THE EFFECT OF TWEEN 80 ON EGGSHELL PERMEABILIZATION IN GALLERIA MELLONELLA (L.) (LEPIDOPTERA, PYRALIDAE)

The development of a species-specific protocol for dechorionation/permeabilization of insect eggs is a necessary prerequisite to cryopreserve the embryos. Here we tested different procedures based on heptane or the surfactant Tween 80 as an alternative to alkane, evaluating their efficacy and toxicity on the early (24 h post-oviposition) and late (75 h post-oviposition) stage embryos. Heptane efficiently permeabilized the eggs of G. mellonella but the hatching rate ranged from 0.1 to 4.2% in the early stage and from 4.3 to 11.2% in the late stage. The embryos treated with 1.25% NaOCl + 0.08% Tween 80 for 2 min showed the same shrinkage and reswelling percentages as eggs exposed to heptane for 10 sec, with a significantly higher hatching percentage in the early (68.2 +/- 1.5%) and late stages (22.4 +/- 3.7%). Thus, 0.08% Tween 80 allows sufficient permeabilization of G. mellonella embryos without the high toxicity of alkane.