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为筛选对虹鳟(Oncorhynchus mykiss)有潜在益生效果的菌株,本研究从健康虹鳟幼鱼肠道中分离到1株潜在益生菌株RT-BS07,采用形态学观察和分子特征分析对该菌株进行了鉴定,并评价了该菌株的抑菌性、耐受性、黏附性和安全性。形态学观察显示,菌株RT-BS07为革兰氏阳性杆菌,有圆形或椭圆形芽孢。16S rRNA比对结果显示,该菌株与枯草芽孢杆菌(Bacillus subtilis)NCBI3610株的同源性为99.86%。结合16S rRNA基因序列分析和形态学观察结果,确定菌株RT-BS07为枯草芽孢杆菌。菌株RT-BS07具备产蛋白酶和纤维素酶能力,其酶活力分别为(0.039±0.002)和(0.393±0.002) U/mg;该菌株可有效抑制温和气单胞菌(Aeromonas sobria)、杀鲑气单胞菌(A.salmonicida)和鲁氏耶尔森氏菌(Yersinia ruckeri)等致病菌。菌株RT-BS07在pH为2～10 LB液体培养基中均可存活;在盐度为2%～8%条件下存活率为5.70%～93.28%,其存活率随盐度升高而降低。体外黏附试验结果表明,菌株RT-BS07可大量黏附在虹鳟前肠黏膜,不破坏肠道组织完整性,对左氟沙星、环丙沙星、氧氟沙星和链霉素等18种抗菌药物敏感。本研究分离筛选得到的枯草芽孢杆菌RT-BS07株具有产消化酶性能,其抗逆性能强,对虹鳟无毒害作用,可作为虹鳟健康养殖及益生菌制剂研发源益生菌候选株。
Abstract：To obtain the potential probiotics, one bacterial strain was isolated by comparing the enzyme production characteristics of the isolates from the intestines of healthy rainbow trout(Oncorhynchus mykiss) juvenile and named RT-BS07. The results of morphological observation showed that the strain with round or oval spores was Gram-positive bacillus, and its colony with fold and hump on the edge was milky white, rough and opaque. According to 16 S rRNA gene sequencing analysis, the gene of RT-BS07 was 99.86% similar to that Bacillus subtilis(NCBI3610). Combined with 16 S rRNA gene sequence analysis, RT-BS07 strain was identified as B. subtilis. The strain had the enzyme production capacity of protease and cellulase, which was(0.039±0.002)and(0.393±0.002) U/mg, respectively. It can effectively inhibit Aeromonas sobria, A. salmonicida and Yersinia ruckeri, and the antibacterial zone area was 5.34, 3.27 and 2.40 cm2, respectively. RT-BS07 strain can survive LB liquid medium at pH 2～10. The survival rate ranged from 5.70% to 93.28% when the salinity varied between 2% and 8%, decreasing when the salinity increased. Adhesion test in vitro showed that RT-BS07 strain can adhere to the anterior intestinal mucosa without destroying the integrity. Antimicrobial susceptibility test indicated that RT-BS07 strain was sensitive to 18 antimicrobial agents such as levofloxacin, ciprofloxacin, ofloxacin, streptomycin among others. These results indicated that the isolated B. subtilis stain RT-BS07 can be used as a candidate strain, providing strain resources for healthy aquaculture of rainbow trout and the development of probiotic preparation.
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基本信息:
中图分类号:S943
引用信息:
[1]樊丹,邓福容,李绍戊,等.一株虹鳟源枯草芽孢杆菌的分离鉴定及生物学特性研究[J],2022,52(02):41-49.
基金信息:
中央级公益性科研院所基本科研业务费专项(HSY202007M);; 现代农业产业技术体系项目(CARS-46);; 冷水性鱼类病害防控创新团队项目(2020TD43)资助~~
2021-03-15
2021
2021-05-12
2021-07-06
2021
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