A mixed amplicon metabarcoding and sequencing approach for surveillance of drug resistance to levamisole and benzimidazole in Haemonchus spp.

• DOI: 10.1016/j.ijpara.2023.07.002
• Publication Date: 2023-08-01T15:16:49Z
• Authors: Emily Kate Francis; Alistair Antonopo...; Mark Edward Westman; Janina McKay-Demeler; Roz Laing; Jan Šlapeta
• Keywords: Levamisole; Parasitology; Amplicon; Pyrosequencing; Veterinary parasitology

Anthelmintic-resistant parasitic nematodes present a significant threat to sustainable livestock production worldwide. The ability detect the emergence of anthelmintic resistance at an early stage, and therefore determine which drugs remain most effective, is crucial for minimising losses. Despite many years research into molecular basis resistance, no molecular-based tools are commercially available diagnosis as it emerges in field settings. We described mixed deep amplicon sequencing approach frequency levamisole (LEV)-resistant single nucleotide polymorphism (SNP) within arc-8 exon 4 (S168T) Haemonchus spp., coupled with benzimidazole (BZ)-resistant SNPs β-tubulin isotype-1 internal transcribed spacer-2 (ITS-2) nemabiome. This constitutes first known multi-drug multi-species diagnostic developed helminths veterinary importance. Of ovine, bovine, caprine camelid Australian isolates we tested, S168T was detected majority spp. populations from sheep goats, but rarely greater than 16%; arbitrary threshold set based on whole genome (WGS) LEV-resistant contortus GWBII. Overall, BZ far more prevalent LEV confirming that still effective class small ruminants New South Wales, Australia. metabarcoding herein paves way towards use large scale surveillance technology field, results can be translated evidence-based recommendations sector.