Lahnsteiner F, Berger B, Weismann T
Institute for Zoology, University of Salzburg, Hellbrunnerstrasse 34, A-5020 Salzburg, Austria.
Theriogenology. 2003 Sep 15;60(5):829-41. doi: 10.1016/s0093-691x(02)01300-6.
To enable cryopreservation of fish semen to become an efficient, routine technique, much more detailed information is required. Therefore, the present study was conducted to investigate various fertilization techniques and media, straw volumes as well as optimal semen volume for cryopreservation. The bleak (Chalcalburnus chalcalburnus) was used as the main model for investigation. Using frozen-thawed semen the fertilization rate was similar up to the morula stage, independent of the fertilization technique. Thereafter, in egg batches fertilized using the wet technique most embryo development stopped. In egg batches fertilized using the dry technique, embryonic development proceeded normally. For cryopreserved semen full activation of sperm motility was obtained at ratios of fertilization media (hatchery water and all tested types of saline solutions) to semen of 10:1. Sperm motility rate was much higher in the saline solutions than in water. In contrast hatching rates were higher when water was used as fertilization medium. Therefore, the requirements necessary for optimal sperm motility and optimal sperm-egg contact were different and so for these parameters optimal levels could not be achieved. When adjusting the freezing and thawing conditions 0.5ml straws as well as larger straws (1.2ml) proved suitable for cryopreservation of cyprinid semen. The highest fertilization rates were obtained with sperm to egg ratios of (1.3-2.5) x 10(6):1 and were 77-92% of fresh semen control. This was also similar for Ch. nasus, R. meidingerii, B. barbatus and C. carpio and suggests that the cryopreservation requirements of spermatozoa are not species specific.
为使鱼类精液的冷冻保存成为一种高效的常规技术，还需要更多详细信息。因此，开展本研究以探究各种受精技术和介质、细管体积以及精液冷冻保存的最佳体积。欧鳊（Chalcalburnus chalcalburnus）被用作主要研究模型。使用冻融精液，直至桑椹胚阶段，受精率与受精技术无关，均相似。此后，在采用湿法受精的卵批次中，大多数胚胎发育停止。在采用干法受精的卵批次中，胚胎发育正常进行。对于冷冻保存的精液，当受精介质（孵化场水和所有测试类型的盐溶液）与精液的比例为10:1时，精子活力可完全激活。盐溶液中的精子活力率远高于水中的。相比之下，当用水作为受精介质时孵化率更高。因此，最佳精子活力和最佳精卵接触所需的条件不同，所以这些参数无法达到最佳水平。调整冷冻和解冻条件时，0.5毫升细管以及更大的细管（1.2毫升）被证明适用于鲤科鱼类精液的冷冻保存。精子与卵子比例为(1.3 - 2.5)×10(6):1时受精率最高，为新鲜精液对照组的77 - 92%。这对于鼻吻鮈、麦氏红鲌、巴氏棒花鱼和鲤鱼也是相似的，表明精子的冷冻保存要求并非物种特异性的。
