Purpureocillium lilacinum (Thom) Luangsa-ard, Houbraken, Hywel-Jones & Samson (Hypocreales: Ophiocordycipitaceae) is a fungus commonly used for controlling nematodes, and also has been reported as an insect pathogen. However, little is known about its effects on insects. Here, the pathogenicity of 9 isolates and the virulence and sublethal effects of 2 isolates were evaluated to control adult Anastrepha ludens (Loew) (Diptera: Tephritidae). The pathogenicity assays demonstrated that the 9 isolates were pathogenic, with mortality percentages ranging from 28.8 to 52.4% and LT₅₀ values were 18 d or more. The strain CFFSUR-A53 was more virulent than CFFSUR-A60, with LC50 values of 7.62 × 10⁶ and 5.2 × 10⁹ conidia per mL, respectively. The isolates reduced the life expectancy of the flies by 65 and 37%, decreased fecundity by 78 and 36%, and reduced egg hatching by 31.4 and 18.5%, respectively.