Martins-Bessa Ana, Rocha António, Mayenco-Aguirre A
Department of Veterinary Sciences and CECAV, UTAD, Quinta de Prados, 5001-801 Vila Real, Portugal.
Theriogenology. 2006 Dec;66(9):2047-55. doi: 10.1016/j.theriogenology.2006.06.004. Epub 2006 Aug 1.
The objective of this work was to evaluate the possibility of substituting glycerol (G) for ethylene glycol (EG) when cryopreserving dog semen. A total of 15 ejaculates from 13 dogs was pooled into five samples and frozen in egg-yolk Tris extenders with variable ethylene glycol and glycerol concentrations, with or without Equex STM Paste. Two widely used glycerol extenders (Uppsala Equex II and Norwegian) were utilized as controls. Semen quality parameters assessed after thawing were total subjective motility (TSM), computer assisted sperm analysis (CASA), eosin-nigrosin staining, and flow cytometry (FC) after staining with the PI/Fitc-PSA (fluorescein isotiocianate conjugated with the agglutinin of Pisum sativum, PSA) fluorochromes. No advantages were seen in using EG to replace G when freezing dog semen or combining EG and G in the freezing medium. The Uppsala Equex II provided the best overall post-thaw parameters, followed by the egg-yolk Tris experimental extender with 5% EG and Equex STM Paste. The extender with 4% EG produced similar results to the Norwegian extender. High correlations (r>0.98) were obtained between eosin-nigrosin staining and FC, as well as between subjective and computerized motility assessment (r>0.90).
这项工作的目的是评估在冷冻犬精液时用甘油（G）替代乙二醇（EG）的可能性。将来自13只犬的总共15份射精样本合并为5个样品，并在含有不同乙二醇和甘油浓度的蛋黄Tris稀释液中冷冻，添加或不添加Equex STM Paste。使用两种广泛使用的甘油稀释液（乌普萨拉Equex II和挪威稀释液）作为对照。解冻后评估的精液质量参数包括总主观活力（TSM）、计算机辅助精子分析（CASA）、伊红-黑色素染色以及用PI/Fitc-PSA（异硫氰酸荧光素与豌豆凝集素PSA偶联）荧光染料染色后的流式细胞术（FC）。在冷冻犬精液或将EG和G组合在冷冻培养基中时，未发现用EG替代G有任何优势。乌普萨拉Equex II提供了最佳的总体解冻后参数，其次是含有5% EG的蛋黄Tris实验稀释液和Equex STM Paste。含有4% EG的稀释液产生的结果与挪威稀释液相似。伊红-黑色素染色与FC之间以及主观和计算机化活力评估之间获得了高度相关性（r>0.98），主观和计算机化活力评估之间的相关性也很高（r>0.90）。
