Comparison of staining protocols to assess sperm viability in cockatiels (Nymphycus hollandicus)

Theriogenology wild Pub Date : 2025-01-01 DOI:10.1016/j.therwi.2025.100129

D. Fischer , A. Bublat , A. Wehrend , M. Lierz

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Abstract

The assessment of sperm viability belongs to the core elements of semen analysis in human and veterinary andrology. In avian medicine various live/dead stains and protocols have been evaluated in a number of commercial poultry and companion bird species. However, viability results are known to differ largely between avian species and the used stains and protocols. Therefore, species specific comparisons of live/dead stains are required, which are absent in numerous bird orders. The aim of this study was to compare the conventional live/dead stains eosin blue 2% (EB), eosin yellow-nigrosin 10% (EYN), eosin blue-nigrosin 5% (EBN), bromphenol blue nigrosin (BBN) and the fluorescence stain SYBR green - propidium iodide (SYBR-PI) in spermatozoa of cockatiels as model for psittacine species. The latter comparison was performed conventionally via cell count as well as using a computer-assisted semen analyzer (CASA). Moreover, all viability stains were correlated to sperm motility values of the same sample, as this has been suggested as suitable method to validate sperm viability. Pairwise comparison between the motility values and the viability values of EB, EYN, EBN, BBN, SYBR-PI were performed immediately, 24 hours and 48 hours after semen collection. Progressive motility values (PMOT) correlated significantly (p < 0.05) with all viability results immediately after semen collection, but PMOT after 24 hours and general motility values only with EB and SYBR/PI. In microscopy EB and SYBR were judged as best suited, because alive and damaged spermatozoa could be effectively distinguished. Conventional and computer-assisted analysis using SYBR/PI demonstrated significant and strong correlations (r_s = 0.939; p < 0.0001). The results of this study, suggest EB and SYBR/PI as suitable stains for viability assessment in cockatiels, while EYN and EBN is not recommendable and BBN seems not suitable at all in psittacine spermatozoa.

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评价荷兰凤尾鱼精子活力的染色方法比较

精子活力的评估是人类和兽医男科精液分析的核心内容。在禽类医学中，已经对许多商业家禽和伴侣鸟类进行了各种活/死染色和方案的评估。然而，已知的生存能力结果在鸟类物种和使用的染色剂和方案之间存在很大差异。因此，需要对活斑和死斑进行物种特异性比较，而这在许多鸟类目中是不存在的。本研究的目的是比较传统的2%伊红蓝（EB）、10%伊红黄黑素（EYN）、5%伊红蓝黑素（EBN）、溴酚蓝黑素（BBN）和SYBR绿色碘化丙啶荧光染色（SYBR- pi）对鹦鹉精的影响。后者的比较通常通过细胞计数以及使用计算机辅助精液分析仪（CASA）进行。此外，所有活力染色都与同一样品的精子活力值相关，因为这被认为是验证精子活力的合适方法。取精后即刻、24 h和48 h两两比较EB、EYN、EBN、BBN、SYBR-PI的活力值和活力值。精液采集后，进行性活力值（PMOT）与所有活力值显著相关（p <； 0.05），但24 小时后的PMOT和一般活力值仅与EB和SYBR/PI相关。在显微镜下，EB和SYBR被认为是最合适的，因为可以有效地区分活精子和受损精子。使用SYBR/PI进行常规分析和计算机辅助分析显示出显著和强相关性(rs = 0.939；p & lt; 0.0001)。本研究结果表明，EB和SYBR/PI染色适合用于鹦鹉尾精子的活力评估，而EYN和EBN不推荐，BBN似乎根本不适合用于鹦鹉尾精子的活力评估。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Theriogenology wild Animal Science and Zoology, Veterinary Science

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审稿时长

56 days