3 resultados para Fragment Theory

em eResearch Archive - Queensland Department of Agriculture


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Data from the eradication of the incursion of Bactrocera papayae Drew and Hancock (Dipt.: Tephritidae) in Australia (1995-1998) are used to assess the significance of various aspects of invasion theory, including the influence of towns on establishment, influence of propagule pressure on the pattern of establishment, and the existence of source-sink dynamics. Because there were no sentinel traps in place, considerable spread had occurred before the eradication campaign started. The distribution of fly density around the epicentre in the town of Cairns and a transect along the main traffic routes to the north and south fitted a Cauchy model with a tail having the same slope as a power model with an exponent of -2.4 extending to 160 km. The Cauchy model indicated that 50% of the flies on the transect would have occurred within 3.2 km of the epicentre, 90% within 13.2 km, and 99% within 60 km. The two major satellites at Mareeba (35 km from the epicentre in Cairns) and Mossman (65 km) were not used for the transect data and had respectively 15 and 30 times the density predicted by the model. The proportion of traps that caught flies (a measure of site occupancy) fell with distance from the epicentre. B. papayae was trapped consistently on only three of the 16 rainforest transects that were surveyed and these were relatively close to urban areas where eradication efforts were intense. Despite there being no eradication effort in the rainforest, the trends to extinction were similar to those in adjacent areas. The strategy of initially concentrating eradication efforts on the core and major satellites while maintaining a quarantine barrier at the airport and the boundaries of the infested area appears to be the key to the containment and rapid eradication of the incursion.

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The highly variable flagellin-encoding flaA gene has long been used for genotyping Campylobacter jejuni and Campylobacter coli. High-resolution melting (HRM) analysis is emerging as an efficient and robust method for discriminating DNA sequence variants. The objective of this study was to apply HRM analysis to flaA-based genotyping. The initial aim was to identify a suitable flaA fragment. It was found that the PCR primers commonly used to amplify the flaA short variable repeat (SVR) yielded a mixed PCR product unsuitable for HRM analysis. However, a PCR primer set composed of the upstream primer used to amplify the fragment used for flaA restriction fragment length polymorphism (RFLP) analysis and the downstream primer used for flaA SVR amplification generated a very pure PCR product, and this primer set was used for the remainder of the study. Eighty-seven C. jejuni and 15 C. coli isolates were analyzed by flaA HRM and also partial flaA sequencing. There were 47 flaA sequence variants, and all were resolved by HRM analysis. The isolates used had previously also been genotyped using single-nucleotide polymorphisms (SNPs), binary markers, CRISPR HRM, and flaA RFLP.flaA HRM analysis provided resolving power multiplicative to the SNPs, binary markers, and CRISPR HRM and largely concordant with the flaA RFLP. It was concluded that HRM analysis is a promising approach to genotyping based on highly variable genes.

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Poultry are considered a major source for campylobacteriosis in humans. A total of 1866 Campylobacter spp. isolates collected through the poultry processing chain were typed using flaA-restriction fragment length polymorphism to measure the impact of processing on the genotypes present. Temporally related human clinical isolates (n = 497) were also typed. Isolates were obtained from whole chicken carcass rinses of chickens collected before scalding, after scalding, before immersion chilling, after immersion chilling and after packaging as well as from individual caecal samples. A total of 32 genotypes comprising at least four isolates each were recognised. Simpson's Index of Diversity (D) was calculated for each sampling site within each flock, for each flock as a whole and for the clinical isolates. From caecal collection to after packaging samples the D value did not change in two flocks, decreased in one flock and increased in the fourth flock. Dominant genotypes occurred in each flock but their constitutive percentages changed through processing. There were 23 overlapping genotypes between clinical and chicken isolates. The diversity of Campylobacter is flock dependant and may alter through processing. This study confirms that poultry are a source of campylobacteriosis in the Australian population although other sources may contribute.