Mohammadi Aliakbar, Sabrivand Adel, Hajarian Hadi
Faculty of Veterinary Medicine, Tabriz University, Tabriz, Iran.
Animal Science Department, Agriculture and Natural Resources Campus, Razi University, Kermanshah, Iran.
Vet Med Sci. 2025 Mar;11(2):e70237. doi: 10.1002/vms3.70237.
In trying to protect honey bee species and maintain genetic diversity, employing effective approaches for drone sperm conservation is crucial. Based on literature, drone sperm cryopreservation extenders and methods have not been fully optimized. Our research aim was to enhance drone bee sperm preservation by incorporating royal jelly (RJ) in the extender of the cryoprotectant-free vitrification method. Drone sperm was collected from adult drone bees (36 days old) using the manual inversion method. Different concentrations of RJ (0%, 0.5%, 1%, 2.5% and 5%) were added to the extender. Diluted sperm were cryopreserved using a cryoprotectant-free vitrification technique where 10 µL aliquots of the diluted sperm were directly dropped into the liquid nitrogen and then stored. Data were analysed based on a completely randomized design with ten replications. Sperm quality parameters, including motility, viability and DNA damage, were evaluated in vitro. Queens were artificially inseminated to measure the ability of motile sperm to reach the spermathecae. The results showed that 1% and 2.5% of RJ supplementation significantly enhanced sperm motility and viability and reduced DNA fragmentation compared to control and higher RJ concentrations. Specifically, the 1% RJ group resulted in the highest sperm viability, while both the 1% and 2.5% groups maintained lower DNA fragmentation rates. Queens inseminated with sperm treated with 1% and 2.5% RJ showed a notably higher number of motile sperm in their spermathecae. In conclusion, supplementation of 1% RJ to the cryoprotectant-free vitrification media may improve drone sperm quality parameters post-warming. Our findings provide valuable insights into optimizing drone bee sperm preservation, contributing to the conservation of these vital pollinators.
在试图保护蜜蜂物种并维持遗传多样性的过程中，采用有效的雄蜂精子保存方法至关重要。根据文献，雄蜂精子冷冻保存稀释液和方法尚未得到充分优化。我们的研究目标是通过在无冷冻保护剂玻璃化方法的稀释液中加入蜂王浆（RJ）来提高雄蜂精子的保存效果。使用手动反转法从成年雄蜂（36日龄）采集雄蜂精子。将不同浓度的RJ（0%、0.5%、1%、2.5%和5%）添加到稀释液中。使用无冷冻保护剂玻璃化技术对稀释后的精子进行冷冻保存，即将10微升稀释后的精子等分试样直接滴入液氮中，然后储存。基于完全随机设计进行数据分析，重复十次。在体外评估精子质量参数，包括活力、存活率和DNA损伤。对蜂王进行人工授精，以测量活动精子到达受精囊的能力。结果表明，与对照组和更高RJ浓度相比，添加1%和2.5%的RJ显著提高了精子活力和存活率，并减少了DNA片段化。具体而言，1%RJ组的精子存活率最高，而1%和2.5%组的DNA片段化率均较低。用1%和2.5%RJ处理的精子进行授精的蜂王，其受精囊中活动精子的数量明显更多。总之，在无冷冻保护剂玻璃化培养基中添加1%RJ可能会改善解冻后雄蜂精子的质量参数。我们的研究结果为优化雄蜂精子保存提供了有价值的见解，有助于保护这些重要的传粉者。
