抑制引起腹泻致病菌的益生菌筛选及机制初步研究

发布时间：2018-01-05 19:34

  本文关键词：抑制引起腹泻致病菌的益生菌筛选及机制初步研究　出处：《哈尔滨工业大学》2010年硕士论文　论文类型：学位论文

  更多相关文章： 益生菌筛选 蛋白质纯化 抑制黏附 致病性大肠杆菌

【摘要】： 腹泻是发展中国家导致儿童死亡的主要原因,每年有大约140万-250万的生命死于腹泻。腹泻除了直接引起健康的损伤外,还引起人体营养失调,生长速度减缓,智力发育等问题。尽管在发达国家腹泻导致死亡的的情况并不经常发生,但是腹泻仍然是引起普通疾病的主要原因,而且还消耗了大量的医疗费用。我们筛选能够抑制引起腹泻致病菌的益生菌,这些益生菌对于预防腹泻发挥重要的作用,益生菌配合药物治疗腹泻既能够使腹泻快速康复,又能够保护肠道菌群免受药物的破坏。用益生菌预防和治疗腹泻还能够有效地降低医疗费用,减轻医疗负担。 从健康成人肠道中分离肠道益生菌。采集粪便样本,稀释之后用RB、CAN和LAM培养基分离出39株益生菌,其中15株双歧杆菌、14株乳酸球菌和10株乳酸杆菌。然后以致病性大肠杆菌(E.coli)作为指标菌株对这39株益生菌进行抑菌筛选,发现乳酸球菌对于致病性大肠杆菌没有抑制作用,大部分双歧杆菌和乳杆菌有抑菌作用。对于剩余的具有抑菌能力的益生菌菌液上清进行中和,发现大部分益生菌丧失抑菌能力,仅有三株益生菌具有较强的抑菌活性,最后通过过氧化氢酶试验和蛋白酶检测试验确定1321、0533和1333这三株菌,不仅能够代谢产生各种酸抑菌,还能够产生抗菌肽抑制致病菌的生长。最后通过蛋白质纯化仪对于益生菌代谢产物中的抗菌肽进行分离纯化。 筛选出能够抑制致病菌的益生菌后还要对益生菌的耐酸、耐胆盐和黏附能力进行筛选,因为只有益生菌能够耐受人体胃和肠道的复杂环境,顺利黏附到人体小肠上皮细胞上才能发挥作用。最后对于益生菌抑制致病菌黏附进行了初步研究,验证了益生菌通过代谢产物抑制黏附抑制腹泻的机制。
[Abstract]:Diarrhoea is the leading cause of death among children in developing countries, with about 1.4 million-2.5 million deaths per year due to diarrhoea, which, in addition to direct health damage, causes nutritional disorders in the human body. Problems such as slower growth and mental development. Although diarrhoea is not a common cause of death in developed countries, diarrhoea is still the leading cause of common diseases. We screen probiotics that inhibit the bacteria that cause diarrhoea, which play an important role in preventing diarrhea. Probiotics combined with drugs to treat diarrhea can not only make rapid recovery of diarrhea, but also protect intestinal flora from drug damage. Probiotics to prevent and treat diarrhea can also effectively reduce the cost of medical treatment. Lighten the burden of medical care. Intestinal probiotics were isolated from the intestinal tract of healthy adults. Fecal samples were collected, and 39 strains of probiotics were isolated by dilution with RBCAN and LAM medium, of which 15 strains were bifidobacterium. 14 strains of Lactococcus lactis and 10 strains of Lactobacillus were used as indicator strains of E. coli to screen 39 strains of probiotics. It was found that Lactococcus had no inhibitory effect on pathogenic Escherichia coli and most of Bifidobacterium and Lactobacillus had bacteriostasis. The supernatant of the remaining probiotic bacteria with bacteriostasis was neutralized. It was found that most probiotics lost their bacteriostatic ability, and only three probiotics had strong bacteriostatic activity. Finally, 1321 was determined by catalase test and protease test. The three strains, 0533 and 1333, can not only metabolize and produce a variety of acid antibacterial bacteria. Antimicrobial peptides can also be produced to inhibit the growth of pathogenic bacteria. Finally, the antimicrobial peptides in the metabolites of probiotics were separated and purified by protein purification apparatus. After screening probiotics which can inhibit pathogenic bacteria, we have to screen the acid resistance, bile salt tolerance and adhesion ability of probiotics, because only probiotics can tolerate the complex environment of human stomach and intestine. Finally, a preliminary study on the inhibition of bacterial adhesion by probiotics was carried out to verify the mechanism of the inhibition of diarrhea by metabolites of probiotics.
【学位授予单位】：哈尔滨工业大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R371

【共引文献】

中国期刊全文数据库 前10条

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