金丽假交替单胞菌的分离鉴定与益生活性检测  

Isolation,Identification and Probiotic Activity of Pseudoalteromonas flavipulchra

作　　者：沈煜 张皓杰 张文娟 王丽荣 蔡秀磊[1] 单虎[1] SHEN Yu;ZHANG Hao-jie;ZHANG Wen-juan;WANG Li-rong;CAI Xiu-lei;SHAN Hu(College of Veterinary Medicine,Qingdao Agricultural University,Qingdao,Shandong,266109,China;Shandong Bilan Biotechnology Co.,Ltd,Tai'an,Shandong,271411,China)

机构地区：[1]青岛农业大学动物医学院,山东青岛266109 [2]山东碧蓝生物科技有限公司,山东泰安271411

出　　处：《动物医学进展》2022年第12期67-72,共6页Progress In Veterinary Medicine

基　　金：2020年度山东省发展和改革委员会新旧动能转换重大工程重大课题攻关项目(鲁发改动能办[2020]1220号);青岛农业大学高层次人才科研基金(663/1119004)。

摘　　要：为探索海洋微生物资源在动物细菌性疾病防控中的应用,从健康斑节对虾分离纯化到1株具有抑菌活性的菌株DL1,通过细菌分离培养、16S rRNA测序和系统发育分析对其进行了初步鉴定,并采用牛津杯法、平板法分别测定了其抑菌活性和群体感应淬灭活性,测定了不同培养时间对菌株DL1的生物量、胞外蛋白含量和抑菌活性的影响。结果显示,菌株DL1的16S rRNA核苷酸序列与金丽假交替单胞菌NCIMB 2033 T同源性为100%,系统发育分析显示二者聚为一簇,亲缘关系最近,鉴定为金丽假交替单胞菌;该菌对金黄色葡萄球菌、鳗弧菌、东方弧菌和大肠埃希氏菌均具有较强的抑菌活性,且在培养48 h时生物量和蛋白量最高、菌株的抑菌活性最强;菌株DL1可以淬灭3-oxo-C6-HSL、C8-HSL、C10-HSL、C12-HSL、3-oxo-C14-HSL等密度感应信号分子,具有密度感应淬灭活性。结果表明,成功分离到1株金丽假交替单胞菌DL1,并测定了其抑菌活性和密度感应淬灭活性,为后续深入开展菌株DL1及其抑菌活性产物的研究与应用奠定了基础。To explore the application of marine microbial resources in the prevention and control of animal bacterial diseases,a strain DL1 with probiotic activity was obtained from healthy spotted shrimp.The strain DL1 was preliminarily identified after cultured based on the 16S rRNA sequencing and phylogenetic analysis.Oxford cup method and plate method were used to determine the antibacterial activity and quorum quenching activity.In addition,the biomass,extracellular protein content and antibacterial activity of different culture stages were determined and compared.The results of 16S rRNA sequence and phylogenetic analysis show that the strain DL1 was closely related with Pseudoalteromonas flavipulchra NCIMB 2033 T,sharing 100%similarity.DL1 could significantly inhibit the growth of Staphylococcus aureus,Vibrio anguillarum,Vibrio orientalis and Escherichia coli.Furthermore,the biomass,protein amount and antibacterial activity were highest and strongest cultured till 48 h.From the results of plate test,DL1 showed strong quenching activity on the signal molecules 3-oxo-C6-HSL,C8-HSL,C10-HSL,C12-HSL and 3-oxo-C14-HSL.This study lays the foundation for further research and application of strain DL1 and its antibacterial products.

关 键 词：金丽假交替单胞菌 分离鉴定 抑菌活性 密度感应淬灭 

分 类 号：S852.617[农业科学—基础兽医学]