Conidiobolus lunulus, a newly discovered entomophthoralean species, pathogenic and specific to leaf-cutter ants

[Display omitted] •Conidiobolus lunulus kills > 85% leaf-cutter ants in 1–2 days under optimal conditions.•Fungal virulence is highest at 24 °C, 88% RH, and when using 2-day-old fungal cultures.•C. lunulus reduces other naturally occurring entomopathogens.•C. lunulus is specific to leaf-cutter ants.•C. lunulus seems a promising candidate for leaf-cutter ant biocontrol. Conidiobolus lunulus is a recently described entomophthoralean species isolated from leaf-cutter ants. This fungus discharges not only primary but also secondary conidia and microconidia of different shapes. Because nothing was known about the biology of the fungus, and its interactions with hosts, we first evaluated if its pathogenicity against leaf-cutter ants changes with the fungal age (time grown in vitro), and if it is related to the conidial structures produced. Afterwards, we tested its virulence at three combinations of temperature and relative humidity. In addition, we noted all visible causes of death by recovering different microorganisms from the dead, non-sterilized, ants to evaluate C. lunulus virulence when pathogens carried naturally by the ants were present. Finally, we used the conditions that lead to the highest mortality to evaluate fungal virulence to other host species, including non-leaf-cutter ants. Results indicated that C. lunulus was pathogenic from a culture age of 1 to 5 days, with a peak at 2-days-old, from which we registered median lethal times of 1–2 days and 85% of the cadavers with fungal conidiation. Our results suggest that primary conidia and moon-shaped microconidia were infective. Evaluations of mortality using 2-days-old cultures on several leaf-cutter ant colonies showed 1) significantly faster mortality of C. lunulus inoculated ants in comparison to controls, 2) significantly greater and faster mortality at 23.7 °C than at 21.2 °C, 3) significantly higher and faster mortality at 88% than at 57% RH, and 4) a significant reduction of other pathogens in C. lunulus inoculated ants in comparison to controls. C. lunulus was highly specific to leaf-cutter ants, as hardly any increase in mortality was observed on inoculated ants, and no conidia were recorded on cadavers of the other three non-leaf-cutter ant species tested. Our results highlight that C. lunulus is a very promising biological control agent against leaf-cutter ants.