TITLE:
First Estimation of Drosophila EPS Solution for Permeabilizing Lepidoptera Galleria mellonella Embryos
AUTHORS:
Muhamad Abidalla, Pio Federico Roversi
KEYWORDS:
Permeabilization, G. mellonella, Drosophila, D-Limonene, Eggshell
JOURNAL NAME:
Advances in Entomology,
Vol.6 No.4,
July
31,
2018
ABSTRACT: The increased importance of the G. mellonella for wide range of scientific research and commercial sides will need to create a germplasm resource banking by cryopreservation. Impermeability is a fundamental limiting factor for the successful cryopreservation of arthropods embryos. The successful permeability of Drosophila embryo by using an embryo permeabilization solvent (EPS) solution encouraged this trial on G. mellonella embryos (stage of 24 hours Post-oviposition (h PO)). Permeability assessment with Rhodamine B and crystal violet dyes showed that G. mellonella embryos can be permeabilized by EPS of D-limonene that has 3 mol ethoxylated alcohol. The permeabilization for 30 sec exposure time was resulted 61.5% ± 5.8% survival rate, 31.7% ± 3.1% uptakes dyes and 40.5% ± 0.3% was the survival rate post loading in 12% Ethylene glycol (EG). The low viability after immersion in liquid nitrogen (LN) (0.6% ± 0.08%) is due to the dual toxicity of EPS and cryoprotectant (CPA) solutions. However, fluorescence images showed sufficient permeability that confirms the possibility to increase the permeability of G. mellonella embryos with EPS solution, and to have the opportunity to improve the viability after LN by improving procedures of loading and dehydration with various CPAs and exposure times, which decrease the toxicity effect.