大黄鱼病原哈维氏弧菌(Vibrio harveyi)拮抗菌的筛选及益生菌特征的分析  

Selection of antagonistic bacteria against pathogenic Vibrio harveyi of Larimichthys crocea and analysis of probiotic characteristics

作　　者：王梦霞[1] 朱晓玲[1] 王君如[1] 刘嘉佩 刘雪珠[1] 

机构地区：[1]浙江海洋学院海洋科学与技术学院,浙江舟山316022

出　　处：《饲料工业》2015年第2期52-58,共7页Feed Industry

基　　金：国家自然科学基金面上项目[31270160];浙江省自然科学基金项目[LY12C03003];国家级大学生创新训练项目[201310340001]

摘　　要：用点种法定性初筛,从65株由海洋生物体上获得的未知菌株中筛选出18株病原哈维氏弧菌的拮抗菌,用酶标法和牛津杯法定量复筛后,确定2株保持较好拮抗效果的菌株X14、X15为目的拮抗菌。在对两菌株益生菌特征分析的研究中,抗菌谱及生长曲线结果显示菌株X14、X15具有较广的抗菌性,且两株拮抗菌在相同的条件下可能较两株病原菌更具生长优势。药物敏感性试验结果表明,分离菌株菌对四环素、环丙沙星、庆大霉素等10种药物敏感。16S r DNA基因同源性分析的结果显示与X14、X15同源性达99%以上的菌株均来自链霉菌属(Streptomyces),进一步建立系统发育树,结果显示菌株X15与灰平链霉菌(Streptomyces griseoplanus)相似性最高,但结合形态特征及理化性质不能将两菌株确定到种。筛选得到的菌株X14、X15在拮抗病原哈维氏弧菌、抗菌谱、生长优势及药敏性等方面均初步满足益生菌的筛选条件。By spotting to qualitatively preliminary screen, we selected 18 strains of antagonistic bacte-ria to etiology of Vibrio harveyi from 65 unknown strains of marine organisms. After quantitatively sec-ond screening by the enzyme-labelled immunosorbent assay（ELISA） and the oxford cup method, wecan determine the two strains X14 and X15 which maintained good antagonism effects as the purpose.By analyzing the characteristics of two strains of probiotics, the results of the antibacterial spectrumand growth curve showed that strains X14 and X15 had wide antimicrobial properties, and further-more, in the same conditions, the growth advantages of the two strains of antagonistic bacterias may bemore than that of the two strains of pathogens. The drug sensitivity test result showed that the separat-ed strains of bacteria were sensitive to 10 kinds of drugs such as tetracycline, ciprofloxacin, gentamy-cin. The results of 16 S rDNA gene homology analysis showed that strains above 99% homology withX14 and X15 were Streptomyces, and we further established a phylogenetic tree of which showed thatwere in the same branch, but combining with themorphological characteristics and physical andchemical properties, it cannot be specificallyidentified to species. The strains X14 and X15 filtered initially met the criteria of probiotics interms of the antagonism to etiology of Vibrio har-veyi, antibacterial spectrum, growth advantageand drug susceptibility, etc.

关 键 词：拮抗菌 抗菌谱 药敏试验 生长曲线 16S RDNA 

分 类 号：S963.736[农业科学—水产养殖]