A group of 30 healthy larvae of G. mellonella of approximately similar size was selected for each treatment.
Aspergillus strains (Table 1) were revived from silica gel preservation by growth on Aspergillus complete agar medium [22] at 30 ºC
Inocula were prepared by harvesting the spores in saline Tween 80 and
the spore number was adjusted.
Each larva was injected with 105 spores directly into the haemocoel through
the last pro-leg [23].
The spores of highly virulent strains were also injected after heating at 70 ºC for 45 min
(heat-killed spores).
The number of colony forming units (CFU) was determined by plating out the known
dilution of spore suspension in triplicate. Inoculated larvae were incubated at 37 ºC in Petri dish
One group of unmanipulated larvae and the other injected with 10 μl saline Tween 80 only were also incubated as controls
Larvae were scored daily for their survival up to 168 h post-inoculation.