Aims:To compare PCR combined with enrichment media with the standard microbiological techniques (SMT) and to determine the most sensitive method for the detection of Salmonella and the identification of Salm. typhimurium (ST), Salm. enteritidis (SE), Salm. gallinarum (SG) and Salm. pullorum (SP). Methods and Results: We analysed 87 samples from poultry using PCR and SMT, PCR being performed from non-selective (NS) and Rappaport-Vassiliadis (RV) media. PCR-NS was less sensitive than PCR-RV and SMT for the detection and identification of Salmonella. PCR-RV detected more positive samples of Salmonella sp. than SMT but both these methods showed similar sensitivity regarding the identification of Salmonella serovars. Conclusions: PCR-RV was more sensitive and decreased the time necessary to detect and identify Salmonella. Significance and Impact of the Study: PCR-RV is a powerful tool for the rapid and accurate detection and identification of Salmonella and can be implemented in diagnostic and food analysis laboratories.
RESUMO.-[Genes associados à patogenicidade deEscherichia coli patogênica para aves (APEC) isoladas de frangos de corte com sintomatologia clínica respiratória.] Os mecanismos de virulência das amostras de Escherichia coli potencialmente patogênicas para aves (APEC) têm sido continuamente estudados e acredita-se ser multifatorial. Certas propriedades são associadas primariamente a amostras virulentas e vêm sendo identificadas em amostras de E. coli isoladas de aves. Neste estudo um total de 61 amostras de E. coli, isoladas de frangos de corte com problemas respiratórios, foram testadas através da Reação em Cadeia da Polimerase (PCR), para a presença dos genes responsáveis pela capacidade de adesão, fimbria P (papC) e fimbria F11 (felA), produção de colicinas (cvaC), presença de aerobactina (iutA), resistência sérica (iss), hemaglutinina temperatura sensível (tsh) e presença de dos antígenos capsulares K1 e K5 (kpsII). O gene iss foi detectado em 73,8%, tsh em 55,7%, iutA em 45,9%, felA em 39,3%, papC em 24,3%, cvaC em 23% e kpsII em 18%.
The current systems of breeding poultry, based on high population density, increase the risk of spreading pathogens, especially those causing respiratory diseases and those that have more than one host. Fowl Cholera (FC) is one such pathogen, and even though it represents one of several avian diseases that should be considered in the differential diagnosis of notifiable diseases that present with sudden death, the pathogenesis and virulence factors involved in FC are still poorly understood. The objective of this study was to investigate twelve genes related to virulence in 25 samples of Pasteurella multocida isolated from FC cases in the southern region of Brazil through the development of multiplex PCR protocols. The protocols developed were capable of detecting all of the proposed genes. The ompH, oma87, sodC, hgbA, hgbB, exBD-tonB and nanB genes were present in 100% of the samples (25/25), the sodA and nanH genes were present in 96% (24/25), ptfA was present in 92% (23/25), and pfhA was present in 60% (15/25). Gene toxA was not identified in any of the samples studied (0/25). Five different genetic profiles were obtained, of which P1 (negative to toxA) was the most common. We concluded that the multiplex-PCR protocols could be useful tools for rapid and simultaneous detection of virulence genes. Despite the high frequency of the analyzed genes and the fact that all samples belonged to the same subspecies of P. multocida, five genetic profiles were observed, which should be confirmed in a study with a larger number of samples.
Dermal squamous cell carcinoma (DSCC; avian keratoacanthoma) is a neoplastic skin lesion of broiler chickens of unknown aetiology. In previous studies, the possibility of the involvement of pox viruses in the cause of DSCC was considered. In this work, a sensitive and specific nested polymerase chain reaction (PCR) protocol was developed that could amplify a 419 base pair DNA fragment of fowlpox virus with a detection limit of less than one infectious unit. Fowlpox virus DNA was always detected in skin samples with fowlpox lesions while it was not detected in samples of unrelated diseases such as cowpox, Marek's disease or infectious laryngotracheitis. Some macroscopically normal skin samples from vaccinated and non-vaccinated birds also produced PCR-positive results, corroborating previous studies on the possibility that a latent or chronic form of fowlpox occurs. Fowlpox virus DNA was consistently detected from DSCC skin lesions, and this finding is discussed.
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