Ascarids exposed: A method for in vitro drug exposure and gene expression analysis of anthelmintic naïve Parascaris spp

J. A. Scare, P. Dini, J. K. Norris, A. E. Steuer, K. Scoggin, H. S. Gravatte, D. K. Howe, P. Slusarewicz, M. K. Nielsen

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Ascarid parasites infect a variety of hosts and regular anthelmintic treatment is recommended for all species. Parascaris spp. is the only ascarid species with widespread anthelmintic resistance, which allows for the study of resistance mechanisms. The purpose of this study was to establish an in vitro drug exposure protocol for adult anthelmintic-naïve Parascaris spp. and report a preliminary transcriptomic analysis in response to drug exposure. Live worms were harvested from foal necropsies and maintained in RPMI-1640 at 37 °C. Serial dilutions of oxibendazole (OBZ) and ivermectin (IVM) were prepared for in vitro drug exposure, and worm viability was monitored over time. In a second drug trial, worms were used for transcriptomic analysis. The final drug concentrations employed were OBZ at 40.1 μm (10 μg mL-1) and IVM at 1.1 μm (1 μg mL-1) for 24 and 3 h, respectively. The RNA-seq analysis revealed numerous differentially expressed genes, with some being potentially related to drug detoxification and regulatory mechanisms. This report provides a method for in vitro drug exposure and the phenotypic responses for Parascaris spp., which could be extrapolated to other ascarid parasites. Finally, it also provides preliminary transcriptomic data following drug exposure as a reference point for future studies of Parascaris spp.

Original languageEnglish
Pages (from-to)659-666
Number of pages8
JournalParasitology
Volume147
Issue number6
DOIs
StatePublished - May 1 2020

Bibliographical note

Funding Information:
This research was funded by the Gluck Equine Research Foundation and the Burroughs Wellcome Fund Collaborative Research Travel Award.

Publisher Copyright:
© Cambridge University Press 2020.

Keywords

  • Anthelmintic
  • Parascaris
  • ascarid
  • in vitro
  • transcriptome

ASJC Scopus subject areas

  • Parasitology
  • Animal Science and Zoology
  • Infectious Diseases

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