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Abstract：
This work studied the performance of the artificial digestion method in terms of recovery and viability of Anisakis simplex third-stage larvae (L3) when previous treatments given to the infected fish muscle may accidentally render viable larvae. For that: a) hake mince was spiked with 10 L3/75g mince, frozen at -10, -15, -20, and -30 °C and immediately thawed, or stored for 12 or 24 h, and subjected to pepsin digestion; b) the mince was spiked under the same conditions, frozen at the above temperatures and thawed immediately. After manual recovery, L3 were assessed for viability, used to spike again the minced fish and subjected to pepsin digestion; c) the mince was spiked with 10 L3 which were: i) living (i.e. chilled), ii) freeze-surviving (live L3 had been previously recovered after freezing at -10 °C), or iii) dead (frozen at -30 °C or - 80 °C), and then subjected to pepsin digestion. Results showed that the artificial digestion method kills a significant number of larvae that may have survived freezing and thus may underestimate the number of viable larvae in a given batch. The method may also underestimate the infection level of fish batches containing dead larvae. It is suggested to take these limitations into account when designing digestion protocols for specific applications, especially when there is a risk of insufficiently treated or cooked fish batches or ready-to-eat foods.
摘要：
这项工作研究了人工消化方法在Anisakissimplex第三阶段幼虫（L3）的恢复和生存能力方面的性能，当时先前对感染的鱼肌肉进行的处理可能会意外使幼虫存活。为此：a）加装10L3/75g碎肉的无须鱼碎肉，在-10，-15，-20和-30°C下冷冻并立即解冻，或储存12或24小时，并进行胃蛋白酶消化；b)在相同条件下加标肉末，在上述温度下冷冻并立即解冻。手动恢复后，对L3进行了生存能力评估，用于再次刺入切碎的鱼并进行胃蛋白酶消化；c）用10L3加标：i）生活（即冷藏），ii)冷冻存活(活L3在-10°C冷冻后先前已恢复),或iii）死亡（在-30°C或-80°C下冷冻），然后进行胃蛋白酶消化。结果表明，人工消化方法杀死了大量可能在冷冻中存活的幼虫，因此可能低估了给定批次中存活幼虫的数量。该方法还可能低估含有死幼虫的鱼批次的感染水平。建议在设计特定应用的消化方案时考虑这些限制，特别是当存在不充分处理或煮熟的鱼批次或即食食品的风险时。
