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Comparison of Methods for Isolating Entomopathogenic Fungi from Soil Samples
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Comparing two methods for quantifying soil-borne Entomophaga maimaiga resting spores.

Ann E Hajek1, Ruth C Plymale, James R Reilly

  • 1Department of Entomology, Cornell University, Ithaca, NY 14853-2601, USA. aeh4@cornell.edu

Journal of Invertebrate Pathology
|August 16, 2012
PubMed
Summary

Quantifying soil-borne fungal spores like those from Entomophaga maimaiga is crucial for pest management. This study refined two methods, finding both effective for detecting gypsy moth pathogen resting spores in soil.

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Area of Science:

  • * Environmental science
  • * Mycology
  • * Soil ecology

Background:

  • * Entomophthoralean fungi, including Entomophaga maimaiga, produce environmentally persistent resting spores (azygospores) crucial for pathogen survival and disease dynamics.
  • * Accurate quantification of these soil-borne spores is essential for understanding pathogen ecology and developing effective pest management strategies for insects like the gypsy moth.
  • * Existing methods for quantifying resting spores in soil can be cumbersome, necessitating improvements in usability and reliability.

Purpose of the Study:

  • * To modify and evaluate two distinct methods for improved quantification of environmentally persistent entomophthoralean resting spores in soil.
  • * To assess the efficacy of these modified methods in recovering resting spores of the gypsy moth pathogen, Entomophaga maimaiga.
  • * To provide regression equations for estimating true resting spore densities from direct counts obtained by the tested methods.

Main Methods:

  • * Modification and comparative testing of two established methods for resting spore quantification: a modified Weseloh and Andreadis (2002) method and a modified Percoll density gradient method.
  • * Application of these methods to soil samples spiked with known concentrations of Entomophaga maimaiga resting spores.
  • * Statistical analysis to compare recovery rates and assess the ability to estimate true spore densities from counts.

Main Results:

  • * Both modified methods demonstrated effectiveness in recovering resting spores at concentrations greater than 100 resting spores/g dry soil.
  • * The modified Weseloh and Andreadis method recovered a higher number of resting spores compared to the modified Percoll density gradient method.
  • * The capacity to estimate true resting spore densities from counts was comparable between both methods, with regression equations yielding R(2) values of ≥0.90 for both.

Conclusions:

  • * The refined methods offer improved usability for quantifying soil-borne resting spores of entomophthoralean pathogens.
  • * Both tested methods are reliable for detecting and estimating densities of Entomophaga maimaiga resting spores in soil at relevant concentrations.
  • * The provided regression equations enhance the accuracy of spore density estimations, aiding ecological and management studies.