Viability and virulence of freeze-dried Beauveria bassiana, Metarhizium acridum, and Hirsutella thompsonii isolates under non-refrigerated conditions in Ghana

The necessity of cold storage for mycoinsecticides poses a logistical challenge in tropical Africa, where maintaining consistent refrigeration is a major obstacle. To investigate alternative preservation methods, our study evaluates conidial-based freeze-dried formulations of Beauveria bassiana (IMI 389521), Metarhizium acridum (IMI 330189), and mycelial-based freeze-dried formulations of Hirsutella thompsonii (IMI 327488 and IMI 391722). Propagules were cryoprotected with lactose, mannitol, and raffinose, encapsulated, and freeze-dried in plastic and glass vials. Bioformulations in glass vials were cryoprotected with mannitol only. Viability (CFU/g each day) of bioformulations, following 16-weeks-storage, and their virulence (at 1 x 10(7) conidia mL(-1) in six days) against key pests of maize and cassava in Africa (larger grain borer, variegated grasshopper and cassava green mite) were determined under ambient laboratory conditions in Ghana. Our results show that non-freeze-dried conidia of B. bassiana and M. acridum had significantly higher viability and efficacy than freeze-dried versions, when stored under non-refrigerated conditions (averaging 27.7 degrees C, 78.8% RH). Freeze-dried H. thompsonii mycelia were neither viable nor virulent against cassava green mite. Freeze-dried B. bassiana and M. acridum in plastic vials remained viable for 12 weeks, while in glass vials they were viable for seven weeks. Freeze-dried B. bassiana in both plastic and glass vials caused 38-43% mortality in larger grain borer, and M. acridum caused 45-82% mortality in variegated grasshopper. Further, conidia cryoprotected with lactose had significantly higher viability and mortality than raffinose and mannitol. Our research contributes to mycoinsecticide development, particularly in the context of storage and application challenges in tropical climates.