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首页> 外文期刊>Harmful Algae >Stability of the intra- and extracellular toxins of Prymnesium parvum using a microalgal bioassay
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Stability of the intra- and extracellular toxins of Prymnesium parvum using a microalgal bioassay

机译：微藻生物测定法检测小球藻细菌内和细胞外毒素的稳定性

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Prymnesium parvum produces a variety of toxic compounds, which affect other algae, grazers and organisms at higher trophic levels. Here we provide the method for development of a sensitive algal bioassay using a microalgal target, Teleaulax acuta, to measure strain variability in P. parvum toxicity, as well as the temporal stability of both the intracellular and the extracellular lytic compounds of P. parvum. We show high strain variation in toxicities after 3 h incubation with LC_(50)s ranging from 24 to 223 × 10~3 cells ml~(-1). Most importantly we prove the necessity of testing physico-chemical properties of P. parvum toxins before attempting to isolate and characterize them. The extracellular toxin in the supernatant is highly unstable, and it loses significant lytic effects after 3 days despite storage at -20 ℃ and after only 24 h stored at 4 ℃. However, when stored at -80 ℃, lytic activity is more easily maintained. Reducing oxidation by storing the supernatant with no headspace in the vials significantly slowed loss of activity when stored at 4 ℃. We show that the lytic activity of the intracellular toxins, when released by sonication, is not as high as the extracellular toxins, however the stability of the intracellular toxins when kept as a cell pellet at -20 ℃ is excellent, which proves this is a sufficient storage method for less than 3 months. Our results provide an ecologically appropriate algal bioassay to quantify lytic activity of P. parvum toxins and we have advanced our knowledge of how to handle and store the toxins from P. parvum so as to maintain biologically relevant toxicity.
机译：小疫霉会产生多种有毒化合物，它们会以较高的营养水平影响其他藻类，放牧者和生物。在这里，我们提供了使用微藻靶标Teleaulax acuta开发灵敏藻类生物测定方法的方法，以测量细小杆球菌毒性的变异性，以及细小杆球菌细胞内和细胞外裂解化合物的时间稳定性。我们发现，LC_（50）s孵育3 h后，毒性的变异很大，范围从24到223×10〜3个细胞ml〜（-1）。最重要的是，我们证明了在尝试分离和表征小球藻毒素的理化特性之前，有必要对其进行测试。上清液中的细胞外毒素高度不稳定，尽管在-20℃下储存3天后和在4℃下储存24小时后仍失去明显的裂解作用。但是，在-80℃下保存时，裂解活性更容易保持。通过将没有顶空的上清液储存在小瓶中来减少氧化，从而显着减慢了在4℃储存时的活性损失。我们发现，当通过超声释放时，细胞内毒素的裂解活性不如细胞外毒素高，但是当细胞沉淀物在-20℃保存时，细胞内毒素的稳定性非常好，这证明这是一种足够的存储方法少于3个月。我们的结果提供了一种生态学上合适的藻类生物测定方法，可以定量分析小菜蛾毒素的裂解活性，并且我们已经掌握了如何处理和储存小菜蛾毒素以保持生物学相关毒性的知识。

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