摘要
从患溃疡病养殖刺参（Apostichopus japonicus）病灶处分离出3株优势菌（RH1、RH2、RH3）,以浸浴、体腔注射、体壁肌肉注射等方式进行感染实验,证实菌株RH2为养殖刺参溃疡病的病原菌,其对刺参的LD50（半数致死量）每尾为5.68×10^6CFU,并证实浸浴、体腔注射的方式无法感染刺参,该菌可能通过体表创伤侵入的方式感染刺参。经形态学观察、生理生化特性分析、16SrRNA和HSP60基因测序确定菌株RH2为溶藻弧菌（Vibrio alginolyticus）。基于16S rRNA基因构建的系统发育树显示,菌株RH2与溶藻弧菌（AF513447）亲缘关系最近,置信度为50%;HSP60基因序列分析表明RH2与溶藻弧菌（AY332570、AF230931）聚为一个分支,且置信度高达99%。菌株RH2的最适生长盐度为25～35,最适生长pH为7.2～8.0,最适生长温度为14～22℃。对18种药物的敏感实验证实菌株RH2对诺氟沙星、复方新诺明、壮观霉素等较为敏感。
Three bacterial strains named RH1, RH2 and RH3 were isolated from the cultured sea cucumber （Apostichopus japonicus ） with skin ulceration. The strain RH2 was proved to be a pathogen by soaking with suspension of bacteria, coelome injection and muscle injection, the LD50 dose was 5.68 × 10^6 CFU per sea cucumber, and it was proved that soaking with suspension of bacteria and coelome injection could not infect the sea cucumber successfully. By methods of morphological observation, physiological and biochemical characteristic analysis, RH2 was identified as Vibrio alginolyticus. 16S rRNA gene and 60 ku heat shock protein （HSP60） gene sequence detection further proved that strain RH2 belonged to V. alginolyticus. Phylogenetic tree of Vibrio based on 16S rRNA gene and HSP60 gene both indicated that strain RH2 showed the highest level of similarity to V. alginolyticus, and its bootstrap was 50% and 99 %, respectively. The optimum salinity and temperature for strain RH2 was 25 - 35 and 14 - 22 ℃, the optimum pH was 7.2 - 8.0. The results of sensitivity to 18 kinds of antibacterial agents showed that strain RH2 was sensitive to norfloxacin,sulfamethoxazolum and spectinomycine.
出处
《水产学报》
CAS
CSCD
北大核心
2007年第4期504-511,共8页
Journal of Fisheries of China
基金
国家自然科学基金(30271016)
山东省教育厅科技计划(032070104)
教育部海水养殖重点实验室开放基金(200212)
中国海洋大学资助项目(1400-813764)
关键词
刺参
溃疡病
溶藻弧菌
16SRRNA基因
HSP60基因
生物学特性
Apostichopus japonicus
skin ulceration
Vibrio alginolyticus
16S rRNA gene
HSP60 gene
biological characteristic
作者简介
杨嘉龙（1982-），男，山东蓬莱人，硕士研究生，主要从事水产病害与免疫学研究。E-mail：jlyang1220@gmail．com
通讯作者：周丽，Tel：0532—82031862。E-mail：zhouli@ouc．edu．cn
