Belala Réda, Briand-Amirat Lamia, Martinot Aline, Thorin Chantal, Michaud Sandrine, Desherces Serge, Youngs Curtis R, Bencharif Djemil
Laboratory of Biotechnology and Reproductive Pathology, ONIRIS: The National Veterinary, Food agriculture, and food hygiene school of Loire Atlantique, Nantes, France.
Laboratory of Biotechnology of Animal Reproduction, University of Blida1, Blida, Algeria.
Reprod Domest Anim. 2019 Aug;54(8):1131-1138. doi: 10.1111/rda.13476. Epub 2019 Jun 23.
The current study aimed to explore the potential usefulness of liquid or lyophilized egg yolk plasma (EYP) as a substitute for low-density lipoproteins (LDL) for cryopreservation of canine spermatozoa. In the first experiment, a total of 20 ejaculates harvested from six Beagles were frozen in extenders containing 6% LDL (control) or liquid or lyophilized EYP at one of three concentrations (20%, 40% or 60%). Motility parameters were assessed 10 min after thawing using computer-assisted sperm analysis. For both liquid and lyophilized EYP, the 40% concentration yielded motility similar (p > 0.05) to that observed with the control extender. In the second experiment, 12 ejaculates collected from the same six dogs were frozen in 6% LDL (Control), 40% liquid EYP or 40% lyophilized EYP extenders. Spermatozoal membrane integrity (hypo-osmotic swelling test [HOSt] and SYBR14/propidium iodide [PI] staining), acrosome integrity (FITC-Pisum sativum agglutinin staining) and DNA integrity (acridine orange staining) characteristics were evaluated 10 min after thawing. Both liquid and lyophilized 40% EYP-based extenders successfully preserved all assessed integrity parameters as efficiently as the control. Results of this study suggest that lyophilized EYP is a viable alternative to LDL in freezing extenders for dog semen.
本研究旨在探索液体或冻干蛋黄血浆(EYP)作为低密度脂蛋白(LDL)替代品用于犬精子冷冻保存的潜在效用。在第一个实验中,从6只比格犬采集的总共20份精液在含有6%LDL(对照)或三种浓度(20%、40%或60%)之一的液体或冻干EYP的稀释液中冷冻。解冻10分钟后,使用计算机辅助精子分析评估活力参数。对于液体和冻干EYP,40%浓度产生的活力与对照稀释液观察到的活力相似(p>0.05)。在第二个实验中,从相同的6只犬采集的12份精液在6%LDL(对照)、40%液体EYP或40%冻干EYP稀释液中冷冻。解冻10分钟后,评估精子膜完整性(低渗肿胀试验[HOSt]和SYBR14/碘化丙啶[PI]染色)、顶体完整性(异硫氰酸荧光素-豌豆凝集素染色)和DNA完整性(吖啶橙染色)特征。基于40%液体和冻干EYP的两种稀释液都能像对照一样有效地成功保存所有评估的完整性参数。本研究结果表明,冻干EYP在犬精液冷冻稀释液中是LDL的可行替代品。