水产品中创伤弧菌的分离、鉴定及毒性研究

Studies on Identification and Toxicity of Vibrio vulnificus Isolated from Aquatic Products

通过溶血试验和小鼠急性毒性实验,评价市售水产品中创伤弧菌和标准创伤弧菌ATCC27562菌株毒性大小。从市售水产品中分离获得的6株疑似创伤弧菌菌株,经生化鉴定和溶细胞素基因扩增、基因测序后证实,除2号分离菌株基因测序未测出外,其它5株确定为创伤弧菌。创伤弧菌标准菌株ATCC27562菌株扩增出400 bp左右的基因条带,而6株分离株扩增出222 bp左右的典型创伤弧菌溶细胞素基因条带。在溶血试验中,创伤弧菌ATCC27562菌株的溶血圈与菌落直径之比达到3.0,1-6号分离株分别为2.3,2.0,2.8,2.0,2.5和2.1。在小鼠急性毒性实验中,创伤弧菌ATCC27562菌株的半致死量（LD50）是3.776×10^7CFU/m L,而3号分离株的LD50为2.218×10^8CFU/m L。创伤弧菌ATCC27562菌株为较强毒株,而3号分离株为弱毒株。

Vibrio vulnificus（V.v） isolated from aquatic product have been evaluated about toxicity by hemolysis test and acute toxicity, compared with standard V.v strain ATCC27562. It has been confirmed through biochemistry tests, cytolysin gene amplification and gene sequencing that six suspected strains isolated from aquatic product are all V.v strains, except from No.2 isolated strain. Six isolated strains have all amplified gene bands about 222bp; while ATCC27562 V.v strain has amplified gene bands about 400 bp. In the hemolysis test, the ratio of diameter between hemolysis circle and colony of standard strain is 3, while the ratio of isolated strains are 2.3,2.0,2.8,2.0,2.5 and 2.1from No.1 strain to No.6 strain, respectively. In the test of acute toxicity, the LD50 of standard strain is 3.776×10^7CFU/m L, while the LD50 of No.3 strain is 2.218×10^8CFU/m L. In conclusion, standard V.v strain ATCC27562 is moderate virulent strain, while No.3 isolated V.v strain is low virulent strain.