and brownish), involvement of adults (4/24) and vaccinated dogs (12/24), extensive involvement of the small intestine (8/20) and the presence of pulmonary edema (7/24) and convulsions (3/24). Feces and intestinal fragments submitted to PCR for the CPV-2 VP2 gene and to virus isolation in cell culture yielded positive results in 100% and 58.3% (14/24) of the cases, respectively. Nucleotide sequencing revealed a high nucleotide identity in VP2 (99.4 to 100%) and a consistent mutation at amino acid 426 (asparagine to glutamic acid), considered a signature of CPV-2c. These results confirm the involvement of CPV-2c in the described cases and demonstrate the importance of CPV-2c infection among Brazilian dogs, calling attention of veterinarians to correctly diagnose the disease, mainly considering the frequent atypical presentations.

RESUMO: O parvovírus canino tipo 2c (CPV-2c) surgiu na Europa no início do ano 2000 e rapidamente se espalhou pelas populações de cães ao redor do mundo. Dados clínicos e moleculares demonstraram a sua circulação em cães brasileiros, porém descrições detalhadas desses casos ainda são escassas. Este artigo descreve os aspectos epidemiológicos, clínicos e patológicos de 24 casos de doença gastroentérica associada com a infecção pelo CPV-2c em cães atendidos em clínicas veterinárias e submetidos ao diagnóstico laboratorial no Sul do Brasil (2014-2016). A maioria dos cães afetados apresentaram sinais e/ou lesões sugestivas de enterite por parvovírus: diarreia, vômitos, hiperemia e hemorragia na membrana serosa do intestino delgado, granulação segmentar difusa, atrofia das vilosidades, necrose e fusão de criptas, metaplasia escamosa e sincícios epiteliais. Alguns casos apresentaram características divergentes das apresentações clássicas, incluindo uma grande variação na cor das fezes (avermelhada e/ou amarelada, marrom-claro, marrom-alaranjada ou amarronzada), a participação dos adultos (4/24) e cães vacinados (12/24), um amplo envolvimento do intestino delgado (8/20), a presença de edema pulmonar (7/24) e convulsões (3/24). As fezes e fragmentos intestinais foram submetidos ao teste de PCR para o gene VP2 do CPV-2, e ao isolamento do vírus em cultura de células produziram resultados positivos em 100% e 58,3% (14/24) dos casos, respectivamente. O sequenciamento dos nucleótidos revelou uma alta identidade de nucleótidos na VP2 (99,4-100%) e uma mutação no aminoácido 426 (asparagina para ácido glutâmico), considerada uma assinatura de CPV-2c. Estes resultados confirmam o envolvimento do CPV-2c nos casos descritos e demonstra a importância da infecção pelo CPV-2c entre os cães do Brasil, chamando a atenção de veterinários para diagnosticar corretamente a doença, principalmente considerando-se as apresentações atípicas frequentes.

Molecular characterization of canine parvovirus and canine enteric coronavirus in diarrheic dogs on the island of St. Kitts: First report from the Caribbean region.

Although canine parvovirus (CPV) and canine enteric coronavirus (CCoV) are important enteric pathogens of dogs and have been studied extensively in different parts of the world, there are no reports on these viruses from the Caribbean region. During 2015-2016, a total of 104 diarrheic fecal samples were collected from puppies and adult dogs, with or without hemorrhagic gastroenteritis, on the Caribbean island of St. Kitts (KNA). By PCR, 25 (24%, n=104) samples tested positive for CPV. Based on analysis of the complete deduced VP2 amino acid sequences, 20 of the KNA CPV strains were assigned to new CPV-2a (also designated as CPV-2a-297A). On the other hand, the VP2 genes of the remaining 5 strains were partially characterized, or could not be sequenced. New CPV-2a was the predominant CPV variant in St. Kitts, contrasting the molecular epidemiology of CPV variants reported in most studies from nearby North and South American countries. By RT-PCR, CCoVs were detected in 5 samples (4.8%, n=104). Based on analysis of partial M-protein gene, the KNA CCoV strains were assigned to CCoV-I genotype, and were closely related to CCoV-I strains from Brazil. To our knowledge, this is the first report on detection and genetic diversity of CPV and CCoV in dogs from the Caribbean region, and underscores the importance of similar studies in the other Caribbean islands.

Percepción de la calidad de vida de los médicos residentes de neurologíay neuropediatría en Colombia / Perception of quality of life in neurology and neuropediatrics residents in Colombia

Tanto las entidades públicas en salud como las asociaciones médicas se preocupan por mejorar la calidad deprestación de sus servicios olvidando que uno de los factores condicionantes básicos de la productividad es elgrado de complacencia que los profesionales tienen con su trabajo.Fue un estudio observacional descriptivo, de corte transversal, realizado a través del Cuestionario de calidadde vida profesional (CVP-35), validado en español, que consta de treinta y cinco ítems.Se llevó a cabo un total de veinticinco encuestas, edad media: 29 años. Los resultados en la encuesta de calidadde vida revelan en el ítem “Trabajo que tengo” que el 100% está en valores > 5 (Bastante a Mucho); en “Satisfaccióncon el tipo de trabajo” el 72% está distribuido en valores > 5 (Bastante a Mucho); en “Satisfaccióncon el sueño”, 72% fueron calificados con valores < 5 (Algo o Nada); en “Calidad de vida percibida” 4% lacalificó como “Nada”, 40% “Algo”, 44% “Bastante” y 12% “Mucho”; y en la variable “Estrés emocional”,76% puntuó valores > 5 (Bastante a Mucho).El estudio de calidad de vida profesional medida por el cuestionario CVP-35 mostró que la mayoría de residentesviven en estrato socioeconómico 3 y 68% no tienen vivienda propia. Una gran cantidad de residentes trabajanmás de cuarenta y ocho horas semanales. Llama la atención que el 40% reportó sufrir de alguna patología. Lashoras de sueño diarias son siete en promedio, lo cual es aceptable, sin embargo 72% no están satisfechos conlas horas de sueño; pese a que 76% refieren tener una gran cantidad de trabajo y presentar estrés emocional,72% están satisfechos con el tipo de trabajo y más de la mitad de los residentes consideran tener una buenacalidad de vida.

Both public health institutions and medical associations concerned about improving prevision of heathcare, forgetting that one of the basic conditioning factors of productivity is the degree of complacency that professionals have with their workIt was a descriptive study, cross-sectional, conducted through the questionnaire of quality of professional life (CVP-35), validated in spanish, consisting of thirty- five items.A total of twenty survey was conducted, mean age: 29 years. The results in quality of life survey reveal in item "Work I have" that 100% values is > 5 (Quite to Much); In "Satisfaction with the type of work" 72% is distributed in values > 5 (Quite to Much); In "Satisfaction with sleep" 72 % were rated with values < 5 (Something or Nothing); In "Perceived quality of life" 4% qualify as "Nothing", 40% "Something", 44% "Quite" and 12% "Much"; and in the variable "Emotional stress" 76 % scored values > 5 (Quite to Much).The study of professional quality of life measured by the CVP -35 questionnaire showed that most residents live in socioeconomic stratum 3 and 68% not have their own housing. A lot of residents work more than forty- eight hours per week. It is striking that 40 % reported suffering from some pathology. The average of daily hours of sleep is seven, which it is acceptable, however 72 % are dissatisfied with the sleep hours. Although 76 % reported having a lot of work and present emotional stress, 72 % are satisfied with the type of work and more than half of residents believe they have a good quality of life.

Enteritis caused by type 2c canine parvovirus in a 5-year-old dog

Background: Canine parvovirosis, caused by canine parvovirus type 2 (CPV-2), emerged in the 1970s as an important disease affecting dogs, causing severe hemorrhagic gastroenteritis and death. It can occur in any breed, gender, and age; however, puppies of 4 to 12 weeks of age are most commonly afflicted. In 2000 a new variant of the virus, called CPV- 2c, was discovered, and has been related to hemorrhagic gastroenteritis in dogs with up to 2 years of age, although some cases have been described in older animals with a full vaccination history. This paper reports a case of enteritis by canine parvovirus type 2c (CPV-2c) in a 5-year-old dog. Case: At necropsy a pallid oral and conjunctival mucosae were observed. The small intestine showed a very reddish and wrinkled serosa, the wall was thickened, the mucosae was diffusely wrinkled and yellowed with evidenced Peyer plaques and there was no content in the final portion of the intestine. The mesenteric lymph nodes were enlarged and reddish. Multiple suffusions on the serosa of the stomach, and petechiae and subepicardial suffusions in the heart were observed. The histological findings were, collapse of the lamina propria of the small intestine, and fusion of the villi, necrosis of enterocytes, atrophy and the disappearance of crypts, with dilation of remaining crypts showing large rounded nuclei with one or two evident nucleoli...(AU)

Identification of canine papillomavirus type 1 (CPV1) DNA in dogs with cutaneous papillomatosis / Identificação do DNA de Papillomavirus canino tipo 1 em cães com papilomas cutâneos no Brasil

Canine oral papillomavirus (COPV), also known as Canine Papillomavirus type 1 (CPV1), induces papillomas at the mucous membranes of the oral cavity and at the haired skin of dogs. The classification of Papillomavirus (PV) types is based on the L1 capsid protein and nucleotide sequence; so far, 14 CPV types have been described in several countries, but the molecular characterization of CPV in Brazil is lacking. This study investigated the presence of the PV in seven papillomas from four mixed breed dogs from Londrina/PR, Southern Brazil, by partial sequencing of the L1 gene. Seven exophytic cutaneous lesions were surgically removed and processed for histopathological and molecular characterization. Histopathology confirmed the lesions as viral papillomas due to typical histological features. Polymerase Chain Reaction (PCR) assay using the FAP59 and FAP64 primers targeted the L1 gene followed by sequence analysis of the amplicons identified CPV1 in all evaluated papilloma samples. This study represents the first description of CPV1 DNA associated with canine papillomatosis in Brazil.

O papilomavírus oral canino (COPV), também denominado Papillomavirus canino tipo 1 (CPV1), tem a capacidade de induzir papilomas na mucosa da cavidade oral e também em pele de cães. A classificação dos tipos de papilomavírus (PV) é baseada na proteína L1 do capsídeo e na sequência de nucleotídeos que a codifica. Atualmente são descritos 14 tipos de CPV, no entanto, ainda faltam estudos moleculares relacionados à identificação dos tipos de CPV no Brasil. O objetivo deste estudo foi investigar a presença de PV em fragmentos de papilomas obtidos de quatro cães sem raça definida, provenientes de Londrina/PR, região sul do Brasil, e definir o tipo viral por meio da análise da sequência parcial de nucleotídeos do gene L1. Sete lesões cutâneas foram cirurgicamente removidas e processadas ​​para a caracterização histopatológica e molecular. O exame histopatológico confirmou as lesões como papilomas. Foi realizada reação em cadeia de polimerase (PCR), utilizando os primers FAP59 FAP64 para a amplificação parcial do gene L1, seguida por análise das sequências dos produtos amplificados, que confirmou a presença do CPV1 em todas as amostras avaliadas. Este estudo representa a primeira identificação do DNA de CPV1 associado com papilomatose canina no Brasil.

Enteritis caused by type 2c canine parvovirus in a 5-year-old dog

Background: Canine parvovirosis, caused by canine parvovirus type 2 (CPV-2), emerged in the 1970s as an important disease affecting dogs, causing severe hemorrhagic gastroenteritis and death. It can occur in any breed, gender, and age; however, puppies of 4 to 12 weeks of age are most commonly afflicted. In 2000 a new variant of the virus, called CPV- 2c, was discovered, and has been related to hemorrhagic gastroenteritis in dogs with up to 2 years of age, although some cases have been described in older animals with a full vaccination history. This paper reports a case of enteritis by canine parvovirus type 2c (CPV-2c) in a 5-year-old dog. Case: At necropsy a pallid oral and conjunctival mucosae were observed. The small intestine showed a very reddish and wrinkled serosa, the wall was thickened, the mucosae was diffusely wrinkled and yellowed with evidenced Peyer plaques and there was no content in the final portion of the intestine. The mesenteric lymph nodes were enlarged and reddish. Multiple suffusions on the serosa of the stomach, and petechiae and subepicardial suffusions in the heart were observed. The histological findings were, collapse of the lamina propria of the small intestine, and fusion of the villi, necrosis of enterocytes, atrophy and the disappearance of crypts, with dilation of remaining crypts showing large rounded nuclei with one or two evident nucleoli...

Identification of canine papillomavirus type 1 (CPV1) DNA in dogs with cutaneous papillomatosis / Identificação do DNA de Papillomavirus canino tipo 1 em cães com papilomas cutâneos no Brasil

Canine oral papillomavirus (COPV), also known as Canine Papillomavirus type 1 (CPV1), induces papillomas at the mucous membranes of the oral cavity and at the haired skin of dogs. The classification of Papillomavirus (PV) types is based on the L1 capsid protein and nucleotide sequence; so far, 14 CPV types have been described in several countries, but the molecular characterization of CPV in Brazil is lacking. This study investigated the presence of the PV in seven papillomas from four mixed breed dogs from Londrina/PR, Southern Brazil, by partial sequencing of the L1 gene. Seven exophytic cutaneous lesions were surgically removed and processed for histopathological and molecular characterization. Histopathology confirmed the lesions as viral papillomas due to typical histological features. Polymerase Chain Reaction (PCR) assay using the FAP59 and FAP64 primers targeted the L1 gene followed by sequence analysis of the amplicons identified CPV1 in all evaluated papilloma samples. This study represents the first description of CPV1 DNA associated with canine papillomatosis in Brazil.(AU)

O papilomavírus oral canino (COPV), também denominado Papillomavirus canino tipo 1 (CPV1), tem a capacidade de induzir papilomas na mucosa da cavidade oral e também em pele de cães. A classificação dos tipos de papilomavírus (PV) é baseada na proteína L1 do capsídeo e na sequência de nucleotídeos que a codifica. Atualmente são descritos 14 tipos de CPV, no entanto, ainda faltam estudos moleculares relacionados à identificação dos tipos de CPV no Brasil. O objetivo deste estudo foi investigar a presença de PV em fragmentos de papilomas obtidos de quatro cães sem raça definida, provenientes de Londrina/PR, região sul do Brasil, e definir o tipo viral por meio da análise da sequência parcial de nucleotídeos do gene L1. Sete lesões cutâneas foram cirurgicamente removidas e processadas ​​para a caracterização histopatológica e molecular. O exame histopatológico confirmou as lesões como papilomas. Foi realizada reação em cadeia de polimerase (PCR), utilizando os primers FAP59 FAP64 para a amplificação parcial do gene L1, seguida por análise das sequências dos produtos amplificados, que confirmou a presença do CPV1 em todas as amostras avaliadas. Este estudo representa a primeira identificação do DNA de CPV1 associado com papilomatose canina no Brasil.(AU)

High local genetic diversity of canine parvovirus from Ecuador.

Canine parvovirus (CPV) comprises three antigenic variants (2a, 2b, and 2c) that are distributed globally with different frequencies and levels of genetic variability. CPVs from central Ecuador were herein analyzed to characterize the strains and to provide new insights into local viral diversity, evolution, and pathogenicity. Variant prevalence was analyzed by PCR and partial sequencing for 53 CPV-positive samples collected during 2011 and 2012. The full-length VP2 gene was sequenced in 24 selected strains and a maximum-likelihood phylogenetic tree was constructed using both Ecuadorian and worldwide strains. Ecuadorian CPVs have a remarkable genetic diversity that includes the circulation of all three variants and the existence of different evolutionary groups or lineages. CPV-2c was the most prevalent variant (54.7%), confirming the spread of this variant in America. Ecuadorian CPV-2c strains clustered in two lineages, which represent the first evidence of polyphyletic CPV-2c circulating in South America. CPV-2a strains constituted 41.5% of the samples and clustered in a single lineage. The two detected CPV-2b strains (3.8%) were clearly polyphyletic and appeared related to Ecuadorian CPV-2a or foreign CPV-2b strains. Besides the substitution at residue 426 that is used to identify the variants, two amino acid changes occurred in Ecuadorian strains: Val139Iso and Thr440Ser. Ser(440) occurred in a biologically relevant domain of VP2 and is here described for the first time in CPV. The associations of Ecuadorian CPV-2c and CPV-2a with clinical symptoms indicate that dull mentation, hemorrhagic gastroenteritis and hypothermia occurred more frequently in infection with CPV-2c than with CPV-2a.