Complex toxin profile of French Mediterranean Ostreopsis cf. ovata strains, seafood accumulation and ovatoxins prepurification.
Complex toxin profile of French Mediterranean Ostreopsis cf. ovata strains, seafood accumulation and ovatoxins prepurification.
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DOI：
10.3390/md12052851
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发表时间：
2014-05-13
期刊：
影响因子：
5.4
通讯作者：
Amzil Z
中科院分区：
文献类型：
--
作者：
Brissard C;Herrenknecht C;Séchet V;Hervé F;Pisapia F;Harcouet J;Lémée R;Chomérat N;Hess P;Amzil Z
牡蛎cf.卵子产生孢子虫毒素类似物，包括卵毒素(OVTXs)和推定的孢子虫毒素(p-PLTX)，它们可以在海洋生物中积累，并可能导致食物中毒。然而，纯化的卵毒素并不广泛获得，其毒性仍不清楚。本研究的目的是为了提高对河口牡蛎生理生态的认识。卵子及其毒素的产生，以及卵毒素纯化工艺的优化。2011年和2012年在法国西北部地中海滨海的维尔弗利奇水华期间，采集了微藻附生细胞和海洋生物，并用LC-MS/MS和溶血试验对其进行了分析。这两种方法得到的结果具有可比性，表明卵毒素具有溶血特性。在贝类中平均检出相当于全肉223μg·kg−1的孢子毒素，超过了欧洲食品安全局推荐的30μg·kg−1的限值。Ostreopsis细胞在原位和实验室培养中显示出相同的毒素谱，其中卵毒素-a(OVTX-a)是含量最丰富的类似物(~50%)，其次是OVTX-b(~15%)、p-PLTX(12%)、OVTX-d(8%)、OVTX-c(5%)和OVTX-e(4%)。牡蛎cf.每L培养一次，卵子可产生2g生物量，最高浓度为300pgPLTX当量细胞−-1。因此，用该菌株10个L可产生约10 mg的PLTX类毒素。从采集的生物质中提取的毒素提取液采用液-液分配或大小排除等不同的技术进行纯化。其中，Sephadex LH20柱层析分离效果最好，净化效率为93%，回收率约为85%，毒素含量提高了13倍。因此，这一提纯步骤应纳入未来的分离练习中。
Ostreopsis cf. ovata produces palytoxin analogues including ovatoxins (OVTXs) and a putative palytoxin (p-PLTX), which can accumulate in marine organisms and may possibly lead to food intoxication. However, purified ovatoxins are not widely available and their toxicities are still unknown. The aim of this study was to improve understanding of the ecophysiology of Ostreopsis cf. ovata and its toxin production as well as to optimize the purification process for ovatoxin. During Ostreopsis blooms in 2011 and 2012 in Villefranche-sur-Mer (France, NW Mediterranean Sea), microalgae epiphytic cells and marine organisms were collected and analyzed both by LC-MS/MS and hemolysis assay. Results obtained with these two methods were comparable, suggesting ovatoxins have hemolytic properties. An average of 223 μg·kg−1 of palytoxin equivalent of whole flesh was found, thus exceeding the threshold of 30 μg·kg−1 in shellfish recommended by the European Food Safety Authority (EFSA). Ostreopsis cells showed the same toxin profile both in situ and in laboratory culture, with ovatoxin-a (OVTX-a) being the most abundant analogue (~50%), followed by OVTX-b (~15%), p-PLTX (12%), OVTX-d (8%), OVTX-c (5%) and OVTX-e (4%). Ostreopsis cf. ovata produced up to 2 g of biomass per L of culture, with a maximum concentration of 300 pg PLTX equivalent cell−1. Thus, an approximate amount of 10 mg of PLTX-group toxins may be produced with 10 L of this strain. Toxin extracts obtained from collected biomass were purified using different techniques such as liquid-liquid partition or size exclusion. Among these methods, open-column chromatography with Sephadex LH20 phase yielded the best results with a cleanup efficiency of 93% and recovery of about 85%, representing an increase of toxin percentage by 13 fold. Hence, this purification step should be incorporated into future isolation exercises.
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影响因子：
2.8
作者：
Aligizaki, Katerina;Katikou, Panagiota;Diogene, Jorge
通讯作者：
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影响因子：
2.8
作者：
GLEIBS, S;MEBS, D;WERDING, B
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影响因子：
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影响因子：
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作者：
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通讯作者：
Pistocchi, Rossella
DOI：
10.1016/j.jasms.2007.11.001
发表时间：
2008-01-01
影响因子：
3.2
作者：
Ciminiello, Patrizia;Dell'Aversano, Carmela;Melchiorre, Nunzia
通讯作者：
Melchiorre, Nunzia