费氏丙酸杆菌费氏亚种生长特性及对致病菌和益生菌作用探讨

目的对1株费氏丙酸杆菌费氏亚种生长特性及发酵代谢物的抑菌和促生长作用进行研究。方法将费氏丙酸杆菌费氏亚种无细胞发酵上清液分别与5株致病菌和3株益生菌添加至24孔板中进行培养,通过测定A_(600 nm)值的变化来研究发酵代谢物的抑菌和促生长作用。结果发酵代谢物中的抑菌成分含量随发酵时间的延长而增加,到第6天后增长趋于平缓。当发酵上清液添加量达到400μL时,溶血性链球菌被完全抑制,达到600μL时,金黄色葡萄球菌、沙门菌、单核细胞增生李斯特菌和大肠埃希菌实验组A_(600 nm)值较对照组分别下降76.1%、68.5%、60.2%和43.9%,而长双歧杆菌和婴儿双歧杆菌实验组A_(600 nm)值较对照组分别上升17.1%和41.5%。结论费氏丙酸杆菌费氏亚种发酵代谢物对5株致病菌均表现出不同程度的抑菌效果,对2株双歧杆菌的生长有较明显的促进作用。     

酪酸梭菌与双歧杆菌对肠道致病菌的体外生物拮抗作用

目的：探讨酪酸梭菌和婴儿型双歧杆菌对某些肠道致病菌的拮抗作用,方法：将酪酸梭菌和婴和型双歧杆菌单株菌及两株菌混合后分别与几株肠道致病菌混合接种于GAM液体培养其中进行厌氧培养,通过平板菌落计数法计算肠道致病菌的菌数,结果：经混合培养后肠道致病菌的菌数明显下降,结论：酪酸梭菌LCL166株和婴儿型双歧杆菌LCL172株在体外能明显抑制几株肠道致病菌的生长繁殖。     

随机比较中国大连地区和日本大阪地区健康成年人 及老年人肠内细菌结构的研究

实验应用复合式肠内细菌群检测法,随机比较了中国大连地区和日本大阪地区６ 名健康成年人（ 简称：ＣＡ,ＪＡ） 及４ 名老年人（ 简称：ＣＯＭ,ＪＯＭ） 的肠内细菌群结构。ＣＡ 肠内细菌总数为１０．２１ ±５．５６（ 每克粪便中的对数值） ,较ＪＡ 的肠内细菌总数１０．８７ ±２．４５ 低,但统计学检查差异不显著;ＣＡ 的双歧杆菌数为９．０１ ±０．１３ ,ＪＡ 的双歧杆菌数为９．９８ ±１．２４ ,统计学检查表明差异显著,而且,ＪＡ 的双歧杆菌占有率为１２．８８ ％ ±３．４５ ％ 明显高于ＣＡ 的６．２９ ％ ±４．５５ ％ ,统计学检查差异显著。ＣＡ 的乳酸菌数为５．４５ ±１．１２ ,ＪＡ 的乳酸菌数为５．５２ ±１．４４ ;在腐败梭菌的菌数上,ＣＡ 为６．４５ ±２．２１ 比ＪＡ 的５．２２ ±２．４５ 略高,但统计学检查差异不明显。ＣＯＭ 肠内细菌总数为１０．４５ ±４．６６ ,较ＪＯＭ 的肠内细菌总数１０．９７ ±５．２５ 低,但统计学检测差异不显著;ＣＯＭ的双歧杆菌数为８．７４ ±２．１３ ,ＪＯＭ 的双歧杆菌数为９．８５ ±０．３４ ,统计学检查表明差异显著,而且,ＪＯＭ 的双歧杆菌占有率为７．５９ ±５．１５ ％ 明显高于ＣＯＭ 的１．１３ ％ ±０     

异麦芽寡糖促进肠道分离菌生长的体外实验

目的 观察添加１％异麦芽寡糖对从正常人阑尾标本中分离双歧杆菌、乳杆菌和类杆菌的体外促生长作用。方法 分别在培养前和培养１２、２４、３６ｈ和４８ｈ,用７２１分光光度计和ｐＨ计测定培养液ＯＤ值和ｐＨ值变化。结果 接种双歧杆菌的培养液ＯＤ值从０．１２８增加到１．７,ｐＨ值由７．１降到４．２,增殖效果与添加１％葡萄糖相当。接种乳杆菌的培养液ＯＤ值从０．１７增加到０．８５,ｐＨ值由７．０降到４．８,增殖效果低于添加１％葡萄糖。接种类杆菌的培养液ＯＤ值从０．１０５增加到１．６５,ｐＨ值由７．１降到５．１,增殖效果高于添加１％葡萄糖。结论 添加１％民麦芽寡糖具有促进肠道分离菌双歧杆菌、乳杆菌和类杆菌生长的作用。     

螺旋藻在体外对双歧杆菌及乳杆菌增殖的促进作用

通过用改良的ＴＰＹ和ＧＡＭ培养基作为螺旋藻的基质,在试管内证实了螺旋藻对双歧杆菌、乳杆菌生长的促进作用。结果显示,钝顶螺旋藻在实验浓度下,对双歧杆菌和乳杆菌的增殖作用与对照组相比有非常显著的差异,这一结果也说明了钝顶螺旋藻在调整肠道菌群,改善肠道内环境方面的作用。     

灭活双歧杆菌调整小鼠抗生素相关性菌群失调

目的：观察灭活的双歧杆菌及其耗尽培养上清液（SCS）对小鼠肠道生理菌群的影响。方法：应用腹腔注射青霉素造成肠菌群失调动物模型,分别以灭活的双歧杆菌菌液,耗尽培养上清液以及活菌菌液对菌群失调小鼠进行灌胃治疗。结果：活菌组、死菌组及SCS组同自然恢复组的肠道生理菌群相比差异均有显著性,死菌组与SCS组相比,差异也有显著性。结论：灭活的双歧杆菌及其SCS对小鼠肠道菌群失调的恢复具有调整作用,尤其对双歧杆菌和乳酸杆菌有更明显的扶持作用。     

昂立植物乳杆菌及其抑菌物质的特性研究

该文研究了昂立植物乳杆菌(LP-Onlly)菌体及其代谢产物的抑菌性能,并对其代谢产物中的抑菌物质进行了部分理化特性的考察,发现LP-Onlly菌体对部分肠道有害菌有抑制作用,代谢产物中的抑菌物质对常见的肠道致病菌和食品腐败微生物具有广谱抑菌作用,对嗜酸乳杆菌及双歧杆菌等益生菌无抑制作用.该物质具有热稳定性,但抑菌活性受pH值的影响较大.     

双歧杆菌对人大肠癌细胞粘附作用的初步探讨

双歧杆菌是人肠道的正常菌群之一。为探讨双歧杆菌对肠道粘膜上皮细胞的粘附机制,本文观察了双岐杆菌ＤＭ９２２７对培养的大肠癌细胞系ＣＣＬ－１８７细胞的粘附作用以及影响粘附的因素,初步建立了体外双歧杆的的粘附模型。结果还发现：在粘附近程中,孵育培养基的环境ｐＨ值在６．０－７．０左右时,钙离子浓度在２．０ｍｍｏｌ／Ｌ时,双歧杆菌ＤＭ９２２７对大肠癌ＣＣＬ一１８７细胞的粘附效果达到最佳。     

体外乳酸菌素促双歧杆菌生长实验初步观察

目的 在体外观察乳酸菌素是否有促双歧杆菌生长的作用。方法 将浓度为１００ｍｇ／ｍｌ、１０ｍｇ／ｍｌ、１ｍｇ／ｍｌ的乳酸菌素液按三角形对等距离分别滴种在接种了双歧杆菌（１０＾６ＣＦ／ｍｌ）的ＢＨＩ培养基上,然后置３５℃厌氧箱培养２ｄ后观察细菌生长情况。结果 不同浓度的乳酸菌素液促进双歧杆菌生长能力不同。浓度为１００ｍｇ／ｍｌ、１０ｍｇ／ｍｌ、１ｍｇ／ｍｌ的乳酸菌素液促双歧杆菌生长的阳性率（％）分别为：９６．７、５６．７、１．０。经统计学秩和检验方法将３种浓度乳酸菌素液促双歧杆菌生长作用进行两两比较得知：１００ｍｇ／ｍｌ与１０ｍｇ／ｍｌ,１００ｍｇ／ｍｌ与１ｍｇ／ｍｌ之间Ｐ〈０．０１,１０ｍｇ／ｍｌ与１ｍｇ／ｍｌ比较Ｐ〈０．０５。结论 乳酸菌素有促进益生菌－－双歧杆生长的作用,而且与其浓度高低有极密切关系,高浓度的     

双歧杆菌活菌制剂对慢性乙肝病人细胞免疫功能影响的观察

在应用双歧杆菌活菌制剂治疗慢乙肝期间,重点观察了Ｔ细胞亚群（ＣＤ３,ＣＤ４,ＣＤ８）、ＮＫ细胞（ＣＤ（１６））、白细胞介素Ⅱ（ＩＬ－２）分泌细胞、肿瘤坏死因子（ＴＮＦ）等细胞免疫指标治疗前后的动态变化,同时观察了病人血内毒素水平的动态变化和乙肝病毒标志物（ＨＢＶＭ）的改变。结果表明：（１）与对照组比较,双歧杆菌活菌制剂可使慢乙肝病人ＣＤ３＋,ＣＤ４＋数目明显增多,而对ＣＤ８＋细胞数目无明显影响;（２）双歧杆菌活菌制剂可使ＣＡＨ组的ＣＤ１６＋细胞显著增多（ｐ＜０．０５）;使ＣＡＨ组和ＣＰＨ组的ＩＬ－２分泌细胞均有非常显著和显著增加（分别ｐ＜０．０１和ｐ＜０．０５）;（３）ＣＡＨ组病人血中内毒素和ＴＮＦ水平在双歧杆菌活菌制剂治疗后,匀出现非常显著降低（ｐ＜０．０１）;ＣＰＨ组ＴＮＦ水平较对照组无显著变化,但内毒素水平较对照组显著降低（ｐ＜０．０５）;（４）满疗程后（６０天）ＣＡＨ组有６例,ＣＰＨ组有５例ＨＢｅＡｇ阴转（分别为２６．０６％和２５．０％）,而对照组仅２例阴转（１３．３３％）,两治疗组与对照组比较有显著性差异（ｐ＜０．０５）。     

Cholesterol assimilation by lactic acid bacteria and bifidobacteria isolated from the human gut

The objective of this study was to evaluate the effect of human gut-derived lactic acid bacteria and bifidobacteria on cholesterol levels in vitro. Continuous cultures inoculated with fecal material from healthy human volunteers with media supplemented with cholesterol and bile acids were used to enrich for potential cholesterol assimilators among the indigenous bacterial populations. Seven potential probiotics were found: Lactobacillus fermentum strains F53 and KC5b, Bifidobacterium infantis ATCC 15697, Streptococcus bovis ATCC 43143, Enterococcus durans DSM 20633, Enterococcus gallinarum, and Enterococcus faecalis. A comparative evaluation regarding the in vitro cholesterol reduction abilities of these strains along with commercial probiotics was undertaken. The degree of acid and bile tolerance of strains was also evaluated. The human isolate L. fermentum KC5b was able to maintain viability for 2 h at pH 2 and to grow in a medium with 4,000 mg of bile acids per liter. This strain was also able to remove a maximum of 14.8 mg of cholesterol per g (dry weight) of cells from the culture medium and therefore was regarded as a candidate probiotic.     

培菲康双歧三联活菌制剂中三株菌对抗菌药物敏感性检测

目的：检测培菲康活菌制剂中双歧杆菌,嗜酸乳杆菌与粪链球菌三株菌对30种抗菌药物的敏感性。方法：用纸片琼脂扩散法（disc agar diffusion,即Kirby-Bauer法）和微量稀释法（microdilution bro9th method)进行检测。结果：这三株菌对大多数抗菌药物敏感,对少数抗菌药物耐受。与文献报导相似,这三株菌与相应的国际参考菌株及临床菌株比较,没有发现明显耐药变异。结论：为避免抗菌药物对双歧三联活菌制剂功效的影响。每天两药宜分开时间服用。服用双歧三联活菌不会引起耐药性的扩散。     

Cholesterol Assimilation by Lactic Acid Bacteria and Bifidobacteria Isolated from the Human Gut

The objective of this study was to evaluate the effect of human gut-derived lactic acid bacteria and bifidobacteria on cholesterol levels in vitro. Continuous cultures inoculated with fecal material from healthy human volunteers with media supplemented with cholesterol and bile acids were used to enrich for potential cholesterol assimilators among the indigenous bacterial populations. Seven potential probiotics were found: Lactobacillus fermentum strains F53 and KC5b, Bifidobacterium infantis ATCC 15697, Streptococcus bovis ATCC 43143, Enterococcus durans DSM 20633, Enterococcus gallinarum, and Enterococcus faecalis. A comparative evaluation regarding the in vitro cholesterol reduction abilities of these strains along with commercial probiotics was undertaken. The degree of acid and bile tolerance of strains was also evaluated. The human isolate L. fermentum KC5b was able to maintain viability for 2 h at pH 2 and to grow in a medium with 4,000 mg of bile acids per liter. This strain was also able to remove a maximum of 14.8 mg of cholesterol per g (dry weight) of cells from the culture medium and therefore was regarded as a candidate probiotic.     

Lipoquinones of some spore-forming rods, lactic-acid bacteria and actinomycetes.

The respiratory quinones of 73 strains of Gram-positive bacteria including spore-forming rods, lactic-acid bacteria and actinomyctes were examined. Menaquinones with seven isoprenoid units (MK-7) were the main quinone type found in representatives of the genus Bacillus and in Sporolactobacillus inulinus. However, a strain of B. thuringiensis produced MK-8 in addition to MK-7, and strains of B. lentus and B. pantothenticus appeared to produce MK-9 and MK-8, respectively, with no MK-7. In the clostridia and lactic-acid bacteria, no quinones were found, except in Pediococcus cerevisiae NCTC 8066 and Lactobacillus casei subsp. rhamnosus ATCC 7469, which contained menaquinones, and Streptococcus faecalis NCTC 775 and HIM 478-1, which contained demethylmenaquinones, in relatively low concentrations. Menaquinones were also found in the actinomycetes (except Actinomyces odontolyticus and Bifidobacterium bifidum which did not produce any quinones) and in Protaminobacter alboflavus ATCC 8458, the so-called Actinobacillus actinoides ATCC 15900 and Noguchia granulosis NCTC 10559.     

Antagonistic activity of probiotic lactobacilli and bifidobacteria against entero- and uropathogens

AIM: To develop in vitro assays for comparing the antagonistic properties and anti-oxidative activity of probiotic Lactobacillus and Bifidobacterium strains against various entero- and urinary pathogens. METHODS AND RESULTS: The antagonistic activity of five probiotic lactobacilli (Lactobacillus rhamnosus GG, Lactobacillus fermentum ME-3, Lactobacillus acidophilus La5, Lactobacillus plantarum 299v and Lactobacillus paracasei 8700:2) and two bifidobacteria (Bifidobacterium lactis Bb12, Bifidobacterium longum 46) against six target pathogens was estimated using different assays (solid and liquid media, anaerobic and microaerobic cultivation) and ranked (low, intermediate and high). Bacterial fermentation products were determined by gas chromatography, and the total anti-oxidative activity of probiotics was measured using linolenic acid test. Pyelonephritic Escherichia coli was highly suppressed by GG and both bifidobacteria strains. Lactobacilli strains 8700:2, 299v and ME-3 were the most effective against Salmonella enterica ssp. enterica in microaerobic while ME-3 and both bifidobacteria expressed high activity against Shigella sonnei in anaerobic milieu. Lact. paracasei, Lact. rhamnosus and Lact. plantarum strains showed intermediate antagonistic activity against Helicobacter pylori under microaerobic conditions on solid media. The highest anti-oxidative activity was characteristic for Lact. fermentum ME-3 (P < 0.05). No efficient antagonist against Clostridium difficile was found. The positive correlations between the pH, lactic acid production and anti-microbial activity for all tested probiotics were assessed. CONCLUSIONS: Developed experimental assays enable to compare the anti-microbial and -oxidative activity of Lactobacillus and/or Bifidobacterium probiotics, which have been claimed to possess the ability of suppressing the growth of various enteric and urinary pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Screening Lactobacillus and Bifidobacterium sp. strains according to their activity in various environmental conditions could precede the clinical efficacy studies for adjunct treatment with probiotics in cure of different gastrointestinal and urinary tract infections.     

Detection of Bifidobacterium species by enzymatic methods

The properties of Bifidobacterium strains of human origin were examined by three enzymic tests and the amounts of acetic and lactic acids produced were also quantified. It was evident that two strains of the American Type Culture Collection (ATCC) did not belong to the genus. Moreover, at least one strain of Bifidobacterium added to some milk preparations did not show distinctive characteristics of the genus. It was also shown that most of bifidobacteria studied produced alpha-galactosidase (EC 3.2.1.22) and alpha-glucosidase (EC 3.2.1.20). The presence of alpha-galactosidase could afford a rapid differentiation of bifidobacteria used in some dairy products since this enzyme was not detected in Lactobacillus strains studied.     

A fluorescence-based method for the detection of adhesive properties of lactic acid bacteria to Caco-2 cells

AIMS: The ability of probiotic micro-organisms to adhere to the intestinal surface is regarded as a substantial advantage in terms of bacteria persistence in the gastrointestinal tract. The aim of the present study was the development of a method based on fluorescent staining of bacteria and subsequent spectrofluorimetric detection to quantify the adhesion of several strains of Lactobacillus and Bifidobacterium to Caco-2 cells. METHODS AND RESULTS: Lactic acid bacteria strains were subjected to fluorescent staining using the viable probe carboxyfluorescein diacetate and subsequently incubated on Caco-2 monolayers. The adhesion of the micro-organisms was determined by spectrofluorimetry following the lysis of the attached bacterial cells and expressed as adhesion percentage. The values obtained for the micro-organisms tested ranged from 4% for Bifidobacterium infantis Bi1 to 10% for a Bifidobacterium mixture containing three different strains. CONCLUSIONS: In the present study we successfully applied fluorescent labelling and fluorimetric detection to investigate the adhesive properties of some Lactobacillus and Bifidobacterium strains and a Bifidobacterium mixture to Caco-2 cells. SIGNIFICANCE AND IMPACT OF THE STUDY: The results proved that fluorescent labelling is suitable for adhesion studies and provides a reliable and safer alternative to radioactive labelling.     

Characterization of the properties of human- and dairy-derived probiotics for prevention of infectious diseases in fish

The present study aimed to investigate the potential probiotic properties of six lactic acid bacteria (LAB) intended for human use, Lactobacillus rhamnosus ATCC 53103, Lactobacillus casei Shirota, Lactobacillus bulgaricus, L. rhamnosus LC 705, Bifidobacterium lactis Bb12, and Lactobacillus johnsonii La1, and one for animal use, Enterococcus faecium Tehobak, for use as a fish probiotic. The strains for human use were specifically chosen since they are known to be safe for human use, which is of major importance because the fish are meant for human consumption. The selection was carried out by five different methods: mucosal adhesion, mucosal penetration, inhibition of pathogen growth and adhesion, and resistance to fish bile. The adhesion abilities of the seven LAB and three fish pathogens, Vibrio anguillarum, Aeromonas salmonicida, and Flavobacterium psychrophilum, were determined to mucus from five different sites on the surface or in the gut of rainbow trout. Five of the tested LAB strains showed considerable adhesion to different fish mucus types (14 to 26% of the added bacteria). Despite their adhesive character, the LAB strains were not able to inhibit the mucus binding of A. salmonicida. Coculture experiments showed significant inhibition of growth of A. salmonicida, which was mediated by competition for nutrients rather than secretion of inhibitory substances by the probiotic bacteria as measured in spent culture liquid. All LAB except L. casei Shirota showed tolerance against fish bile. L. rhamnosus ATCC 53103 and L. bulgaricus were found to penetrate fish mucus better than other probiotic bacteria. Based on bile resistance, mucus adhesion, mucus penetration, and suppression of fish pathogen growth, L. rhamnosus ATCC 53103 and L. bulgaricus can be considered for future in vivo challenge studies in fish as a novel and safe treatment in aquaculture.     

具有降胆固醇功能的益生菌的筛选

采用半选择性培养基MRS从传统发酵食品中筛选出2株细菌,经鉴定一株为双歧杆菌,另一株为乳酸杆菌.按照益生菌标准对上述2株菌的耐酸、耐胆盐、抗药性、抑菌等性能进行评价,结果表明其均可作为益生菌用于降胆固醇产品.     

Intra-specific variation of Lactobacillus plantarum and Lactobacillus pentosus in sensitivity towards various bacteriocins

Fifty-two strains belonging to the Lactobacillus plantarum species group were identified and typed. They represented 32 clones of Lactobacillus plantarum and 7 clones of Lactobacillus pentosus. Sensitivity of all strains towards bacteriocins of four different producer strains was investigated using a deferred inhibition test (DIT). Substantial intra-specific variation in sensitivity of clones was observed towards bacteriocinogenic lactic acid bacteria producing nisin ( Lactococcus lactis ATCC 11454) or pediocin PA-1 ( Pediococcus acidilactici PAC-1.0), while none of the strains were sensitive towards the two remaining bacteriocin producers. The minimum inhibitory concentration (MIC) of nisin towards selected strains confirmed the DIT results. No correlation between the susceptibility of fourteen selected strains towards nisin and an array of antibiotics was found. The present study indicates that the variation in bacteriocin-sensitivity within target species might be a potential limitation for the application of bacteriocins as biopreservatives.