In Vitro Inactivation of Kudoa septempunctata Spores Infecting the Muscle of Olive Flounder Paralichthys olivaceus
In Vitro Inactivation of Kudoa septempunctata Spores Infecting the Muscle of Olive Flounder Paralichthys olivaceus
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DOI：
10.1089/fpd.2015.2003
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发表时间：
2016-01-01
影响因子：
2.8
通讯作者：
Kobayashi, Shoko
中科院分区：
文献类型：
--
作者：
Yokoyama, Hiroshi;Funaguma, Naoko;Kobayashi, Shoko
七星虫是一种寄生在牙鲆躯干肌肉上的粘孢子虫，最近被报道为一种引起人类食物中毒的病原体。患者在摄入受感染的比目鱼的生肉后出现急性腹泻和呕吐。最近食物中毒病例的增加促使我们制定了这种寄生虫的控制策略。本研究用荧光染料Hoechst 33342和碘化丙啶(PI)进行活体染色，比较了几种温度和化学处理对七星克雷伯菌孢子的灭活效果。试验结果表明，25%乙醇作用5min，80℃作用10s，柠檬烯10L/mL作用5min，盐度为千分之0和160‰作用5min，杀灭孢子效果较好。为了验证处理后七星草孢子的毒性损失，用Caco-2细胞进行了紧密连接屏障完整性检测。Caco-2检测结果与Hoechst 33342-PI染色检测结果一致。考虑到处理技术在养殖牙鲆生产过程中的应用，还需要进一步研究确定一种实用的灭活孢子的处理程序。
Kudoa septempunctata, a myxosporean parasite infecting the trunk muscles of olive flounder (Paralichthys olivaceus), has been recently reported to be the causative agent of a type of food poisoning in humans. Patients exhibited acute diarrhea and vomiting after ingestion of the raw flesh of infected flounder. A recent increase in the number of food-poisoning cases has prompted us to develop a control strategy of this parasite. In this study, we evaluated the efficacy of several temperature and chemical treatments for inactivating K. septempunctata spores in vitro using the vital staining assay with the fluorescent dyes Hoechst 33342 and propidium iodide (PI). Screening tests of treatment methods against K. septempunctata suggested that 25% ethanol for 5min, 80 degrees C for 10s, limonene at 10L/mL for 5min, and salinities at 0 parts per thousand and 160 parts per thousand for 5min were effective for killing spores. To verify toxicity loss in K. septempunctata spores after the treatments, tight junction barrier integrity assays with Caco-2 cells were conducted. The results of the Caco-2 assays corresponded well with those of the Hoechst 33342-PI staining assay. Further studies are required to determine a practical treatment procedure for inactivating spores considering the treatment application in the production process of cultured olive flounder.