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P-glycoproteins (Pgp) have been proposed as contributors to the widespread macrocyclic lactone (ML) resistance in several nematode species including a major pathogen of foals,  Parascaris univalens . Using new and available RNA-seq data, ten different genomic loci encoding Pgps were identified and characterized by transcriptome-guided RT-PCRs and Sanger sequencing. Phylogenetic analysis revealed an ascarid-specific Pgp lineage, Pgp-18, as well as two paralogues of Pgp-11 and Pgp-16. Comparative gene expression analyses in  P. univalens  and  Caenorhabditis elegans  show that the intestine is the major site of expression but individual gene expression patterns were not conserved between the two nematodes. In  P. univalens ,  Pun Pgp-9,  Pun Pgp-11.1 and  Pun Pgp-16.2 consistently exhibited the highest expression level in two independent transcriptome data sets. Using RNA-Seq, no significant upregulation of any Pgp was detected following in vitro incubation of adult  P. univalens  with ivermectin suggesting that drug-induced upregulation is not the mechanism of Pgp-mediated ML resistance. Expression and functional analyses of  Pun Pgp-2 and  Pun Pgp-9 in  Saccharomyces cerevisiae  provide evidence for an interaction with ketoconazole and ivermectin, but not thiabendazole. Overall, this study established reliable reference gene models with significantly improved annotation for the  P. univalens  Pgp repertoire and provides a foundation for a better understanding of Pgp-mediated anthelmintic resistance.

The P-glycoprotein repertoire of the equine parasitic nematode Parascaris univalens