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摘要:
目的 评估普通拟杆菌（Bacteroides vulgatus）Bv46的生物特性和益生潜能。 方法 模拟胃肠道的酸性和胆盐环境，评估普通拟杆菌Bv46耐逆性特点；通过溶血、药敏和明胶酶定性实验评估其安全性；基于其自聚集、疏水性和对HT-29细胞的黏附情况评估其黏附能力；利用琼脂扩散、色谱和吸光度测定，评估Bv46对致病菌的拮抗、产酸和抗氧化功能。 结果 分离自健康人群的普通拟杆菌Bv46在模拟胃液和0.3%胆盐中的存活率分别为59.55%和63.76%；无溶血现象和明胶酶活性，除对氨苄西林和青霉素G耐药外，对其他所检测抗生素均敏感；Bv46具有中等自聚集能力、高疏水性和对HT-29细胞的中等黏附性；对致病性大肠埃希菌和鼠伤寒沙门菌的生长有抑制作用；此外，Bv46可以产生短链脂肪酸，并且具有抗氧化作用。 结论 普通拟杆菌Bv46是潜在的益生菌。 Abstract:Objective To evaluate probiotic potential of Bacteroides vulgatus Bv46. Methods The Bv46 strain was tested on their tolerance to artificial gastric juices and bile salts. The Bv46 was assessed the hemolytic activity, antimicrobial susceptibility and gelatinase activity. Moreover, the probiotic properties of Bv46 were evaluated about auto-aggregation, hydrophobicity, adhesion to HT-29 cells, antimicrobial activity against pathogens, acid production and antioxidant properties. Results It was 59.55% and 63.76% to the survival rates of Bv46 isolated from healthy people in simulated gastric juice and 0.3% bile salts respectively. Bv46 showed no haemolytic and gelatinase activities, and were only resistant to ampicillin and penicillin G. Bv46 showed medium auto-aggregation ability, but high hydrophobicity and medium adhesion to HT-29 cells. Moreover, Bv46 inhibited the growth of pathogenic Escherichia coli and Salmonella typhimurium. Furthermore, Bv46 produced short-chain fatty acids, and had antioxidant effects. Conclusion The Bv46 is a potential probiotic. -
Key words:
- Bacteroides vulgatus /
- Adhesion /
- Probiotic /
- Short chain fatty acids
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图 1 普通拟杆菌Bv46溶血和明胶酶活性检测结果
注：A. 普通拟杆菌Bv46溶血实验阴性；B. 普通拟杆菌Bv46明胶酶实验阴性；C. 金黄色葡萄球菌（ATCC 25923）作为明胶实验阳性对照［9］
Figure 1. Haemolytic and gelatinase activity of Bacteroides vulgatus Bv46
图 2 普通拟杆菌Bv46黏附HT-29细胞的结果
注：1 000倍光学显微镜下观察普通拟杆菌Bv46黏附HT-29细胞的情况。不加菌的HT-29细胞作为黏附的空白对照，植物乳杆菌HT121作为阳性对照
Figure 2. Adhesion of Bacteroides vulgatus Bv46 to HT-29 cell
表 1 普通拟杆菌Bv46对酸和0.3%胆盐耐受性
Table 1. Acid and 0.3% bile salt resistance of Bacteroides vulgatus Bv46
菌株名称 酸耐受性a
（存活率,%）0.3%胆盐耐受性
（存活率,%）普通拟杆菌Bv46 59.55±0.18 63.76±0.85 注：a. pH值3.0的含有3 g/L胃蛋白酶的BHI液体培养基 
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表 2 用 E-test试纸条检测普通拟杆菌Bv46的抗生素敏感性
Table 2. Antibiotic susceptibility test of Bacteroides vulgatus Bv46 by E-test strips
抗生素名称 普通拟杆菌Bv46 氨苄西林 耐药 青霉素G 耐药 阿莫西林–克拉维酸 敏感 头孢曲松 敏感 美罗培南 敏感 四环素 敏感 克林霉素 敏感 氯霉素 敏感 甲硝唑 敏感
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表 3 普通拟杆菌Bv46自聚集、疏水性和DPPH自由基清除率结果
Table 3. The self-aggregation, hydrophobicity and DPPH free radical scavenging rate of Bacteroides vulgatus Bv46
菌株名称 自聚集
（%）疏水性
（%）DPPH自由基
清除率（%）普通拟杆菌Bv46 38.53±1.32 69.27±1.56 81.43±2.01 植物乳杆菌HT121 73.32±1.45 64.70±1.78 98.87±1.64
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表 4 普通拟杆菌Bv46对致病菌的抑制作用
Table 4. Antimicrobial activity against pathogens of Bacteroides vulgatus Bv46
致病菌名称 大肠埃希菌
（CICC 24186）大肠埃希菌
（ATCC 43895）鼠伤寒沙门菌
（ATCC 14028）金黄色葡萄球菌
（ATCC 25923）李斯特菌
（ ATCCBAA-697）抑菌情况 ++ ++ ++ − − 注：符号与直径的对应关系：−. 无抑菌圈；+. 2～6 mm抑菌圈；++. 7～11 mm抑菌圈
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表 5 普通拟杆菌Bv46培养上清液的pH监测及短链脂肪酸含量测定
Table 5. pH value and short chain fatty acids in the culture supernatents of Bacteroides vulgatus Bv46
上清液pH监测 上清中短链脂肪酸含量测定（μg/mL） 0 h pH 24h pH 乙酸 丙酸 异丁酸 丁酸 7.40±0.03 5.88±0.09 1486.21 173.75 6.07 0.95
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