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Screening for lactobacilli with probiotic properties in the infant gut microbiota

机译:在婴儿肠道菌群中筛选具有益生菌性质的乳酸菌

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Lactobacilli are believed to be beneficial for the human hosts and are currently being evaluated as potentially probiotic bacteria. In this study, Lactobacillus strains were isolated from infant faeces and were examined invitro for potential probiotic properties. Faecal specimens from 63 healthy, full-term infants were collected at 4, 30 and 90 days after delivery. Seventy-four Lactobacillus strains were isolated and one or more different phenotypes from each infant (n=44) were selected for further testing. The bacterial isolates were identified mainly as L.gasseri, L.crispatus, Lactobacillus paracasei, L.salivarius, L.fermentum after amplification and sequencing of 16s rRNA gene. The strains were examined for acid and bile tolerance, adhesion to Caco-2 cells, antibiotic susceptibility and antimicrobial activity against selected enteric pathogens. The great majority of the isolated lactobacilli were susceptible to ampicillin, amoxicillin/clavulanic acid, tetracycline, erythromycin, cephalothin, chloramphenicol and rifampicin. Resistance to vancomycin or bacitracin was detected to 34% of the strains. Twenty strains out of forty-four exhibited significant tolerance to bile salts. Those strains were subsequently tested for resistance to low pH conditions (pH 2 and 3). Interestingly, 85% (17 strains) of the tested lactobacilli remained unaffected at pH 3 after 3h of incubation, 6 strains were found resistant at pH 2 after 1.5h and only 2 strains found resistant after 3h of incubation. Two of the strains were able to adhere to Caco-2 cells. In conclusion, two isolates fulfilled the invitro probiotic criteria and are good candidates for further invivo evaluation.
机译:乳酸杆菌被认为对人类宿主有益,并且目前被评估为潜在的益生菌。在这项研究中,乳酸杆菌菌株是从婴儿粪便中分离出来的,并在体外进行了潜在益生菌特性的检测。在分娩后第4、30和90天收集了63名健康足月婴儿的粪便标本。分离出74株乳酸杆菌菌株,并从每个婴儿(n = 44)中选择一种或多种不同的表型进行进一步测试。经过16s rRNA基因的扩增和测序,细菌分离物主要鉴定为加氏乳杆菌,crispatus,副干酪乳杆菌,唾液乳杆菌,发酵乳杆菌。检查了菌株的酸和胆汁耐受性,对Caco-2细胞的粘附性,抗生素敏感性和对选定的肠道病原体的抗菌活性。绝大部分分离出的乳杆菌对氨苄西林,阿莫西林/克拉维酸,四环素,红霉素,头孢菌素,氯霉素和利福平敏感。对34%的菌株检测到对万古霉素或杆菌肽的抗性。四十四个菌株中有二十个对胆盐表现出显着的耐受性。随后测试这些菌株对低pH条件(pH 2和3)的抗性。有趣的是,在孵育3h后,pH值为3的被测乳杆菌中有85%(17个菌株)不受影响,在孵育1.5h后,pH值为2时有6个菌株具有抗性,孵育3h后只有2个菌株具有抗性。其中两个菌株能够粘附至Caco-2细胞。总之,两个分离株符合体外益生菌标准,是进一步进行体内评价的良好候选者。

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