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)

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.

Characterization and evaluation of apoptotic potential of double gene construct pVIVO.VP3.NS1.

Viral gene oncotherapy, targeted killing of cancer cells by viral genes, is an emerging non-infectious therapeutic cancer treatment modality. Chemo and radiotherapy in cancer treatment is limited due to their genotoxic side effects on healthy cells and need of functional p53, which is mutated in most of the cancers. VP3 (apoptin) of chicken infectious anaemia (CIA) and NS1 (Non structural protein 1) of Canine Parvovirus-2 (CPV-2) have been proven to have oncolytic potential in our laboratory. To evaluate oncolytic potential of VP3 and NS1 together these genes needed to be cloned in a bicistronic vector. In this study, both these genes were cloned and characterized for expression of their gene products and its apoptotic potential. The expression of VP3 and NS1 was studied by confocal microscopy and flowcytometry. Expression of VP3 and NS1 in pVIVO.VP3.NS1 transfected HeLa cells in comparison to mock transfected cells indicated that the double gene construct expresses both the products. This was further confirmed by flowcytometry where there was increase in cells expressing VP3 and NS1 in pVIVO.VP3.NS1 transfected group in comparison with the mock control group. The apoptotic inducing potential of this characterized pVIVO.VP3.NS1 was evaluated in human cervical cancer cell line (HeLa) by DNA fragmentation assay, TUNEL assay and Hoechst staning. This double construct was observed to induce apoptosis in HeLa cells.

Isolation and identification of canine parvovirus type 2b in Korean dogs / 대한수의학회지

Canine parvovirus (CPV) is a major diarrhea-causing agent in puppies. Since CPV type 2 (CPV-2) emerged in 1978, new antigenic variants including CPV-2a, CPV-2b, and CPV-2c have been identified in many countries. Two puppies died suddenly at a veterinary clinic in Gyeonggi province, South Korea. Two viruses were isolated in A72 cells, confirmed as CPV strains based on a CPV rapid kit and an indirect fluorescence test and designated QIACP1403 and QIACP1404. The nucleotide sequences of complete VP2 genes of QIACP1403 and QIACP1404 were determined, and the corresponding amino acid sequences were deduced. Molecular analyses revealed that the QIACP1403 and QIACP1404 isolates were type CPV-2b. Several mutated amino acids were detected on VP2 gene residues of the two isolates. Phylogenetic analyses showed that the two isolates were most closely related to strain CPV-BM11, which was isolated from Chinese dogs in 2011. Our results suggest that these isolates may be a candidate for a vaccine to prevent CPV infection in dogs after conducting passages of the isolates in an in vitro culture system.

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.

Serologic Survey of Rabies Virus, Canine Distemper Virus and Parvovirus in Wild Raccoon Dogs (Nyctereutes procyonoides koreensis) in Korea

Oral rabies vaccination (ORV) program for the wild animals in rabies risk regions of Korea has been conducted since 2000. Evaluation of ORV program under field condition and information concerning the incidence of exposure to canine distemper and canine parvovirus (CPV) are needed in wild raccoon dogs (Nyctereutes procyonoides koreensis). Ninety four sera of wild raccoon dogs were screened for antibodies against rabies, canine distemper virus (CDV) and CPV in Korea. The overall prevalence of antibodies against rabies virus (RABV), CDV and CPV in wild raccoon dogs was 35.1%, 89.4% and 24.5%, respectively. Comparisons of sero-prevalences of RABV, CDV and CPV were assayed in two regions (Gyeonggi-do and Gangwon-do). The Gyeonggi-do (36.4%) showed higher sero-positive rate against CPV than Gangwon-do (20.8%). In contrast, Gangwon-do (41.7% and 97.2%) showed higher sero-positive rates against RABV and CDV than Gyeonggi-do (13.6% and 63.6%). These results indicate that there was severe circulation of CDV and CPV among wild raccoon dogs in the two regions of Korea. Furthermore, raccoon dogs showing a protective antibody titer (0.5 IU/ml) were 15.9%, suggesting that new rabies control program such as trap-vaccination-release (TVR) should be launched urgently in rabies risk regions.

Isolation and characterization of canine parvovirus type 2c circulating in Uruguay / Isolamento e caracterização da cepa tipo 2c do parvovirus canino circulante no Uruguai

This research reports the first CPV-2c isolation in cell culture (canine fibroma cell line A-72) in Uruguay. The isolates were obtained from 13 rectal swabs of Uruguayan dogs with parvovirosis. Samples were submitted to PCR with two sets of primers, restriction fragment length polymorphism (RFLP), partial sequencing of the gene encoding for VP2 capsid protein and phylogenetic characterization. The strain isolated was confirmed as CPV-2c. These results contribute to a better knowledge of CPV strains circulating in Uruguay and promote an evaluation of the efficacy of heterologous vaccines used to protect against the circulating strains.

Este trabalho relata o primeiro isolamento do CPV-2c em cultura de células (linhagem celular de fibroma canino A-72), no Uruguai. Os isolados foram obtidos a partir de 13 suabes retais de cães uruguaios com parvovirose. As amostras foram submetidas à reação em cadeia da polimerase (PCR) com dois pares de primers, polimorfismo de comprimento de fragmentos de restrição (RFLP), sequenciamento parcial do gene que codifica a proteína capsidial VP2 e caracterização filogenética. A cepa isolada foi confirmada como CPV-2c. Os resultados contribuem para um melhor conhecimento das cepas do CPV circulantes no Uruguai e incitam uma maior investigação sobre a eficácia das vacinas produzidas com cepas heterólogas utilizadas atualmente para proteger contra cepas circulantes.

Partial VP2 sequencing of canine parvovirus (CPV) strains circulating in the state of Rio de Janeiro, Brazil: detection of the new variant CPV-2c

Canine parvovirus (CPV) is the most important enteric virus for dogs and it seems to be undergoing continuous evolution, generating new genetic and antigenic variants throughout the world. The aim of this study was to analyze the distribution of CPV variants from 1995 to 2009 and to investigate the circulation of the new variant CPV-2c in Rio de Janeiro, Brazil. In addition, the clinical features of CPV infection were also reported. After CPV laboratorial confirmation by HA/HI and PCR, thirty-two fecal samples were analyzed by sequencing a 583-bp fragment of the VP2 gene. One sample, collected in 2008 was typed as the new type CPV-2c. All samples from 1995 to 2003 were identified as "new CPV-2a". From 2004 to 2006, both "new CPV-2a" and CPV-2b were observed. From 2006 to 2009, most of the samples were characterized as CPV-2b. The classical signs of CPV enteritis were observed in 16/18 CPV-2a and 5/13 CPV-2b infected puppies. These results show that continuous epidemiological surveillance of CPV strain distribution is essential for studying the patterns of CPV-2a and 2b spread and for determining whether the new variant CPV-2c has become permanently established in Brazilian canine population.

Exposure of pampas fox (Pseudalopex gymnocercus) and crab-eating fox (Cerdocyon thous) from the Southern region of Brazil to Canine distemper virus (CDV), Canine parvovirus (CPV) and Canine coronavirus (CCoV)

The exposure of 13 Brazilian free-ranging nondomestic canids (five pampas fox - Pseudalopex gymnocercus and eight crab-eating fox -Cerdocyon thous) from Southern region of Brazil, to Canine distemper virus (CDV), canine parvovirus (CPV) and Canine coronavirus (CCoV) was investigated. Antibodies against CDV were detected in 38.5 percent (5/13) of the samples. There were anti-CDV antibodies in 60 percent (3/5) of P. gymnocercus and in 25 percent (2/8) of C. thous. The frequency was higher among the adults and males. Eleven canids (84.6 percent) presented antibodies against CPV, 80 percent (4/5) were from P. gymnocercus and 87.5 percent (7/8) were from C. thous. There was no difference in positivity rate against CPV between gender and age. Antibodies against CCoV were detected in 38.5 percent (5/13) of the samples, with 60 percent (3/5) of positivity in P. gymnocercus and 25 percent (2/8) in C. thous. The frequency of antibodies against CCoV was higher among the adults and males. The study showed that these canids were exposed to CDV, CPV and CCoV.

Foi investigada a ocorrência de exposição em 13 canídeos não domésticos de vida livre (cinco graxains-do-campo - Pseudalopex gymnocercus e oito graxains-do-mato - Cerdocyon thous) da região sul do Brasil ao vírus da cinomose canina (CDV), parvovírus canino (CPV) e coronavírus canino (CCoV). Anticorpos contra o CDV foram detectados em 38,5 por cento (5/13) das amostras. Haviam anticorpos anti-CDV em 60 por cento (3/5) dos P. gymnocercus e em 25 por cento (2/8) dos C. thous. A freqüência foi maior entre machos e adultos. Para CPV, 11 canídeos (84,6 por cento) apresentaram anticorpos, 80 por cento (4/5) eram da espécie P. gymnocercus e 87,5 por cento (7/8) eram C. thous. Não houve diferença de positividade para o CPV entre sexos e idades. Anticorpos contra o CCoV foram detectados em 38,5 por cento (5/13) das amostras, sendo 60 por cento (3/5) de positividade entre os P. gymnocercus e 25 por cento (2/8) entre os C. thous. A freqüência de anticorpos para CCoV foi maior entre os machos e adultos. O estudo revelou que estes canídeos foram expostos ao CDV, CPV e CCoV.

Development of a Polyclonal Antibody-based AC-ELISA and Its Comparison with PCR for Diagnosis of Canine Parvovirus Infection / 中国病毒学·英文版

A polyclonal antibody-based antigen-capture ELISA (AC-ELISA) has been developed for detection of Canine parvovirus (CPV) antigens in faecal samples of dogs. The assay uses rabbit anti-CPV polyclonal antibody as the capture antibody, guinea pig anti-CPV polyclonal antibody as tracing antibody and anti-guinea pig HRPO conjugate as the detection system. The optimum dilution of the capture antibody and the tracing antibody capable of detecting the CPV-2 antigens was found to be 1:1 600 and 1:400, respectively, in the check-board titration. In this study, a total of 152 samples (129 faecal samples and 23 cell culture supernatant) were tested both by AC-ELISA and by polymerase chain reaction (PCR). Of the samples tested, 69 and 78 samples were found positive by AC-ELISA and PCR, respectively. The AC-ELISA had relative sensitivity, relative specificity and accuracy of 88.4%, 100.0% and 91.4% respectively. The analytical sensitivity of AC-ELISA was estimated to be 102.8 TCID50/mL whereas PCR sensitivity was 100.8 TCID50/mL. The AC-ELISA is a simple, quick and reliable method for screening large numbers of faecal samples of dogs suspected of CPV infection.

Exposición infantil a plastificantes potencialmente tóxicos en productos de uso oral / Infant exposure to potentially toxic plasticizers in products for oral use

OBJETIVO: Determinar la prevalencia en el uso de productos infantiles orales entre menores de tres años de edad y medir su concentración de ftalatos, sustancias potencialmente tóxicas. MATERIAL Y MÉTODOS: Se realizó, en 1999, una entrevista domiciliaria a 199 madres de niños del área metropolitana de la ciudad de Toluca. Por cromatografía de gases se identificaron y cuantificaron diversos ftalatos de productos de uso oral empleados por los niños participantes y se estimó la contribución de estas fuentes a la ingesta diaria de ftalatos. RESULTADOS: La prevalencia de uso de estos productos fue de 13 por ciento, siendo mayor entre los niños, menores de 18 meses de edad, pertenecientes al estrato socioeconómico bajo. Las concentraciones variaron desde trazas hasta 67.0 por ciento del peso. La exposición media calculada proveniente de los productos manufacturados con policloruro de vinilo y ftalatos fue de 13.94 µg/ kg de peso/día, IC 95 por ciento (9.08, 18.89). CONCLUSIONES: La exposición a ftalatos proveniente de productos para chupar o morder se encuentra dentro de los límites reportados en otros países; sin embargo, otras fuentes pueden incrementarla. Dado que algunos ftalatos han mostrado ser tóxicos en el sistema reproductivo, y este potencial efecto es plausible en el hombre, es necesaria la investigación de otras fuentes y determinar la exposición total a través de biomarcadores.

OBJECTIVE: To estimate the prevalence of oral product use in children less than three years of age, and to measure the concentration of phthalates as potentially toxic products. MATERIAL AND METHODS: In 1999, 199 mothers of children living in the city of Toluca agreed to household interviews. Samples of oral products used by the children were taken and analyzed by gas chromatography to identify and quantify phthalate concentrations, to estimate the daily intake of phthalates from this source. RESULTS: The prevalence of oral product use was 13 percent. Male infants less than 18 months of age of low socioeconomic level used them more frequently. The concentrations ranged from traces to 67 percent weight. The mean exposure to products manufactured with polyvinyl chloride and phthalates was 13.94 µg/kg/day (95 percent CI 9.08, 18.89). CONCLUSIONS: The daily dose of phthalate intake from products for infants to suck or bite did not exceed the recommended limit established in other countries. Nevertheless, other sources can contribute to increase the total dose. Since some phthalates are harmful to the reproductive system in animals and this potential effect may also be expected in humans, it is necessary to assess other sources and determine exposure using biomarkers.

Preparation of canine multivalence serum and its application / 中国免疫学杂志

Objective:To prepare the canine multivalence serum for emergent prevation and theatmentor canines infection disease.Methods:Selecting 20 healthy Kunming dog,which is more than one years old.The canines were immuned by regulary with Canine distemper virus(CDV),Canine parvovirus virus(CPV),Infectious canine hepatitis virus(ICHV),Canine coronavirus(CCV),Canine parainflu-enza virus(CPIV).The blood were draw in freedom germ from neck's artery at canines and then blood was centrifugate for isolating serum.To check antibody's tite, micro serum neutrolization(SN) and hemagglutination inhibition(HI) were used.Results:Canine multivalence serum is successfully preparation and good curative effects in clinic.Conclusion:Canine multivalence serum may be of great value in economics and society and contribute to curative disease.