In spotted wolffish Anarhichas minor aquaculture, cryopreservation is used to secure sperm availability throughout the entire spawning season. Under current protocols, sperm is cryopreserved in 0.5-mL straws. This implies thawing a considerable number of straws for insemination with cryopreserved sperm. In this work, we scale up the spotted wolffish sperm cryopreservation procedure through the development of a protocol for sperm cryopreservation in 5-mL cryovials. Different freezing (distances from the liquid nitrogen surface) and thawing rates were tested. The best results were obtained with cryovials frozen at a distance of 1.5 cm from the liquid nitrogen surface and thawed either at 15 or 10 °C for 4 and 6 min, respectively. Under these conditions, similar percentage of motile cells, sperm velocity and percentage of viable cells were obtained in comparison with the sperm cryopreserved in the traditional 0.5-mL straws. This protocol will facilitate the process of insemination with cryopreserved sperm in the spotted wolffish hatcheries.

在斑点狼鱼Anarhichas minor 中水产养殖，冷冻保存用于确保整个产卵季节的精子可用性。根据目前的协议，精子被冷冻保存在 0.5 毫升的细管中。这意味着解冻大量的吸管，用冷冻保存的精子进行授精。在这项工作中，我们通过制定 5 毫升冷冻管中精子冷冻保存的协议来扩大斑点狼鱼精子冷冻保存程序。测试了不同的冷冻（距液氮表面的距离）和解冻速率。最好的结果是在距液氮表面 1.5 厘米处冷冻并在 15 或 10°C 下分别解冻 4 和 6 分钟的冷冻管获得了最佳结果。在这些条件下，类似百分比的运动细胞，与传统 0.5 mL 吸管中冷冻保存的精子相比，获得了精子速度和活细胞百分比。该协议将促进在斑点狼鱼孵化场中用冷冻保存的精子进行授精的过程。