乳酸杆菌细胞裂解物对金黄色葡萄球菌、大肠埃希菌的抑制作用

目的探讨乳杆菌DM8909裂解物在体内外对金黄色葡萄球菌、大肠埃希菌的抑制作用。方法通过对乳杆菌超声波破碎制成裂解物,分别用乳杆菌裂解物原液、裂解物稀释液、发酵上清液、乳杆菌活菌制剂进行体内、体外实验,观察乳杆菌各成分对金黄色葡萄球菌、大肠埃希菌的抑制作用。结果德氏乳酸杆菌裂解物对金黄色葡萄球菌、大肠埃希菌的抑制作用与乳杆菌活菌制剂的抑制作用相近。结论德氏乳酸杆菌裂解物在体内外对金黄色葡萄球菌、大肠埃希菌均有较强的抑制作用。     

地衣芽胞杆菌对实验性家兔阴道炎影响的研究

目的通过探讨地衣芽胞杆菌活菌制剂对实验性家兔细菌性阴道病的影响,恢复家兔阴道微生态平衡和正常菌群环境。方法（1）采用注射用氨苄西林和甲硝唑生理盐水溶液注入家兔阴道进行冲洗,建立家兔阴道脱污染动物模型。（2）取40只阴道脱污染家兔,其中20只接种大肠埃希菌,20只接种金黄色葡萄球菌,建立家兔阴道感染模型。（3）地衣芽胞杆菌对家兔阴道菌群失调的调整作用：采用不同浓度的地衣芽胞杆菌菌液（10^6CFU／ml、10^8CFU／ml、10^9CFU／ml）对感染家免阴道进行接种,分析和考察地衣芽胞杆菌对家兔阴道菌群失调的影响以及对家兔阴道黏膜的影响。结果（1）动物经过金黄色葡萄球菌感染,通过地衣芽胞杆菌治疗后,动物阴道内芽胞杆菌、肠杆菌、乳杆菌数量明显上升,葡萄球菌数量明显下降,自细胞数量减少,黏膜红肿减轻、分泌物减少,治疗作用明显。（2）大肠埃希菌感染动物经地衣芽胞杆菌治疗后,动物阴道内芽胞杆菌、乳杆菌数量明显上升,肠杆菌数量明显降低,白细胞数量减少,黏膜红肿消失、分泌物减少。结论地衣芽胞杆菌对实验性家兔金黄色葡萄球菌和大肠埃希菌阴道炎的治疗有效。地衣芽胞杆菌与乳杆菌的作用相似,具有维持阴道菌群平衡的作用。     

地衣芽胞杆菌对白色念珠菌等的拮抗作用

目的了解地衣芽胞杆菌在试管内与阴道正常菌群共生关系的情况。方法将地衣芽胞杆菌菌液分别与葡萄球菌、大肠埃希菌、白色念珠菌、德氏乳杆菌混合培养,定量计数各菌在不同时间内单独培养和混合培养时各菌的活菌数。结果地衣芽胞杆菌生长不受金黄色葡萄球菌、白色念珠菌和大肠埃希菌的影响,金黄色葡萄球菌和白色念珠菌在有地衣芽胞杆菌存在的情况下,其生长受到明显的抑制（P〈0.05）;乳杆菌在12-48 h内,有显著的抑制地衣芽胞杆菌生长的作用,而乳杆菌的生长不受地衣芽胞杆菌的存在与否而正常生长。结论地衣芽胞杆菌对金黄色葡萄球菌及白色念珠菌在体外具有明显的拮抗作用,地衣芽胞杆菌对大肠埃希菌、乳杆菌无明显的体外拮抗作用。     

乳酸杆菌体外对大肠埃希菌O-78拮抗作用试验

目的：研究乳酸杆菌体外对大肠埃希菌O-78的拮抗作用。方法：将乳酸杆菌与大肠埃希菌O-78体外共培养观察拮抗情况及用电镜观察形态变化。结果：等量同时间接种,24h内乳杆菌就将大肠埃氏菌O-78全部抑杀;乳酸杆菌早24h接种,4-8h内将大肠埃希菌O-78全部抑杀;大肠埃希菌O-78早24h接种,在12-24h内全部被抑杀。电镜观察发现,经乳酸杆菌培养液处理,大肠埃希菌O-78细胞膜形态发生变化。结论：本试验中所用乳酸杆菌体外具有抑制杀死大肠埃希菌O-78的作用。     

乳酸杆菌代谢产物对引起阴道炎常见细菌的抑制作用的初步探讨

目的探讨乳酸杆菌代谢产物对临床常见引起阴道炎的大肠埃希菌、铜绿假单胞菌、金黄色葡萄球菌、白色念珠菌、伤寒杆菌和肠球菌的抑菌作用。方法采用营养琼脂平板培养基定量涂菌,国际标准药敏杯给药的药敏试验法,检测乳酸杆菌代谢产物对大肠埃希菌、铜绿假单胞菌、金黄色葡萄球菌、白色念珠菌、伤寒杆菌和肠球菌的抑菌环的大小。结果乳酸杆菌代谢产物对大肠埃希菌、铜绿假单胞菌、金黄色葡萄球菌和伤寒杆菌有明显的抑菌作用,对肠球菌、白色念珠菌无抑菌作用。结论在临床上可应用乳酸杆菌及其制剂调节阴道微生态平衡,治疗细菌性阴道炎。     

两种生物状态肠道益生菌的粘附和粘附拮抗效应的对比研究

目的:研究活菌和灭活菌两种生物状态的肠道主要益生菌--德氏乳杆菌、双歧杆菌和肠球菌对肠黏膜上皮细胞粘附性及其对肠道几种常见病原菌的粘附拮抗效应.方法:用光镜和电镜技术分析了两种生物状态的三种益生菌对肠黏膜上皮细胞的粘附指数,通过排除实验、竞争实验和替代实验研究了两种生物状态益生菌对侵袭性大肠埃希菌、产毒性大肠埃希菌和痢疾志贺菌的粘附拮抗效应,应用平板扩散法观察了三种益生菌的代谢乏液对上述肠道病原菌的抑制能力.结果:德氏乳杆菌和肠球菌的灭活状态较活菌状态对肠黏膜上皮细胞的粘附性显著增高,双歧杆菌经灭活后对细胞的粘附性与活菌相比差异无显著性,两种生物状态的三种益生菌对肠道致病菌均具有粘附拮抗作用.滤过后的德氏乳杆菌、双歧杆菌和肠球菌的代谢乏液对侵袭性大肠埃希菌、产毒性大肠埃希菌和痢疾志贺菌均具有较明显的抑制作用,经42℃、65℃和100℃加热不影响德氏乳杆菌和双歧杆菌代谢乏液的抑菌作用.结论:灭活状态的德氏乳杆菌、双歧杆菌和肠球菌是具有潜在开发价值的微生态制剂.     

乳酸杆菌代谢产物对一些常见菌黏附阴道上皮细胞的抑制作用的初步探讨

目的 探讨乳酸杆菌代谢产物对大肠埃希菌、铜绿假单胞菌、金黄色葡萄球菌和白色念珠菌黏附阴道上皮细胞的抑制作用.方法 刮取健康妇女阴道上皮细胞进行体外培养,观察在乳酸杆菌代谢产物的干预下大肠埃希菌、铜绿假单胞菌、金黄色葡萄球菌和白色念珠菌黏附阴道上皮细胞的情况.结果 和结论 乳酸杆菌代谢产物能够明显抑制大肠埃希菌、铜绿假单胞菌、金黄色葡萄球菌和白色念珠菌对阴道上皮细胞的黏附.     

乳酸杆菌对大鼠细菌性阴道感染动物模型疗效观察

目的探讨乳酸杆菌对大鼠阴道菌群抑制及阴道菌群的调理作用。方法通过大鼠阴道混合感染葡萄球菌和大肠埃希菌的动物模型,以阴道分泌物的菌群分析和阴道黏膜炎症的治愈程度为指标,观察乳酸杆菌治疗组的治疗效果,用乳酸杆菌菌液冲洗治疗来去除大肠埃希菌和葡萄球菌在大鼠阴道的定植。结果通过各组大鼠阴道冲洗液检出的平均对数值比较,乳酸杆菌治疗组的大鼠阴道内大肠埃希菌和葡萄球菌的数量明显较对照组的数量减少（P〈0．05）,而且治疗组黏膜炎症的治愈程度显著好于对照组。结论乳酸杆菌能有效地去除大肠埃希菌和葡萄球菌的大鼠阴道定植和改善大鼠外阴的炎症症状。     

片球菌素PediocinPA-1抑菌活性检测

目的检测通过基因工程获得的片球菌素Pediocin PA-1抑菌活性。方法采用琼脂扩散法检测片球菌素Pediocin PA-1对单核细胞增生李斯特杆菌、金黄色葡萄球菌、铜绿假单胞菌、沙门菌和大肠埃希菌O157的抑菌活性。结果片球菌素Pediocin PA-1对单核细胞增生李斯特杆菌、金黄色葡萄球菌、沙门菌、铜绿假单胞菌和大肠埃希菌O157等均有抑制作用。其中对单核细胞增生李斯特杆菌、沙门菌、大肠埃希菌和金黄色葡萄球菌的抑制作用效果明显,对铜绿假单胞菌有微弱的抑制作用。结论通过基因工程获得的片球菌素Pediocin PA-1具有抑菌活性。     

2株乳酸菌对人工诱发鸡大肠埃希菌病的预防性实验

目的观察植物乳杆菌和粪链球菌2株乳酸菌预防鸡大肠埃希菌病的效果。方法把2株乳酸菌添加到肉鸡的饲料中饲喂,至14日龄时用鸡源致病性大肠埃希菌人工诱发鸡大肠埃希菌病,10 d后统计发病率、死亡率和有效预防率。结果成功诱发出鸡大肠埃希菌病,植物乳杆菌和粪链球菌预防鸡大肠埃希菌病的有效率非常高。结论植物乳杆菌和粪链球菌可以用于预防鸡大肠埃希菌病。     

全子宫切除术对阴道菌群的影响

目的研究全子宫切除术后阴道菌群及pH的变化,寻求适当的治疗措施,以提高术后患者的生活质量。方法分别观察术后未发生阴道炎、发生老年性阴道炎及正常近龄妇女3组共90例,进行菌群分析、pH测定,并使用乳杆菌DM9811代谢物脂肪酸组分对发生老年性阴道炎者进行治疗。结果术后妇女检出各种菌的数量少,乳杆菌的数量更少,阴道内没有占绝对优势的菌种;发生阴道炎组乳杆菌数量下降更明显,葡萄球菌及大肠埃希菌数量相对增多。术后阴道pH在5.9~7.3,发生阴道炎组较未发生阴道炎组高;以乳杆菌DM9811代谢物脂肪酸组分阴道用药治疗老年性阴道炎,治愈率为82.14%。结论全子宫切除术后阴道乳杆菌数量下降,pH升高,易发生老年性阴道炎。治疗以乳杆菌DM9811代谢物脂肪酸组分阴道用药效果佳。     

Production of cytokines by vaginal epitheliocytes in the process of interaction with dominant and associative microsymbionts

AIM: Study the expression of cytokines by vaginal epitheliocytes in the process of interaction with dominant and associative microsymbionts. MATERIALS AND METHODS: IL-8, IL-6, IL-1beta and TNFalpha expression in response to interaction with heat inactivated Lactobacillus spp., Staphylococcus aureus, Escherichia coli, Corynebacterium spp. or their secretory products in comparison with basal expression of cytokines by vaginal epitheliocytes was studied. Results. Lactobacilli secretory products were shown not to influence the expression of IL-8 and IL-1beta and moderately stimulated IL-6 and TNFalpha expression. Contact of epitheliocytes with heat inactivated lactobacilli increased secretion of IL-8, IL-6 and IL-1beta and reduced TNFalpha production. Secretory products of S. aureus and E. coli caused stimulation of IL-6, IL-1beta production and practically did not change the expression of IL-8 and TNFalpha. Contact of epitheliocytes with heat inactivated S. aureus sup pressed TNFalpha production and had no influence on IL-8, IL-6 and IL-1beta expression, contact with E. colistimulated TNFalpha and IL-1beta expression and suppressed IL-6 expression. Changes in cytokine expression during interaction of epitheliocytes with corynebacteria were largely similar to the results of interaction with lactobacilli except IL-6 production that was markedly stimulated by corynebacteria secretory products. Conclusion. In epithelial-bacterial interactions dominant and associative microorganisms have a differential effect on functional status of mucosal epitheliocytes manifesting in production of cytokines that could be the basis of mucosal immunity regulation.     

人胃粘膜乳酸杆菌与幽门螺杆菌感染的相关性

目的:探讨胃内正常菌群乳酸杆菌对幽门螺杆菌(Helicobacter pylori,HP)的影响。方法:51例胃粘膜活检标本均取自于行胃镜检查的胃炎患者,分离培养乳酸杆菌,通过扩增其16S rRNA基因并测序来鉴定乳酸杆菌的种类;胃炎程度及活动度的分类依据悉尼分类系统,运用改良Gimesa染色鉴定HP感染。结果:胃粘膜中共分离出9种乳酸杆菌,分离阳性率为49.0%;乳酸杆菌阳性病人与阴性病人的HP感染率、胃炎程度的差异及胃炎活动度的差异均无统计学意义(P>0.05);HP阳性病人的胃炎程度较HP阴性病人更严重(P<0.05);有益生菌作用的乳酸杆菌与非益生菌类乳酸杆菌的HP感染率差异无统计学意义(P>0.05)。结论:胃内乳酸杆菌的存在对HP感染无影响。     

Influence of probiotic vaginal lactobacilli on in vitro adhesion of urogenital pathogens to vaginal epithelial cells

AIMS: Lactobacilli, the predominant micro-organisms of the vaginal microbiota, play a major role in the maintenance of a healthy urogenital tract by preventing the colonization of pathogenic bacteria. The aim of the present study was to assess the ability of four vaginal Lactobacillus strains, previously selected for their probiotic features, to block in vitro the adherence of three human urogenital pathogens to vaginal epithelial cells (VEC). METHODS AND RESULTS: Three types of assays were performed in order to determine the inhibitory effect of lactobacilli on adhesion of urogenital pathogens to VEC: blockage by exclusion (lactobacilli and VEC followed by pathogens), competition (lactobacilli, VEC and pathogens together) and displacement (pathogens and VEC followed by the addition of lactobacilli). Bacterial adhesion to VEC was quantified by microscopy (x1000) after Gram's stain. All the strains were able to inhibit by exclusion and competition the adhesion of Staphylococcus aureus to VEC but none was able to decrease the attachment of Escherichia coli by neither of the mechanisms assayed. Only Lactobacillus acidophillus CRL 1259 and Lactobacillus paracasei CRL 1289 inhibited the attachment of Group B streptococci (GBS) to VEC by exclusion and competition respectively. CONCLUSIONS: Lactobacillus of vaginal origin were able to inhibit the attachment of genitouropathogenic Staph. aureus and GBS to the vaginal epithelium. SIGNIFICANCE AND IMPACT OF THE STUDY: The results support the probiotic potential of these Lactobacillus strains as anti-infective agents in the vagina and encourage further studies about their capacity to prevent and manage urogenital tract infections in females.     

The effect of Lactobacillus spp. on the attachment of enterotoxigenic Escherichia coli to isolated porcine enterocytes

A total of 43 strains of lactobacilli were isolated from the gastrointestinal tract of piglets at the time of weaning. Isolates, grown on solid media, were allocated to strongly adherent or non/weakly adherent groups on the basis of numbers attaching to isolated porcine enterocytes. Strains of Lactobacillus fermentum were disproportionally represented amongst the strongly-adherent strains and Lact.acidophilus and Lact. salivarius amongst the non/weakly-adherent group. Lactobacilli showed significantly better attachment ability when grown on agar than when grown in broth culture. Strongly adherent strains were not found to effect the attachment of enterotoxigenic Escherichia coli to porcine enterocytes, tested under the conditions of exclusion (lactobacilli added to the enterocytes before E. coli ), competition (lactobacilli and E. coli added simultaneously) and displacement ( E. coli added before lactobacilli). Tests were made with [¹⁴C]-labelled E. coli. Suspensions of bacteria and enterocytes were passed through a filter selected to retain enterocytes but pass free bacterial cells. Counts (dpm) obtained from filters after solubilization were taken as a measure of E. coli attachment. Some strains of lactobacilli coaggregated with enterotoxigenic E. coli with K88 fimbriae, but not with a K88-negative mutant strain. These were excluded from the competitive exclusion experiments. In the apparent absence of a direct effect on the association of E. coli with host tissue, removal of potential gut pathogens by aggregation could contribute to the probiotic properties ascribed to lactic acid bacteria.     

Effect of lactobacilli onE. coli adhesion to caco-2 cellsin Vitro

Inhibitory effect of various lactobacilli against pathogenic strains of E. coli in model system Caco2 cells was determined by enumerating the number of adhering E. coli after pre-incubation (exclusion), post-incubation (displacement) or co-incubation (competition) with lactobacilli. Porcine E. coli strain F107 (F18ab, Stx2v) in the competition assay with porcine lactobacillus strain P10 gave bacterial counts 7.25 (log CFU per well); in the exclusion test it was only 7.05 while in displacement test it reached 7.29. The lowest E. coli counts adhering to Caco-2 cells were in exclusion assay (pre-incubation, Lactobacillus inoculated as the first). Pre-treatment of E. coli with our lactobacilli strains reduced the cultivable E. coli numbers.