Studies on paralytic toxins in a ribbon worm adherent to cultured oysters in Hiroshima Bay, Hiroshima Prefecture, Japan

日本广岛县广岛湾附着在养殖牡蛎上的带状蠕虫麻痹毒素的研究

• 批准号: 11660205
• 负责人: ASAKAWA Manabu
• 金额: $ 0.58万
• 依托单位: Hiroshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11660205/
• 关键词: Hiroshima Bay; ribbon worm; tetrodotoxin; oyster; テトロドトキシン; 付着生物

In 1998, in Hiroshima Bay, Hiroshima Prefecture, Japan, specimens of the ribbon worm, "himomushi" adherent to the shells of cultured oysters were found to contain toxins which showed strong paralytic action in mice. The specimens were identified as an undescribed species of the genus Procephalothrix (formerly Cephalothrix). The maximum toxicity (as tetrodotoxin, TTX) was 14,734 MU/g whole body. The "himomushi" toxin (HMT) was extracted from the worm with 80% methanol acidified with acetic acid and the extract defatted with dichloromethane. The aqueous layer was chromatographed on activated charcoal and the unbound and bound toxic fractions were analyzed by high-performance liquid chromatography and gas chromatography-mass spectrometry. It was rather unexpectedly revealed from these results that HMT was comprised of TTX, 4-epiTTX, anhydro TTX and unidentified toxins. The main toxic component of HMT was isolated from a pooled specimen by a method which consisted of treatment with activat … More ed charcoal, chromatography on Bio-Gel P-2 and Bio-Rex 70 (H^+ form), and finally crystallization from an acidified methanolic solution. The recrystallized toxin showed a specific toxicity of 3,520 MU/mg. This toxin showed (M+H)^+ and (M+H-H_2O)^+ ion peaks at m/z 320 and 302, respectively by electrospray ionization-mass spectrometry(ESI-MS). The molecular weight thus determined was 319 for TTX in accordance with its reported molecular weight. The absorption band at 3353, 3235, 1666, 1612 and 1076cm^<-1> were observed in infrared spectrum of this toxin. This spectrum was indistinguishable from that of TTX.The ^1H-NMR spectrum for the recrystallized toxin was the same as that for TTX.The pair of doublets centered at 2.33 (J=10.0 Hz) and 5.48ppm(J=10.0 Hz), which are characteristic of tetrodotoxin, were shown to be coupled by double irradiation. By GC-MS analysis, the alkali-hydrolyzate of this toxin, like TTX, showed the quinazoline skeleton (C_9-base).These findings are the first direct evidence of the existence of TTX in the ribbon worms found on the surface of the shells of oysters hanging onto floating oyster culture rafts in Hiroshima Bay, Hiroshima Prefecture. Less

1998年，在日本广岛县的广岛湾，发现附着在养殖牡蛎壳上的带状蠕虫“himomushi”样本含有毒素，对老鼠有强烈的麻痹作用。这些标本被鉴定为原发属（原头发属）的一个未描述的物种。最大毒性（以河豚毒素计）为14，734 MU/g全身。用80%甲醇醋酸酸化提取血吸虫毒素，二氯甲烷脱脂。在活性炭上对水层进行色谱分析，并通过高效液相色谱法和气相色谱-质谱法对未结合和结合的毒性组分进行分析。从这些结果中相当意外地揭示出HMT由TTX、4-epiTTX、脱水TTX和未鉴定的毒素组成。HMT的主要毒性成分是通过一种方法从混合的标本中分离出来的，该方法包括用活化剂处理， ...更多信息 艾德活性炭，在Bio-Gel P-2和Bio-Rex 70（H^+型）上层析，最后从酸化甲醇溶液中结晶。重结晶的毒素显示出3，520 MU/mg的比毒性。电喷雾质谱（ESI-MS）分析表明，该毒素在m/z320和302处分别有（M+H）^+和（M+H-H_2 O）^+离子峰。根据TTX的报道分子量，由此测定的TTX分子量为319。在红外光谱中，在3353、3235、1666、1612和1076 cm ~ 2处有明显的吸收带<-1>。重结晶毒素的~ 1H-NMR谱与TTX相同，在2.33（J= 10.0Hz）和5.48ppm（J= 10.0Hz）处有一对双重峰，这是河豚毒素的特征峰。经GC-MS分析，该毒素的碱解产物与TTX一样，都具有喹唑啉骨架（C_9-base），这是广岛县广岛湾牡蛎养殖浮筏上发现的带状蠕虫中存在TTX的第一个直接证据。少

项目成果

Manabu Asakawa: "Paralytic toxins in a ribbon worm Cephalothrix species (Nemertean) adherent to cultured oysters in HiroshimaBay, Hiroshima Prefecture, Japan"Toxicon. 38. 763-773 (2000)

Manabu Asakawa：“日本广岛县广岛湾养殖牡蛎上附着的带状蠕虫头丝虫（纽虫）中的麻痹毒素”Toxicon。

Manabu Asakawa: "Paralytic toxins in a ribbon worm Cephalothrix species (Nemertean) adherent to cultured oysters in Hiroshima Bay, Hiroshima Prefecture, Japan"Toxicon. 38. 763-773 (2000)

Manabu Asakawa：“日本广岛县广岛湾养殖牡蛎上附着的带状蠕虫头丝虫（纽虫）中的麻痹毒素”Toxicon。