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从剧毒海洋腹足纲动物半褶织纹螺中分离出的细菌的毒素筛选与鉴定

Toxin-screening and identification of bacteria isolated from highly toxic marine gastropod Nassarius semiplicatus.

作者信息

Wang Xiao-Jie, Yu Ren-Cheng, Luo Xuan, Zhou Ming-Jiang, Lin Xiang-Tian

机构信息

Key Laboratory of Marine Ecology and Environmental Sciences, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.

出版信息

Toxicon. 2008 Jul;52(1):55-61. doi: 10.1016/j.toxicon.2008.04.170. Epub 2008 May 29.

Abstract

Bacteria isolated from a highly toxic sample of gastropod Nassarius semiplicatus in Lianyungang, Jiangsu Province in July 2007, were studied to probe into the relationship between bacteria and toxicity of nassariid gastropod. The toxicity of the gastropod sample was 2 x 10(2)mouse unit (MU) per gram of tissue (wet weight). High concentration of tetrodotoxin (TTX) and its analogues (TTXs) were found in the digestive gland and muscle of the gastropod, using high performance liquid chromatography coupled with mass chromatography (LC-MS). Bacterial strains isolated from the digestive gland were cultured and screened for TTX with a competitive ELISA method. Tetrodotoxin was detected in a proportion of bacterial strains, but the toxin content was low. Partial 16S ribosomal DNA (rDNA) of the TTX-producing strains was then sequenced and compared with those published in the GenBank to tentatively identify the toxic strains. It was found that most of the toxic strains were closely affiliated with genus Vibrio, and the others were related to genus Shewanella, Marinomonas, Tenacibaculum and Aeromonas. These findings suggest that tetrodotoxin-producing bacteria might play an important role in tetrodotoxin accumulation/production in N. semiplicatus.

摘要

对2007年7月从江苏省连云港市采集的高毒性织纹螺样本中分离出的细菌进行了研究,以探讨细菌与织纹螺毒性之间的关系。该织纹螺样本的毒性为每克组织(湿重)2×10²小鼠单位(MU)。采用高效液相色谱-质谱联用(LC-MS)法,在织纹螺的消化腺和肌肉中发现了高浓度的河豚毒素(TTX)及其类似物(TTXs)。从消化腺中分离出的细菌菌株进行培养,并用竞争性酶联免疫吸附测定法筛选TTX。在部分细菌菌株中检测到了河豚毒素,但毒素含量较低。然后对产生TTX的菌株的部分16S核糖体DNA(rDNA)进行测序,并与GenBank中公布的序列进行比较,以初步鉴定有毒菌株。结果发现,大多数有毒菌株与弧菌属密切相关,其他的则与希瓦氏菌属、海单胞菌属、黄杆菌属和气单胞菌属有关。这些发现表明,产河豚毒素的细菌可能在织纹螺河豚毒素的积累/产生中起重要作用。

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