BackgroundThe pine processionary moth Thaumetopoea wilkinsoni Tams, 1926 (Lepidoptera: Notodontidae), causes severe skin reactions to animals and humans. The insect also destroys pine ecosystems by feeding on pine leaves at its larval stage. Instead of chemical control, eco-friendly biological control methods should be preferred to combat this species.ResultsThe purpose of this study was to evaluate the efficacy of Beauveria bassiana isolate (GOPT-331) and Metarhizium brunneum isolates (ORP-13 and ORP-18) against the second and fourth larval instars of T. wilkinsoni under laboratory conditions. T. wilkinsoni eggs were collected from pine trees at Ondokuz Mayis University in Samsun, Turkey, in 2021, and the second and fourth larval instars were used for the experiment. Two milliliters of a spray of the three fungal isolates was applied to the larvae for each concentration (1 x 10(5)-1 x 10(8) conidia ml(-1)). The mortality rates of GOPT-331, ORP-13, and ORP-18 were changed between 91.1 and 100% for the second-instar larvae and between 86.6 and 97.7% for the fourth-instar larvae at 1 x 10(8) conidia ml(-1). The ORP-13 isolate showed the lowest LC50 values.ConclusionIn conclusion, it is suggested that all the three isolates were virulent to T. wilkinsoni and can be used for biological control of this species. The promising results from the study were obtained from trials conducted under controlled laboratory conditions, and it will be critical to explore the potential of these promising entomopathogenic fungi in field conditions.