Cryopreservation of spermatozoa of the terrestrial Puerto Rican frog, Eleutherodactylus coqui

Published source details Michael S.F. & Jones C. (2004) Cryopreservation of spermatozoa of the terrestrial Puerto Rican frog, Eleutherodactylus coqui. Cryobiology, 48, 90-94.
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This study is summarised as evidence for the following.

	Action	Category
	Amphibians: Freeze sperm or eggs for future use Action Link
	Freeze sperm or eggs for future use Action Link

Amphibians: Freeze sperm or eggs for future use
Freeze sperm or eggs for future use

A replicated study of captive Puerto Rican frogs Eleutherodactylus coqui in the USA (Michael & Jones 2004) found that cryopreservation of sperm was successful with a cryoprotectant and fetal bovine serum (FSB). FBS alone resulted in only 8% viability. However, sperm viability was significantly higher with addition of sucrose or glycerol to FBS (sucrose: 28%; glycerol: 30%; dimethyl sulfoxide: 20%). Viability did not differ significantly with dimethyl sulfoxide. Prior to freezing sperm had a viability of 56% and so normalized viabilities were: 14% for FBS alone and 35%, 50% and 54% with added dimethyl sulfoxide, sucrose and glycerol respectively. Testes of wild caught frogs were macerated in solution. Sperm was then mixed with a cryoprotectant solution (six replicates/treatment): heat inactivated FBS alone, FBS with 2M sucrose, FBS with 2M glycerol or FBS with 2M dimethyl sulfoxide. Mixtures were frozen at −80°C for 24 hours and then thawed rapidly in a 20°C water bath. Fluorescent dye was used to examine sperm.