The cooling and warming rates within straw were measured with a copper constance thermocouple inserted into a straw with the cryosolution diluted semen. The straws were cooled at 8°C/min from 4°C down to -80°C, and 360°C/min down to LN2 temperature. The straws were thawed for 20 s at 580°C/min up to -2°C and 133°C/min up to 20°C in a water bath at 28°C. Motility was evaluated by a skilled operator on a subjective scale (from 0 to 10, 0:no motion, 10:fast movement of 90-100% sperms). Milt samples were placed on a slide under a dark field microscope and activated with filtered and sterilised seawater (3.9% salinity) to yield a final 1:100 dilution (milt to other liquids); no coverslip was added to the sample. For the motility profiles, scores were attributed to samples 0,30,60,120,180,240 and 300 s after activation. The best fertility data available to date (obtained in 1999) are for the fecundation of S.aurata oocytes with thawed D.puntazzo sperm, with an average result of 33.7% (best batch 47.1%, worst 11.2%) fertility rate 72 h after fecundation; egg-sperm ratio was 1/237000 and contact time 3 min; control with fresh S.aurata sperm gave 61.3%.
Notes on Diplodus puntazzo sperm cryopreservation.
ABELLI, Luigi;
2008
Abstract
The cooling and warming rates within straw were measured with a copper constance thermocouple inserted into a straw with the cryosolution diluted semen. The straws were cooled at 8°C/min from 4°C down to -80°C, and 360°C/min down to LN2 temperature. The straws were thawed for 20 s at 580°C/min up to -2°C and 133°C/min up to 20°C in a water bath at 28°C. Motility was evaluated by a skilled operator on a subjective scale (from 0 to 10, 0:no motion, 10:fast movement of 90-100% sperms). Milt samples were placed on a slide under a dark field microscope and activated with filtered and sterilised seawater (3.9% salinity) to yield a final 1:100 dilution (milt to other liquids); no coverslip was added to the sample. For the motility profiles, scores were attributed to samples 0,30,60,120,180,240 and 300 s after activation. The best fertility data available to date (obtained in 1999) are for the fecundation of S.aurata oocytes with thawed D.puntazzo sperm, with an average result of 33.7% (best batch 47.1%, worst 11.2%) fertility rate 72 h after fecundation; egg-sperm ratio was 1/237000 and contact time 3 min; control with fresh S.aurata sperm gave 61.3%.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
