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Simple and Effective Methods of Freezing Capercaillie (Tetrao urogallus L.) Semen

机译:冷冻Capercaillie(Tetrao urogallus L.)精液的简单有效方法

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摘要

A continuous decline in the number and range of capercaillie (Tetrao urogallus L.) in many European countries can be observed, mostly due to habitat destruction by human activity, unecological forestry management, and increased density of natural predators. Ex situ in vitro gene banks provide a unique opportunity to preserve the genetic material for future generations. Simple and effective cryopreservation methods for capercaillie semen are discussed. Semen was collected from seven males kept in the Capercaillie Breeding Centre at Forestry Wisła in Poland. Within five minutes after collection, ejaculates were diluted with EK diluent, then divided into two parts, and subjected to two freezing procedures: in pellets and in straws. In fresh semen, ejaculate clearness, viscosity, color and volume, as well as sperm concentration, motility and morphology, were evaluated, while in frozen-thawed semen only motility and morphology of sperm were determined. Fertilizing ability of thawed semen was examined for samples frozen in straws. Significant (P<0.05) differences between individual males were found in relation to the majority of fresh semen traits: ejaculate volume averaged 102.1 µL (varying from 49.0 to 205.0); average sperm concentration was 632.5 x106 mL-1 (178.8–1257.1); percentage of live normal cells varied from 39.2 to 70.3% (58.7% on an average); percentage of motile cells ranged from 76.0 to 85.7%) and motility parameters were male dependent, as well. Both cryopreservation methods had a negative effect on morphology and motility of frozen-thawed semen; however, the straw method yielded 60.7% and the pellet method 42.5% of live cells in total in thawed semen (P<0.05), while the number of live normal (intact) cells was similar (22.4 and 22.2%, respectively). Egg fertility varied between 77.8 and 91.7% (average 84.4%). Both freezing procedures seem to be effective in obtaining acceptable viability and high fertilizing potency of thawed sperm and can be used to create a gene bank of capercaillie semen.
机译:在许多欧洲国家中,可以看到Capercaillie(Tetrao urogallus L.)的数量和范围持续下降,这主要是由于人类活动破坏了栖息地,非生态林业管理以及自然捕食者密度增加。异地体外基因库为保存子孙后代的遗传物质提供了独特的机会。讨论了一种简单而有效的冷冻Capercaillie精液的方法。精液是从波兰林业WisłaCapercaillie繁育中心饲养的七只雄性中采集的。收集后五分钟内,将射精用EK稀释剂稀释,然后分成两部分,并进行两次冷冻程序:在小球中和在稻草中。在新鲜精液中,评估了射精的透明度,粘度,颜色和体积,以及精子的浓度,活力和形态,而在冷冻融化的精液中,仅确定了精子的活力和形态。对于在秸秆中冷冻的样品,检查了解冻精液的受精能力。与大多数新鲜精液性状相关,各个雄性之间存在显着差异(P <0.05):射精量平均为102.1 µL(从49.0到205.0)。平均精子浓度为632.5 x10 6 mL -1 (178.8–1257.1);正常活细胞的百分比在39.2%至70.3%之间(平均58.7%);运动细胞的百分比在76.0%至85.7%之间,运动参数也取决于男性。两种冷冻保存方法均对冻融精液的形态和运动性产生负面影响。然而,在融化后的精液中,秸秆法产生的活细胞总数为60.7%,沉淀法产生的活细胞总数为42.5%(P <0.05),而正常(完整)活细胞的数量相似(分别为22.4%和22.2%)。鸡蛋的受精率介于77.8和91.7%之间(平均84.4%)。两种冷冻程序似乎都可以有效地获得融化的精子的可接受的生存力和高受精力,并且可以用于创建Capercaillie精液的基因库。

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