Loomis P R, Amann R P, Squires E L, Pickett B W
J Anim Sci. 1983 Mar;56(3):687-93. doi: 10.2527/jas1983.563687x.
The fertility of frozen-thawed semen was compared with that obtained using fresh semen extended in skim milk. Semen for freezing was obtained in June from four stallions of unknown fertility; two ejaculates were collected 1 to 2 h apart every 3 or 4 d. The gel-free fraction of the ejaculate was mixed 1:1 with a glucose-EDTA solution (disodium ethylenediaminetetraacetic acid) and centrifuged at 650 x g for 15 min. The spermatozoa were resuspended in an EDTA-lactose-egg yolk extender containing 5% glycerol. Semen was frozen in .5-ml French straws containing 250 x 10(6) progressively motile spermatozoa before freezing. Only for 31% of the 54 ejaculates frozen was post-thaw spermatozoal motility greater than or equal to 50% of the percentage of progressively motile spermatozoa observed during evaluation of the neat semen. Spermatozoa in second ejaculates apparently were more susceptible to the adverse effects of dilution and centrifugation than spermatozoa in first ejaculates. Only samples containing greater than 30% progressively motile spermatozoa after freezing and thawing (at 38 C) were used for insemination. In June and July, 101 mares were inseminated daily with semen from one of three stallions beginning on d 2 and continuing through the end of estrus for one cycle. Mares were inseminated with semen in one straw or with 250 x 10(6) progressively motile spermatozoa extended in 10 ml of skim milk. Because of the poor survival of spermatozoa that had been frozen and thawed, mares inseminated with frozen-thawed semen received 100 to 130 x 10(6) progressively motile spermatozoa as compared with 250 x 10(6) progressively motile spermatozoa for mares inseminated with fresh semen. One cycle pregnancy rates, based on rectal palpation 50 to 60 d after ovulation, were 29% using frozen-thawed semen and 66% using fresh semen (P less than .05). Values for individual stallions were 19, 24 and 47% with frozen semen and 47, 61 and 67% with fresh semen. Routine use of frozen stallion semen is not recommended at this time.
将冻融精液的生育能力与使用在脱脂乳中稀释的新鲜精液所获得的生育能力进行了比较。用于冷冻的精液于6月从4匹生育能力未知的种马采集;每隔3或4天,每隔1至2小时采集2次射精。将射精的无凝胶部分与葡萄糖-乙二胺四乙酸溶液(乙二胺四乙酸二钠)按1:1混合,并以650×g离心15分钟。精子重悬于含有5%甘油的乙二胺四乙酸-乳糖-蛋黄稀释液中。精液在冷冻前被装入含有250×10⁶条进行性运动精子的0.5毫升法式细管中。在冷冻的54次射精中,只有31%的解冻后精子活力大于或等于在评估原精液时观察到的进行性运动精子百分比的50%。第二次射精中的精子显然比第一次射精中的精子更容易受到稀释和离心的不利影响。只有冻融(38℃)后含有超过30%进行性运动精子的样本用于授精。在6月和7月,101匹母马从第2天开始,每天用3匹种马之一的精液进行授精,持续到发情期结束,为期一个周期。母马用一管精液或用在10毫升脱脂乳中稀释的250×10⁶条进行性运动精子进行授精。由于冻融精子的存活率低,用冻融精液授精的母马接受100至130×10⁶条进行性运动精子,而用新鲜精液授精的母马接受250×10⁶条进行性运动精子。基于排卵后50至60天直肠触诊的一个周期妊娠率,使用冻融精液为29%,使用新鲜精液为66%(P<0.05)。使用冷冻精液时,各匹种马的值分别为19%、24%和47%,使用新鲜精液时分别为47%、61%和67%。目前不建议常规使用冷冻种马精液。