Metagenome of parasites in Cambodian dog feces from Boeung Kork 05
Total DNA from faecal dog samples from Cambodia was subject to three independent real-time PCR amplifications. ITS-2 rDNA (ITS-2) was amplified by the NC-1 and NC-2 primers designed by Gasser et al. (1993), and two -tubulin isotype 1 regions were amplified using assay BZ167 and BZ200 developed by Jimenez Castro et al. (2019) and include key diagnostic SNPs conferring benzimidazole resistance. All primers were adapted for Illumina metabarcoding using Next Generation Sequencing (NGS). The ITS-2 target was amplified using 32-36 amplification cycles, while 40 amplification cycles were used for -tubulin isotype-1 (BZ167, BZ200). Amplicons were submitted for amplicon sequencing using Illumina NGS at the Ramaciotti Centre for Genomics, University of New South Wales, Sydney, Australia and sequenced on MiSeq using MiSeq Reagent Kits v2 (250PE, Illumina). The single run included 384 samples including metabarcoded samples of hookworm amplicons from Australia and New Zealand, Australian ruminant nematodes and Australian Angiostrongylus.