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牦牛源产细菌素屎肠球菌的分离鉴定和益生特性
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西南民族大学引进人才科研启动金资助项目(RQD2021074)


Isolation, identification, and probiotic properties of bacteriocin-producing Enterococcus faecium from yak
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    摘要:

    【背景】抗生素的滥用导致牦牛肠道常见病原菌耐药性增加,益生菌作为对抗耐药性细菌的新型武器,应用前景广阔。【目的】获取益生特性优良的牦牛源益生菌。【方法】将20份牦牛粪便样本在含0.5% CaCO3的MRS培养基上分离纯化,以大肠杆菌和金黄色葡萄球菌为指示菌,用牛津杯法筛选有抑菌活性的菌株;排除酸和过氧化氢后,经耐酸耐热试验和蛋白酶敏感试验筛选产细菌素菌株,用形态学和16S rRNA基因序列分析鉴定;通过对大肠杆菌、沙门氏菌等腹泻病原菌体外抑菌试验、耐模拟胃肠液、测定自聚集能力和疏水性及抗生素敏感试验分析益生特性。【结果】从20份牦牛粪便样本中共分离出11株产生溶钙圈的菌株,其中6株对大肠杆菌和金黄色葡萄球菌抑菌效果显著,经复筛得到2株产细菌素的乳酸菌SC6和SC9,经鉴定均为屎肠球菌(Enterococcus faecium)。其中SC9对腹泻病原菌抑菌效果明显,有良好的耐受性和肠道黏附能力,对5种常用抗生素均敏感。【结论】屎肠球菌SC9有一定的抗逆性和潜在的益生能力,具备作为益生菌的潜力。

    Abstract:

    [Background] The abuse of antibiotics leads to the increase in drug resistance of common pathogenic bacteria in the intestinal tract of yaks. While probiotics have a promising application against drug-resistant bacteria.[Objective] To obtain yak probiotics with excellent probiotic properties is the purpose. [Methods] Twenty yak fecal samples were isolated and purified on MRS medium containing 0.5% CaCO3, and strains with bacterial inhibitory activity were screened out by the Oxford cup method using Escherichia coli and Staphylococcus aureus as indicator bacteria. After acid and hydrogen peroxide were discharged, the bacteriocin-producing strains were screened out by acid and heat resistance test and protease sensitivity test, and identified by morphology and 16S rRNA sequence analysis. The probiotic properties were analyzed by in vitro bacteriostatic test against diarrhea pathogenic such as E. coli and Salmonella spp., measuring self-aggregation ability and hydrophobicity, tolerance to simulated gastrointestinal juice test, and antibiotic sensitivity test. [Results] Eleven strains of calcium soluble circles were isolated from 20 yak fecal samples, six of which had significant inhibitory effects on E. coli and S. aureus. Two strains of bacteriocin-producing lactic acid bacteria (SC6 and SC9) were obtained by re-screening, both of which were identified as Enterococcus faecium. The strain SC9 had obvious bacteriostatic effect on the bacteria of yak diarrhea, with good tolerance and intestinal adhesion ability in the simulated gastric and intestinal fluid, and it was sensitive to five commonly used antibiotics. [Conclusion] E. faecium SC9 has certain stress resistance and the potential as a probiotic.

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王朝,冉旋,雷江英,陈德纯. 牦牛源产细菌素屎肠球菌的分离鉴定和益生特性[J]. 微生物学通报, 2023, 50(8): 3454-3466

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  • 收稿日期:2022-10-21
  • 最后修改日期:
  • 录用日期:2023-01-07
  • 在线发布日期: 2023-08-08
  • 出版日期: 2023-08-20
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