Epizootiology of canine distemper in naturally infected dogs in Goiânia, Brazil / Epizootiologia da cinomose em cães naturalmente infectados em Goiânia, Brasil

Although the epizootiological profile of canine distemper in Goiânia is unknown, there is clinical evidence for a high incidence of canine distemper virus (CDV) infection among dogs. Therefore, this study determined the epizootiological characteristics of canine distemper in naturally infected dogs. Data of 46 dogs that tested positive for the CDV based on immunochromatography or reverse transcription-polymerase chain reaction were collected. Data on the sex, breed, age, and vaccination status were obtained from these dogs, and extraneural and neural sign analyses were performed. Although, the infected dogs belonged to both sexes, different breeds, and different age groups, a greater proportion of cases were seen in adults (1-6 years), undefined breeds, and unvaccinated dogs. Among the CDV-positive dogs, 10.87% had been vaccinated. In addition, 4.35% showed neural signs, 8.69% showed extraneural signs, and 86.96% showed both. High lethality was observed, with viral antigen and/or DNA detected in 82.61% dead dogs. Only 8.70% of the total CDV-infected dogs remained alive at the time of their assessment.

Embora o perfil epizootiológico da cinomose canina em Goiânia seja desconhecido, há evidencia clínica para alta incidência da infecção pelo vírus da cinomose (CDV) nos cães. Este estudo objetivou determinar as características epizootiológicas da cinomose em cães naturalmente infectados. Dados de 46 cães positivos por imunocromatografia ou reação em cadeia da polimerase via transcriptase reversa para o CDV foram coletados. Dados sobre sexo, raça, idade, estado vacinal foram obtidos desses cães, e os sinais extraneurais e neurais foram analisados. Animais de ambos os sexos, diferentes raças e idades foram acometidos. A maior proporção de casos foi vista em adultos (de um a seis anos), sem raça definida e não vacinados. Dentre os cães positivos, 10,87% haviam sido vacinados. Em adição, 4,35% apresentaram sinais neurais, 8,69% sinais extraneurais e 86,96% mostraram ambos. Alta letalidade foi observada, com o antígeno viral e/ou DNA identificado em 82,61% dos cães que foram a óbito. Apenas 8,7% dos cães infectados permaneceram vivos até o momento da avaliação.

Padronização de um teste ELISA indireto para diagnóstico da cinomose canina / Standardization of an indirect ELISA test for canine diagnosis diagnosis

A cinomose é uma enfermidade causada pelo vírus Canine Distemper Virus (CDV). Essa doença afeta principalmente cães, mas também acomete outras espécies domésticas e selvagens. A imunidade do animal está relacionada ao grau que a esse patógeno vai atingir o organismo do indivíduo. Ela afeta a respiração do animal, pode causar vômito, diarreia, convulsões, podendo levar o animal à óbito. O objetivo do presente trabalho foi padronizar um teste ELISA indireto com antígeno de superfície para o diagnostico cinomose utilizando amostras de soro canino. Para padronização da técnica, fez-se necessário o estudo da diluição do antígeno para identificar a melhor concentração para sensibilização da placa. O teste foi aplicado primeiramente com diferentes diluições do antígeno para detecção do melhor desempenho do antígeno. Feito isso, foi testado em um banco de soro de 45 animais comprovadamente positivos no teste ELISA comercial e em soro de 45 animais comprovadamente negativos no teste ELISA comercial, posteriormente foi calculado o ponto de corte, especificidade e sensibilidade do teste. O teste ELISA indireto se mostrou com excelência como um teste de diagnóstico para a cinomose canina, obtendo-se ponto de corte de densidade óptica de 0,229, sensibilidade de 95,5% e especificidade de 84,4%.

Distemper is a disease or the disease by the CDV virus, Distemper Virus. This disease mainly affects dogs, but also affects other domestic and wild species. The animal's immunity is related to the degree to which it will reach the individual's organism. It affects the animal's breathing, can cause vomiting, diarrhea, convulsions, and can lead to death. The aim of the present work test was to standardize an indirect ELISA for distemper diagnosis in experiments using a surface antigen. For the study of technical identification, it was necessary to specify the antigen for the best concentration of plaque sensitization. The test was initially applied with different dilutions of the antigen to detect the best performance of the antigen. This was tested in a serum bank of 45 animals proven positive in the commercial ELISA test and in the serum of 45 animals proven negative in the commercial ELISA test, later it was tested on the cut-off point, specificity and sensitivity of the test. The indirect ELISA test proved to be excellent as a diagnostic test for canine distemper, with an optical density cut-off of 0.229, sensitivity of 95.5% and specificity of 84.4% being obtained.

Antigenic and immunogenic properties of the canine distemper virus nucleocapsid protein expressed in Escherichia coli employing codon optimized synthetic gene / Propriedades antigênicas e imunogênicas da proteína do nucleocapsídeo do virus da cinomose canina expressa em Escherichia coli empregando gene sintético com codons otimizados

Despite common occurrence and importance of canine distemper disease the majority of tests currently available for diagnosis are hampered by either low sensitivity or specificity. In this study it was evaluated antigenic and immunogenic characteristics of a conserved region of nucleocapsid protein of canine distemper virus (rCDV NP) expressed in Escherichia coli employing a codon optimized synthetic gene. The expression of rCDVNP in Star strain (mean 300μg/mL, purified) was confirmed by SDS-PAGE and Western blot analysis by using His-Tag monoclonal antibodies. Western blot and ELISA, employing positive and negative control dog sera, demonstrated the rCDVNP antigenicity. The rCDVNP was inoculated in hens and immunoglobulin Y (IgY) was purified from the egg yolk. The mean yield of IgY was 28.55mg/mL. IgY reacted with the recombinant protein as demonstrated by Western blot and ELISA assays. In summary, our findings demonstrated that rCDVNP is antigenic since CDV positive dog sera recognized the protein in vitro. Additionally, the rCDVNP proved to be immunogenic in hens being possible to isolate a high concentration of specific IgY antibodies from the egg yolk. Taken together, these results indicate that the rCDVNP along with the specific IgY could be useful tools for development of the canine distemper immunodiagnostic assays.(AU)

Apesar da ocorrência comum e importância da cinomose canina, a maioria dos testes atualmente disponíveis para diagnóstico são prejudicados pela baixa sensibilidade ou especificidade. Neste estudo foram avaliadas características antigênicas e imunogênicas de uma região conservada da proteína do nucleocapsídeo do virus da cinomose canina (rCDV NP) expressa em Escherichia coli empregando um gene sintético e codons otimizados. A expressão na cepa Star (média de 300μg/mL, purificada) foi confirmada por SDS-PAGE e Western blot utilizando anticorpos monoclonais anti-His-Tag. A antigenicidade da rCDVNP foi demonstrada por western blot e ELISA empregando soros de cães positivos e negativos. A rCDVNP foi inoculada em galinhas e imunoglobulina Y (gY) foi obtida e purificada a partir da gema. A produção média de IgY foi 28.55mg/mL. Anticorpos IgY reagiram com a proteína recombinante, quando analisados por Western blot e ELISA. Em resumo, nossos achados demonstram que a rCDVNP produzida é antigênica, uma vez que os anticorpos de soro de cães positivos para CDV reconheceram a proteína in vitro. Além disso, a rCDVNP foi imunogênica em galinhas, sendo possível isolar anticorpos IgY específicos a partir da gema do ovo em altas concentrações. Tomados em conjunto, estes resultados indicam que a rCDVNP juntamente com a IgY específica podem ser ferramentas úteis para elaborar ensaios de imunodiagnóstico de cinomose canina.(AU)

Antigenic and immunogenic properties of the canine distemper virus nucleocapsid protein expressed in Escherichia coli employing codon optimized synthetic gene / Propriedades antigênicas e imunogênicas da proteína do nucleocapsídeo do virus da cinomose canina expressa em Escherichia coli empregando gene sintético com codons otimizados

Despite common occurrence and importance of canine distemper disease the majority of tests currently available for diagnosis are hampered by either low sensitivity or specificity. In this study it was evaluated antigenic and immunogenic characteristics of a conserved region of nucleocapsid protein of canine distemper virus (rCDV NP) expressed in Escherichia coli employing a codon optimized synthetic gene. The expression of rCDVNP in Star strain (mean 300μg/mL, purified) was confirmed by SDS-PAGE and Western blot analysis by using His-Tag monoclonal antibodies. Western blot and ELISA, employing positive and negative control dog sera, demonstrated the rCDVNP antigenicity. The rCDVNP was inoculated in hens and immunoglobulin Y (IgY) was purified from the egg yolk. The mean yield of IgY was 28.55mg/mL. IgY reacted with the recombinant protein as demonstrated by Western blot and ELISA assays. In summary, our findings demonstrated that rCDVNP is antigenic since CDV positive dog sera recognized the protein in vitro. Additionally, the rCDVNP proved to be immunogenic in hens being possible to isolate a high concentration of specific IgY antibodies from the egg yolk. Taken together, these results indicate that the rCDVNP along with the specific IgY could be useful tools for development of the canine distemper immunodiagnostic assays.(AU)

Apesar da ocorrência comum e importância da cinomose canina, a maioria dos testes atualmente disponíveis para diagnóstico são prejudicados pela baixa sensibilidade ou especificidade. Neste estudo foram avaliadas características antigênicas e imunogênicas de uma região conservada da proteína do nucleocapsídeo do virus da cinomose canina (rCDV NP) expressa em Escherichia coli empregando um gene sintético e codons otimizados. A expressão na cepa Star (média de 300μg/mL, purificada) foi confirmada por SDS-PAGE e Western blot utilizando anticorpos monoclonais anti-His-Tag. A antigenicidade da rCDVNP foi demonstrada por western blot e ELISA empregando soros de cães positivos e negativos. A rCDVNP foi inoculada em galinhas e imunoglobulina Y (gY) foi obtida e purificada a partir da gema. A produção média de IgY foi 28.55mg/mL. Anticorpos IgY reagiram com a proteína recombinante, quando analisados por Western blot e ELISA. Em resumo, nossos achados demonstram que a rCDVNP produzida é antigênica, uma vez que os anticorpos de soro de cães positivos para CDV reconheceram a proteína in vitro. Além disso, a rCDVNP foi imunogênica em galinhas, sendo possível isolar anticorpos IgY específicos a partir da gema do ovo em altas concentrações. Tomados em conjunto, estes resultados indicam que a rCDVNP juntamente com a IgY específica podem ser ferramentas úteis para elaborar ensaios de imunodiagnóstico de cinomose canina.(AU)

Detection of respiratory viruses in shelter dogs maintained under varying environmental conditions

Three dog shelters in Rio Grande do Sul were investigated for associations between the occurrence of respiratory viruses and shelter environmental conditions. Nasal secretions randomly collected during the cold season were tested via PCR, and this data collection was followed by nucleotide sequencing of the amplicons. In shelter /1 (poor sanitary and nutritional conditions, high animal density and constant contact between dogs), 78% (58/74) of the nasal samples were positive, 35% (26/74) of which were in single infections and 44% (32/74) of which were in coinfections. Shelters /2 and /3 had satisfactory sanitary and nutritional conditions, outdoors exercise areas (/2) and animal clustering by groups (/3). In shelter /2, 9% (3/35) of the samples were positive for Canine parainfluenza virus (CPIV), and 6% (2/35) were positive for Canid herpesvirus 1 (CaHV-1). In shelter /3, 9% (7/77) of the samples were positive for Canine adenovirus type 2 (CAdV-2), and 1% (1/77) were positive for Canine distemper virus (CDV). The amplicon sequences (CPIV and CDV nucleoprotein gene; CAdV-2 E3 gene; CaHV-1 glycoprotein B gene) showed 94-100% nucleotide identity with GenBank sequences. Our results demonstrate that CPIV, CAdV-2 and CDV are common in dog shelters and that their frequencies appear to be related with environmental and nutritional conditions. These results indicate the need for control/prevention measures, including vaccination and environmental management, to minimize these infections and improve dog health.(AU)

CARACTERIZAÇÃO IMUNO-HISTOQUÍMICA DE ANTÍGENOS DO VÍRUS DA CINOMOSE E ADENOVÍRUS CANINO TIPO 2 EM LONTRAS Lontra longicaudis (Olfers, 1818)

Um estudo histopatológico retrospectivo com amostras de tecido (pulmão, estômago, rins, fígado e pele) de seis lontras neotropicais de vida livre (Lontra longicaudis) foi realizado para investigar a possível ocorrência de padrões de doença associados a agentes de doenças infecciosas virais comuns dos cães domésticos. Os ensaios de imuno-histoquímica (IHQ) foram projetados para identificar antígenos de tecido intralesional do vírus da Cinomose (CDV) e adenovírus canino -1 (CAdV-1) e -2 (CAdV-2). Os padrões histopatológicos mais frequentes diagnosticados foram pneumonia intersticial (83,33%; 5/6) e degeneração vacuolar hepatocelular (50%; 3/6). Os IHQ identificaram imunorreatividade intralesional intracitoplasmática para antígenos de CDV em todas as lontras avaliadas, com imunomarcação positiva ocorrendo nas células epiteliais dos pulmões, estômago, rins, fígado e pele. Antígenos CAdV-2 intracitoplasmáticos foram identificados nas células epiteliais das glândulas peribrônquicas em quatro lontras com pneumonia intersticial. Esses achados resultaram em infecções singulares e mistas nestas lontras neotropicais, representam o primeiro relato de infecção concomitante por CDV e CAdV-2 em lontras neotropicais de vida livre do Hemisfério Sul e sugerem que esta espécie de mamífero é suscetível a agentes etiológicos virais comuns aos cães domésticos e pode desenvolver padrões de doença histopatológica semelhantes.

A retrospective histopathological study with tissue samples (lung, stomach, kidneys, liver and skin) from six free-living neotropical otters (Otter longicaudis) was conducted to investigate the possible occurrence of disease patterns associated with common viral infectious disease agents of domestic dogs. Immunohistochemistry (IHQ) assays were designed to identify antigens of intralesional tissue of the Cinomosis virus (CDV) and canine adenovirus -1 (CAdV-1) and -2 (CAdV-2). The most frequent histopathological patterns diagnosed were interstitial pneumonia (83.33%; 5/6) and hepatocellular vacuolar degeneration (50%; 3/6). IHQ identified incytoplasmic intralesional immunoreactivity for CDV antigens in all otters evaluated, with positive immunosmarcation occurring in the epithelial cells of the lungs, stomach, kidneys, liver and skin. Incytoplasmic CAdV-2 antigens were identified in the epithelial cells of the peribronchial glands in four otters with interstitial pneumonia. These findings resulted in single and mixed infections in these neotropical otters, representing the first report of concomitant CDV and CAdV-2 infection in neotropical free-living otters from the Southern Hemisphere and suggest that this mammal species is susceptible to viral etiological agents common to domestic dogs and may develop similar histopathological disease patterns.

CARACTERIZAÇÃO IMUNO-HISTOQUÍMICA DE ANTÍGENOS DE MORBILLIVÍRUS CANINO EM FELÍDEOS NEOTROPICAIS DO SUL DO BRASIL

O morbillivirus canino (CDV) é considerado um patógeno emergente em animais silvestres e tem demonstrado alta capacidade de infectar diferentes espécies animais. Entretanto, relatos na literatura brasileira descrevendo a infecção de felídeos neotropicais são escassos. Consequentemente, o objetivo do trabalho foi identificar partículas virais de CDV em tecidos de felídeos neotropicais do Sul do Brasil utilizando um protocolo imuno-histoquímico (IHQ) para identificar a presença intralesional de antígenos do CDV, podendo então fazer a associação com os achados histopatológicos. Foram utilizados tecidos provenientes de três espécies de felídeos neotropicais: puma (Puma concolor, n=1), maracajá (Leopardus wideii, n=1) e gato- mourisco (Herpailurus yagouaroundi). Todos os felídeos eram mantidos no Santuário Biológico de Bela Vista. Fragmentos de pulmão e vesícula urinária de todos os animais foram coletados e processados para avaliação histopatológica de rotina a fim de identificar possíveis padrões de lesões associados ao CDV, como ocorrido em cães domésticos. Com base no padrão da doença, identificado pela histopatologia, foram realizados ensaios de IHC para identificação de antígenos do CDV. A análise histopatológica revelou pneumonia intersticial nos pulmões do maracajá e gato- mourisco, e degeneração balonosa do urotélio da bexiga do puma. Ambas as alterações revelaram imunorreatividade positiva para antígenos de CDV. Em conclusão, antígenos intralesionais de CDV foram identificados em tecidos de felídeos neotropicais, confirmando infecção ativa nesses animais. Esses achados adicionam às poucas descrições de infecções induzidas por CDV em hospedeiros mamíferos que não o cão doméstico do Brasil.

Canine morbillivirus (CDV) is considered an emerging pathogen in wild animals and has demonstrated a high capacity to infect different animal species. However, reports in the Brazilian literature describing the infection of neotropical felids are scarce. Consequently, the aim of the study was to identify CDV viral particles in tissues of neotropical felids from southern Brazil using an immunohistochemical protocol (IHC) to identify the intralesional presence of CDV antigens, and then be able to make the association with histopathological findings. Tissues from three species of Neotropical felids were used: cougar (Puma concolor, n = 1), margay (Leopardus wideii, n = 1) and jaguarundi (Herpailurus yagouaroundi). All felids were kept in the Bela Vista Biological Sanctuary. Lung and urinary bladder fragments from all animals were collected and processed for routine histopathological evaluation in order to identify possible patterns of CDV- associated injuries, as occurred in domestic dogs. Based on the disease pattern, identified by histopathology, IHC assays were performed to identify CDV antigens. Histopathological analysis revealed interstitial pneumonia in the margay and jaguarundi lungs and ballooning degeneration of the cougars urinary bladder urothelium. Both findings revealed positive immunoreactivity for CDV antigens. In conclusion, intralesional antigens of CDV were identified in tissues of neotropical felids, confirming active infection in these animals. These findings add to the few descriptions of CDV-induced infections in mammalian hosts other than Brazil's domestic dog.

Canine distemper virus: detection of viral RNA by Nested RT-PCR in dogs with clinical diagnosis / Vírus da cinomose canina: detecção do RNA viral pelo Nested RT-PCR em cães com diagnóstico clínico

Canine distemper virus (CDV) is a pathogen which affects dogs and causes severe disease leading to death. Dogs infected with CDV can be diagnosed by RNA detection by Nested PCR technique. The following study proposed to evaluate CDV RNA in blood, urine and saliva samples. The Nested-PCR technique was able to detect CDV RNA in different types of biologic samples. The higher number of positive results was obtained in urine samples.(AU)

vírus da cinomose canina (CDV) é um patógeno que afeta cães, causando doença grave e que pode levar a morte. Os cães infectados pelo CDV podem ser diagnosticados pela detecção do RNA utilizando-se a técnica de Nested-PCR. O presente estudo teve como objetivo avaliar o RNA do CDV no sangue, urina e saliva em cães com diagnóstico clínico de cinomose. A técnica de Nested-PCR foi capaz de detectar o RNA em diferentes tipos de amostras biológicas. Obteve-se um maior número de resultados positivos em amostras de urina.(AU)

ANÁLISE FILOGENÉTICA DO GENE DE FUSÃO DE CEPAS BRASILEIRAS DO VÍRUS DA CINOMOSE CANINA EM CÃES

A cinomose canina é uma doença viral sistêmica altamente contagiosa, de distribuição mundial e que afeta uma grande variedade de carnívoros terrestres. Este estudo teve como objetivo analisar o gene F completo de cepas de campo brasileiras do vírus da cinomose canina (CDV) e compará-las com outros isolados de CDV. As amostras biológicas caninas (n=15) utilizadas neste estudo foram coletadas entre 2003-2004 (n=6) e 2013-2016 (n=9) e submetidas a ensaio de RT-PCR para amplificação do gene F completo do CDV. A análise das sequências com 2.426 bp de 14 cepas brasileiras de CDV foram classificadas na linhagem EU1 / SA1, com um agrupamento temporal entre amostras antigas (2003-2004) e contemporâneas (2013-2016), independentemente do status vacinal dos animais amostrados. Uma cepa brasileira de CDV agrupou-se na linhagem Rockborn-like, apresentando alta similaridade (98,5%) com a cepa vacinal Rockborn. Para todas as cepas brasileiras, a região Fsp apresentou a maior variação nas sequências de aminoácidos (67,4% - 96,2%). As cepas brasileiras apresentaram-se mais divergentes em relação às cepas vacinais classificadas como NA1 (24,5% - 36,3%) quando comparadas com a também cepa vacinal Rockborn (11,2% - 14,9%). Dezessete resíduos de cisteína foram encontrados no gene F completo e quatro sítios de glicosilação não conservados foram identificados na região Fsp das cepas brasileiras de CDV. Os resultados sugerem a circulação da linhagem EU1 / SA1 por 25 anos no Brasil e a atual cocirculação de cepas antigas e contemporâneas de CDV. A região Fsp demonstrou ser adequada para estudos evolutivos. Este é o primeiro estudo que realizou a análise do gene F completo de cepas de camplo brasileiras de CDV contribuindo com informações relacionadas com a epidemiologia molecular do vírus.

Canine distemper is a highly contagious systemic viral disease of worldwide distribution and that affects a wide variety of terrestrial carnivores. This study aimed to analyze the whole canine distemper virus (CDV) F gene of Brazilian field strains isolated in dogs and compared it with other CDV isolates. Canine biological samples (n = 15) that were collected among 2003-2004 (n=6) and 2013-2016 (n=9) were subjected to RT-PCR assays for the amplification of full-length CDV F gene. The sequence analysis of 2,426 bp length of 14 Brazilian CDV field strains were classified as EU1/SA1 lineage, with a temporal clustering into past (2003-2004) and contemporaneous (2013-2016) strains, regardless of the vaccination status of animals sampled. One Brazilian strain clustered into the Rockborn-like lineage, showing high similarity (98.5%) with the Rockborn vaccine isolate. For all the Brazilian strains, the Fsp region showed high aa variation (67.4% 96.2%). The Brazilian strains were more Fsp-divergent of the NA1 (24.5% 36.3%) than to the Rockborn (11.2% 14.9%) vaccine strains. Seventeen cysteine residues in the full-length F gene and four non-conserved glycosylation sites were found in the Fsp region of Brazilian CDV strains. The results suggest a 25-year circulation of EU1/SA1 lineage in Brazil and reveal a currently co-circulation of past and contemporaneous CDV strains. The Fsp-coding region of CDV genome was shown to be suitable for evolutionary studies. This is the first study dedicated to the whole CDV F gene analysis from Brazilian field strains, complementing the knowledge on molecular epidemiology of the virus.

A hematologic and electrophoretic study in puppies vaccinated against canine distemper virus and canine parvovirus

Background: Canine distemper is a contagious multisystemic viral disease that affects canines and others carnivores. Canine parvovirus infection is one of the most important viral diseases in young dogs. Side effects of vaccine generally include fever, lethargy and local inflammation. Complementary exams are important to evaluate the strenght of immungenic stimulation. This study was aimed at evaluating hematological and electrophoretic alterations in puppies after inoculation of live attenuated vaccine against canine distemper virus and canine parvovirus. Materials, Methods & Results: Five non-breeding newborn dogs of the same litter were used. Animals received three subcutaneous injection of 1mL (at days 0, 21 and 42). Blood was collected at day 0 (day of vaccination) and for three times for each dose: at days 7, 14 and 21 (first dose); at days 28, 35 and 42 (second dose); and at days 49, 56 and 63 (third dose). Blood containing ethylenediaminetetraacetic acid as anticoagulant was used for hematological evaluation. The total serum protein were determined by the biuret method, using commercial reagent, according to fabricant instructions. Serum was used for protein fractionation by using cellulose acetate strip electrophoresis. A decrease in platelet count was observed at days 7 and 28 post-vaccination. Lymphocyte number increased 88.4%, as well as the level of the protein fractions alpha-1 globulin (68%) and alpha-2 globulin (41.4%) at day 7. Moreover, a 5-fold increase in the fibrinogem concentration and in the number of eosinophils was observed at day 14. Thereafter, the platelet count decreased by 27.3% and the number of monocytes increased 5-fold at day 28. Discussion: Mild to moderate thrombocytopenia is often observed in dogs 3-5 days post-vaccination with live attenuated vaccines, mainly those against CDV and CPV. Besides the platelet damage caused by the CDV per se, infected animals showed secondary immune-mediated thrombocytopenia and decreased platelet production due to direct viral megakaryocyte infection. The increase in alpha-1 globulin may be related to the augment in the synthesis of alpha-1 antitrypsin, the main protein of the alpha-1 globulin region, in response to the vaccine-induced acute inflammatory process. The alpha-2 globulin region includes haptoglobin, alpha-2 macroglobulin and ceruloplasmin, and the increase observed in this fraction suggested that both haptoglobin and ceruloplasmin levels were augmented, following acute inflammatory response pattern. Fibrinogen is a soluble plasma glycoprotein that is converted by thrombin into fibrin during blood clotting. Despite the increase in fibrinogen concentration be the best indicator of inflammation in large animals, the hyperfibrinogenemia observed suggests that the inflammatory process was adequate to stimulate synthesis of this acute phase protein (P < 0,05). Absence of lymphocytosis observed at days 49, 56 and 63 associated to the progressive increase of the gamma globulin fraction, although not statistically significant, suggested an augment of B lymphocytes. The eosinophilia was observed in highlighting the presence of inflammation. Moreover, an increase in monocyte count indicating the presence of subacute or chronic inflammation after the second dose of the vaccine.

A hematologic and electrophoretic study in puppies vaccinated against canine distemper virus and canine parvovirus / A hematologic and electrophoretic study in puppies vaccinated against canine distemper virus and canine parvovirus

Background: Canine distemper is a contagious multisystemic viral disease that affects canines and others carnivores. Canine parvovirus infection is one of the most important viral diseases in young dogs. Side effects of vaccine generally include fever, lethargy and local inflammation. Complementary exams are important to evaluate the strenght of immungenic stimulation. This study was aimed at evaluating hematological and electrophoretic alterations in puppies after inoculation of live attenuated vaccine against canine distemper virus and canine parvovirus.Materials, Methods & Results: Five non-breeding newborn dogs of the same litter were used. Animals received three subcutaneous injection of 1mL (at days 0, 21 and 42). Blood was collected at day 0 (day of vaccination) and for three times for each dose: at days 7, 14 and 21 (first dose); at days 28, 35 and 42 (second dose); and at days 49, 56 and 63 (third dose). Blood containing ethylenediaminetetraacetic acid as anticoagulant was used for hematological evaluation. The total serum protein were determined by the biuret method, using commercial reagent, according to fabricant instructions. Serum was used for protein fractionation by using cellulose acetate strip electrophoresis. A decrease in platelet count was observed at days 7 and 28 post-vaccination. Lymphocyte number increased 88.4%, as well as the level of the protein

Background: Canine distemper is a contagious multisystemic viral disease that affects canines and others carnivores. Canine parvovirus infection is one of the most important viral diseases in young dogs. Side effects of vaccine generally include fever, lethargy and local inflammation. Complementary exams are important to evaluate the strenght of immungenic stimulation. This study was aimed at evaluating hematological and electrophoretic alterations in puppies after inoculation of live attenuated vaccine against canine distemper virus and canine parvovirus.Materials, Methods & Results: Five non-breeding newborn dogs of the same litter were used. Animals received three subcutaneous injection of 1mL (at days 0, 21 and 42). Blood was collected at day 0 (day of vaccination) and for three times for each dose: at days 7, 14 and 21 (first dose); at days 28, 35 and 42 (second dose); and at days 49, 56 and 63 (third dose). Blood containing ethylenediaminetetraacetic acid as anticoagulant was used for hematological evaluation. The total serum protein were determined by the biuret method, using commercial reagent, according to fabricant instructions. Serum was used for protein fractionation by using cellulose acetate strip electrophoresis. A decrease in platelet count was observed at days 7 and 28 post-vaccination. Lymphocyte number increased 88.4%, as well as the level of the protein

Avaliação prospectiva do soro hiperimune na terapia de cães com vírus da cinomose canina / Prospective evaluation of the immunoglobulin serum in the treatment of dogs with canine distemper virus / La evaluación prospectiva de suero hiperinmune en la terapia de perros con el virus del moquillo canino

A cinomose canina é uma doença infecto-contagiosa de alta mortalidade e prognóstico reservado devido o acometimento neurológico irreversível e limitada opção de tratamento. O presente estudo avaliou a eficácia do uso de soro hiperimune no tratamento de cães com cinomose na fase sistêmica da infecção. Os animais foram diagnosticados através de teste comercial imunocromatográfico. Após a constatação da infecção, foram aplicados 2 ml/kg de soro hiperimune pela via subcutânea. A evolução da terapia foi acompanhada clinicamente e por análises hematológicas, e ao fim de sete dias pós-tratamento, novo teste imunocromatográfico foi realizado. Os resultados demonstraram que 80% dos animais após o tratamento sobreviveram e se recuperaram clinicamente, sendo que, o grupo de animais não vacinados apresentou melhor desempenho. O produto mostrou-se eficaz no tratamento quando introduzido no início do quadro clínico.(AU)

Canine distemper (CDV) is an infectious disease of high mortality and guarded prognosis due to the irreversible neurological damage and lack of therapeutic protocols. The aim of this study was to evaluate the efficacy of immunoglobulin serum therapy in dogs with systemic phase of CDV infection. The animals were diagnosed by immunochromatographic test and positive dogs were treated with 2 ml/kg of immunoglobulin sera. Follow up procedures was made by laboratorial exams and clinical evaluation every day during 7 days pos-treatment. Immunochromatographic evaluation for CDV antigens was repeated in the 7th day after treatment. The results showed that 80% of the treated dogs' survived, and the group of unvaccinated animals showed better performance. The product was effective in the treatment when introduced at the beginning of the clinical condition.(AU)

El moquillo canino es una enfermedad infecto-contagiosa de alta mortalidad y pronóstico reservado debido el acometimiento neurológico irreversible y limitada opción de tratamiento. El presente estudio evaluó la eficacia del uso de suero hiperinmune en el tratamiento de canes con moquillo en la fase sistémica de la infección. Los animales fueron diagnosticados a través de test comercial inmunocromatográfico. Después de la constatación de la infección, fueron aplicados 2 ml/kg de suero hiperinmune por la vía subcutánea. La evolución de la terapia fue acompañada clínicamente y por análisis hematológicos, y al fin de siete días póst-tratamiento, nuevo test inmunocromatográfico fue realizado. Los resultados demostraron que 80% de los animales después de tratamiento sobrevivieron y se recuperaron clínicamente, dado que, el grupo de animales no vacunados presentó mejor desempeño. El producto se mostró eficaz en el tratamiento cuando es introducido en el inicio del cuadro clínico.(AU)

Avaliação prospectiva do soro hiperimune na terapia de cães com vírus da cinomose canina / Prospective evaluation of the immunoglobulin serum in the treatment of dogs with canine distemper virus / La evaluación prospectiva de suero hiperinmune en la terapia de perros con el virus del moquillo canino

A cinomose canina é uma doença infecto-contagiosa de alta mortalidade e prognóstico reservado devido o acometimento neurológico irreversível e limitada opção de tratamento. O presente estudo avaliou a eficácia do uso de soro hiperimune no tratamento de cães com cinomose na fase sistêmica da infecção. Os animais foram diagnosticados através de teste comercial imunocromatográfico. Após a constatação da infecção, foram aplicados 2 ml/kg de soro hiperimune pela via subcutânea. A evolução da terapia foi acompanhada clinicamente e por análises hematológicas, e ao fim de sete dias pós-tratamento, novo teste imunocromatográfico foi realizado. Os resultados demonstraram que 80% dos animais após o tratamento sobreviveram e se recuperaram clinicamente, sendo que, o grupo de animais não vacinados apresentou melhor desempenho. O produto mostrou-se eficaz no tratamento quando introduzido no início do quadro clínico.

Canine distemper (CDV) is an infectious disease of high mortality and guarded prognosis due to the irreversible neurological damage and lack of therapeutic protocols. The aim of this study was to evaluate the efficacy of immunoglobulin serum therapy in dogs with systemic phase of CDV infection. The animals were diagnosed by immunochromatographic test and positive dogs were treated with 2 ml/kg of immunoglobulin sera. Follow up procedures was made by laboratorial exams and clinical evaluation every day during 7 days pos-treatment. Immunochromatographic evaluation for CDV antigens was repeated in the 7th day after treatment. The results showed that 80% of the treated dogs' survived, and the group of unvaccinated animals showed better performance. The product was effective in the treatment when introduced at the beginning of the clinical condition.

El moquillo canino es una enfermedad infecto-contagiosa de alta mortalidad y pronóstico reservado debido el acometimiento neurológico irreversible y limitada opción de tratamiento. El presente estudio evaluó la eficacia del uso de suero hiperinmune en el tratamiento de canes con moquillo en la fase sistémica de la infección. Los animales fueron diagnosticados a través de test comercial inmunocromatográfico. Después de la constatación de la infección, fueron aplicados 2 ml/kg de suero hiperinmune por la vía subcutánea. La evolución de la terapia fue acompañada clínicamente y por análisis hematológicos, y al fin de siete días póst-tratamiento, nuevo test inmunocromatográfico fue realizado. Los resultados demostraron que 80% de los animales después de tratamiento sobrevivieron y se recuperaron clínicamente, dado que, el grupo de animales no vacunados presentó mejor desempeño. El producto se mostró eficaz en el tratamiento cuando es introducido en el inicio del cuadro clínico.

Expression of pro-and-anti-apoptotic antigens in the cerebellum of dogs naturally infected with canine distemper virus

Canine distemper virus (CDV) may induce multifocal demyelination in the central nervous system of infected dogs. The present work investigated apoptosis in white and gray matter (granular layer) in the cerebellum of naturally infected dogs by the analysis of the expression of the pro-apoptotic antigens caspase 2 and 3 , b(terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL-staining) positivity, annexin-V immunodetection, and the presence of the anti-apoptotic antigens, BCl-2 and p53. Cerebellum specimens were obtained from the Laboratory of Animal Pathology, from 1995 to 2009, and the 5-µm thick fragments were stained both with hematoxylin-eosin and Shorr. All samples were diagnosed as positive for CDV genome by reverse transcriptase polymerase chain reaction targeting the nucleocapsid gene. The anti-apoptotic pathways evidenced in this study were BCl-2 and p53 proteins that were intensively detected in cerebellum of CDV positive slides (40-80% of labeled cells/mm2). In addition, the apoptosis markers annexin-V and TUNEL are directly correlated among the same samples (80 and 40% of labeled cells, respectively). This is the first description of p53 and annexin-V expression, characterized as anti-apoptotic and apoptotic proteins, involvement in canine natural cases of CDV infections. (AU)

Expression of pro-and-anti-apoptotic antigens in the cerebellum of dogs naturally infected with canine distemper virus

Canine distemper virus (CDV) may induce multifocal demyelination in the central nervous system of infected dogs. The present work investigated apoptosis in white and gray matter (granular layer) in the cerebellum of naturally infected dogs by the analysis of the expression of the pro-apoptotic antigens caspase – 2 and – 3 , b(terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL-staining) positivity, annexin-V immunodetection, and the presence of the anti-apoptotic antigens, BCl-2 and p53. Cerebellum specimens were obtained from the Laboratory of Animal Pathology, from 1995 to 2009, and the 5-µm thick fragments were stained both with hematoxylin-eosin and Shorr. All samples were diagnosed as positive for CDV genome by reverse transcriptase polymerase chain reaction targeting the nucleocapsid gene. The anti-apoptotic pathways evidenced in this study were BCl-2 and p53 proteins that were intensively detected in cerebellum of CDV positive slides (40-80% of labeled cells/mm2). In addition, the apoptosis markers annexin-V and TUNEL are directly correlated among the same samples (80 and 40% of labeled cells, respectively). This is the first description of p53 and annexin-V expression, characterized as anti-apoptotic and apoptotic proteins, involvement in canine natural cases of CDV infections.

A hematologic and electrophoretic study in puppies vaccinated against canine distemper virus and canine parvovirus / A hematologic and electrophoretic study in puppies vaccinated against canine distemper virus and canine parvovirus

Background: Canine distemper is a contagious multisystemic viral disease that affects canines and others carnivores. Canine parvovirus infection is one of the most important viral diseases in young dogs. Side effects of vaccine generally include fever, lethargy and local inflammation. Complementary exams are important to evaluate the strenght of immungenic stimulation. This study was aimed at evaluating hematological and electrophoretic alterations in puppies after inoculation of live attenuated vaccine against canine distemper virus and canine parvovirus.Materials, Methods & Results: Five non-breeding newborn dogs of the same litter were used. Animals received three subcutaneous injection of 1mL (at days 0, 21 and 42). Blood was collected at day 0 (day of vaccination) and for three times for each dose: at days 7, 14 and 21 (first dose); at days 28, 35 and 42 (second dose); and at days 49, 56 and 63 (third dose). Blood containing ethylenediaminetetraacetic acid as anticoagulant was used for hematological evaluation. The total serum protein were determined by the biuret method, using commercial reagent, according to fabricant instructions. Serum was used for protein fractionation by using cellulose acetate strip electrophoresis. A decrease in platelet count was observed at days 7 and 28 post-vaccination. Lymphocyte number increased 88.4%, as well as the level of the protein

Background: Canine distemper is a contagious multisystemic viral disease that affects canines and others carnivores. Canine parvovirus infection is one of the most important viral diseases in young dogs. Side effects of vaccine generally include fever, lethargy and local inflammation. Complementary exams are important to evaluate the strenght of immungenic stimulation. This study was aimed at evaluating hematological and electrophoretic alterations in puppies after inoculation of live attenuated vaccine against canine distemper virus and canine parvovirus.Materials, Methods & Results: Five non-breeding newborn dogs of the same litter were used. Animals received three subcutaneous injection of 1mL (at days 0, 21 and 42). Blood was collected at day 0 (day of vaccination) and for three times for each dose: at days 7, 14 and 21 (first dose); at days 28, 35 and 42 (second dose); and at days 49, 56 and 63 (third dose). Blood containing ethylenediaminetetraacetic acid as anticoagulant was used for hematological evaluation. The total serum protein were determined by the biuret method, using commercial reagent, according to fabricant instructions. Serum was used for protein fractionation by using cellulose acetate strip electrophoresis. A decrease in platelet count was observed at days 7 and 28 post-vaccination. Lymphocyte number increased 88.4%, as well as the level of the protein