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从油橄榄微生物群中筛选和鉴定新型益生菌候选菌株的体外研究

In vitro selection and characterization of new probiotic candidates from table olive microbiota.

作者信息

Botta Cristian, Langerholc Tomaz, Cencič Avrelija, Cocolin Luca

机构信息

Department of Forestry, Agriculture and Food Sciences, University of Torino, Torino, Italy.

Department of Microbiology, Biochemistry, Molecular Biology and Biotechnology, Faculty of Agriculture and Life Sciences, University of Maribor, Maribor, Slovenia.

出版信息

PLoS One. 2014 Apr 8;9(4):e94457. doi: 10.1371/journal.pone.0094457. eCollection 2014.

Abstract

To date, only a few studies have investigated the complex microbiota of table olives in order to identify new probiotic microorganisms, even though this food matrix has been shown to be a suitable source of beneficial lactic acid bacteria (LAB). Two hundred and thirty eight LAB, belonging to Lactobacillus plantarum, Lactobacillus pentosus and Leuconostoc mesenteroides species, and isolated from Nocellara Etnea table olives, have been screened in this survey through an in vitro approach. A simulation of transit tolerance in the upper human gastrointestinal tract, together with autoaggregation and hydrophobicity, have been decisive in reducing the number of LAB to 17 promising probiotics. None of the selected strains showed intrinsic resistances towards a broad spectrum of antibiotics and were therefore accurately characterized on an undifferentiated and 3D functional model of the human intestinal tract made up of H4-1 epithelial cells. As far as the potential colonization of the intestinal tract is concerned, a high adhesion ratio was observed for Lb. plantarum O2T60C (over 9%) when tested in the 3D functional model, which closely mimics real intestinal conditions. The stimulation properties towards the epithelial barrier integrity and the in vitro inhibition of L. monocytogenes adhesion and invasion have also been assessed. Lb. plantarum S1T10A and S11T3E enhanced trans-epithelial electrical resistance (TEER) and therefore the integrity of the polarized epithelium in the 3D model. Moreover, S11T3E showed the ability to inhibit L. monocytogenes invasion in the undifferentiated epithelial model. The reduction in L. monocytogenes infection, together with the potential enhancement of barrier integrity and an adhesion ratio that was above the average in the 3D functional model (6.9%) would seem to suggest the Lb. plantarum S11T3E strain as the most interesting candidate for possible in vivo animal and human trials.

摘要

迄今为止,尽管食用橄榄这种食物基质已被证明是有益乳酸菌的合适来源,但仅有少数研究调查了食用橄榄的复杂微生物群,以鉴定新的益生菌微生物。在本次调查中,通过体外方法对从埃特纳火山诺塞拉食用橄榄中分离出的238株乳酸菌进行了筛选,这些乳酸菌属于植物乳杆菌、戊糖乳杆菌和肠系膜明串珠菌。在上消化道中模拟转运耐受性以及自聚集和疏水性,对于将乳酸菌数量减少到17种有前景的益生菌起了决定性作用。所选菌株均未表现出对广谱抗生素的固有抗性,因此在由H4-1上皮细胞组成的人肠道未分化3D功能模型上进行了精确表征。就肠道潜在定植而言,在3D功能模型中进行测试时,植物乳杆菌O2T60C的黏附率很高(超过9%),该模型紧密模拟了真实的肠道条件。还评估了对上皮屏障完整性的刺激特性以及对单核细胞增生李斯特菌黏附和侵袭的体外抑制作用。植物乳杆菌S1T10A和S11T3E增强了跨上皮电阻(TEER),从而增强了3D模型中极化上皮的完整性。此外,S11T3E在未分化上皮模型中表现出抑制单核细胞增生李斯特菌侵袭的能力。单核细胞增生李斯特菌感染的减少,加上屏障完整性的潜在增强以及3D功能模型中高于平均水平(6.9%)的黏附率,似乎表明植物乳杆菌S11T3E菌株是体内动物和人体试验最有前景的候选菌株。

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