Recognition of thermostable direct hemolysin-producing strains of V.parahaemolyticus by C.retropictus

逆行念珠菌对副溶血弧菌耐热直接产溶血素菌株的识别

• 批准号: 05660365
• 负责人: KUMAZAWA Norichika
• 金额: $ 1.41万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05660365/
• 关键词: V.parahaemolyticus; C.retropictus; colonization; plasma proteins; 血漿; SDS-PAGE; 抗原性; 耐熱性溶血毒

Thermostable direct hemolysin (TDH)-producing strains of Vibrio parahaemolyticus were detected from an estuarine Neritid gastropod, Clithon retropictus. Adult C.retropictus contains mature hemocytes in the hemolymph while juvenile one contains immature hemocytes which is attracted chemotactically to V.parahaemolyticus and phagocytize it only in the presence of molluscan plasma. These evidences suggest that V.parahaemolyticus could colonize in the alimentary tract of the juveniles even in the presence of hemocytes. IN this study, I analyzed the levels of V.parahaemolyticus strains to colonize in the alimentary tract of the gastropod and also composition of plasma proteins of the gastropod with SDS-PAGE and gel diffusion.Adult and juvenile C.retropictus ingested with TDH-producing strain D-3 and TDH-non-producing strains R-13 and N-18at a level of 10^5 cfu/g were maintained in UV-irradiated recirculating artificial seawater with a salinity of 20* at 20ﾟC for up to 21 days. In adults, all strains decreased to a level of 10^0 cfu/g within 21 days. In juveniles, strain D-3 survived at a level of 10^4-10^5 cfu/g for at least 21 days while atrains R-13 and N-18 decreased to the level of 10^2-10^1 cfu/g at the 21st day. Thus, TDH-producing strains would be concentrated in estuaries by the repeated passes through the alimentary tract of the juveniles.Plasmas of C.retropictus and two marine Neritids, Nerita albicilla and Heminerita japonica, exclusive of V.parahaemolyticus, were adapted to SDS-PAGE and gel diffusion with antisera of respective plasma. Main bands common to 3 gastropods and those different each other were seen in SDS-PAGE.Antigens specific to each gastropod and those common in three gastropods were seen in gel diffusion.

从一种河口型的Neritid腹足类动物Clithon retropictus中分离到一株产热稳定直接溶血素（TDH）的副溶血弧菌。成年C.retropictus在血淋巴中含有成熟的血细胞，而幼年C. retropictus含有未成熟的血细胞，其仅在软体动物血浆存在下被副溶血性弧菌趋化吸引并吞噬。这些证据表明，副溶血性弧菌即使在血细胞存在的情况下也能在幼鱼的消化道中定殖。在这项研究中，用10^5 cfu/g的TDH产生菌D-3和非产生菌R-13、N-18对反纹隐翅虫成虫和幼虫进行紫外线（UV）-紫外线（UV）-琼脂糖凝胶电泳（SDS-PAGE）和琼脂糖凝胶电泳（琼脂糖凝胶电泳）分析，结果表明，D-3和R-13、N-18对反纹隐翅虫成虫和幼虫的血浆蛋白组成有显著影响。盐度为20* 的辐照再循环人工海水在20 ° C下持续长达21天。在成虫中，所有菌株在21天内均降至10^0 cfu/g的水平。在幼鱼中，菌株D-3在10^4-10^5 cfu/g的水平下存活至少21天，而菌株R-13和N-18在第21天下降到10^2-10^1 cfu/g的水平。将反纹隐杆藻（C.retropictus）和两种海洋Neritids（Nerita albicilla和Heminerita japonica）（不包括副溶血性弧菌）的血浆进行SDS-PAGE和凝胶扩散。SDS-聚丙烯酰胺凝胶电泳显示了3种腹足类共有的主要条带和彼此不同的条带，凝胶扩散法显示了3种腹足类共有的抗原和各腹足类特异的抗原。