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Envenomation and inoculation procedures

OP Olga V. Polenogova
MK Marsel R. Kabilov
MT Maksim V. Tyurin
UR Ulyana N. Rotskaya
AK Anton V. Krivopalov
VM Vera V. Morozova
KM Kseniya Mozhaitseva
NK Nataliya A. Kryukova
TA Tatyana Alikina
VK Vadim Yu. Kryukov
VG Viktor V. Glupov
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Two H. hebetor females were placed for 10 h in 90 mm Petri dishes with ten fifth-instar larvae of G. mellonella for envenomation. Then, eggs of the parasitoid were washed off of paralyzed larvae with distilled water. Envenomated and nonenvenomated larvae were dipped in either a water-Tween suspension of B. bassiana (1.0 × 106 conidia/mL), or in a conidia-free water-Tween solution for 10 s. Then, the larvae were placed in 90 mm Petri dishes (10 larvae per dish) with 80 mm discs of filter paper moistened by sterile water (1 mL per disk). Petri dishes with insects were maintained in a climatic chamber (KBWF 240, Binder, Germany) at 28 °C and 75% RH. Larval mortality was estimated over 12 days. Cadavers were placed in moisture chambers to confirm mortality due to B. bassiana.

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