Preliminary validation of a novel high-resolution melt-based typing method based on the multilocus sequence typing scheme of Streptococcus pyogenes

The major limitation of current typing methods for Streptococcus pyogenes, such as emm sequence typing and T typing, is that these are based on regions subject to considerable selective pressure. Multilocus sequence typing (MLST) is a better indicator of the genetic backbone of a strain but is not widely used due to high costs. The objective of this study was to develop a robust and cost-effective alternative to S. pyogenes MLST. A 10-member single nucleotide polymorphism (SNP) set that provides a Simpson’s Index of Diversity (D) of 0.99 with respect to the S. pyogenes MLST database was derived. A typing format involving high-resolution melting (HRM) analysis of small fragments nucleated by each of the resolution-optimized SNPs was developed. The fragments were 59–119 bp in size and, based on differences in G+C content, were predicted to generate three to six resolvable HRM curves. The combination of curves across each of the 10 fragments can be used to generate a melt type (MelT) for each sequence type (ST). The 525 STs currently in the S. pyogenes MLST database are predicted to resolve into 298 distinct MelTs and the method is calculated to provide a D of 0.996 against the MLST database. The MelTs are concordant with the S. pyogenes population structure. To validate the method we examined clinical isolates of S. pyogenes of 70 STs. Curves were generated as predicted by G+C content discriminating the 70 STs into 65 distinct MelTs.

The birds of the Bush Heritage, Cravens Peak Reserve

Bird communities were studied in two subregional areas of Cravens Peak, the Toko Plains and the Simpson-Strzelecki Dunefields, using the point counts method. A total of 42 2ha 20 minute surveys, 46 five-hundred metre radius area surveys and 170 5km drive through area surveys were conducted and observations made. Bird species were identified, counted and recorded. The data were compared in the two subregions and, as a whole, considering species groups according to land system on which the ecosystem occurs, the specific ecosystem and according to their general feeding habits (insectivore, omnivore, frugivore, granivore, nectarivore and carnivore). Species richness and species relative abundance were compared using Simpson’s Diversity Index and the data revealed that species are distributed largely on the basis of habitat. In general, areas with a greater number of vegetation strata recorded greater species diversity. Overall, the Tall Open Acacia georginae Shrubland on alluvial floodplains has a greater diversity of birds in a 2ha area (0.87, Simpson’s Index of Diversity 1-D) compared to the other survey sites.