Viability assessment is an important part of semen analysis, and various live/dead staining protocols have been used in semen of avian species. Results of live/dead count differed between dyes, staining protocols and bird species, impeding comparability between studies and requiring species‐specific comparisons of viability stains. In raptor semen, similar comparisons are absent. Thus, the aim of the present study was to compare eight conventional viability stains. Eosin blue 2% [EB], eosin blue 2% with the addition of 3% sodium citrate [EB2], eosin blue–nigrosin 5% [EBN5], eosin yellow–nigrosin 5% [EYN5], eosin yellow–nigrosin 10% [EYN10], eosin blue–aniline blue [EBA], eosin yellow–aniline blue [EYA] and bromophenol blue–nigrosin [BBN] were evaluated in comparison with the fluorescence stain SYBR^® Green–propidium iodide [SYBR‐PI] in spermatozoa of falcons. The comparison was performed using conventional light microscopy which is applicable in breeding centres, veterinary practices and field studies. Additionally, live/dead stains were correlated to motility values of the same samples to validate sperm viability. Light microscopy using EB and using SYBR‐PI enabled an effective and clear differentiation between alive and dead spermatozoa of falcons. Motility values correlated significantly and strongly with EB only (r = .629; p < .001), but not with any other stain used in the study. Therefore, our results suggest EB as the most suitable stain for viability assessment in the semen of large falcons.

中文翻译：

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使用各种染色方案对大型猎鹰 (Falco spp.) 中的精子进行活力评估。

活力评估是精液分析的重要组成部分，各种活/死染色方案已用于禽类精液。活/死计数结果因染料、染色方案和鸟类物种而异，阻碍了研究之间的可比性，并需要对生存力染色进行物种特异性比较。在猛禽精液中，没有类似的比较。因此，本研究的目的是比较八种传统的活力染料。曙红蓝 2% [EB]、曙红蓝 2% 添加 3% 柠檬酸钠 [EB2]、曙红蓝-苯胺黑 5% [EBN5]、曙红黄-苯胺黑 5% [EYN5]、曙红黄-苯胺黑 10% [EYN10]、曙红蓝-苯胺蓝 [EBA]、曙红黄-苯胺蓝 [EYA] 和溴酚蓝-苯胺黑 [BBN] 与荧光染料 SYBR ^®进行比较评估猎鹰精子中的绿色碘化丙啶 [SYBR-PI]。比较是使用适用于繁殖中心、兽医实践和实地研究的传统光学显微镜进行的。此外，活/死污渍与相同样本的运动值相关联，以验证精子活力。使用 EB 和使用 SYBR-PI 的光学显微镜能够有效且清晰地区分猎鹰的活精子和死精子。运动性值仅与 EB 显着且强烈相关（r  = .629；p  < .001），但与研究中使用的任何其他染色无关。因此，我们的结果表明 EB 是最适合评估大型猎鹰精液活力的染色剂。