A single entomopathogenic nematode infection assay for Drosophila melanogaster larvae

IF 1.6 Q2 MULTIDISCIPLINARY SCIENCES

MethodsX Pub Date : 2025-01-06 DOI:10.1016/j.mex.2025.103157

Sreeradha Mallick, Eric Kenney, Jacob Rashap, Ioannis Eleftherianos

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Abstract

The entomopathogenic nematodes (EPNs) Steinernema carpocapsae and Steinernema hermaphroditum can efficiently infect the fruit fly, Drosophila melanogaster. The EPN infective juvenile (IJ) stage is the free-living and non-feeding stage that seeks out suitable insects to infect. While previous studies have described successful infection of D. melanogaster larvae with a standard amount of 100 IJs, the pathogenicity of a single IJ nematode towards insects remains poorly understood. Given the variability in pathogenesis among individual IJs, investigating the ability of a single IJ to cause infection in fly larvae addresses a significant gap in our understanding of host-parasite interactions. This protocol aims to evaluate the infection efficiency of a single IJ of S. carpocapsae and S. hermaphroditum. This information contributes towards a better understanding of the EPN-insect interactions and promises to improve the use of EPNs in pest management strategies. The method is summarized below:

•
Individual third instar wild-type D. melanogaster larvae are infected with either a single IJ or 100 IJs of either nematode species. Uninfected control larvae are treated with sterile water.
•
Larval survival is monitored over time.
•
Survival curves are generated, and results are statistically analyzed to determine the effect of a single nematode on the fly larval mortality.

Abstract Image