Embryo surface disinfection is utilized in aquaculture to decrease the risk of pathogen introduction into established colonies. Zebrafish embryos are commonly disinfected with unbuffered sodium hypochlorite at 25–50 ppm for 10 min with or without concurrent treatment with chemicals, including pronase (Pron), sodium thiosulfate, and/or methylene blue; however, the impact of these chemicals on embryo survival and development has not been evaluated. In this study, AB and casper embryos were exposed to disinfection protocols that used Pron, sodium thiosulfate, and/or methylene blue (given alone, in various combinations, or all three combined) with 50 and 100 ppm sodium hypochlorite performed 6 and 24 h postfertilization (HPF). All groups were evaluated for survival, hatching, and malformations at 5 days postfertilization. Maximal survival (69%–97%) and hatching rates (66%–94%) were generally observed with sodium hypochlorite disinfection followed by exposure to both Pron and sodium thiosulfate and maintenance in standard embryo medium without methylene blue. Methylene blue had variable effects on survival and hatching. Higher survival and hatching rates were seen in AB embryos disinfected at 6 HPF and casper embryos disinfected at 24 HPF. Susceptibility to sodium hypochlorite toxicity differed by strain, emphasizing the need to test disinfection protocols on small embryo cohorts.

中文翻译：

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链霉蛋白酶、硫代硫酸钠和亚甲蓝组合对次氯酸钠表面消毒斑马鱼 (Danio rerio) 胚胎发育和存活的影响

胚胎表面消毒用于水产养殖，以降低病原体引入已建立菌落的风险。斑马鱼胚胎通常用 25-50 ppm 的无缓冲次氯酸钠消毒 10 分钟，同时使用或不使用化学物质进行处理，包括链霉蛋白酶 (Pron)、硫代硫酸钠和/或亚甲蓝；然而，尚未评估这些化学品对胚胎存活和发育的影响。在本研究中，AB 和casper胚胎暴露于使用 Pron、硫代硫酸钠和/或亚甲蓝（单独给药，以各种组合或所有三种组合给药）与 50 和 100 ppm 次氯酸钠的消毒方案中，在受精后 6 和 24 小时进行。在受精后 5 天评估所有组的存活率、孵化率和畸形率。在次氯酸钠消毒后，通常可以观察到最大存活率 (69%–97%) 和孵化率 (66%–94%)，然后暴露于 Pron 和硫代硫酸钠，并在不含亚甲蓝的标准胚胎培养基中维持。亚甲蓝对存活和孵化有不同的影响。在 6 HPF 和casper消毒的 AB 胚胎中观察到更高的存活率和孵化率胚胎在 24 HPF 下消毒。对次氯酸钠毒性的敏感性因菌株而异，强调需要在小胚胎群体上测试消毒方案。