传统发酵乳制品和肉制品中乳酸菌的益生菌潜力:体外试验和分子表征的评估

F. Pavli, A. Argyri, Olga S. Papadopoulou, G. Nychas, N. Chorianopoulos, C. Tassou
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引用次数: 32

摘要

本研究的目的是评估从希腊传统发酵产品中分离的乳酸菌(LAB)的益生菌潜力。通过一系列体外试验,包括在模拟胃肠道条件下的生存(对低pH、胆汁盐抗性和胆汁盐水解的抗性)和安全性评估(对抗生素的耐药性、溶血和抗菌活性),以鼠李糖乳杆菌GG和干酪乳杆菌Shirota作为参考菌株,选择潜在的候选益生菌。最初,共有255株乳酸菌分离株被恢复并在模拟胃肠道条件下进行了生存筛选,133株在这些测试中表现出中等或良好的行为,随后用分子工具在物种水平上进行了分化和表征。菌株鉴定采用脉冲场凝胶电泳技术,物种鉴定采用扩增的16S rRNA基因限制性内切分析。采用针对recA基因的特异性多重PCR方法对属植物芽孢杆菌(Lb. plantarum)群的分离物进行了种级鉴定。从133株分离株中,回收到47株不同的菌株,分别分为:酿酒乳杆菌(14株)、弯曲乳杆菌(4株)、肠系膜乳杆菌(4株)、乳酸乳球菌(4株)、干酪乳杆菌(1株)、短乳杆菌(1株)、植物芽孢杆菌(10株)、戊酸芽孢杆菌(7株)和副植物芽孢杆菌(2株)。选取经鉴定的胃肠道试验表现良好的菌株进行安全性评价。总之,鉴定的47株菌株中有19株在模拟胃肠道条件下表现良好,也被认为是安全的,因此具有与参考菌株相似或更好的理想体外益生菌特性。这些菌株可能被认为是在体内和原位研究中进一步研究的良好候选者,以评估它们潜在的健康益处以及它们作为新型益生菌起始物或辅助培养物的性能。
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Probiotic Potential of Lactic Acid Bacteria from Traditional Fermented Dairy and Meat Products: Assessment by In Vitro Tests and Molecular Characterization
The aim of the present study was to evaluate the probiotic potential of lactic acid bacteria (LAB) isolated from Greek traditional fermented products. A series of In vitro tests that included survival in simulated gastrointestinal conditions (resistance to low pH, bile salts resistance and bile salts hydrolysis) and safety assessment (resistance to antibiotics, haemolytic and antimicrobial activity) were performed to select potential probiotic candidates, while Lactobacillus rhamnosus GG and Lactobacillus casei Shirota were used as reference strains. Initially, a total of 255 isolates of LAB have been recovered and screened for their survival in simulated gastrointestinal tract conditions and 133 isolates that exhibited moderate or good behavior in these tests were subsequently differentiated and characterized at species level with molecular tools. Pulsed Field Gel Electrophoresis was applied for strain differentiation, while species differentiation was based on restriction analysis of the amplified 16S rRNA gene. Specific multiplex PCR assay targeting the recA genes was applied to resolve the species level of the isolates, belonged to Lb. plantarum group. From the 133 isolates, 47 different strains were recovered and were assigned to Lactobacillus sakei(14), Lactobacillus curvatus (4), Leuconostoc mesenteroides (4), Lactococcus lactis (4), Lactobacillus casei group (1), Lactobacillus brevis (1), Lb. plantarum (10), Lb. pentosus (7) and Lb. paraplantarum (2). The identified strains with good behavior to the gastrointestinal tract tests were selected and further evaluated for their safety aspect. In conclusion, 19 out of the 47 identified strains were assessed as well-behaved, under simulated gastrointestinal conditions and also considered as safe, possessing thus desirable In vitro probiotic properties similar or better to that of the reference strains. These strains may be considered as good candidates for further investigation at in vivo and in situ studies to assess their potential health benefits and their performance as novel probiotic starters or adjunct cultures.
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