caa a22 4500 397567278 CHVBK 20180308164831.0 cr unu---uuuuu 161202e199607 xx s 000 0 eng 10.3382/ps.0750915 doi (NATIONALLICENCE)oxford-10.3382/ps.0750915 Cryopreservation of Rooster Sperm Using Methyl Cellulose [Elektronische Daten] [J. J. PHILLIPS, R. K. BRAMWELL, J. K. GRAHAM] Experiments were designed to determine when, during the cryopreservation process, sperm lose fertilizing capacity and whether the cryoprotectant, methyl cellulose (MC), could be used in combination with glycerol to cryopreserve sperm and remain in the inseminate without reducing fertility. Semen diluted in Minnesota Avian extender (MNA) and inseminated immediately had greater fertility (75%) than semen processed for cryopreservation (12 to 60%). The largest decreases in fertility were due to addition of glycerol to sperm and to cryopreservation. In another experiment, fertility of inseminates containing 0, 1, and 2% glycerol were 82, 29, and 21% , respectively, for eggs collected 2 to 5 d after insemination. When 0.5% MC was added to the same three treatments, fertility rates were 88, 63, and 69%, respectively. Semen cryopreserved in MNA containing 9% glycerol; MC + 3% glycerol; MC + 4% glycerol; MC + 9% glycerol; or 9% glycerol with the cryoprotectant removed post-thaw by dilution and subsequent centrifugation exhibited 59, 30, 35, 60, and 69% viable cells, respectively; and 65, 38, 46, 69, and 65% motile sperm, respectively. Sperm cryopreserved with MC and either 4 or 9% glycerol exhibited similar numbers of sperm binding to chicken perivitelline layers in vitro as did fresh sperm, whereas sperm frozen with MC and 3% glycerol bound oocytes with only 31% efficiency (P < 0.05). The extent to which cryopreserved sperm penetrated the perivitelline layer in vitro was independent of glycerol concentration, but was four times more efficient than that of fresh sperm (P < 0.05). The fertility rates of fresh semen, semen frozen in 9% glycerol with the cryoprotectant removed after thawing, and semen frozen in MC with either 3 or 4% glycerol were 87.4, 27.6, 0.8, and 0.5%, respectively (P < 0.05). The MC reduces the contraceptive effects of glycerol when inseminated with fresh sperm, but does not maintain fertilizing capacity in frozen-thawed sperm when used in combination with 3 or 4% glycerol. © 1996 Poultry Science Association, Inc. Physiology and Reproduction nationallicence cryopreservation nationallicence rooster nationallicence spermatozoa nationallicence methyl cellulose nationallicence PHILLIPS J. J. aut BRAMWELL R. K. aut GRAHAM J. K. 2To whom correspondence should be addressed. aut Poultry Science Oxford University Press 75/7(1996-07), 915-923 0032-5791 75:7<915 1996 75 ps https://doi.org/10.3382/ps.0750915 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.3382/ps.0750915 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- PHILLIPS J. J. aut NATIONALLICENCE 700 1- BRAMWELL R. K. aut NATIONALLICENCE 700 1- GRAHAM J. K. 2To whom correspondence should be addressed aut NATIONALLICENCE 773 0- Poultry Science Oxford University Press 75/7(1996-07), 915-923 0032-5791 75:7<915 1996 75 ps Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford