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Chemical pesticides have been used for pest control for many decades, but they cause serious problems, including insecticide resistance, secondary pest resurgence, and negative environmental impacts. Therefore, sustainable alternatives to chemical pesticides are necessary for pest control. Entomopathogenic fungi (EPF) are natural epizootic pathogens of insects, and some of them have been used as microbial biocontrol agents. Herein, we attempted to construct an entomopathogenic fungal library (EFLib) via the “ Tenebrio molitor  pathogenicity-based fungal collection method (TmPC)” to select EPF for control of the serious agricultural pest  Spodoptera litura . A total of 172 soil samples were collected in northern and central Taiwan for the EFLib construction. The isolation efficiency of TmPC was 64.02%. The EFLib consisted of 101 isolates and was designated as the National Chung Hsing University (NCHU) EFLib. Among these isolates, 26 showed high virulence (mortality = 100%) to  T. molitor  larvae. Based on the results of molecular identification, the highly virulent isolates belonged to seven genera, including  Beauveria, Clonostachys, Fusarium, Cordyceps, Penicillium, Purpureocillium , and  Metarhizium . To evaluate the potential of these isolates for  Spodoptera litura  control, 12 isolates were selected for pathogenicity screening against  S. litura  larvae. A total of six EPF isolates belonging to the genera  Beauveria  and  Metarhizium  showed rapid eradication of the  S. litura  larvae. To rank the potential of these fungal strains for pest management, the six isolates were subjected to thermotolerance and conidial production assays, and a novel effective conidia number (ECN) formula was applied. The results indicated that the ECN index of  Beauveria australis  (NCHU-113) was much higher than that of  Metarhizium  isolates. However, among the  Metarhizium  isolates, NCHU-95 showed the highest ECN index. Altogether, NCHU-69 and NCHU-113 should be further tested in field trials. To our knowledge, this is the first attempt to integrate pathogenicity or virulence and ECN data into EPF screening and ranking, providing a baseline for mass selection of potential EPF strains for further applications.

Construction and Selection of an Entomopathogenic Fungal Library From Soil Samples for Controlling Spodoptera litura