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<p>Optimizing sperm quality is essential in aquaculture, especially for producing all-female <i>Oncorhynchus mykiss</i> populations using sex-reversed males. Because sex-reversed males lack sperm ducts, semen is collected by gonadal maceration, a step that may expose spermatozoa to thermal and mechanical stress. We evaluated how post-maceration storage temperature (2, 4, 12&#xa0;°C for 24&#xa0;h) is associated with sperm performance in a within-male design, using an immediate post-extraction control (T0, 0&#xa0;h) as a descriptive baseline for comparison. Sperm kinematics were quantified by computer-assisted sperm analysis, and cellular readouts were assessed by flow cytometry for viability (% SYTOX-negative), mitochondrial membrane potential (ΔΨm/% JC-1 red), superoxide anion production (DHE/% DHE-high), and DNA fragmentation (% TUNEL-positive). After 24&#xa0;h, the warmer condition (12&#xa0;°C) was associated with lower total and progressive motility, lower mitochondrial membrane potential and viability, and higher superoxide levels, relative to cold conditions (2–4&#xa0;°C). Fertilization was analyzed as binomial data (k/n); proportions were lower at 12&#xa0;°C, but should be interpreted with caution given the small number of eggs assessed per replicate. Correlations between cellular biomarkers and motility/fertilization were generally weak, although a moderate negative association was observed between mitochondrial status and superoxide anion production. Overall, these results suggest that sperm from sex-reversed males is sensitive to post-maceration handling temperature, supporting the practical approach of maintaining tissues, tools and homogenates at ~ 0–4&#xa0;°C throughout the collection and storage workflow.</p>

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Post-maceration storage temperature affects sperm quality and fertilization in sex-reversed rainbow trout

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年4月11日

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  • Sebastián Ávila,
  • Osvaldo Merino,
  • Leydy Sandoval-Vargas,
  • Francisco Estay,
  • Miguel Colihueque Mancilla,
  • Paola Niedmann,
  • Eduardo Rojas,
  • Wellison Amorim Pereira,
  • Iván Valdebenito Isler,
  • Elías Figueroa-Villalobos

摘要

Optimizing sperm quality is essential in aquaculture, especially for producing all-female Oncorhynchus mykiss populations using sex-reversed males. Because sex-reversed males lack sperm ducts, semen is collected by gonadal maceration, a step that may expose spermatozoa to thermal and mechanical stress. We evaluated how post-maceration storage temperature (2, 4, 12 °C for 24 h) is associated with sperm performance in a within-male design, using an immediate post-extraction control (T0, 0 h) as a descriptive baseline for comparison. Sperm kinematics were quantified by computer-assisted sperm analysis, and cellular readouts were assessed by flow cytometry for viability (% SYTOX-negative), mitochondrial membrane potential (ΔΨm/% JC-1 red), superoxide anion production (DHE/% DHE-high), and DNA fragmentation (% TUNEL-positive). After 24 h, the warmer condition (12 °C) was associated with lower total and progressive motility, lower mitochondrial membrane potential and viability, and higher superoxide levels, relative to cold conditions (2–4 °C). Fertilization was analyzed as binomial data (k/n); proportions were lower at 12 °C, but should be interpreted with caution given the small number of eggs assessed per replicate. Correlations between cellular biomarkers and motility/fertilization were generally weak, although a moderate negative association was observed between mitochondrial status and superoxide anion production. Overall, these results suggest that sperm from sex-reversed males is sensitive to post-maceration handling temperature, supporting the practical approach of maintaining tissues, tools and homogenates at ~ 0–4 °C throughout the collection and storage workflow.