嗜酸乳杆菌与嗜热链球菌共发酵互生机理的研究

目的:对嗜酸乳杆菌和嗜热链球菌共发酵时两者的互生作用机理进行研究。方法:以质量浓度100gL脱脂乳为培养基,将嗜酸乳杆菌主要代谢产物—氨基酸,如赖氨酸、精氨酸、缬氨酸分别以0.0083mgml、0.0036mgml、0.0053mgml的量加入含嗜热链球菌脱脂乳培养基中,40℃培养,测定凝乳时间;将嗜热链球菌的代谢产物-乳酸、甲酸分别为0.1332mgml和0.075mgml的量加入含嗜酸乳杆菌脱脂乳培养基中,37℃培养,测定其发酵乳的凝乳时间。结果:嗜热链球菌发酵乳凝乳时间由12h缩短到4h,pH为4.52～4.62,吉尔涅尔度为54.23～64.74°T;嗜酸乳杆菌发酵乳的凝乳时间由16h缩短为5h,pH为4.61～4.65;且嗜酸乳杆菌在CO2环境中发酵时,发酵时间明显缩短。结论:嗜酸乳杆菌和嗜热链球菌共发酵时具有互生关系。     

复方嗜酸乳杆菌对轻中度溃疡性结肠炎诱导和维持缓解作用的临床研究

目的:观察复方嗜酸乳杆菌对轻中度溃疡性结肠炎诱导和维持缓解作用的疗效.方法:第一阶段将102例轻、中度溃疡性结肠炎患者随机分成A组(34例)、B组(34例)和C组(34例),分别服用美沙拉嗪片、复方嗜酸乳杆菌、美沙拉嗪+复方嗜酸乳杆菌治疗12周后观察3组临床疗效,缓解率及缓解时间.第二阶段将各组缓解者随访1年,观察3组维持时间及复发率.结果:3组临床疗效评价,临床缓解时间、缓解率比较无显著性差异见(P＞0.05),3组患者维持临床缓解时间、复发率比较差异无显著性意义(P＞0.05).所以由中国株嗜酸乳杆菌,日本株嗜酸乳杆菌、粪链球菌和枯草杆菌等四种菌粉组成的复方片剂是值得推荐的维持治疗缓解期UC的有效药物;两者联合用药并未增效,联合使用复方嗜酸乳杆菌片和美拉沙嗪作为溃疡性结肠炎的联合维持治疗是没有必要的.结论:复方嗜酸乳杆菌对轻中度溃疡性结肠炎诱导和维持缓解作用的疗效与美拉沙嗪比较无差异,没有必要联合使用复方嗜酸乳杆菌片和美拉沙嗪作为溃疡性结肠炎的联合维持治疗.     

复方嗜酸乳杆菌对轻中度溃疡性结肠炎诱导和维持缓解作用的临床研究

目的:观察复方嗜酸乳杆菌对轻中度溃疡性结肠炎诱导和维持缓解作用的疗效。方法:第一阶段将102例轻、中度溃疡性结肠炎患者随机分成A组(34例)、B组(34例)和C组(34例),分别服用美沙拉嗪片、复方嗜酸乳杆菌、美沙拉嗪+复方嗜酸乳杆菌治疗12周后观察3组临床疗效,缓解率及缓解时间。第二阶段将各组缓解者随访1年,观察3组维持时间及复发率。结果:3组临床疗效评价,临床缓解时间、缓解率比较无显著性差异见(P>0.05),3组患者维持临床缓解时间、复发率比较差异无显著性意义(P>0.05)。所以由中国株嗜酸乳杆菌,日本株嗜酸乳杆菌、粪链球菌和枯草杆菌等四种菌粉组成的复方片剂是值得推荐的维持治疗缓解期UC的有效药物;两者联合用药并未增效,联合使用复方嗜酸乳杆菌片和美拉沙嗪作为溃疡性结肠炎的联合维持治疗是没有必要的。结论:复方嗜酸乳杆菌对轻中度溃疡性结肠炎诱导和维持缓解作用的疗效与美拉沙嗪比较无差异,没有必要联合使用复方嗜酸乳杆菌片和美拉沙嗪作为溃疡性结肠炎的联合维持治疗。     

嗜酸乳杆菌黏附鸡胚肠黏膜上皮细胞能力的研究

目的建立肠黏膜上皮细胞模型和测定嗜酸乳杆菌的黏附能力。方法将嗜酸乳杆菌用胃蛋白酶和HCl-H2O低pH以及反复冻融、灭活等方法处理后测定其黏附能力。结果成功地建立了鸡胚肠黏膜上皮细胞模型;经胃蛋白酶和HCl-H2O低pH以及灭活处理后,嗜酸乳杆菌的黏附能力和对照组差异有显著性。反复冻融后的嗜酸乳杆菌黏附能力与对照组差异无显著性。结论胃蛋白酶和HCl-H2O pH2.0会使嗜酸乳杆菌黏附肠黏膜上皮细胞能力下降,热灭活可使嗜酸乳杆菌的黏附能力提高,反复冻融对嗜酸乳杆菌的黏附能力无明显影响。     

家兔腹泻生态疗法的实验观察

根据动物微生态学理论,将3株正常菌：嗜酸乳杆菌、两歧双歧杆菌、粪链球菌研制成复方生态制剂,对家免细菌性腹泻进行治疗试验。共治疗75例,治愈率为96．00％,比痢特灵对照组治愈率提高16．00％。该制剂价格低廉,无毒、副作用,使用安全可靠,是一种防治家免细菌性腹泻的较理想制剂。     

双歧杆菌四联活菌片中嗜酸乳杆菌的分离及生长动力学研究

目的以双歧杆菌四联活菌片为实验材料,利用酸化的MRS培养基筛选分离得到嗜酸乳杆菌,对其进行进一步的生长动力学研究,确定嗜酸乳杆菌的生长数学模型。方法通过浓度梯度稀释法利用改良MRS培养基对双歧杆菌四联活菌片中的嗜酸乳杆菌进行分离,利用分光光度仪和平板菌落计数两种不同的方法测定嗜酸乳杆菌在发酵过程中不同发酵时间的细胞浓度的动态变化,经软件处理后拟合出嗜酸乳杆菌细胞生长的Logistic数学模型。结果Logistic方程能很好地拟合嗜酸乳杆菌细胞生长的动态变化,并得到嗜酸乳杆菌在本实验条件下的数学模型,为进一步研究、利用嗜酸乳杆菌生长能力、产酸能力和产香能力等具有重要的理论指导意义。     

嗜酸乳杆菌YIT2004株的抗生素敏感性 及对万古霉素耐药机制

摘要：目的 研究嗜酸乳杆菌YIT2004株的抗生素敏感性及其耐药机制。方法 最低抑菌浓度法检测嗜酸乳杆菌YIT2004株对抗生素的敏感性;提取YIT2004株基因组,用特异性引物对耐药基因进行PCR扩增。结果 嗜酸乳杆菌YIT2004株对青霉素、氨苄西林、亚胺培南、庆大霉素、红霉素和克林霉素敏感,对万古霉素耐药;YIT2004株基因组中不存在vanA、vanB耐药基因,但存在高度相似的aad、ddl万古霉素耐药基因。该耐药为固有耐药,不具备传递性。结论 嗜酸乳杆菌YIT2004株对青霉素、氨苄西林、亚胺培南、庆大霉素、红霉素和克林霉素敏感,对万古霉素固有耐药且其耐药性不可传递。嗜酸乳杆菌YIT2004株的使用具有安全性。     

乳酸菌固体制剂计数方法研究

从培养基、稀释剂的角度对乳酸菌固体制剂样品进行计数方法比较。昂立1号○R优菌多颗粒样品中嗜酸乳杆菌在改良MC培养基上培养(72±3)h后菌落形态非常小,很难进行计数,在改良MC培养基中添加0.1%吐温80将会刺激嗜酸乳杆菌的生长,利于计数;采用MRS液体溶解复水10~15 m in、0.1%蛋白胨盐水稀释的计数结果明显高于直接采用生理盐水复水、稀释方法,两者差异有非常显著性。     

嗜酸乳杆菌LA85对小鼠免疫机能的研究

目的：研究嗜酸乳杆菌LA85对小鼠免疫机能的影响。方法：将160只雄性小鼠随机分为4组,即对照组、低、中、高三个剂量组。采用灌胃的方式给予实验小鼠不同剂量的嗜酸乳杆菌LA85。通过对小鼠平均体重、脾脏和胸腺免疫指数、细胞免疫、体液免疫、单核-吞噬性细胞和自然杀伤细胞（NK细胞）活性的评价,探讨嗜酸乳杆菌LA85对小鼠免疫机能的影响机制。结果：与对照组相比,嗜酸乳杆菌LA85低、中、高剂量组能够显著提升小鼠的胸腺指数,嗜酸乳杆菌LA85高剂量组能够显著提升小鼠的脾脏指数。同时能够显著提高小鼠的淋巴细胞增殖分化能力、半数溶血值、巨噬细胞的吞噬能力、NK细胞的细胞活性;嗜酸乳杆菌对小鼠的体重没有影响。结论：给予小鼠口服嗜酸乳杆菌LA85可以通过改善细胞免疫和体液免疫功能,从而提高免疫力。     

嗜酸乳杆菌产细菌素生物学特性的研究

嗜酸乳杆菌在MRS培养基中培养得到的细菌素经30、60、90、121℃处理20min后,活性几乎不变,对以金黄色葡萄球菌为代表的革兰阳性菌、大肠埃希菌为代表的革兰阴性菌有明显的抑制作用,Tricine—SDS—PAGE和不同pH值抑菌实验测定结果证明,嗜酸乳杆菌细菌素是一组低分子量,并在pH2—4时有抑菌作用的活性物质。     

嗜酸乳杆菌LB10.16及枯草芽孢杆菌BS7.29安全性评价

目的:评价枯草芽孢杆菌BS7.29和嗜酸乳杆菌LB10.16的安全性,为其应用研究提供技术支持。方法:本文通过对小鼠灌服、急性攻毒、腹腔注射、小鼠最大耐受剂量、急性攻毒等试验方法进行枯草芽孢杆菌BS7.29和嗜酸乳杆菌LB10.16的安全性研究。结果:1×108cfu/kg.bw的LB10.16和5×108cfu/kg.bw的BS7.29能够明显提高肠道推动力,改善小鼠肠道蠕动性。结论:两个菌种急性攻毒试验结果表明属于无毒物质,安全性比较高。     

嗜酸乳杆菌LB10.16及枯草芽孢杆菌BS7.29安全性评价

目的：评价枯草芽孢杆菌BS7.29和嗜酸乳杆菌LB10.16的安全性,为其应用研究提供技术支持。方法：本文通过对小鼠灌服、急性攻毒、腹腔注射、小鼠最大耐受剂量、急性攻毒等试验方法进行枯草芽孢杆菌BS7.29和嗜酸乳杆菌LB10.16的安全性研究。结果：1×108cfu/kg.bw的LB10.16和5×108cfu/kg.bw的BS7.29能够明显提高肠道推动力,改善小鼠肠道蠕动性。结论：两个菌种急性攻毒试验结果表明属于无毒物质,安全性比较高。     

Evidence for the involvement of thiocyanate in the inhibition of Candida albicans by Lactobacillus acidophilus

Lactobacillus acidophilus has been found to inhibit Candida albicans when grown on MRS agar plates. Attempts to isolate an active factor responsible for this inhibition from liquid culture and agar plates were not successful. The addition of sodium thiocyanate to the agar was found to increase the inhibition offered by the lactobacillus. The results indicate that hydrogen peroxide produced by the lactobacillus is being used to convert the thiocyanate to hypothiocyanate which is more toxic. The involvement of a lactobacillus peroxidase in this conversion is postulated.     

Lactobacillus acidophilus expressing recombinant K99 adhesive fimbriae has an inhibitory effect on adhesion of enterotoxigenic Escherichia coli

The most common enteric colibacillosis in neonatal and newborns is caused by enterotoxigenic Escherichia coli(ETEC). Colonization of ETEC in the small intestine is associated with adhesions using fimbriae, which is known as a specific adhesion factor and provides highly specific means for anchoring and prerequisite for an infectious agent. In the present study we have engineered Lactobacillus acidophilus to produce recombinant K99 fimbriae, which is used for the colonization to the intestine of pigs. The expression of K99 fimbrial protein was confirmed using SDS-PAGE, immunoblot and agglutination analyses. To evaluate a function of the K99 fimbrial protein, inhibition and competition tests were performed on pre-screened intestinal brush border from pigs. The tests showed that recombinant L. acidophilus, not control L. acidophilus, had a significant inhibitory effect to and competition against K99+ E. coli in a dose dependent manner. In conclusion, we demonstrated that recombinant K99 fimbriae producing L. acidophilus was able to prevent E. coli binding to intestinal brush border.     

The determination of the antagonistic activity of Solco lactobacteria (Lactobacillus acidophilus Lat 11/83) using gnotobiotic technology

Lactobacillus acidophilus strain Lat 11/83 has been used for the study of its antagonistic activity with respect to pathogenic microorganisms in experiments on conventional germ-free animals. The results of these experiments indicate that the above strain may be recommended as a highly active antagonist for the treatment and prophylaxis of intestinal dysbacteriosis of different etiology.     

乳酸菌抑制葡聚糖硫酸钠诱导的结肠黏膜氧化应激损伤的研究

目的研究乳酸菌抗氧化活性与缓解葡聚糖硫酸钠(DSS)诱导结肠炎的关系。方法评价5株乳酸菌体外抑制大鼠肝细胞脂质过氧化活性,用DSS诱导小鼠结肠炎模型研究抗氧化性不同的菌株体内抗氧化活性;小鼠分成7组,分别是Control组、DSS组、L.plantarum Fn008+DSS组、L.casei Fn012+DSS组、L.acidophilus Fn022+DSS组、L.sakei Fn034+DSS组和L.acidophilus Fn037+DSS组。结果体外实验表明,Fn022和Fn034的丙二醛(MDA)抑制率高于LGG,但差异无统计学意义,Fn008和Fn037的MDA抑制率显著低于LGG,Fn012的MDA抑制率最低。体内实验表明,DSS可导致小鼠氧化应激,升高结肠内容物的自由基水平,降低结肠抗氧化能力;Fn008和Fn037预处理能显著降低DSS诱导结肠内容物的自由基水平,Fn008能显著降低结肠组织的髓过氧化物酶(MPO)水平,提高结肠抗氧化能力,Fn022和Fn034的干预能力居中,Fn012的干预能力最弱。相关性分析表明乳酸菌体外MDA清除率与体内MPO有负相关趋势(r=-0.862,P=0.06)。结论乳酸菌抗氧化活性与其在肠道抑制炎性细胞激活的作用有关,本实验中Fn008和Fn037具有显著的体内抗氧化作用。     

The effect of a preparation made from Solco lactobacteria on the survival rate and intestinal microflora of irradiated animals

Lactobacillus acidophilus strain Lat 11/83 Solco has been used for the normalization of intestinal microflora in experimental post-irradiation intestinal dysbacteriosis in mice. The results of these experiments indicate that live Solco lactobacteria, introduced intragastrically, facilitate the enhancement of the survival rate of irradiated animals and the normalization of microflora in their gastrointestinal tract. The strain can be used as bacterial preparation for the regulation of intestinal microbiocenosis.     

The effect of "Solco" lactobacteria (Lactobacillus acidophilus) on the survival and microflora of mice with a Salmonella infection

The Solco lactobacterial strain L. acidophilus Lat 11/83 has been used for the normalization of intestinal microflora in experimental post-infectious intestinal dysbacteriosis in mice. The results of experiments indicate that the intragastric administration of live Solco lactobacteria contributes to an increase in the survival rate of infected animals and the normalization of their gastrointestinal microflora. This strain may be used as a bacterial preparation for the regulation of intestinal microbiocenosis.     

Derivation of DNA probes for enumeration of a specific strain of Lactobacillus acidophilus in piglet digestive tract samples.

Four DNA probes were derived that hybridized specifically to DNA from Lactobacillus acidophilus O. The probes were constructed by randomly cloning lactobacillus DNA in plasmid vector pBR322. Two of the probes (pSR1 and pSR2) were composed of vector and plasmid DNA inserts (3.6 and 1.6 kb, respectively); the others (pSR3 and pSR4) were composed of vector and chromosomally derived inserts (6.9 and 1.4 kb, respectively). The probes were used to enumerate, by colony hybridization, strain O in digestive tract samples collected from piglets inoculated 24 hours previously with a culture of the strain. The probes did not hybridize to DNA from lactobacilli inhabiting the digestive tract of uninoculated piglets. Strain O made up about 10% of the total lactobacillus population of the pars esophagea and about 20% of the population in other digestive tract samples.