Abstract
The management of Culex pipiens (Diptera: Culicidae), key vectors of arboviruses like West Nile virus, necessitates sustainable alternatives to chemical insecticides. This study screened indigenous entomopathogenic fungi (EPF) from forest soils in Achaia, Greece, for their larvicidal efficacy against Cx. pipiens biotype molestus. Fifteen fungal isolates were obtained via insect baiting and identified as Beauveria and Metarhizium species. A comprehensive bioassay at 1 × 10(8) conidia mL(-1) revealed significant variation in pathogenicity after 72 h. Two isolates, Beauveria bassiana (BB) (Hypocreales: Cordycipitaceae) and Metarhizium anisopliae (K3(1)) (Hypocreales: Clavicipitaceae), exhibited the highest virulence among the tested isolates, each causing 60% mortality with a rapid median lethal time (LT(50)) of ~18.5 h. Survival analysis, Cox modeling, and non-linear kinetic modeling (Gompertz/Richards) classified three distinct virulence clusters: high/rapid, moderate/consistent, and low/delayed. A pathogenicity network analysis and a composite virulence index further validated BB and K3(1) as the most effective candidates. These results demonstrate the high isolate specificity of fungal efficacy and underscore the importance of screening local fungal diversity. The identified high-virulence isolates represent promising, environmentally sound candidates for the development of targeted biopesticides. Future research should focus on formulation for aquatic environments and integration into resistance-resilient integrated vector management programs.