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首页> 外文期刊>Harmful Algae >Gonyaulax Spinifera From The Adriatic Sea: Toxin Production And Phylogenetic Analysis
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Gonyaulax Spinifera From The Adriatic Sea: Toxin Production And Phylogenetic Analysis

机译：亚得里亚海的Gonyaulax Spinifera：毒素产生和系统发育分析

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摘要

The north-western coasts of the Adriatic sea have long been subjected to recurring cases of harmful algae blooms and concomitant mussel contamination. In 1995, yessotoxins (YTXs) were detected for the first time in Adriatic mussels and their presence was subsequently linked to the presence of Protoceratium reticulatum and occasionally Lingulodinium polyedrum in seawater. Since then, farmed molluscs are yearly found to contain YTXs for long periods with a severe impact on aquaculture activities. In 2004, unusually high amounts of homoYTXs were detected in mussels. In this period, a careful look at the phytoplankton composition highlighted that P. reticulatum and L. polyedrum were nearly absent in seawater whereas the dinoflagellate Gonyaulax spinifera was present at high densities. C. spinifera has been recently associated with YTX production in New Zealand. The present work was performed in order to ascertain whether Adriatic G. spinifera was a toxic species. Determination of the toxin content of natural samples (mussels and net haul plankton samples) as well as monitoring of the dinoflagellate species present in seawaters allowed us to correlate the presence of C. spinifera to that of homoYTX in mussels. Cultures of C. spinifera were set up which gave us the opportunity to investigate toxin content and profile of the dinoflagellate, for the first time, by liquid chromatography-mass spectrometry (LC-MS) and to perform a molecular characterization of the species through rDNA sequencing. Two G. spinifera isolates, sampled in different years, were analyzed and resulted to be different in both toxin content and gene sequence: the isolate sampled in 2005 produced only low levels of YTX (5.4 pg cell~(-1)) while that sampled in 2006 contained higher toxin amounts, namely 33.4 pg cell~(-1) of homoYTX and 3.6 pg cell~(-1) of YTX; the two isolates also differed in 111 and 176 nucleotide positions, respectively, in the SSU and partial LSU rDNA genes. Comparing similar sequences of G. spinifera strains contained in GenBank, divergences among the Adriatic strains and strains of different geographical origin emerged; on the contrary, a good similarity was evidenced (1.3%) between the Adriatic isolate sampled in 2006 and two New Zealand strains; the similarity was confirmed also by the presence in all strains of YTXs, although the New Zealand strains were analyzed by ELISA which could not distinguish among the different YTX analogues. Due to an originally confusing description of the species and to the availability of very low number of deposited sequences the doubt arises whether the studied G. spinifera strains belong to the same species. However, although the taxonomy of the species complex G. spinifera remains largely unresolved, this study allowed to unambiguously link the presence of G. spinifera to recurring mussel toxicity episodes.
机译：长期以来，亚得里亚海的西北海岸屡屡发生有害藻类繁殖和贻贝污染的案例。 1995年，在亚得里亚贻贝中首次检测到了毒素毒素（YTXs），随后它们的存在与网状Protoceratium reticulatum的存在以及偶尔在海水中的Lingulodinium polyedrum的存在有关。从那时起，每年都会发现养殖的软体动物长期含有YTX，严重影响了水产养殖活动。 2004年，在贻贝中发现了异常大量的homoYTX。在此期间，仔细观察浮游植物的组成可以看出，海水中几乎不存在网状疟原虫和多角藻，而高密度存在鞭毛藻。梭状芽胞杆菌最近已与新西兰的YTX生产相关。进行本工作是为了确定亚得里亚棘孢菌是否是有毒物种。天然样品（贻贝和长途浮游生物样品）中毒素含量的测定以及海水中存在的鞭毛鞭毛藻物种的监测，使我们能够将贻贝中梭状芽胞杆菌与homoYTX的存在相关联。建立了棘孢梭状芽孢杆菌的培养物，这使我们有机会首次通过液相色谱-质谱法（LC-MS）来研究重鞭毛藻的毒素含量和特征，并通过rDNA对该物种进行分子表征排序。分析了两种在不同年份取样的棘孢假单胞菌分离物，其毒素含量和基因序列均不同：2005年取样的分离物仅产生低水平的YTX（5.4 pg cell〜（-1）），而取样时在2006年，毒素含量更高，分别是homoYTX的33.4 pg细胞〜（-1）和YTX的3.6 pg细胞〜（-1）。这两个分离株在SSU和LSU rDNA部分基因中的111和176个核苷酸位置也不同。比较GenBank中的棘孢假单胞菌菌株的相似序列，发现亚得里亚菌株与不同地理起源的菌株之间存在差异。相反，在2006年采样的亚得里亚分离物与两种新西兰菌株之间，有很好的相似性（1.3％）。尽管所有的YTX菌株都通过ELISA进行了分析，但不能区分不同的YTX类似物，但所有菌株均存在相似性。由于最初对该物种的描述令人困惑，并且由于保藏序列的数量极少，因此人们对所研究的刺叶松柏菌株是否属于同一物种提出了疑问。但是，尽管物种复合体棘孢菌的分类学在很大程度上仍未解决，但这项研究可以明确地将棘孢菌的存在与贻贝复发毒性发作联系起来。

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