While there was no visible relationship between geography or body site of infection, there was a clear separation between the koala and non-koala strains (Figure 4). As ancestral relationships are not being inferred between the koala and non-koala hosts, unrooted phylogenetic CCI-779 research buy trees were used to illustrate this data. Figure 3 Phylogenetic tree of omp A sequences from koala C. pecorum isolates, with previously published sequence information. Unrooted; inferred by the neighbour-joining

method with bootstrapping support (1000 replicates). Figure 4 Phylogenetic tree of the koala C. pecorum isolates sequenced, with previously published sequence information. Unrooted; constructed using concatenated sequences of

ompA, incA, and ORF663 using the neighbour-joining method with bootstrapping support (1000 replicates). Genotypic analysis of the ompA, incA, tarP, and ORF663 genes To highlight the GNS-1480 discriminatory power of ompA, incA, tarP, and ORF663, C. pecorum-specific GW-572016 in vivo genotypes were established based on their level of nucleotide dissimilarity and aligned with the phylogenetic gene trees outlined above (Figure 1). The ompA gene was able to separate the koala samples into four genotypes, the incA gene produced three genotypes, the tarP gene separated the clinical samples into two genotypes, while ORF663 was able to discriminate between seven distinct genotypes. Recombination Each of the four shortlisted genes (ompA, incA, ORF663, tarP) was tested for evidence of recombination by the RDP. All sequences were found to deviate from clonality by all six recombination tests (P < 0.001), which is consistent with previous reports regarding ompA and ORF663 [19, 53]. Discussion The current study revealed three novel and significant characteristics

of the evolution and genetic diversity of C. pecorum infections in the koala: (1) the ompA gene has a phylogenetic history that is congruent with other gene targets in the C. pecorum genome, yet is phylogenetically-insufficient for use as a single gene marker; (2) the tarP and ORF663 genes are potentially useful in representing C. pecorum Resveratrol genomic diversity and evolution, and (3) koala C. pecorum infections appear to be monophyletic, possibly suggesting a limited number of cross-host transmission events between koalas and non-koala hosts. The ompA gene is one of the most polymorphic genes across all Chlamydia species [23] and as a result, was previously selected as the molecular marker of choice in epidemiological and genotyping studies of C. pecorum infections of the koala. This increased nucleotide diversity is reported to be due to the antigenicity of MOMP and the selective pressure of the host’s immune response [54]. Early C. trachomatis studies and more recent C.