Azaspiracids (AZA) are a group of lipophilic polyether compounds which have been implicated in shellfish poisoning incidents around Europe. They are produced by a few species of the dinophycean genera Azadinium and Amphidoma (Amphidomataceae). The presence of AZA toxins in Norway is well documented, but knowledge of the distribution and diversity of Azadinium and other Amphidomataceae along the Norwegian coast is rather limited and poorly documented. On a research survey along the Norwegian coast in 2015 from the Skagerrak in the South to Trondheimsfjorden in the North, plankton samples from 67 stations were analysed for the presence of Azadinium and Amphidoma and their respective AZA by on-board live microscopy, real-time PCR assays specific for Amphidomataceae, and liquid chromatography-tandem mass spectrometry (LC–MS/MS).

Microscopy using live samples and positive real-time PCR assays using a general family probe and two species specific probes revealed the presence of Amphidomataceae distributed throughout the sampling area. Overall abundance was low, however, and was in agreement with a lack of detectable AZA in plankton samples. Single cell isolation and morphological and molecular characterisation of established strains revealed the presence of 7 amphidomatacean species (Azadiniun spinosum, Az. poporum, Az. obesum, Az. dalianense, Az. trinitatum, Az. polongum, Amphidoma languida) in the area. Azaspiracids were produced by the known AZA producing species Az. spinosum, Az. poporum and Am. languida only. LC–MS/MS analysis further revealed that Norwegian strains produce previously unreported AZA for Norway (AZA-11 by Az. spinosum, AZA-37 by Az. poporum, AZA-38 and AZA-39 by Am. languida), and also four novel compounds (AZA-50, -51 by Az. spinosum, AZA-52, -53 by Am. languida), whose structural properties are described and which now can be included in existing analytical protocols. A maximum likelihood analysis of concatenated rDNA regions (SSU, ITS1-ITS2, partial LSU) showed that the strains of Az. spinosum fell in two well supported clades, where most but not all new Norwegian strains formed the new Ribotype B. Ribotype differentiation was supported by a minor morphological difference with respect to the presence/absence of a rim around the pore plate, and was consistently reflected by different AZA profiles. Strains of Az. spinosum from ribotype A produce AZA-1, -2 and -33, whereas the new strains of ribotype B produce mainly AZA-11 and AZA-51. Significant sequence differences between both Az. spinosum ribotypes underline the need to redesign the currently used qPCR probes in order to detect all AZA producing Az. spinosum.

The results generally underline the conclusion that for the Norwegian coast area it is important that amphidomatacean species are taken into account in future studies and monitoring programs.

氮杂spiracids（AZA）是一组亲脂性聚醚化合物，与欧洲各地的贝类中毒事件有关。它们是由附生植物Azadinium和Amphidoma（Amphidomataceae）的几种物种产生的。挪威存在AZA毒素的文献已被充分记载，但有关沿挪威海岸的Azadinium和其他两栖纲科的分布和多样性的知识却十分有限，文献记载也很少。在2015年从南部的Skagerrak到北部的Trondheimsfjorden沿挪威海岸进行的一项研究调查中，分析了来自67个站点的浮游生物样本中是否存在Azadinium和Amphidoma以及通过板载活体显微镜，两栖动物科特有的实时PCR测定法和液相色谱-串联质谱法（LC-MS / MS）对其各自的AZA进行分析。

使用活样品的显微镜检查和使用普通家族探针和两种物种特异性探针的阳性实时PCR分析揭示了存在于整个采样区域的两性科。但是，总体丰度较低，并且与浮游生物样品中缺乏可检测的AZA一致。单细胞分离，并建立菌株的形态学和分子表征揭示的7个amphidomatacean物种（存在Azadiniun棘，了Az。poporum，了Az。obesum，了Az。dalianense，了Az。trinitatum，了Az。polongum，Amphidoma languida中的区域）。氮杂螺菌酸是由已知的AZA产生物种产生的阿兹 棘，了Az。poporum和Am。仅限languida。LC-MS / MS分析进一步揭示了挪威菌株产生挪威先前未报告AZA（AZA-11通过了Az。棘通过，AZA-37了Az。poporum，AZA-38和由AZA-39 PM。languida），并且还有四个新型化合物（AZA-50，-51通过了Az。棘，AZA-52，-53通过PM。languida），其结构特性被描述并且现在可以被包括在现有的分析方案。串联rDNA区域（SSU，ITS1-ITS2，部分LSU）的最大似然分析表明，Az菌株。刺糖落入两个支撑良好的进化枝中，其中大多数但不是所有新的挪威菌株形成了新的RibotypeB。关于在孔板周围是否存在边缘的微小形态差异，支持了Ribotype分化，并且始终如一地反映出不同的AZA配置文件。Az株。来自核糖型A的刺糖产生AZA-1，-2和-33，而新的核糖型B菌株主要产生AZA-11和AZA-51。两个Az之间的显着序列差异。spinosum核糖型强调需要重新设计当前使用的qPCR探针以检测所有产生AZA的Az。菠菜。

结果总体上强调了以下结论：对于挪威沿海地区，在未来的研究和监测计划中必须考虑两栖类动物物种。