Shazada Nururshopa Eskander, Siddique Mohammad Abdul Momin, Zhang Songpei, Ma Zhijun, Rodina Marek, Linhart Otomar
Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Research Institute of Fish Culture and Hydrobiology, University of South Bohemia in České Budějovice, Zátiší 728/II, Vodňany, 38925, Czech Republic.
Department of Biotechnology and Genetic Engineering, Noakhali Science and Technology University, Noakhali, 3814, Bangladesh.
Fish Physiol Biochem. 2025 Feb;51(1):14. doi: 10.1007/s10695-024-01413-7. Epub 2024 Dec 2.
Short-term storage and management of sperm in vitro is an easy and economical process in which suitable extenders can be utilized to extend the storage period and prevent sperm function impairment. Therefore, the current study aimed to evaluate the effect of suitable extenders during the short-term storage of sterlet sperm and determine their fertilizing capacity and hatching success. Three extenders containing a composition of 16, 20, and 24 mM NaCl, 1 mM KCl, 0.1 mM CaCl, 10 mM Tris, pH 8.0 with osmolarity of 46, 55, and 62 mOsm/kg, were used to dilute the sperm of four sexually mature sterlet males (n = 4). Using a CASA system, the motility and velocity of undiluted and diluted sperm with extenders (E1 - E3) were assessed over 6 days at 0-2 °C. The short-term stored diluted sperm was then used in the fertilization and hatching assay, and undiluted fresh and stored sperm was used as a control. A two-way factorial analysis of variance (ANOVA) model confirmed significant effects on sperm motility, curvilinear velocity (VCL), and straight-line velocity (VSL) (P < 0.001), as well as their interaction with the extender. The model was decomposed into a one-way ANOVA to examine the impacts of extenders and storage time. With increasing storage periods, the sperm motility and velocity gradually decreased for diluted sperm with three extenders (E1-E3) but sharply decreased for undiluted sperm (Control). The motility of undiluted sperm was found 3.77 ± 4.09% at 4 days, whereas sperm diluted with extenders showed 57.57 ± 12.33% (E1), 64.34 ± 11.86% (E2), and 61.40 ± 12.41% (E3) motility at 6 days. This study explored extenders optimized with higher osmolarity (39-62 mOsm/kg) and lower K (1 mmol/L) as the most suitable medium for storing sterlet sperm for 6 days. After 6 days post storage, sperm diluted with extenders E1-E3 achieved a fertilization rate of 31.29 ± 14.2%, 31.66 ± 8.84%, and 30.67 ± 10.02%, respectively, and hatching success of 29.58 ± 13.4%, 30.50 ± 7.89%, and 27.95 ± 9.62%, respectively with freshly ovulated eggs.
体外短期储存和管理精子是一个简单且经济的过程，在此过程中可利用合适的稀释液来延长储存期并防止精子功能受损。因此，本研究旨在评估合适的稀释液在小体鲟精子短期储存期间的作用，并确定其受精能力和孵化成功率。使用三种含有16、20和24 mM氯化钠、1 mM氯化钾、0.1 mM氯化钙、10 mM Tris（pH 8.0）且渗透压分别为46、55和62 mOsm/kg的稀释液，对四只性成熟的小体鲟雄性的精子（n = 4）进行稀释。使用计算机辅助精子分析（CASA）系统，在0至2°C下对未稀释的精子以及用稀释液（E1 - E3）稀释后的精子的活力和速度进行了6天的评估。然后将短期储存的稀释精子用于受精和孵化试验，并将未稀释的新鲜精子和储存精子用作对照。双向析因方差分析（ANOVA）模型证实了对精子活力、曲线速度（VCL）和直线速度（VSL）有显著影响（P < 0.001），以及它们与稀释液之间的相互作用。该模型被分解为单向ANOVA，以检验稀释液和储存时间的影响。随着储存时间的增加，用三种稀释液（E1 - E3）稀释的精子的活力和速度逐渐下降，但未稀释的精子（对照）则急剧下降。未稀释精子在4天时的活力为3.77±4.09%，而用稀释液稀释的精子在6天时的活力分别为57.57±12.33%（E1）、64.34±11.86%（E2）和61.40±12.41%（E3）。本研究探索了渗透压较高（39 - 62 mOsm/kg）且钾含量较低（1 mmol/L）的优化稀释液，作为储存小体鲟精子6天的最合适培养基。储存6天后，用稀释液E1 - E3稀释的精子与刚排卵的卵子受精率分别为31.29±14.2%、31.66±8.84%和30.67±10.02%，孵化成功率分别为29.
