黑鲷肠炎病原恶臭假单胞菌的分离和鉴定  

Isolation and characterization of Pseudomonas putida,a causative agent of gastroenteritis in black sea bream,Sparus macrocephalus

作　　者：毛芝娟[1] 王美珍[2] 陈吉刚[1] 杨季芳[1] 

机构地区：[1]浙江万里学院生物与环境学院,宁波315100 [2]华中农业大学水产学院,武汉430070

出　　处：《渔业科学进展》2010年第3期23-28,共6页Progress in Fishery Sciences

基　　金：宁波市科技局择优委托项目(2007C10037)资助

摘　　要：2008年3月,浙江省象山港养殖黑鲷暴发了严重的肠炎病情,病鱼肠道肿胀,充满黄色黏液。以ZoBell2216E平板自病鱼肠道黏液分离到4株菌BB01、BB02、BB03和BB04。通过人工感染试验验证了前3株为病原菌,其中BB01菌株96h的半数致死浓度LD50为4.61×104CFU/g鱼体重。3株分离菌革兰氏染色阴性,短杆状,氧化酶试验阳性,氧化葡萄糖,不产气,4℃生长,精氨酸双水解酶阳性,利用麦芽糖、柠檬酸盐,硝酸盐降解阳性,DNA酶、乙酰胺酶阴性;与已发表的恶臭假单胞菌16SrRNA序列的同源性达99%以上;经形态和理化特性鉴定及16SrDNA序列测定,此3株菌同属于恶臭假单胞菌。药敏试验结果表明,菌株BB01对头孢类等多数抗生素种类敏感,而对氧氟沙星耐药。An outbreak of serious mortality among the cultured black sea bream Sparus macrocephalus,characterized by a swollen intestine containing yellow fluid,occurred in early spring of 2008 in Xiangshan Bay,Zhejiang Province.Four strains of bacteria,BB01,BB02,BB03 and BB04 were isolated from the intestinal yellow fluid of the moribund black sea bream with Zobell2216E agar and/or thiosulfate citrate bile salt sucrose agar.The former three were identified as causative agents via artificial challenge.The 96h LD50 value of BB01 was 4.61×104 CFU/g bream body weight.The former three strains were characterized as Pseudomonas putida through morphological,physical and biochemical profiles tests and 16S rRNA sequencing.The three strains were Gram-negative,short-rod-shaped,oxidase and glucose oxidation positive,gas negative,maltose and citrates utilization positive,arginine dihydrolase positive,nitrate reduction positive,DNA enzyme and acetamidase negative.The 16S rRNA sequence of BB01 shared the identity of over 99% with those of P.putida strains.Drug sensitivity tests showed that the strain BB01 was susceptible to most of the tested antibiotics while resistant to ofloxacin.

关 键 词：黑鲷 病原 恶臭假单胞菌 16S RDNA 

分 类 号：S941.42[农业科学—水产养殖]