Díaz Ruiz Esther, González Ariza Antonio, León Jurado José Manuel, Arando Arbulu Ander, Bermúdez Oria Alejandra, Fernández Prior África, Delgado Bermejo Juan Vicente, Navas González Francisco Javier
Department of Genetics, Faculty of Veterinary Medicine, University of Cordoba, 14014 Cordoba, Spain.
Agropecuary Provincial Centre, Diputación Provincial de Córdoba, 14071 Cordoba, Spain.
Animals (Basel). 2023 Oct 2;13(19):3079. doi: 10.3390/ani13193079.
Sperm cryopreservation is effective in safeguarding genetic biodiversity in avian species. However, during this process, spermatozoa are very susceptible to plasma membrane peroxidation in the presence of high concentrations of reactive oxygen species (ROS). To mitigate this effect, the addition of exogenous antioxidants, such as hydroxytyrosol (3,4-dihydroxyphenylethanol; HT), an antioxidant derived from olive oil, to the cryopreservation sperm diluent, could be useful. To verify this, a cryopreservation diluent was supplemented with different concentrations (0 μg/mL, 50 μg/mL, 100 μg/mL, and 150 μg/mL) of HT. For this, semen was collected in 10 replicates from 16 roosters of the Utrerana avian breed, and a pool was prepared with the optimum quality ejaculates in each replicate. After cryopreservation, spermatozoa were thawed and different in vitro semen quality parameters were evaluated. A discriminant canonical analysis (DCA) was carried out and revealed that total motility (TM; Lambda = 0.301, F = 26,173), hypo-osmotic swelling test (HOST; Lambda = 0.338, F = 22,065), and amplitude of lateral head displacement (ALH, Lambda = 0.442; F = 14,180) were the variables with the highest discriminant power. Finally, a chi-squared automatic interaction detection (CHAID) decision tree (DT) was performed excluding fresh semen samples and ROS was found to be the most valuable variable to discriminate between the different established freezing groups. Samples in the absence of HT or with low concentrations of this antioxidant showed less desirable ROS values in cryopreserved rooster semen. The present study could lead to the improvement of cryopreservation techniques for the genetic material of local poultry breeds and optimize the conservation programs of endangered native avian breeds.
精子冷冻保存对于保护禽类物种的遗传生物多样性是有效的。然而，在此过程中，在高浓度活性氧（ROS）存在的情况下，精子极易受到质膜过氧化的影响。为减轻这种影响，向冷冻保存精子稀释液中添加外源性抗氧化剂，如从橄榄油中提取的抗氧化剂羟基酪醇（3,4 - 二羟基苯乙醇；HT），可能会有所帮助。为验证这一点，在冷冻保存稀释液中添加了不同浓度（0 μg/mL、50 μg/mL、100 μg/mL和150 μg/mL）的HT。为此，从16只乌特雷纳品种公鸡中采集精液，每个重复采集10份，然后将每个重复中质量最佳的射精精液混合制备样本。冷冻保存后，解冻精子并评估不同的体外精液质量参数。进行了判别式典型分析（DCA），结果显示总活力（TM；Lambda = 0.301，F = 26,173）、低渗肿胀试验（HOST；Lambda = 0.338，F = 22,065）和头部侧向位移幅度（ALH，Lambda = 0.442；F = 14,180）是判别能力最强的变量。最后，进行了卡方自动交互检测（CHAID）决策树（DT）分析，排除新鲜精液样本后，发现ROS是区分不同既定冷冻组的最有价值变量。在冷冻保存的公鸡精液中，未添加HT或添加低浓度这种抗氧化剂的样本显示出不太理想的ROS值。本研究可能会改进当地家禽品种遗传物质的冷冻保存技术，并优化濒危本土禽类品种的保护计划。
