Motility and cryopreservation of spermatozoa of European common frog, Rana temporaria

Published source details Mansour N., Lahnsteiner F. & Patzner R.A. (2010) Motility and cryopreservation of spermatozoa of European common frog, Rana temporaria. Theriogenology, 74, 724-732.
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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: Use hormone treatment to induce sperm and egg release Action Link
	Use hormone treatment to induce sperm and egg release during captive breeding Action Link

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

A replicated study in 2009 of captive European common frogs Rana temporaria in Austria (Mansour, Lahnsteiner & Patzner 2010) found that the most effective cryopreservation protocol was sperm in motility-inhibiting saline (MIS) with 5% glycerol, 2.5% sucrose and 5% hen egg yolk, frozen 10 cm above liquid nitrogen and thawed at 22 °C for 40 seconds. Sperm motility was maintained following incubation for 40 minutes at 4°C in MIS with 10% dimethyl sulfoxide (71%), 5% glycerol (69%) or 10% methanol (59%), but not 10% propandiol (0%). When frozen, in combination with sucrose, dimethyl sulfoxide resulted in significantly greater sperm motility and viability (10%; 42% respectively) than glycerol (8%; 25%). With MIS, motility and viability was similar with either dimethyl sulfoxide (13%; 27%) or glycerol (10%; 29%). Sperm frozen in MIS with sucrose and methanol had no motility. Sperm frozen 5 cm above liquid nitrogen had no motility, whereas at 10 cm motility was 30–35%. Addition of 5% (vs 10%) egg yolk and 2.5% sucrose to MIS with glycerol significantly increased hatching rate compared to all other treatments (23 vs 2–12%). Motility and viability did not differ. Testes from wild males were macerated (3/treatment). Sperm was frozen in liquid nitrogen. Fertilization was tested using 25–30 eggs.
Amphibians: Use hormone treatment to induce sperm and egg release
Use hormone treatment to induce sperm and egg release during captive breeding

A replicated study in 2009 of captive European common frogs Rana temporaria in Austria (Mansour, Lahnsteiner & Patzner 2010) found that injecting males with human chorionic gonadotrophin increased sperm production. Males stimulated with hormones had greater sperm production than untreated males (0.004 vs 0.002 testis/body weight). The same was true for the sperm cell concentration (80 vs 11 x 10⁶/ml in 1.5 ml motility-inhibiting saline/testes). Males received injections of 150 IU of human chorionic gonadotrophin and were killed after 15 hours. Testes were removed weighed and macerated in motility-inhibiting saline.