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Home / Equine Research Bank / Parasitology / Ascarids exposed: a method for in vitro drug exposure and ge...
Parasitology2020; 147(6); 659-666; doi: 10.1017/S0031182020000189

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

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.
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Publication Date: 2020-02-12 PubMed ID: 32046800PubMed Central: PMC10317622DOI: 10.1017/S0031182020000189Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research focuses on ascarid parasites and their response to anthelmintic treatment. It presents a method for in vitro drug exposure and examines genetic expression in reaction to the treatment, particularly in the Parascaris spp. variety, which shows widespread anthelmintic resistance.

Introduction to Ascarid Parasites

  • The study is centered around ascarid parasites, which are known to infect a variety of hosts. Regular anthelmintic treatment – a category of medication specifically designed to expel parasitic worms – is advised for all species.
  • Particularly, the study pays attention to the Parascaris spp. species. This species is noteworthy because it is the only ascarid species that has developed widespread resistance to anthelmintic therapies.

Purpose of the Study

  • The primary purpose of the study is the development of an in vitro (conducted in a laboratory setting) drug exposure protocol for adult anthelmintic-naïve (not previously exposed to antiparasitic drugs) Parascaris spp. The report also provides a preliminary study of how these parasites’ genes express themselves in response to drug exposure.

Methodology

  • Live worms were retrieved from foal necropsies and kept in a solution called RPMI-1640 at temperatures of 37 degrees Celsius.
  • Serial dilutions of two antiparasitic drugs, oxibendazole (OBZ) and ivermectin (IVM), were prepared for in vitro drug exposure. Throughout this procedure, worm viability was monitored.
  • In a second drug trial, worms were used for transcriptomic analysis. This analysis allows to study the full range of RNA transcripts produced by the genome under specific circumstances, in this case, the exposure to anthelminthic drugs.
  • The final drug concentrations implemented were OBZ at 40.1 μm (10 μg mL-1) and IVM at 1.1 μm (1 μg mL-1) for a span of 24 hours and 3 hours, respectively.

Results and Conclusion

  • The RNA-seq analysis, a technique to survey the presence and quantity of RNA in a biological sample, indicated several differentially expressed genes, some of which might be tied to drug detoxification processes and regulatory systems within the parasite.
  • The study provides a model for the use of in vitro drug exposure and the phenotypic (observable physical traits) responses for Parascaris spp.
  • This method could potentially be extrapolated and applied to the study of other ascarid parasites.
  • Lastly, the study serves as a preliminary reference point for future transcriptomic studies related to Parascaris spp. post-drug exposure, and it points towards the possibility of using this approach to better understand the mechanisms behind drug resistance in parasites.

Cite This Article

APA
Scare JA, Dini P, Norris JK, Steuer AE, Scoggin K, Gravatte HS, Howe DK, Slusarewicz P, Nielsen MK. (2020). Ascarids exposed: a method for in vitro drug exposure and gene expression analysis of anthelmintic naïve Parascaris spp. Parasitology, 147(6), 659-666. https://doi.org/10.1017/S0031182020000189

Publication

Parasitology
ISSN: 1469-8161
NlmUniqueID: 0401121
Country: England
Language: English
Volume: 147
Issue: 6
Pages: 659-666

Researcher Affiliations

Scare, J A
  • Department of Veterinary Science, M.H. Gluck Equine Research Center, University of Kentucky, Lexington, KY, USA.
Dini, P
  • Department of Veterinary Science, M.H. Gluck Equine Research Center, University of Kentucky, Lexington, KY, USA.
Norris, J K
  • Department of Veterinary Science, M.H. Gluck Equine Research Center, University of Kentucky, Lexington, KY, USA.
Steuer, A E
  • Department of Veterinary Science, M.H. Gluck Equine Research Center, University of Kentucky, Lexington, KY, USA.
Scoggin, K
  • Department of Veterinary Science, M.H. Gluck Equine Research Center, University of Kentucky, Lexington, KY, USA.
Gravatte, H S
  • Department of Veterinary Science, M.H. Gluck Equine Research Center, University of Kentucky, Lexington, KY, USA.
Howe, D K
  • Department of Veterinary Science, M.H. Gluck Equine Research Center, University of Kentucky, Lexington, KY, USA.
Slusarewicz, P
  • MEP Equine Solutions, 3905 English Oak Circle, Lexington, KY40514, USA.
Nielsen, M K
  • Department of Veterinary Science, M.H. Gluck Equine Research Center, University of Kentucky, Lexington, KY, USA.

MeSH Terms

  • Animals
  • Anthelmintics / pharmacology
  • Ascaridoidea / drug effects
  • Ascaridoidea / genetics
  • Drug Resistance / genetics
  • Gene Expression
  • Gene Expression Profiling
  • Genes, Helminth
  • In Vitro Techniques
  • RNA-Seq
  • Transcriptome

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