Stanishevskaya Olga, Silyukova Yulia, Pleshanov Nikolai, Kurochkin Anton, Fedorova Elena, Fedorova Zoya, Perinek Oksana, Prituzhalova Anna, Meftakh Inessa
Branch of the L.K. Ernst Federal Science Center for Animal Husbandry, Russian Research Institute of Farm Animal Genetics and Breeding, 196625 Saint-Petersburg, Russia.
Animals (Basel). 2021 Jan 14;11(1):189. doi: 10.3390/ani11010189.
The aim of this study was to create balanced media for the cryopreservation of rooster semen in pellets to maintain the functional state of the sperm after thawing. Fructose was replaced by trehalose in experimental media in proportions of 10% (LCM-T10) and 20% (LCM-T20), while LCM was used as a control. After artificial insemination of the hens, the eggs were incubated (n = 400). To determine the functional safety of spermatozoa in the genital tract of hens after 5, 10, and 15 days from the last insemination, we used a method for assessing the interaction of sperm with the perivitelline membrane. Significantly higher rates of egg fertilization (82-86%) were obtained when using LCM-T10 and LCM-T20 compared to control (79%, < 0.05). Egg fertility on the 5th day from the last insemination with the LCM-T20 diluent reached 100% versus 86% in the control; on the 10th day, the fertility rates were 55% versus 20%, respectively. The best results for fertility duration were obtained by freezing spermatozoa with LCM-T20 medium. The numbers of interaction points of spermatozoa with the perivitelline membrane were as follows: on the 5th day from the last insemination with LCM-T20-461.5 ± 11.5 holes/cm (LCM-control-13.7 ± 2.7 holes/cm), < 0.01; on the 10th day with LCM-T20-319.3 ± 12.9 holes/cm (LCM-control-14.9 ± 3.5 holes/cm); and on the 15th day with LCM-T20-345.2 ± 11.1 holes/cm (LCM-control-0 holes/cm). In conclusion, the use of trehalose in LCM diluent medium can increase the fertility of frozen/thawed sperm and the duration of their fertility in the genital tract of hens.
本研究的目的是制备用于冷冻保存颗粒状公鸡精液的平衡培养基,以维持解冻后精子的功能状态。在实验培养基中,用海藻糖按10%(LCM-T10)和20%(LCM-T20)的比例替代果糖,而LCM用作对照。对母鸡进行人工授精后,将鸡蛋进行孵化(n = 400)。为了确定最后一次授精后5天、10天和15天母鸡生殖道内精子的功能安全性,我们采用了一种评估精子与卵黄膜相互作用的方法。与对照组(79%,P<0.05)相比,使用LCM-T10和LCM-T20时获得了显著更高的卵子受精率(82-86%)。用LCM-T20稀释剂最后一次授精后第5天的卵子受精率达到100%,而对照组为86%;在第10天,受精率分别为55%和20%。用LCM-T20培养基冷冻精子获得了最佳的受精持续时间结果。精子与卵黄膜的相互作用点数如下:最后一次授精后第5天,LCM-T20为461.5±11.5个孔/cm(LCM对照组为13.7±2.7个孔/cm)(P<0.01);第10天,LCM-T20为319.3±12.9个孔/cm(LCM对照组为14.9±3.5个孔/cm);第15天,LCM-T20为345.2±11.1个孔/cm(LCM对照组为0个孔/cm)。总之,在LCM稀释培养基中使用海藻糖可提高冷冻/解冻精子的受精能力及其在母鸡生殖道内的受精持续时间。