Microbial profile of short-beaked echidna: blood with Theileria tachyglossi and Eimeria echidnae
To identify the eukaryotes present in the diagnostic specimen, we used a community profiling based amplification 454 sequencing approach of conserved eukaryotic SSU rDNA. Whole genomic DNA from the blood sample (50-100 uL) was extracted using the Invitrogen PureLink Genomic DNA mini kit (Life Technologies, Australia) according to the manufacturer’s instructions. The gDNA was submitted to the diversity profiling service at the Australian Genomic Research Facility, Australia. The SSU rDNA assay applied was based on primer pair Euk1A (5’-CTGGTTGATCCTGCCAG-3’) and Euk516 (5’-CCAGACTTGCCCTCC-3’), amplifying approximately 500-550 bp of the 5’-end of the eukaryotic SSU rDNA. The barcoded PCR amplicon was pooled and sequenced on the 454 GS‐FLX platform.