Screening and characteristic analysis of Bacillus velezensis from sea cucumber ( Apostichopus japonicus ) ponds

Abstract

PDF HTML阅读 XML下载 导出引用 引用提醒 刺参养殖池塘一株贝莱斯芽孢杆菌的分离及其生理特性 DOI: 作者: 作者单位: 1. 上海海洋大学 水产与生命学院, 上海 201306;2. 中国水产科学研究院黄海水产研究所, 青岛海洋科学与技术国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266071;3. 山东海跃水产科技有限公司, 山东 东营 257500 作者简介: 王成强(1988-),男,研究实习员,主要从事水产动物营养与饲料研究.E-mail:chengqiangwang@126.com 通讯作者: 中图分类号: S947 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）；中央级公益性科研院所基本科研业务费专项资金项目（20603022016008）. Screening and characteristic analysis of Bacillus velezensis from sea cucumber (Apostichopus japonicus) ponds Author: Affiliation: 1. Shanghai Ocean University, Shanghai 201306, China;2. College of Fesheries and Life Science, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences;Qingdao National Laboratory for Marine Science and Technology, Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao 266071, China;3. Shandong Haiyue Aquatic Science and Technology Co., Ltd, Dongying 257500, China Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:2014年5月，在东营的刺参（）养殖池塘环境中对水样和底泥样品进行了细菌分离和益生菌的筛选，共获得66株可培养细菌。以刺参“腐皮综合征”主要致病菌为指示菌进行拮抗作用实验，使用选择培养基对菌株的产淀粉酶和蛋白酶能力进行测定，筛选获得一株潜在益生菌株——DY-6，并进行了生理生化实验、16S rDNA基因序列分析、菌株生长特性及其对刺参的安全性研究。结果表明，DY-6对其指示菌假交替单胞菌（Vibrio parahaemolyticus）和溶藻弧菌（）均有良好的抑制作用，抑菌圈直径分别达到24 mm、22 mm、27 mm和37 mm，对淀粉和蛋白培养基的水解直径分别为28 mm和20 mm。利用该菌对刺参进行高浓度胁迫实验测试其安全性，实验期间浸浴组和投喂组刺参状态良好，没有发病和死亡现象。利用细菌生理生化和16S rDNA基因序列分析表明，DY-6与贝莱斯芽孢杆菌（ CP015911）的相似性为99%。因此，初步鉴定该菌株为贝莱斯芽孢杆菌。同时，将该菌株与商品化微生态制剂产品中分离的4株益生菌进行了比较，DY-6在温度20~35℃、盐度0~35范围内生长较快，优于上述4株菌，4 h进入对数生长期，10 h达到生长高峰。筛选的贝莱斯芽孢杆菌具有良好的抑菌能力，生长速度快，并且具有广温广盐的优点，不仅能够降低疾病发生率，还可更好地适应黄河口地区夏季水温偏高、盐度波动大的特点，由此具有良好的益生菌商业化开发潜力。 Abstract:To screen local probiotic strains, 66 cultivable bacterial strains were isolated from water and sediment samples from cultured ponds of sea cucumber () in Dongying in May 2014. Bacterial antagonistic experiments were conducted using V. parahaemolyticus and , which are main pathogenic bacteria of ‘skin ulcer syndrome’ for . In addition, 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 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, respectively. 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 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. Moreover, 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. 参考文献 相似文献 引证文献