No abstract

Canine distemper virus (CDV) is the cause of a severe and highly contagious disease in dogs. Practical diagnosis of canine distemper based on clinical signs and laboratory tests are required to confirm CDV infection. The present study aimed to develop a molecular assay to detect and differentiate field and vaccine CDV strains. Reverse transcription followed by nested real time polymerase chain reaction (RT-nqPCR) was developed, which exhibited analytical specificity (all the samples from healthy dogs and other canine infectious agents were not incorrectly detected) and sensitivity (all replicates of a vaccine strain were positive up to the 3125-fold dilution - 10(0.7) TCID50). RT-nqPCR was validated for CDV detection on different clinical samples (blood, urine, rectal and conjunctival swabs) of 103 animals suspected to have distemper. A total of 53 animals were found to be positive based on RT-nqPCR in at least one clinical sample. Blood resulted in more positive samples (50 out of 53, 94.3%), followed by urine (44/53, 83.0%), rectal (38/53, 71%) and conjunctival (27/53, 50.9%) swabs. A commercial immunochromatography (IC) assay had detected CDV in only 30 conjunctival samples of these positive dogs. Nucleoprotein (NC) gene sequencing of 25 samples demonstrated that 23 of them were closer to other Brazilian field strains and the remaining two to vaccine strains. A single nucleotide sequences difference, which creates an Msp I restriction enzyme digestion, was used to differentiate between field and vaccine CDV strains by restriction fragment length polymorphism (RFLP) analysis. The complete assay was more sensitive than was IC for the detection of CDV. Blood was the more frequently positive specimen and the addition of a restriction enzyme step allowed the differentiation of vaccine and Brazilian field strains.

No abstract

Pesq. Vet. Bras. 32(9) Psittaciformes are one of the most endangered groups of birds, and several Brazilian species are classiϐied between vulnerable and critically endangered. It is thus necessary to identify agents that cause infections in captive wild animals and to assess the risks posed thereof and to design interventions to minimize the possibility of disease outbreaks, leading to the conservation of endangered species. The purpose of this study was to identify enteropathogenic Escherichia coli (EPEC) cloacal isolates from asymptomatic psittacines in captivity and evaluate the distribution of the EPEC pathotype. Cloacal swabs were obtained from 46 asymptomatic birds, and resulting isolates were tested by polymerase chain reaction (PCR) for the presence of the attaching and effacing gene (eae) and bundle-forming pilus structural gene (bfpA) of EPEC. Samples from several species were tested, and three samples were found to be positive for the eae and bfpA genes and characterized as typical EPEC. This is the ϐirst report of this pathotype in asymptomatic psittacines. Although certain E. coli strains are more pathogenic than others, various factors should be considered when determining the potential of E. coli isolates to cause disease in captive psittacines. Birds that are positive for the EPEC (typical) strain could be zoonotic sources of infection, and may have acquired these strains through contact with humans or domestic animals. These ϐindings may also be valuable for the long-term management of endangered species ex situ as one EPEC sample was isolated from a Red-tailed Amazon (Amazona brasiliensis). classiϐicadas desde vulneráveis à criticamente ameaçadas de extinção. Torna-se, portanto, necessário identiϐicar os agentes que causam infecções em animais selvagens em cativeiro e determinar os riscos relacionados de modo a intervir sobre os fatores envolvidos para diminuir a possibilidade de surtos de doenças e promover a conservação de espécies ameaçadas. O objetivo deste estudo foi identiϐicar Escherichia coli Enteropatogência (EPEC) de isolados cloacais de psitacídeos assintomáticos em cativeiro e avaliar a distribuição do patotipo EPEC. Suabes cloacais foram coletados de 46 psitacídeos assintomáticos e os isolados foram testados pela reação em cadeia pela polimerase (PCR) para a presença do gene attaching and effacing (eae) e bundle forming pilus (bfpA) de EPEC. Amostras oriundas de diversas espécies foram testadas e três amostras resultaram positivas para os genes eae e bfp e caracterizadas como EPEC Pesq. Vet. Bras. 32(9):922-926, setembro 2012 923 Molecular detection of enteropathogenic Escherichia coli in asymptomatic captive psittacines típicas. Esse é o primeiro relato em psitacídeos assintomá-ticos para esse patotipo. Apesar de que algumas cepas de E.coli serem mais patogênicas do que outras, diversos fatores devem ser considerados para determinar o potencial de isolados de E.coli de causar doença em psitacídeos em cativeiro. Aves positivas para cepas de EPEC (típicas) poderiam ser fontes de in...

O Brasil possui um considerável número de espécies de psitacídeos catalogados, perfazendo cerca de 80 espécies, sendo que as Araras canindé (Ara ararauna), uma das maiores representantes dessa ordem, podem ser encontradas em florestas nas diversas regiões brasileiras. O Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis (IBAMA) normalizou a comercialização de animais da fauna silvestre provenientes de criadouros e conseqüentemente ocorreu um aumento do número destes animais como de estimação. Atualmente, há pouco conhecimento sobre os parâmetros clínicos e laboratoriais de espécies silvestres em cativeiro. O presente trabalho teve como objetivo determinar parâmetros de bioquímica sangüínea de Araras canindé (Ara ararauna) saudáveis de sexo e faixa etária distintas mantidas em um criatório comercial com alimentação e manejo controlados e padronizados. Foram colhidas amostras de sangue de 35 araras canindé (11 filhotes e 24 adultos) e remetidas ao Laboratório de Análises Clínicas Veterinárias da Universidade de Passo Fundo (UPF) para determinação dos seguintes indicadores bioquímicos: ácido úrico (AU), albumina (ALB), aspartato aminotransferase (AST), cálcio (Ca), colesterol (Col), creatina quinase (CK), fosfatase alcalina (FA), fósforo inorgânico (Pi), frutosamina (Fru), gama glutamil transferase (GGT), proteínas totais (PT) e uréia (UR). Entre as aves adultas, foram encontradas diferenças significativas nos valores de CK (superior nos machos), Ca e AU (superiores nas fêmeas). Entre aves adultas e filhotes foram constatadas diferenças significativas nos valores de AU, Ca, ALB, COL, FA, Pi e FRU. A maioria das alterações detectadas pode ser relacionada à diferença na dieta fornecida e à condição fisiológica de cada categoria de aves. Os dados obtidos podem ser utilizados como parâmetros de referência para as araras canindé brasileiras.

No abstract

Parrots in captivity are frequently affected by Escherichia coli (E. coli) infections. The objective of this study was to collect information on the carrier state for E. coli pathotypes in asymptomatic free-ranging parrots. Cloacal swabs were collected from nestlings of Hyacinth, Lear's macaws and Blue-fronted Amazon parrots and tested by polymerase chain reaction (PCR) for virulence factors commonly found in enteropathogenic, avian pathogenic, and uropathogenic E. coli strains. In total, 44 samples were cultured and E. coli isolates were yielded, from which DNA was extracted and processed by PCR. Genes commonly found in APEC isolates from Blue-fronted Amazon parrots and Hyacinth macaws were expressed in 14 of these 44 samples. One atypical EPEC isolate was obtained from a sample from Lear's macaw. The most commonly found gene was the increased serum survival (iss) gene. This is the first report, that describes such pathotypes in asymptomatic free-living parrots. The findings of this study suggest the presence of a stable host/parasite relationship at the time of the sampling brings a new understanding to the role that E. coli plays in captive and wild parrots. Such information can be used to improve husbandry protocols as well as help conservation efforts of free-living populations.

Tobacco farmers are routinely exposed to complex mixtures of the compounds present in tobacco leaves, including organic and inorganic pesticides. Penetration through skin is the most significant route of uptake in occupational exposure to chemicals, including dust and liquids containing toxic and carcinogenic substances. This study evaluates the genotoxic effect of tobacco leaves with and without dermal exposure to flumetralin in Mus musculus, determining cell damage by the micronucleus test and the Comet assay as well as antioxidant enzyme activities and hematologic parameters. Nicotine was used as positive control. Blood samples were collected for 0, 3, 24 and 48 h exposure periods, and DNA damage by Comet assay and micronucleus test was evaluated for all these periods. Bone marrow and liver cells were also evaluated for the 48 h exposure period. Significant differences between Comet assay results in blood cells from animals exposed to tobacco leaves with and without pesticide were found in 24 and 48 h exposure periods in relation to negative control. Bone marrow cells from the group exposed to leaves with pesticide (48 h) also demonstrated significant increase in DNA damage. Concerning the micronucleus test, only animals exposed to tobacco leaves without pesticide (24 h) showed increase in frequency of micronuclei when compared to the negative control. Oxidative stress activities also were demonstrated for different groups. The results demonstrate the injury effect caused by tobacco leaves in different Mus musculus tissues, suggesting that the effects of dermal exposure to tobacco leaves are caused by complex mixtures present in the plant, but mainly by nicotine.