RESUMO
Feline immunodeficiency virus (FIV) is an important pathogen of domestic and wild felids. Although serological tests suggest the presence of FIV in cats from Colombia, no molecular characterization has been reported. Here, we describe the near-complete genome of FIV subtype A from a Colombian domestic cat.
RESUMO
Few studies have described enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies against porcine circovirus type 2 (PCV2) based on antigens produced in cell culture. Furthermore, few articles have described viral purification techniques for members of the family Circoviridae. This occurs because circoviruses are difficult to isolate, noncytopathogenic, and produce low viral titres in cell culture. Thus, for overcoming these difficulties in the cultivation of PCV2, this study aimed to develop a double-antibody sandwich ELISA based on the cell culture antigen PCV2b for the quantification of anti-PCV2 antibodies. A 20% and 50% discontinuous sucrose cushion was used for viral purification, which enabled the separation of cell culture proteins in the 20% sucrose cushion and a greater viral concentration in the 50% sucrose cushion. Following isopycnic centrifugation, PCV2 was concentrated in the band with density values from 1.330 to 1.395g/cm3. Viral purification was assessed using SDS-PAGE, indirect ELISA and electron microscopy. The standardised ELISA revealed a strong linear correlation (r= 0.826, p < 0.001) when compared with a commercial ELISA kit. The assay exhibited low variability (inter-assay coefficient of variation of 4.24% and intra-assay of 1.80%) and excellent analytical specificity conferred by the capture antibody produced in rabbit. Thus, this ELISA is a rapid, specific and convenient method for the detection of antibodies against PCV2 in studies of experimental and natural infection, and in monitoring the response to vaccination on commercial farms.(AU)
Há poucos relatos na literatura de métodos de ELISA (Enzyme-linked immunosorbent assay), para a detecção de anticorpos contra o circovírus suíno tipo 2 (PCV2), baseados em antígenos produzidos em cultivo celular, bem como uma escassez de trabalhos descrevendo técnicas de purificação viral para os membros da família Circoviridae. Isso ocorre, pois os circovírus são de difícil isolamento, não causam efeito citopático e produzem um baixo título viral em cultivo celular. Assim, para superar essas dificuldades encontradas no cultivo do PCV2, este estudo objetivou desenvolver um sandwich ELISA com duplo anticorpo, baseado no antígeno de PCV2 produzido em cultivo celular, para a quantificação de anticorpos anti-PCV2. Um colchão de sacarose descontínuo a 20% e 50% foi utilizado para a purificação viral, o qual possibilitou a separação das proteínas oriundas do cultivo celular no colchão de sacarose a 20% e uma maior concentração viral no colchão de sacarose a 50%. Com a ultracentrifugação isopícnica, o PCV2 ficou mais concentrado na banda com valores de densidade de 1,330 a 1,395g/cm3. A purificação viral foi avaliada pelas técnicas de SDS-PAGE, ELISA indireto e microscopia eletrônica. Assim, o método de ELISA padronizado revelou uma forte correlação linear (r = 0,826, p < 0,001) quando comparado com um kit de ELISA comercial. O ensaio demonstrou baixa variabilidade (coeficientes de variação inter-teste de 4,24% e intra-teste de 1,80%) e uma excelente especificidade analítica conferida pelo anticorpo de captura produzido em coelho. Portanto, o método de ELISA demonstrou ser rápido, específico e conveniente para a detecção de anticorpos contra o PCV2 em estudos de infecção natural e experimental, além da monitoria da resposta à vacinação contra o PCV2 em granjas comerciais.(AU)
Assuntos
Ensaio de Imunoadsorção Enzimática , Circovirus , Sacarose , Anticorpos , Centrifugação IsopícnicaRESUMO
Few studies have described enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies against porcine circovirus type 2 (PCV2) based on antigens produced in cell culture. Furthermore, few articles have described viral purification techniques for members of the family Circoviridae. This occurs because circoviruses are difficult to isolate, noncytopathogenic, and produce low viral titres in cell culture. Thus, for overcoming these difficulties in the cultivation of PCV2, this study aimed to develop a double-antibody sandwich ELISA based on the cell culture antigen PCV2b for the quantification of anti-PCV2 antibodies. A 20% and 50% discontinuous sucrose cushion was used for viral purification, which enabled the separation of cell culture proteins in the 20% sucrose cushion and a greater viral concentration in the 50% sucrose cushion. Following isopycnic centrifugation, PCV2 was concentrated in the band with density values from 1.330 to 1.395g/cm3. Viral purification was assessed using SDS-PAGE, indirect ELISA and electron microscopy. The standardised ELISA revealed a strong linear correlation (r= 0.826, p<0.001) when compared with a commercial ELISA kit. The assay exhibited low variability (inter-assay coefficient of variation of 4.24% and intra-assay of 1.80%) and excellent analytical specificity conferred by the capture antibody produced in rabbit. Thus, this ELISA is a rapid, specific and convenient method for the detection of antibodies against PCV2 in studies of experimental and natural infection, and in monitoring the response to vaccination on commercial farms.(AU)
Há poucos relatos na literatura de métodos de ELISA (Enzyme-linked immunosorbent assay), para a detecção de anticorpos contra o circovírus suíno tipo 2 (PCV2), baseados em antígenos produzidos em cultivo celular, bem como uma escassez de trabalhos descrevendo técnicas de purificação viral para os membros da família Circoviridae. Isso ocorre, pois os circovírus são de difícil isolamento, não causam efeito citopático e produzem um baixo título viral em cultivo celular. Assim, para superar essas dificuldades encontradas no cultivo do PCV2, este estudo objetivou desenvolver um sandwich ELISA com duplo anticorpo, baseado no antígeno de PCV2 produzido em cultivo celular, para a quantificação de anticorpos anti-PCV2. Um colchão de sacarose descontínuo a 20% e 50% foi utilizado para a purificação viral, o qual possibilitou a separação das proteínas oriundas do cultivo celular no colchão de sacarose a 20% e uma maior concentração viral no colchão de sacarose a 50%. Com a ultracentrifugação isopícnica, o PCV2 ficou mais concentrado na banda com valores de densidade de 1,330 a 1,395g/cm3. A purificação viral foi avaliada pelas técnicas de SDS-PAGE, ELISA indireto e microscopia eletrônica. Assim, o método de ELISA padronizado revelou uma forte correlação linear (r = 0,826, p <0,001) quando comparado com um kit de ELISA comercial. O ensaio demonstrou baixa variabilidade (coeficientes de variação inter-teste de 4,24% e intra-teste de 1,80%) e uma excelente especificidade analítica conferida pelo anticorpo de captura produzido em coelho. Portanto, o método de ELISA demonstrou ser rápido, específico e conveniente para a detecção de anticorpos contra o PCV2 em estudos de infecção natural e experimental, além da monitoria da resposta à vacinação contra o PCV2 em granjas comerciais.(AU)
Assuntos
Anticorpos , Circovirus , Ensaio de Imunoadsorção Enzimática , Sacarose , Centrifugação IsopícnicaRESUMO
Partial nucleotide sequences of ORF72 (glycoprotein D, gD), ORF64 (infected cell protein 4, ICP4) and ORF30 (DNA polymerase) genes were compared with corresponding sequences of EHV-1 reference strains to characterize the molecular variability of Brazilian strains. Virus isolation assays were applied to 74 samples including visceral tissue, total blood, cerebrospinal fluid (CSF) and nasal swabs of specimens from a total of 64 animals. Only one CSF sample (Iso07/05 strain) was positive by virus isolation in cell culture. EHV-1 Iso07/05 neurologic strain and two abortion visceral tissues samples (Iso11/06 and Iso33/06) were PCR-positive for ORF33 (glycoprotein B, gB) gene of EHV-1. A sequence analysis of the ORF72, ORF64 and ORF30 genes from three EHV-1 archival strains (A3/97, A4/72, A9/92) and three clinical samples (Iso07/05, Iso11/06 and Iso33/06) suggested that among Brazilian EHV-1 strains, the amplified region of the gD gene sequence is highly conserved. Additionally, the analysis of ICP4 gene showed high nucleotide and amino acid identities when compared with genotype P strains, suggesting that the EHV-1 Brazilian strains belonged to the same group. All the EHV-1 Brazilian strains were classified as non-neuropathogenic variants (N752) based on the ORF30 analysis. These findings indicate a high conservation of the gD-, ICP4- and ORF30-encoding sequences. Different pathotypes of the EHV-1 strain might share identical genes with no specific markers, and tissue tropism is not completely dependent on the gD envelope, immediate-early ICP4 and DNA polymerase proteins.
Assuntos
Variação Genética , Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 1/classificação , Herpesvirus Equídeo 1/genética , Doenças dos Cavalos/virologia , Animais , Brasil , Análise por Conglomerados , Sequência Conservada , DNA Viral/química , DNA Viral/genética , Genótipo , Infecções por Herpesviridae/virologia , Cavalos , Dados de Sequência Molecular , Fases de Leitura Aberta , Filogenia , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido NucleicoRESUMO
Partial nucleotide sequences of ORF72 (glycoprotein D, gD), ORF64 (infected cell protein 4, ICP4) and ORF30 (DNA polymerase) genes were compared with corresponding sequences of EHV-1 reference strains to characterize the molecular variability of Brazilian strains. Virus isolation assays were applied to 74 samples including visceral tissue, total blood, cerebrospinal fluid (CSF) and nasal swabs of specimens from a total of 64 animals. Only one CSF sample (Iso07/05 strain) was positive by virus isolation in cell culture. EHV-1 Iso07/05 neurologic strain and two abortion visceral tissues samples (Iso11/06 and Iso33/06) were PCR-positive for ORF33 (glycoprotein B, gB) gene of EHV-1. A sequence analysis of the ORF72, ORF64 and ORF30 genes from three EHV-1 archival strains (A3/97, A4/72, A9/92) and three clinical samples (Iso07/05, Iso11/06 and Iso33/06) suggested that among Brazilian EHV-1 strains, the amplified region of the gD gene sequence is highly conserved. Additionally, the analysis of ICP4 gene showed high nucleotide and amino acid identities when compared with genotype P strains, suggesting that the EHV-1 Brazilian strains belonged to the same group. All the EHV-1 Brazilian strains were classified as non-neuropathogenic variants (N752) based on the ORF30 analysis. These findings indicate a high conservation of the gD-, ICP4- and ORF30-encoding sequences. Different pathotypes of the EHV-1 strain might share identical genes with no specific markers, and tissue tropism is not completely dependent on the gD envelope, immediate-early ICP4 and DNA polymerase proteins.(AU)
Assuntos
Animais , Variação Genética , Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 1/classificação , Herpesvirus Equídeo 1/genética , Doenças dos Cavalos/virologia , Brasil , Análise por Conglomerados , Sequência Conservada , DNA Viral/química , DNA Viral/genética , Genótipo , Infecções por Herpesviridae/virologia , Cavalos , Dados de Sequência Molecular , Fases de Leitura Aberta , Filogenia , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido NucleicoRESUMO
Partial nucleotide sequences of ORF72 (glycoprotein D, gD), ORF64 (infected cell protein 4, ICP4) and ORF30 (DNA polymerase) genes were compared with corresponding sequences of EHV-1 reference strains to characterize the molecular variability of Brazilian strains. Virus isolation assays were applied to 74 samples including visceral tissue, total blood, cerebrospinal fluid (CSF) and nasal swabs of specimens from a total of 64 animals. Only one CSF sample (Iso07/05 strain) was positive by virus isolation in cell culture. EHV-1 Iso07/05 neurologic strain and two abortion visceral tissues samples (Iso11/06 and Iso33/06) were PCR-positive for ORF33 (glycoprotein B, gB) gene of EHV-1. A sequence analysis of the ORF72, ORF64 and ORF30 genes from three EHV-1 archival strains (A3/97, A4/72, A9/92) and three clinical samples (Iso07/05, Iso11/06 and Iso33/06) suggested that among Brazilian EHV-1 strains, the amplified region of the gD gene sequence is highly conserved. Additionally, the analysis of ICP4 gene showed high nucleotide and amino acid identities when compared with genotype P strains, suggesting that the EHV-1 Brazilian strains belonged to the same group. All the EHV-1 Brazilian strains were classified as non-neuropathogenic variants (N752) based on the ORF30 analysis. These findings indicate a high conservation of the gD-, ICP4- and ORF30-encoding sequences. Different pathotypes of the EHV-1 strain might share identical genes with no specific markers, and tissue tropism is not completely dependent on the gD envelope, immediate-early ICP4 and DNA polymerase proteins.
Assuntos
Animais , Variação Genética , Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 1/classificação , Herpesvirus Equídeo 1/genética , Doenças dos Cavalos/virologia , Brasil , Análise por Conglomerados , Sequência Conservada , DNA Viral/química , DNA Viral/genética , Genótipo , Cavalos , Infecções por Herpesviridae/virologia , Dados de Sequência Molecular , Fases de Leitura Aberta , Filogenia , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido NucleicoRESUMO
Rabies transmitted by the hematophagous bat Desmodus rotundus represents a public health concern and a burden for the Brazilian livestock industry. Current evidence suggests that rabies occurrence is related to landscape characteristics, topography, hydrography, animal production systems and land use...
A raiva transmitida por morcegos hematófagos da espécie Desmodus rotundus representa uma preocupação de saúde pública e causa de importantes prejuízos para a pecuária brasileira. A evidência atual sugere que a ocorrência de raiva está relacionada às características da paisagem, topografia, hidrografia, sistemas de produção animal e usos da terra...
Assuntos
Animais , Quirópteros/virologia , Vírus da Raiva/genéticaRESUMO
Rabies transmitted by the hematophagous bat Desmodus rotundus represents a public health concern and a burden for the Brazilian livestock industry. Current evidence suggests that rabies occurrence is related to landscape characteristics, topography, hydrography, animal production systems and land use. However, a few studies have analyzed the possible connections among geographic factors and the molecular diversity of the rabies virus, furthering the understanding of the spatial and temporal dynamics of outbreaks. A study reported that the latest rabies epizootics in herbivores reported in the eastern region of São Paulo (close to the Minas Gerais border) occurred in two epidemic waves; the first was before 1998, and the other occurred after 1999. Using this evidence, the aim of the present study was to analyze cases of rabies in herbivores in the southern region of Minas Gerais (2000-2009) and their possible relationship with the aforementioned epidemics, considering the geographic characteristics of the region. Partial sequences of glycoprotein (539 nt) and nucleoprotein genes (414 nt) were obtained from 31 rabies virus isolates from herbivores. A phylogenetic tree was proposed for each genomic region using the Neighbor joining method, fixing the Kimura 2-parameter evolution model with a bootstrap level of 1,000 replications. Genetic sublineages were plotted on maps, considering rabies risk areas for herbivores in São Paulo, as well as topographic characteristics and hydrographic basins, to visualize any apparent distribution pattern influenced by those features. The phylogenetic trees had concordant topologies, suggesting a possible common origin for rabies outbreaks in herbivores along the SP/MG border, surrounding the less elevated portions of the Serra da Mantiqueira and along the hydrographic basins of Piracicaba/Jaguarí, Paranaíba do Sul, Grande, Pardo and Mogi-Guaçu rivers.The co-circulation of several viral lineages was observed in some municipalities, possibly due to an overlapping of rabies outbreaks. Inferred protein sequences of both genes showed synonymous mutations, except among residues 20 to 200, corresponding to the external domain of the glycoprotein. This information prompted cooperation among the animal health services of both states to reinforce rabies control in the border area.(AU)
A raiva transmitida por morcegos hematófagos da espécie Desmodus rotundus representa uma preocupação de saúde pública e causa de importantes prejuízos para a pecuária brasileira. A evidência atual sugere que a ocorrência de raiva está relacionada às características da paisagem, topografia, hidrografia, sistemas de produção animal e usos da terra. Contudo, existem poucos estudos que analisem as possíveis conexões entre fatores geográficos e a diversidade molecular do vírus da raiva, permitindo a compreensão da dinâmica espacial e temporal dos focos de raiva. Um desses trabalhos estabeleceu que a última epizootia de raiva dos herbívoros registrada no leste do estado de São Paulo (na fronteira com Minas Gerais), aconteceu em duas ondas epidêmicas, sendo a primeira em 1998 e, em 1999, a segunda. Considerando esta evidência, o intuito do presente estudo foi analisar casos de raiva em herbívoros na região sudeste de Minas Gerais (2000-2009) e sua possível relação com a epidemia previamente mencionada, incluindo as características geográficas da região. Foram obtidas sequencias parciais dos genes da glicoproteína (539 nt) e da nucleoproteína (414 nt) a partir de 31 isolados de vírus da raiva procedentes de herbívoros. Foi proposta uma árvore filogenética para cada região genômica usando o método de Neighbor joining, fixando o modelo evolutivo Kimura 2 - parâmetros com um nível de bootstrap de 1000 replicações. As sublinhagens genéticas foram localizadas sobre mapas, considerando as áreas de risco para raiva dos herbívoros em São Paulo, assim como as características topográficas e bacias hidrográficas com o intuito de visualizar qualquer padrão aparente de distribuição segundo essas características. As duas árvores filogenéticas mostraram topologias concordantes, sugerindo uma possível origem comum para os surtos que aconteceram ao longo da fronteira SP/MG, ao redor das porções menos elevadas da Serra da Mantiqueira e acompanhando as bacias hidrográficas dos rios Piracicaba/Jaguarí, Paranaíba do Sul, Grande, Pardo e Mogi-Guaçu. Foi possível observar circulação de varias linhagens virais simultaneamente em alguns municípios, possivelmente por causa de sobreposição de surtos. As sequencias de proteína inferidas a partir dos dois genes mostraram mutações sinônimas, excetuando aquelas encontradas entre os resíduos 20 a 200, correspondentes ao domínio externo da glicoproteína. Esta informação salienta a importância da cooperação entre as autoridades sanitárias de ambos os estados para reforçar o programa de controle da doença nas áreas limítrofes.(AU)
Assuntos
Animais , Vírus da Raiva/genética , Quirópteros/virologiaRESUMO
Gastroenteritis is one of the leading causes of morbidity and mortality among young and newborn animals and is often caused by multiple intestinal infections, with rotavirus and bovine coronavirus (BCoV) being the main viral causes in cattle. Given that BCoV is better studied than equine coronaviruses and given the possibility of interspecies transmission of these viruses, this research was designed to compare the partial sequences of the spike glycoprotein (S), hemagglutinin-esterase protein (HE), and nucleoprotein (N) genes from coronaviruses from adult cattle with winter dysentery, calves with neonatal diarrhea, and horses. To achieve this, eleven fecal samples from dairy cows with winter dysentery, three from calves, and two from horses, all from Brazil, were analysed. It could be concluded that the enteric BCoV genealogy from newborn and adult cattle is directly associated with geographic distribution patterns, when S and HE genes are taken into account. A less-resolved genealogy exists for the HE and N genes in cattle, with a trend for an age-related segregation pattern. The coronavirus strains from horses revealed Betacoronavirus sequences indistinguishable from those found in cattle, a fact previously unknown.
Assuntos
Coronavirus Bovino/classificação , Coronavirus Bovino/genética , Genes Virais , Filogenia , Animais , Brasil , Bovinos , Geografia , CavalosRESUMO
Cryptosporidium spp. are cosmopolitan protozoa that infect fishes, reptiles, amphibians, birds and mammals. More than 20 species are recognized within this genus. Rodents are a group of abundant and ubiquitous organisms that have been considered reservoirs of Cryptosporidium for humans and livestock. The aim of this study was to design specific primers for the gene encoding 18S rRNA, potentially capable of amplifying any species or genotype of Cryptosporidium spp. and evaluate the diagnostic attributes of the nested-PCR based on such probes. The primers were designed to amplify the shortest segment as possible to maximize the sensitivity of the test, but preserving the discriminatory potential of the amplified sequences for phylogenetic inferences. The nested-PCR standardized in this study (nPCR-SH) was compared in terms of sensitivity with another similar assay (nPCR-XIAO) that has been largely used for the detection and identification of Cryptosporidium spp. worldwide. We also aimed to molecularly characterize samples of Cryptosporidum spp. isolated from synanthropic rodents using these probes. Forty-five rodents were captured in urban areas of the municipality of Umuarama, Paraná State, Brazil. Fecal samples were submitted to three molecular tests (nested-PCRs), two of them targeted to the 18S rDNA gene (nPCR-SH and nPCR-XIAO) and the third targeted to the gene encoding actin (nPCR-actin). The nPCR-SH was tested positive on samples of Cryptosporidum parvum, Cryptosporidum andersoni, Cryptosporidum meleagridis, Cryptosporidum hominis, Cryptosporidum canis, and Cryptosporidum serpentis. Sixteen samples of rodents were positive by nPCR-SH, six by nPCR-XIAO and five by nPCR-actin. Sequencing of amplified fragments allowed the identification of Cryptosporidum muris in three samples of Rattus rattus, and two genotypes of Cryptosporidium, the genotypes mouse II and III. Cryptosporidium genotype mouse II was found in one sample of Mus musculus and genotype mouse III, in twelve samples, being five from R. rattus and seven from M. musculus. The results of this study demonstrated that the primers designed for detection of Cryptosporidium spp. were more efficient than those used in the nPCR-XIAO. Genotypes or species of Cryptosporidium that can be usually transmitted for human beings and livestock were not found in synanthropic rodents, suggesting that the importance of these animals in zoonotic transmission of cryptosporidiosis should be revisited.
Assuntos
Criptosporidiose/veterinária , Cryptosporidium/isolamento & purificação , Primers do DNA , Camundongos/parasitologia , Ratos/parasitologia , Doenças dos Roedores/parasitologia , Actinas/genética , Animais , Sequência de Bases , Brasil , Criptosporidiose/parasitologia , Criptosporidiose/transmissão , Cryptosporidium/classificação , Cryptosporidium/genética , Primers do DNA/normas , DNA de Protozoário/isolamento & purificação , DNA Ribossômico , Reservatórios de Doenças/parasitologia , Fezes/parasitologia , Dados de Sequência Molecular , Oocistos/classificação , Filogenia , Reação em Cadeia da Polimerase/normas , RNA Ribossômico 18S/genética , Doenças dos Roedores/transmissão , Alinhamento de SequênciaRESUMO
To evaluate the most controversial issue concerning current feline coronavirus (FCoV) virology, the coexisting hypotheses of the intrahost and interhost origins of feline infectious peritonitis virus (FIPV) in regard to the pathogenesis of feline infectious peritonitis (FIP), this study aimed to assess the molecular diversity of the membrane gene FCoVs in 190 samples from 10 cats with signs of FIP and in 5 faecal samples from cats without signs of FIP. All samples from the non-FIP cats and 25.26% of the samples from the FIP cats were positive for the FCoV membrane (M) gene. Mutations in this gene consisted of SNP changes randomly scattered among the sequences; few mutations resulted in amino acid changes. No geographic pattern was observed. Of the cats without FIP that harboured FECoV, the amino acid sequence identities for the M gene were 100% among cats (Cats 1-3) from the same cattery, and the overall sequence identity for the M gene was ≥91%. In one cat, two different lineages of FCoV, one enteric and one systemic, were found that segregated apart in the M gene tree. In conclusion, the in vivo mutation transition hypothesis and the circulating high virulent-low virulent FCoV hypothesis have been found to be plausible according to the results obtained from sequencing the M gene.
Assuntos
Coronavirus Felino/genética , Coronavirus Felino/patogenicidade , Fezes/virologia , Peritonite Infecciosa Felina/genética , Peritonite Infecciosa Felina/virologia , Variação Genética/genética , Animais , Gatos , Coronavirus Felino/isolamento & purificaçãoRESUMO
Infectious diseases in wild animals have been increasing as a result of their habitat alterations and closer contact with domestic animals. Canine distemper virus (CDV) has been reported in several species of wild carnivores, presenting a threat to wildlife conservation. We described the first case of canine distemper virus infection in lesser grison (Galictis cuja). A free-ranging individual, with no visible clinical sigs, presented sudden death after one day in captivity. Molecular diagnosis for CDV infection was performed using whole blood collected by postmortem intracardiac puncture, which resulted positive. The virus phylogeny indicated that domestic dogs were the probable source of infection.(AU)
Doenças infecciosas em animais selvagens têm aumentado devido às alterações em seu habitat e ao maior contato com animais domésticos. A cinomose já foi descrita em diversas espécies de carnívoros selvagens, representando uma ameaça à conservação da vida selvagem. Nesse estudo é descrito o primeiro caso de infecção pelo vírus da cinomose em um furão (Galictis cuja). Um indivíduo de vida livre, sem sinais clínicos aparentes, apresentou morte súbita após um dia em cativeiro. Foi realizado o diagnóstico molecular para detecção do vírus da cinomose canina, sendo o resultado positivo. A filogenia do vírus indicou que cães domésticos foram a provável fonte de infecção.(AU)
Assuntos
Animais , Cães , Mustelidae/virologia , Vírus da Cinomose Canina/isolamento & purificação , Ecossistema/efeitos adversos , Animais Selvagens , Filogenia , EcossistemaRESUMO
Infectious diseases in wild animals have been increasing as a result of their habitat alterations and closer contact with domestic animals. Canine distemper virus (CDV) has been reported in several species of wild carnivores, presenting a threat to wildlife conservation. We described the first case of canine distemper virus infection in lesser grison (Galictis cuja). A free-ranging individual, with no visible clinical sigs, presented sudden death after one day in captivity. Molecular diagnosis for CDV infection was performed using whole blood collected by postmortem intracardiac puncture, which resulted positive. The virus phylogeny indicated that domestic dogs were the probable source of infection.
Doenças infecciosas em animais selvagens têm aumentado devido às alterações em seu habitat e ao maior contato com animais domésticos. A cinomose já foi descrita em diversas espécies de carnívoros selvagens, representando uma ameaça à conservação da vida selvagem. Nesse estudo é descrito o primeiro caso de infecção pelo vírus da cinomose em um furão (Galictis cuja). Um indivíduo de vida livre, sem sinais clínicos aparentes, apresentou morte súbita após um dia em cativeiro. Foi realizado o diagnóstico molecular para detecção do vírus da cinomose canina, sendo o resultado positivo. A filogenia do vírus indicou que cães domésticos foram a provável fonte de infecção.
Assuntos
Animais , Cães , Animais Selvagens , Ecossistema/efeitos adversos , Mustelidae/virologia , Vírus da Cinomose Canina/isolamento & purificação , Ecossistema , FilogeniaRESUMO
The aim of the study was to evaluate rickettsial infection in ticks from wild birds of the Semidecidual and Atlantic Rainforest remnants of three municipalities of the State of Paraná, southern Brazil. Overall, 53 larvae and nymphs collected from birds were checked for the presence of Rickettsia DNA by molecular tests. Five tick species were tested: Amblyomma aureolatum (Pallas), Amblyomma calcaratum Neumann, Amblyomma longirostre (Koch), Amblyomma ovale Koch, and Amblyomma parkeri Fonseca and Aragão. A. longirostre ticks were infected with the spotted fever group agents Rickettsia amblyommii strain AL (32.3% infection rate) and Rickettsia parkeri strain NOD (5.9% infection rate). A new rickettsial genotype was detected in the tick A. parkeri (50% infection rate), which had never been reported to be infected by rickettsiae. Through phylogenetic analysis, this new genotype, here designated as strain ApPR, grouped in a cluster composed by different strains of Rickettsia africae, Rickettsia sibirica, and R. parkeri. We consider strain ApPR to be a new genotype of R. parkeri. This study reports for the first time rickettsial infection in ticks from birds in southern Brazil. The role of migrating birds in the dispersal of these rickettsial strains should be considered in ecological studies of spotted fever group agents in Brazil.
Assuntos
Aves/parasitologia , Ixodidae/microbiologia , Rickettsia/isolamento & purificação , Animais , Brasil , Ecossistema , Genótipo , Filogenia , Rickettsia/classificação , Rickettsia/genéticaRESUMO
BACKGROUND: Porcine circovirus type 2 (PCV2) has been associated with several disease complexes, including reproductive failure. The aim of this study was to identify the subtypes of PCV2 that are associated with reproductive failure in pigs from the State of São Paulo, Brazil and to investigate co-infections with other infectious organisms. FINDINGS: Samples of 168 aborted foetuses or mummified foetuses from five farrow-to-finish swine farms known to be infected with PCV2 and located in the State of São Paulo were tested for PCV2 by polymerase chain reaction (PCR). Positive samples were additionally tested for porcine parvovirus (PPV), Leptospira spp. and Brucella spp. by PCR. PCV2 was detected in 18 of the samples (10.7%). PPV, Brucella spp. and Leptospira spp were found in 2, 10 and 0 cases, respectively. Eleven PCV2 strains were sequenced and determined to be either genotype 2a (n = 1) or 2b (n = 10). CONCLUSIONS: The findings indicate that the frequency of PCV2 infections in aborted porcine foetuses from the State of São Paulo is rather low (10.7%) and that co-infection with other pathogens is common and may be involved in PCV2 associated reproductive failure. No repeatable, characteristic amino acid motifs for regions of the PCV2 capsid protein seemed to be associated with abortion in sows.
Assuntos
Aborto Animal/virologia , Infecções por Circoviridae/veterinária , Circovirus/isolamento & purificação , Coinfecção/veterinária , Doenças dos Suínos/virologia , Aborto Animal/epidemiologia , Aborto Animal/microbiologia , Animais , Brasil/epidemiologia , Brucella/genética , Brucella/isolamento & purificação , Brucelose/epidemiologia , Brucelose/microbiologia , Brucelose/veterinária , Infecções por Circoviridae/epidemiologia , Infecções por Circoviridae/virologia , Circovirus/classificação , Circovirus/genética , Coinfecção/epidemiologia , Coinfecção/microbiologia , Coinfecção/virologia , Feminino , Leptospira/genética , Leptospira/isolamento & purificação , Leptospirose/epidemiologia , Leptospirose/microbiologia , Leptospirose/veterinária , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/veterinária , Infecções por Parvoviridae/virologia , Parvovirus Suíno/genética , Parvovirus Suíno/isolamento & purificação , Filogenia , Reação em Cadeia da Polimerase , Análise de Sequência de DNA/veterinária , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/microbiologiaRESUMO
Free-living adult Amblyomma incisum ticks were collected in an Atlantic rainforest area at Intervales State Park, State of São Paulo, Brazil. From an A. incisum specimen, rickettsiae were successfully isolated in Vero cell culture by the shell vial technique. Rickettsial isolation was confirmed by optical microscopy, transmission electron microscopy, and PCRs targeting portions of the rickettsial genes gltA, htrA, rrs, and sca1 on infected cells. Fragments of 1,089, 457, 1,362, and 443 nucleotides of the gltA, htrA, rrs, and sca1 genes, respectively, were sequenced. By BLAST analysis, the partial sequence of rrs of the A. incisum rickettsial isolate was closest to the corresponding sequence of Rickettsia bellii (99.1% similarity). The gltA partial sequence was closest to the corresponding sequences of "Candidatus Rickettsia tarasevichiae" (96.1% similarity) and Rickettsia canadensis (95.8% similarity). The htrA partial sequence was closest to the corresponding sequence of R. canadensis (89.8% similarity). The sca1 partial sequence was closest to the corresponding sequence of R. canadensis (95.2% similarity). Since our rickettsial isolate was genetically distinct from other Rickettsia species, we propose a new species designated Rickettsia monteiroi sp. nov. Phylogenetic analyses indicated that R. monteiroi belongs to the canadensis group within the genus Rickettsia, together with the species R. canadensis and "Candidatus R. tarasevichiae". Little or no antibody cross-reaction was observed between sera of R. monteiroi-inoculated guinea pigs and R. bellii-, Rickettsia rickettsii-, or R. canadensis-inoculated guinea pigs.
Assuntos
Infecções por Rickettsia/microbiologia , Rickettsia/classificação , Rickettsia/genética , Carrapatos/microbiologia , Animais , Sequência de Bases , Brasil , Chlorocebus aethiops , DNA Bacteriano/genética , Microscopia , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Células VeroRESUMO
Phylogenetic analyses based on mitochondrial 16S rDNA sequences were generated from Rhipicephalus sanguineus group specimens collected in 29 localities among 9 Latin-American countries, plus ticks collected in South Africa, Spain, and Italy. Sequences from Latin America generated six different haplotypes (A, B, C, D, E, and F). Phylogenetic analyses generated trees that segregated our tick sequences into two distinct clades: one is represented by haplotypes A-C, and South African R. sanguineus and Rhipicephalus turanicus ticks; the second clade is represented by haplotypes D-F, and European R. sanguineus and R. turanicus ticks. When haplotypes A-F are plotted in the Latin America map according to their geographical coordinates, it is clearly seen that haplotypes D-F are restricted to the southern portion of this continent, whereas haplotypes A-C are distributed in areas between northern Mexico and Brazil (except for the extreme south of this last country, where haplotype E was present). Hence, our phylogenetic analyses separated New World specimens of R. sanguineus into two distinct clades, one represented by tropical and subtropical populations (haplotypes A-C), here designated as the 'tropical' species. On the other hand, haplotypes D-F are here designated as the 'temperate' species because of their distribution in the southern portion of South America. Until recently, it was assumed that the R. sanguineus group was represented by a single species in the New World, namely R. sanguineus. While the present results coupled with recent studies support the presence of at least two species under the taxon R. sanguineus in the New World, they also show that even in the Old World, the taxon R. sanguineus might be represented by more than one species, since our phylogenetic analysis segregated European and South African R. sanguineus ticks into two distinct clades. The same can be applied for Spanish and South African R. turanicus.
Assuntos
Rhipicephalus sanguineus/classificação , Rhipicephalus sanguineus/genética , Animais , Análise por Conglomerados , DNA Mitocondrial/química , DNA Mitocondrial/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Genótipo , Haplótipos , América Latina , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
The present study evaluated the rickettsial infection in a laboratory colony of cat fleas, Ctenocephalides felis felis (Bouche) in Brazil. All flea samples (30 eggs, 30 larvae, 30 cocoons, 30 males, and 30 females) tested by polymerase chain reaction (PCR) were shown to contain rickettsial DNA. PCR products, corresponding to the rickettsial gltA, htrA, ompA and ompB gene partial sequences were sequenced and showed to correspond to Rickettsia felis, indicating that the flea colony was 100 percent infected by R. felis. The immunofluorescence assay (IFA) showed the presence of R. felis-reactive antibodies in blood sera of 7 (87.5 percent) out of 8 cats that were regularly used to feed the flea colony. From 15 humans that used to work with the flea colony in the laboratory, 6 (40.0 percent) reacted positively to R. felis by IFA. Reactive feline and human sera showed low endpoint titers against R. felis, varying from 64 to 256. With the exception of one human serum, all R. felis-reactive sera were also reactive to Rickettsia rickettsii and/or Rickettsia parkeri antigens at similar titers to R. felis. The single human serum that was reactive solely to R. felis had an endpoint titer of 256, indicating that this person was infected by R. felis.
Assuntos
Humanos , Animais , Gatos , Sequência de Bases , Gatos , Reação em Cadeia da Polimerase , Infecções por Rickettsiaceae , Rickettsia felis/isolamento & purificação , Sifonápteros , Técnicas e Procedimentos Diagnósticos , Imunofluorescência , MétodosRESUMO
We report a clinical case of spotted fever group rickettsiosis acquired in Sao Paulo, Brazil. Definitive diagnosis was supported by seroconversion between acute-phase and convalescent-phase serum samples. Molecular analysis of skin samples indicated the agent was a novel spotted fever group strain closely related to Rickettsia africae, R. parkeri, and R. sibirica.
Assuntos
Rickettsia/classificação , Rickettsia/isolamento & purificação , Febre Maculosa das Montanhas Rochosas , Idoso , Animais , Anticorpos Antibacterianos/sangue , Brasil/epidemiologia , DNA Bacteriano/análise , DNA Bacteriano/isolamento & purificação , Humanos , Masculino , Dados de Sequência Molecular , Rickettsia/genética , Rickettsia/imunologia , Febre Maculosa das Montanhas Rochosas/diagnóstico , Febre Maculosa das Montanhas Rochosas/epidemiologia , Febre Maculosa das Montanhas Rochosas/microbiologia , Febre Maculosa das Montanhas Rochosas/patologia , Análise de Sequência de DNA , Pele/microbiologia , Pele/patologiaRESUMO
The present study evaluated the rickettsial infection in a laboratory colony of cat fleas, Ctenocephalides felis felis (Bouche) in Brazil. All flea samples (30 eggs, 30 larvae, 30 cocoons, 30 males, and 30 females) tested by polymerase chain reaction (PCR) were shown to contain rickettsial DNA. PCR products, corresponding to the rickettsial gltA, htrA, ompA and ompB gene partial sequences were sequenced and showed to correspond to Rickettsia felis, indicating that the flea colony was 100% infected by R. felis. The immunofluorescence assay (IFA) showed the presence of R. felis-reactive antibodies in blood sera of 7 (87.5%) out of 8 cats that were regularly used to feed the flea colony. From 15 humans that used to work with the flea colony in the laboratory, 6 (40.0%) reacted positively to R. felis by IFA. Reactive feline and human sera showed low endpoint titers against R. felis, varying from 64 to 256. With the exception of one human serum, all R. felis-reactive sera were also reactive to Rickettsia rickettsii and/or Rickettsia parkeri antigens at similar titers to R. felis. The single human serum that was reactive solely to R. felis had an endpoint titer of 256, indicating that this person was infected by R. felis.