Susceptibility of Metarhizium spp. and other entomopathogenic fungi to dodine-based selective media

Abstract

The fungicide dodine has been widely used in selective media to isolate entomopathogenic fungi (EF) from contaminating microorganisms, primarily bacteria and non-entomopathogenic fungi. In order to isolate the fungus Metarhizium acridum from soil for grasshopper and Mormon cricket control in the western USA, the susceptibility of M. acridum was compared with two Metarhizium spp. and other EF species. The isolates were inoculated onto mycological media with concentrations of dodine ranging from 0.0001 to 0.03% active ingredient (A.I.). In addition, susceptibilities of five Metarhizium spp. isolates to two sources of dodine, Syllit® commercial fungicide (65% A.I.) and Sigma® reagent grade (99% A.I.), were compared using Czapek agar medium. Responses to the two dodine sources were virtually identical. Accordingly, subsequent experiments used the less expensive Syllit dodine. Three media [Czapek, potato dextrose agar plus yeast extract (PDAY) and oatmeal agar] were evaluated for appropriateness as the base in selective media. Germination of all three of the M. acridum isolates tested was almost completely inhibited by dodine concentrations of 0.002% A.I. in Czapek or 0.006% A.I. in PDAY. On the other hand, M. robertsii and M. anisopliae isolates were considerably more tolerant, with germination not being inhibited until 0.010% A.I. in Czapek or 0.030% A.I. in PDAY. The higher vulnerability of the isolates to low concentrations of dodine in Czapek medium suggests that this medium would be less effective than PDAY in a selective medium. Oatmeal agar greatly improved fungal growth, but the levels of inhibition were lower. Therefore, PDAY was selected as the best selective basal medium. The lowest concentration that inhibited a common soil-inhabiting fungus, Aspergillus nidulans, was 0.001% A.I. Dodine tolerances were highest with M. robertsii, M. anisopliae, and Beauveria bassiana, followed by Isaria fumosorosea and Lecanicillium spp. The least tolerant EF isolates were M. acridum.