兰州大学机构库 >口腔医学院
益生性唾液乳杆菌的筛选及其抑菌物质和粘附作用的初步探究
Alternative TitleProbiotic Lactobacillus salivarius strains’ Screening and a primary research of its antibiotics and
王立鹤
Thesis Advisor杨兰
2016-05-14
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword唾液 唾液乳酸杆菌 抑菌物质 粘附 益生性
Abstract目的:从健康人群口腔中筛选具有益生潜能的唾液乳酸杆菌,为益生菌用于口腔疾病的防治做出初步探究。 方法:1.取健康志愿者的唾液进行选择性培养,经革兰染色、16srDNA测序鉴定得到唾液乳杆菌,将唾液乳酸杆菌的无菌发酵液上清对变形链球菌进行抑制实验,筛选出目标菌株并命名。2.取目标菌株的无菌发酵液上清进行抑菌物质的有效抑菌浓度、对热的稳定性、对蛋白酶的敏感性以及抑菌物质的广谱性等多方面的分析。3.以变形链球菌为指示菌,将其与目标菌株共培养,观察其粘附状况,初步研究目标菌株与变形链球菌的体外粘附能力。 结果:1. 本实验成功筛选出目标菌株并命名为 w22a。2. (1) w22a产出的抑菌物质随着浓度的增加其抑菌作用开始增强。(2)w22a产出的抑菌物质有一定的热稳定性。(3)w22a的抑菌物质对广谱蛋白酶敏感。(4)w22a产出的抑菌物质对多种口腔常见的致病菌有着程度不一的抑制作用。3. w22a在体外有着良好的粘附能力。 结论:本研究筛选出具有口腔益生潜能的唾液乳酸杆菌w22a,其产出的抑菌物质对热稳定,对广谱蛋白酶敏感并能抑制多种口腔常见致病菌。另外,w22a来源于口腔,能够对口腔环境有着良好的粘附能力及适应性。因此,其是一株有口腔益生潜能的唾液乳杆菌,具有一定的研究价值及应用前景。
Other AbstractObjectives: This study aimed to screen the probiotic potential of Lactobacillus salivarius strain from oral cavity of healthy volunteers, and to investigate its probiotic properties against oral diseases for providing a preliminary strategy. Methods: (1) Selecting Volunteers that healthy and have not any oral disease, then gathering their unstimulated saliva to selective culture using the LBS solid medium. Selecting the withe and different size circular bacterial colonies to monoclonal cultivate. Lactobacillus salivarius are selected by the Lactobacillus strains’ Gram staining and 16srDNA sequencing and BLAST in NCBI. Moreover, Lactobacillus salivarius strains are cutruled by liquid MRS medium and gathering their fermentation supernatant by high speed Centrifugation and filtration. Using fermentation supernatant to inhibit the streptococcus mutans(SM) and selecting the aim strains which have the significant ability of bacteriostasis. (2) Gathering the aim strain’s fermentation supernatant and detect bactericidal substance’s effect of bacteriostasis in different concentration. We also should detecte its stability to temperature, the sensitivity to protease, and broad-spectrum antibacterial ability. (3) The aim strains and SM UA159 are co-cultured upon the cover glass by different concentration. Using the light microscope and scanning electron microscope(SEM) to observe their adhesive ability in vitro. Results: 1. We have Selected the 4 Lactobacillus salivarius strains that have different genotype and find they have different bacteriostatic ability for SM UA159. One strain is determined the targeted species which have a significant effect of inhibiting SM UA159 and named w22a. 2. (1) Bacteriostatic ability is not obvious when the w22a’s sterile fermented supernatant diluted 1/3 as stoste. The diameter of inhibition zone have no significant differences compared with controlled group (P>0.05). But The effect of inhibiting streptococci mutans showed an increasing trends with the antibacterial substance’s concentration. (2) The w22a’s sterile fermented supernatant is treated by different temperature between 60℃ to 90℃ and it’s ability of inhibit SM UA159 have no influence, there are no significant differences between each groups(P>0.05). (3) The w22a’s sterile fermented supernatant is treated by catalase and other broad-spectrum proteases and it’s ability of inhibit SM UA159 have a significant destroying. The effect of Proteinase K is more s...
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Language中文
Document Type学位论文
Identifierhttp://ir.lzu.edu.cn/handle/262010/217910
Collection口腔医学院
Recommended Citation
GB/T 7714
王立鹤. 益生性唾液乳杆菌的筛选及其抑菌物质和粘附作用的初步探究[D]. 兰州. 兰州大学,2016.
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