鲤源高黏附力地衣芽孢杆菌HF-07107分离及其益生菌特性研究

Isolation and probiotic characteristics of Bacillus licheniformis HF-07107 with high adhesion from common carp (Cyprinus carpio L.)

  • 摘要: 【目的】分离、鉴定鲤源高黏附力地衣孢杆菌菌株,并综合评价其作为鲤养殖饲料添加剂的潜力,为开发宿主源益生菌在鲤养殖中的应用提供参考。【方法】从健康鲤的肠道及其内容物中分离出潜在益生菌,根据形态学特征、生理生化特性、16S rRNA序列比对分析、菌株系统发育进化树构建等对分离菌株进行鉴定。通过检测分离菌株的自聚集能力、共聚集能力及其在有机溶剂中的疏水性评估其黏附能力,并通过测定菌株的耐酸、耐胆盐和耐高温能力,对嗜水气单胞菌和金黄色葡萄球菌的抑菌能力及对抗生素药敏片的敏感性,明确其是否具有益生菌特性。【结果】从健康鲤的肠道及其内容物中分离出菌株HF-07107,形态学特征显示该菌株能产生芽孢、呈革兰氏阳性、杆状,其生理生化特征与地衣芽孢杆菌一致,其16S rRNA序列与地衣芽孢杆菌ATCC 14580T相似性为99.31%,且与地衣芽孢杆菌聚类在同一分支上,综合鉴定该菌株为地衣芽孢杆菌。菌株HF-07107自聚集能力最高达72.92%,与迟缓爱德华氏菌和嗜水气单胞菌的共聚集能力分别为96.15%和60.23%,在有机溶剂甲苯、二甲苯、三氯甲烷、正己烷中的疏水性值较高(90.00%以上),说明菌株HF-07107有高黏附力。同时,菌株HF-07107能耐受pH 2.0、0.3%胆盐浓度和90 ℃高温环境,抑制嗜水气单胞菌和金黄色葡萄球菌的生长,对复方新诺明、丁胺卡那、环丙沙星、庆大霉素、红霉素、四环素、头孢唑林和诺氟沙星等8种抗生素敏感。【结论】鲤源潜在益生菌地衣芽孢杆菌HF-07107具有高黏附特性,能耐受不良环境,抑制病原菌生长,可作为宿主源益生菌应用于水产养殖。

     

    Abstract: 【Objective】To isolate and characterize Bacillus licheniformis strains with high adhesion from common carp and comprehensively assess its probiotic potential as feed additives for common carp culture,which provided reference for the application of host-derived probiotics in common carp culture.【Method】Potential probiotics were isolated from intestinal tract and its contents of healthy common carp. The isolated strains were identified by morphological characteristics, physiological and biochemical characteristics,16S rRNA sequence comparative analysis and phylogenetic tree construction. The adhesive ability of the isolated strain was evaluated by detecting its self-aggregation ability,co-aggregation ability and hydrophobicity in organic solvents.The acid resistance,bile salt tolerance and high temperature resistance,as well as its antibacterial ability to Aeromonas hydrophila and Staphylococcus aureus and its sensitivity to antibiotic tablets were detected to determine whether the strain had probiotic properties.【Result】Strain HF-07107 were isolated from intestinal tract and its contents of healthy common carp. The morphological characteristics of strain HF-07107 showed that it was spore-producing,Gram-positive,rod-shaped,and its physiological and biochemical identification results were consistent with those of Bacillus licheniformis. The similarity of 16S rRNA sequence between the strain HF-07107 and B. licheniformis ATCC 14580T was 99.31%,and strain HF-07107 was in the same cluster with B. licheniformis in the phylogenetic tree. The strain HF-07107 was comprehensively identified as B. licheniformis. The strain had a high self-aggregation ability of 72.92%,co-aggregation ability with Edwardsiella tarda and A. hydrophila(96.15% and 60.23%)and high hydrophobicity value with toluene,xylene,chloroform and n-hexane(more than 90.00%),indicating strain HF-07107 had high adhesion. At the same time,strain HF-07107 could tolerate pH 2.0,0.3% bile salt and 90 ℃ high temperature environment,inhibit the growth of A. hydrophila and S. aureus and was sensitive to bactrim,amikacin,ciprofloxacin,gentamicin,erythromycin,tetracycline,cefazolin and norfloxacin.【Conclusion】B. licheniformis HF-07107,a potential probiotic from common carp with high adhesion characteristics,can tolerate adverse environment,and inhibit the growth of pathogenic bacteria. Therefore,it can be used as a host-derived probiotic in aquaculture.

     

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