Touzet Nicolas, Franco Jose M, Raine Robin
The Martin Ryan Institute, National University of Ireland, Galway, Ireland.
Appl Environ Microbiol. 2007 May;73(10):3333-42. doi: 10.1128/AEM.02161-06. Epub 2007 Mar 2.
A comparative analysis of the morphology, toxin composition, and ribosomal DNA (rDNA) sequences was performed on a suite of clonal cultures of the potentially toxic dinoflagellate Alexandrium minutum Halim. These were established from resting cysts or vegetative cells isolated from sediment and water samples taken from the south and west coasts of Ireland. Results revealed that strains were indistinguishable, both morphologically and through the sequencing of the D1-D2 domain of the large subunit and the ITS1-5.8S-ITS2 regions of the rDNA. High-performance liquid chromatography fluorescence detection analysis, however, showed that only strains derived from retentive inlets on the southern Irish coast synthesized paralytic shellfish poisoning (PSP) toxins (GTX2 and GTX3), whereas all strains of A. minutum isolated from the west coast were nontoxic. Toxin analysis of net hauls, taken when A. minutum vegetative cells were in the water column, revealed no PSP toxins in samples from Killary Harbor (western coast), whereas GTX2 and GTX3 were detected in samples from Cork Harbor (southern coast). These results confirm the identity of A. minutum as the most probable causative organism for historical occurrences of contamination of shellfish with PSP toxins in Cork Harbor. Finally, random amplification of polymorphic DNA was carried out to determine the degree of polymorphism among strains. The analysis showed that all toxic strains from Cork Harbor clustered together and that a separate cluster grouped all nontoxic strains from the western coast.
对一系列潜在有毒的微小亚历山大藻(Alexandrium minutum Halim)克隆培养物进行了形态学、毒素成分和核糖体DNA(rDNA)序列的比较分析。这些培养物是从爱尔兰南部和西部海岸采集的沉积物和水样中分离出的休眠孢囊或营养细胞建立的。结果显示,无论是在形态上,还是通过对rDNA大亚基的D1-D2结构域以及ITS1-5.8S-ITS2区域进行测序,各菌株之间都无法区分。然而,高效液相色谱荧光检测分析表明,只有源自爱尔兰南部海岸截留入口的菌株能合成麻痹性贝类毒素(PSP毒素,GTX2和GTX3),而从西海岸分离出的所有微小亚历山大藻菌株均无毒。当微小亚历山大藻营养细胞处于水柱中时,对网捞样本进行的毒素分析显示,基拉尼港(西海岸)的样本中未检测到PSP毒素,而在科克港(南部海岸)的样本中检测到了GTX2和GTX3。这些结果证实了微小亚历山大藻是科克港历史上贝类受PSP毒素污染事件最可能的致病生物。最后,进行了随机扩增多态性DNA分析,以确定菌株之间的多态性程度。分析表明,来自科克港的所有有毒菌株聚集在一起,而另一个单独的聚类则将来自西海岸的所有无毒菌株归为一组。