have exact matches with the primer sequence the qiime suite of programs was used to align chimera check cluster and carry out phylogenetics on sequence reads as well as to measure microbial u03b1 diversities and to plot rarefaction curves to determine if sequencing was carried out to sufficient depth u03b2 diversities were calculated on the sequence reads based on weighted unifrac and principal coordinate analysis pcoa performed kingviewer was used to visualise pcoa plots trimmed fasta sequences were assessed using blast against the silva version 100 database the resulting blast output was parsed using megan version 62 3 0 megan assigns reads to ncbi taxonomies by employing the lowest common ancestor algorithm which assigns each rna tag to the lowest common ancestor in the taxonomy from a subset of the best scoring matches in the blast result bit scores were used from within megan for filtering the results prior to tree construction and summarisation absolute cut off blast bit score 86 relative cut off 10 of the top hit the statistical significance of differences in proportions of microbial taxa was determined by the non parametric kruskal wallis test kruskal and wallis 1952 using minitab u00ae statistical package castenholz tye medium was chosen to selectively support the growth of strains from the genus thermus castenholz tye medium was prepared by mixing 5 parts 2 u00d7 castenholz salts with one part 1 tye and 4 parts distilled water castenholz salts 2 u00d7 contained 0 2 g nitrilotriacetic acid 0 12 g caso 4 2h 2 o 0 2 g mgso 4 h 2 o 0 016 g nacl 0 21 g kno 3 1 4 g nano 3 0 22 g na 2 hpo 4 2 0 ml fecl 3 solution 0 03 and.