Method: Cold-storage temperature and dilution effects on kinematics of Brahman bull sperm patterns assessed with a Computer-Assisted Semen Analysis portable device

Animal - Open Space Pub Date : 2026-12-01 Epub Date: 2025-12-17 DOI:10.1016/j.anopes.2025.100117

P. García , F. Sevilla , I. Araya-Zúñiga , P. Cervantes-Acosta , A. Hernández-Beltrán , B. Domínguez-Mancera , R. Molina-Montero , M. Barrientos-Morales , J. Rodríguez , A. Salamanca-Carreño , A. Valverde

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Abstract

Beef cattle profitability depends on bull fertility. To replace traditional subjective bull quality ratings, a portable Computer-Assisted Semen Analysis (CASA) device can be used to generate objective results. The device requires careful control of temperature, dilution rate, and other methodological variables. This work provides a bovine semen analysis adapted methodology using a CASA portable device. It considers the effects of dilution, cooling times, and temperature in farms, where CASA evaluations of sperm quality, motility, and kinematics are frequently carried out. Fifteen ejaculates from five Brahman bulls were collected by electro-ejaculation. Aliquots were extended in Optixcell at 1:1 or 1:40 (v:v), stored at 5 °C and 17 °C, and evaluated after 2, 4, 8, 24, 48, and 72 h. Motility and kinematics were measured with iSperm®, and viability with an eosin–nigrosine stain. An ANOVA determined the effects of dilution rate, temperature, and cooling times, as well as interactions of the same. When effects proved statistically significant, mean comparisons ± SEM were carried out using the Bonferroni test (P < 0.05). The 1:1 dilution precluded CASA tracking, whereas the 1:40 dilution (∼90 × 10⁶ mL⁻¹) produced reliable outputs.

Total and progressive motility declined with storage time (P < 0.05), occurring after 8 h at 17 °C and 6 h at 5 °C. Cooling reduced curvilinear, average-path, and straight-line velocities; yet at 5 °C, the highest absolute speeds were observed, whereas 17 °C yielded the most progressive trajectories, with higher LIN and STR indices. Membrane integrity dropped sharply between 48 and 72 h. Dilution to 1:40 and storage at 17 °C conserved more progressive motility, viability, and membrane integrity for up to 24 h than 5 °C. The protocol is useful for developing field guidelines to preserve sperm quality traits that could be associated with fertility during short-term storage in beef systems using fresh artificial insemination.

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方法：用计算机辅助精液分析便携式装置评估冷藏温度和稀释对婆罗门公牛精子形态运动学的影响

肉牛的盈利能力取决于公牛的生育力。为了取代传统的主观公牛质量评级，可使用便携式计算机辅助精液分析（CASA）设备生成客观结果。该装置需要仔细控制温度、稀释率和其他方法学变量。这项工作提供了一个牛精液分析适应的方法使用CASA便携式设备。它考虑了稀释、冷却时间和农场温度的影响，在那里经常进行CASA精子质量、活力和运动学评估。用电射精法收集了5头婆罗门公牛的15次射精。在Optixcell中按1:1或1:40 （v:v）延长等分，在5°C和17°C保存，并在2、4、8、24、48和72 h后进行评估。用iSperm®测定运动性和运动学，并用伊红-黑氨酸染色法测定活力。方差分析确定了稀释率、温度和冷却时间的影响，以及它们之间的相互作用。当效应证明具有统计学意义时，采用Bonferroni检验进行均值比较±SEM （P < 0.05）。1:1稀释排除了CASA跟踪，而1:40稀释（~ 90 × 106 mL−1）产生可靠的输出。总运动力和进行性运动力随储存时间的延长而下降（P < 0.05），分别发生在17°C下8 h和5°C下6 h。冷却降低了曲线速度、平均路径速度和直线速度；然而，在5°C时，观察到最高的绝对速度，而在17°C时，产生了最先进的轨迹，具有更高的LIN和STR指数。在48至72小时之间，膜的完整性急剧下降。稀释至1:40，并在17°C下保存，与5°C相比，可在24小时内保存更多的进行性、活力和膜的完整性。该方案有助于制定现场指导方针，以在使用新鲜人工授精的牛肉系统中短期储存期间保持精子质量特征，这些特征可能与生育能力有关。

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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Animal - Open Space

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