Assessment of in vitro killing assays for detecting praziquantel-induced death in Posthodiplostomum minimum metacercariae

Exp Parasitol. 2017 Oct:181:70-74. doi: 10.1016/j.exppara.2017.07.008. Epub 2017 Jul 29.

Abstract

Control of parasitic infections may be achieved by eliminating developmental stages present within intermediate hosts, thereby disrupting the parasite life cycle. For several trematodes relevant to human and veterinary medicine, this involves targeting the metacercarial stage found in fish intermediate hosts. Treatment of fish with praziquantel is one potential approach for targeting the metacercaria stage. To date, studies investigating praziquantel-induced metacercarial death in fish rely on counting parasites and visually assessing morphology or movement. In this study, we investigate quantitative methods for detecting praziquantel-induced death using a Posthodiplostomum minimum model. Our results revealed that propidium iodide staining accurately identified praziquantel-induced death and the level of staining was proportional to the concentration of praziquantel. In contrast, detection of ATP, resazurin metabolism, and trypan blue staining were poor indicators of metacercarial death. The propidium iodide method offers an advantage over simple visualization of parasite movement and could be used to determine EC50 values relevant for comparison of praziquantel sensitivity or resistance.

Keywords: Drug screening; Metacercariae; Parasite drug assays; Praziquantel; Trematode.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Animals
  • Anthelmintics / pharmacology*
  • Coloring Agents
  • Fish Diseases / parasitology*
  • Indicators and Reagents / metabolism
  • Iowa
  • Metacercariae / drug effects
  • Oxazines / metabolism
  • Perciformes / parasitology*
  • Ponds
  • Praziquantel / pharmacology*
  • Propidium
  • Spectrophotometry
  • Trematoda / drug effects*
  • Trematode Infections / parasitology
  • Trematode Infections / veterinary*
  • Trypan Blue
  • Xanthenes / metabolism

Substances

  • Anthelmintics
  • Coloring Agents
  • Indicators and Reagents
  • Oxazines
  • Xanthenes
  • resazurin
  • Propidium
  • Praziquantel
  • Adenosine Triphosphate
  • Trypan Blue