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United States Department of Agriculture

Agricultural Research Service

Title: Cryopreservation of Dipteran Ebryos

Authors
item Wang, Wenbo - DEPT PLANT PATH, NDSU
item Leopold, Roger
item Freeman, Thomas - DEPT PLANT PATH, NDSU

Submitted to: Society for Cryobiology Meeting
Publication Type: Abstract Only
Publication Acceptance Date: May 30, 2000
Publication Date: N/A

Technical Abstract: This presentation describes our work on developing protocols for cryo- preservation of embryos of the house fly, Musca domestica, screwworm fly, Cochliomyia hominivorax, and Caribbean fruit fly, Anastrepha suspensa. Our studies on cryopreserving embryos of these insects established that the Drosophila melanogaster embryo cryopreservation protocols were not directly ysuitable for use with these insects. Significant progress was made when a vitrification solution containing ethylene glycol, polyethylene glycol and trehalose was formulated and when cooling and recovery of our protocol included steps which passed the embryos through liquid nitrogen vapor. More than 70% of house fly embryos withstand treatments of dechorionation, permeabilizaton, loading with cryoprotectant and dehydration in vitrification solution, but the cooling and warming steps still cause some reduction in embryo viability. Up to 57% of vitrified M. domestica embryos hatch into larvae and development of these larvae into pupae and fertile adults was observed. With a similar procedure, colonies of two strains of C. hominivorax have been reestablished after liquid nitrogen storage. A mean of 52.5% of C. hominivorax embryos hatched into larvae after storage in liquid nitrogen, 21.9% of these larvae developed into pupae and 74.0% of the pupae emerged as fertile adult flies. Results on hatching, pupal weight and adult emergence of the F1 generation showed that there was no significant difference between the untreated colonies and the groups having cryopreserved parents. Recent work on cryopreservation of A. suspensa embryos, has shown that up to 47% of vitrified embryos hatch into larvae when our protocol is used. About 29% of these larvae developed into pupae, and fertile adults were also obtained from these pupae.

Last Modified: 8/29/2014
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