中国水产科学2018，Vol.25Issue(3)：567-575,9.DOI:10.3724/SP.J.1118.2018.17383
刺参养殖池塘一株贝莱斯芽孢杆菌的分离及其生理特性
Screening and characteristic analysis of Bacillus velezensis from sea cucumber (Apostichopus japonicus) ponds
摘要
Abstract
To screen local probiotic strains, 66 cultivable bacterial strains were isolated from water and sediment samples from cultured ponds of sea cucumber (Apostichopus japonicus) in Dongying in May 2014. Bacterial an-tagonistic experiments were conducted using Pseudoalteromonas nigrifaciens, Vibrio splendidus, V. parahaemo-lyticus and V. alginolyticus, which are main pathogenic bacteria of 'skin ulcer syndrome' for A. japonicus. In addi-tion, selective medium was used to test the amylase production and protease activities of the isolated strains and to isolate a potential probiotic strain, DY-6. Then, its physiological-biochemical experiments and similarity analysis of 16s rDNA sequences as well as the growth characteristics of DY-6 experiments have been conducted. The safety experiment for the sea cucumber was also proceed. The DY-6 strain was successful in inhibiting P. nigrifaciens, V. splendidus, V. parahaemolyticus, and V. alginolyticus, as indicated by inhibition zone diameters of 24, 22, 27, and 37 mm, respectively. Meanwhile, the hydrolysis zone diameters of amylase and protease were 28 and 20 mm, re-spectively. A safety experiment was designed to test the safety of DY-6 for sea cucumber. During the experiment, all sea cucumbers were in good conditions without falling ill. Based on physiological and biochemical experiments and 16s rDNA sequence analysis, strain DY-6 was determined to be 99％ similar to Bacillus velezensis CP015911 and, therefore, preliminarily identified as B. velezensis. When compared to four bacterial strains isolated from commercial probiotic products, DY-6 exhibited superior growth at 20–35℃ and under salinities of 0 to 35. More-over, DY-6 came into the logarithmic growth phase after 4 h of cultivation and reached growth peak after 10 h. In summary, strain DY-6 exhibited superb inhibition of sea cucumber pathogens, rapid growth, and both euryhalinous and euryhaline features. Therefore, DY-6 could be used to reduce the occurrence of sea cucumber disease and is also suitable for use at high temperatures during summer and in fluctuation salinity levels, as in the Yellow River Estuary. Thus, strain DY-6 has potential for commercial development.关键词
黄河口/刺参/贝莱斯芽孢杆菌/抑菌/生理特性/安全性Key words
Yellow River Estuary/Apostichopus japonicus/Bacillus velezensis/antagonistic experiment/physio-logical characteristics/biochemical characteristic/safety分类
农业科技引用本文复制引用
王金燕,李彬,王印庚,廖梅杰,荣小军,张正,牛盈盈,宁鲁光..刺参养殖池塘一株贝莱斯芽孢杆菌的分离及其生理特性[J].中国水产科学,2018,25(3):567-575,9.基金项目
山东省农业良种工程重大课题 （）
山东省自主创新与成果转化专项(2014ZZCX06205) （2014ZZCX06205）
中央级公益性科研院所基本科研业务费专项资金项目(20603022016008). （20603022016008）
