双歧杆菌表面多糖通过免疫调控和抑制病原菌侵染与寄主相互促进

双歧杆菌(Bifidobacterium)是人类胃肠道微生物群落的一个重要组成部分,许多双歧杆菌目前被应用于治疗干预方面,对人类健康具有益生作用。但是,双歧杆菌是如何在人类胃肠道中定殖并起着重要的益生作用还不清楚。最近,卡耐基梅隆大学和东安格里亚大学等研究人员发现,双歧杆菌能够在细胞表面产生一种表皮相关多糖,     

双歧杆菌的分离鉴定及活菌胶囊的研制

从母乳哺育的３～５月龄健康婴儿的粪便中分离到可疑双歧杆菌，通过驯化，获得一株较耐氧的菌株。经属和种的系统生理生化鉴定，确认为两叉双歧杆菌。将它制成活菌胶囊，在本校小范围服用结果表明，该菌对急慢性肠炎、腹泻、便秘等症有显著疗效。     

植物多糖的提取及对双歧杆菌的增殖作用

采用水提法提取枸杞、红枣、甘薯、淮山4种植物原料及花菇多糖,正交设计优化提取条件,并将植物提取多糖用于双歧杆菌的增殖试验。结果表明,植物多糖提取优化条件,加水比例分别为:枸杞、红枣1∶10,花菇1∶15,甘薯1∶3,淮山1∶9;提取温度:80～95℃;提取时间1～3h。5种原料在优化条件下的多糖提取率相对于植物取样质量分别为:枸杞51.8%、红枣53.2%、花菇28.2%、甘薯18.8%、淮山18.3%。5种原料提取多糖对双歧杆菌的增殖作用:菌体在对数生长期生长速率常数(R)提高135%～85%,世代时间(G)缩短56%～44%。     

根癌家杆菌介导的玉米遗传转化体系的建立

用根癌农杆菌菌株LBA4404（pTOK233)转化多种基因型的玉米幼胚。幼胚与农杆菌共培养3d后，检测到了GUS基因的瞬时表达。利用GUS基因的瞬时表达对农杆菌转化玉米的条件进行了优化，共培养后的幼不经含潮霉素的培养基筛选培养两个月后，获得了多种基因型的抗性愈伤组织。其中综3自交系的抗性愈伤组织分化出植株，P9－10、综3自交系R0结实得到后代。Southern杂交分析证实了霉素基因在玉米自交系P9－10基因组中的整合，建立了农杆菌介导的玉米遗传转化体系。     

螺旋藻对抗生素相关性腹泻小鼠大便中菌群的影响

通过对腹泻小鼠灌食螺旋藻,对比给药组、模型组和正常组小鼠大便中菌种的变化,探索螺旋藻对抗生素相关腹泻小鼠肠道菌群的影响。结果表明:抗生素相关性腹泻导致小鼠的大便菌群异常,异常菌群有一定的自我恢复能力,生理盐水对其无明显治疗作用,螺旋藻灌胃可以明显抑制大便菌群的异常增殖,并促进其中双歧杆菌和乳杆菌菌群数量回升,对菌群异常的恢复有显著效果。     

高活菌数复合益生菌发酵乳工艺优化

益生菌发酵乳中的活菌数是保证其功能特性的关键因素,为提高益生菌发酵乳中的活菌总数,以干酪乳杆菌和双歧杆菌复合菌种为试验对象,以发酵过程中的pH值和吸光度平均值值为试验指标,在单因素试验的基础上,利用 Box-Benhnken中心组合试验和响应面分析法研究了接种量、益生菌接种比例、发酵温度、葡萄糖添加量以及大豆多肽添加量对发酵乳pH值和活菌数的影响,并建立了复合益生菌发酵模型。响应面优化试验结果表明复合益生菌发酵的最佳工艺条件是脱脂乳乳固体质量分数为12%,接种量为6%,干酪乳杆菌：双歧杆菌接种体积比为3∶1,葡萄糖添加量为2.9%,大豆多肽添加量为0.8%,发酵温度为34℃,在此条件下预测值为1.076,验证试验得到实际值为1.087,与理论预测值相比,相对误差为1.0%。发酵乳最大活菌数为4.1×1011 cfu/mL,与已有研究相比,活菌数提高了1-2个对数级。在4℃条件下贮藏21 d后,发酵乳的活菌数仍然保持在4.7×1010 cfu/mL。研究结果为工业化生产高活菌数的益生菌饮料提供参考。     

球形红杆菌磷耐受能力和除磷特性的研究

通过纯种培养试验，考察了废水生物除磷优势菌球形红杆菌的磷耐受能力和除磷特性。结果表明，球形红杆菌具有较强的磷耐受能力，且磷酸盐含量会对其除磷特性产生一定的影响。缺磷环境会使菌体产生过量摄磷现象；而在高磷环境下，菌体摄磷量的变化趋势与直接转入富磷培养类似：磷酸盐含量为1％时，菌体摄磷量基本未发生变化，当磷酸盐含量达到2％时，由于菌体的生长受到影响，摄磷量略有下降。结果还表明，球形红杆菌在微好氧培养过程中能达到与厌氧－－微好氧培养过程相似的除磷效果，从而提示只要废水中存在足够的碳源，即使全部是微好氧过程，该菌也能有效地摄取磷酸盐而达到较好的除磷效果，由此可见，将球形红杆菌应用于高浓度含磷废水的实际处理工艺中前景看好。     

寡聚半乳糖醛酸生物活性研究进展

寡聚半乳糖醛酸主要是指由2~20个半乳糖醛酸通过α-1,4键连接而成的一种功能性低聚糖,部分半乳糖醛酸以甲酯化的形式存在。文中介绍了寡聚半乳糖醛酸的结构特点,评述了寡聚半乳糖醛酸对植物体(诱导活性、促进生长发育、抑菌)、动物体(促进双歧杆菌增殖、阻止有毒物质吸附、抗癌、促进微量元素吸收)所产生的生物活性,有利于寡聚半乳糖醛酸的进一步研究。     

山羊板皮炭疽杆菌的辐射灭菌研究

研究了＾６Ｃｏγ射线对不同含菌量山羊板皮中炭疽杆菌的杀灭作用。结果表明：剂量大于２０ｋＧｙ的辐照处理，完全能够将疫皮中的炭疽杆菌彻底杀灭。在１４．６－２７．４ｋＧｙ剂量范围内，辐照处理对经不同时间贮藏的山羊板皮成革的抗张强度、伸长率、含水量等各项理化指标未产生明显影响。     

Bt杀虫基因向水稻内生细菌的转化研究

本研究成功地构建以水稻体内定殖的优势细菌巨大芽孢杆菌为载体菌的工程杀虫内生细菌，这一内生工程杀虫细菌的建成是以水稻内生细菌的动态研究，定殖研究以及重组ＤＮＡ和细菌转化方法研究背景为基础，利用杀虫毒性强，表达苏云金芽孢杆菌δ－内毒素较高的重组质粒为供体，通过改进的ＰＥＧ原生质体转化法及新型高新电脉冲穿孔转化法完成。     

Cordycepin: selective growth inhibitor derived from liquid culture of Cordyceps militaris against Clostridium spp

The growth responses of nine human intestinal bacteria to liquid culture of Cordyceps militaris Link. Pt. (Ascomycotina: Clavicipitaceae) collected from a pupa of Bombyx mori L. (Lepidoptera: Bombycidae) were examined using spectrophotometric and impregnated paper disk methods and compared to those of tetracycline and chloramphenicol, as well as those of Coptis japonica root-derived berberine chloride. The biologically active constituent of the cultures was characterized as cordycepin (3'-deoxyadenosine) by spectroscopic analysis. This compound revealed potent growth-inhibiting activity toward Clostridium paraputrificum and Clostridium perfringens at 10 microgram/disk without adverse effects on the growth of Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium adolescentis, Lactobacillus acidophilus, and Lactobacillus casei, whereas tetracycline and chloramphenicol inhibited the growth of these lactic acid-producing bacteria, clostridia and Escherichia coli. However, C. militaris-derived materials revealed no growth stimulation on the bifidobacteria and lactobacilli. These results may be an indication of at least one of the pharmacological actions of C. militaris. As a naturally occurring antibacterial agent, cordycepin could be useful as a new preventive agent against various diseases caused by clostridia.     

Fermentation of β-glucans derived from different sources by bifidobacteria: evaluation of their bifidogenic effect

β-Glucans obtained from barley, seaweed, bacteria, and mushroom sclerotia were incubated with pure cultures of Bifidobacterium infantis, Bifidobacterium longum, and Bifidobacterium adolescentis for a 24 h batch fermentation to evaluate their bifidogenic effect with inulin as the positive control. The pH value in all culture media was decreased by 0.5-1.5 units. All β-glucans supported the growth of the three bifidobacteria with B. infantis, having a relatively larger increase in populations (3-4 log(10) colony forming units). B. infantis produced almost double the amount of total short-chain fatty acids (SCFAs) than the other two bifidobacteria. The SCFA profile of B. infantis had a relatively higher proportion of propionic and butyric acid but less acetic acid than the other bifidobacteria. The utilization of all the β-glucans isolated from different sources regardless of their differences in glycosidic linkages and molecular weight by all three bifidobacteria was comparable to that of inulin.     

Selective growth-inhibiting effects of compounds identified in Tabebuia impetiginosa inner bark on human intestinal bacteria

The growth-inhibiting activity of anthraquinone-2-carboxylic acid and lapachol identified in the inner bark of taheebo, Tabebuia impetiginosa, toward 10 human intestinal bacteria was evaluated by using a paper disk diffusion bioassay and compared to those of seven lapachol congeners (1,4-naphthoquinone, naphthazarin, menadione, lawsone, plumbagin, juglone, and dichlone) as well as two commercially available antibiotics, chloramphenicol and tetracycline. Anthraquinone-2-carboxylic acid exhibited very strong growth inhibition of Clostridium paraputrificum at 1 microg/disk while 100 microg/disk of lapachol was needed for moderate growth inhibition of the same organism. These two isolates exhibited weak inhibition of Clostridium perfringens and Escherichia coli at 100 microg/disk while no adverse effects were observed on the growth of Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium infantis, Lactobacillus acidophilus, and Lactobacillus casei at 1000 microg/disk. Structure-activity relationships indicate that a methyl group in the C-2 position of 1,4-naphthoquinone derivatives might play an important role in antibacterial activity.     

Growth-inhibiting effects of Coptis japonica root-derived isoquinoline alkaloids on human intestinal bacteria.

The growth-inhibiting activity of Coptis japonica (Makino) root-derived materials toward eight human intestinal bacteria was examined using an impregnated paper disk method and compared to that of four commercially available isoquinoline alkaloids [berberine sulfate (BS), berberine iodide (BI), palmatine chloride (PC), and palmatine sulfate(PS)], as well as that of Thea sinensis leaf-derived epigallocatechin gallate (EGCG). The biologically active constituents of the Coptis extract were characterized as the isoquinoline alkaloids berberine chloride (BC), palmatine iodide (PI), and coptisine chloride (CC) by spectral analysis. The growth responses varied with both chemical and bacterial strain used. In a test using 500 microg/disk, BC and PI produced a clear inhibitory effect against Bifidobacterium longum, Bifidobacterium bifidum, Clostridium perfringens, and Clostridium paraputrificum, whereas weak or no inhibition was observed in Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus casei, and Escherichia coli. At 1000 microg/ disk, CC revealed weak or no growth inhibition toward all test bacteria, whereas EGCG exhibited weak growth inhibition against only C. perfringens and C. paraputrificum. Among various isoquinoline alkaloids, BC exhibited more potent inhibitory activity toward C. perfringens than BI and BS, whereas the inhibitory effect was more pronounced in PI compared to PC and PS. The Coptis root-derived materials did not promote growth of B. longum and C. perfringens.     

Effects of lactulose supplementation on the growth of bifidobacteria and biotransformation of isoflavone glycosides to isoflavone aglycones in soymilk

Bifidobacterium animalis subsp. lactis bb12 and B. longum 20099 were used to hydrolyze isoflavone glycosides (IG) to biologically active forms, which are isoflavone aglycones (IA), in soymilk (SM) prepared from soy protein isolate (SPI) and SM supplemented with 0.5% (w/v) of lactulose (SML). Supplementation of lactulose significantly ( p < 0.05) enhanced the viable counts of B. animalis subsp. lactis bb12 and B. longum 20099 up to 2.34 and 2.15 log CFU/mL, respectively. Bifidobacterium animalis subsp. lactis bb12 and B. longum 20099 utilized 3.32 and 3.75 mg/mL of lactulose at 24 h of incubation, respectively. Supplementation of lactulose also appeared to be a key factor in decreasing the pH of SML. The biotransformation of IG to IA was enhanced significantly by 6.8-17.1% and 12.8-13.5% in SML by B. animalis subsp. lactis bb12 and B. longum 20099, respectively. However, the presence of lactulose in SML showed the stimulating effect on B. longum 20099 only after 12 h of incubation. Isoflavone aglycones ranged from 69.5 to 77.1% of total isoflavone compounds in SML after incubation.     

Evaluation of enhanced hygroscopicity, bifidogenicity, and anticariogenicity of enzymatically synthesized beta-galactosyl-trehalose oligosaccharides

beta-Galactosyl-trehalose oligosaccharides (beta-GTOs) were enzymatically prepared as a mixture of 6-beta-galactosyl-trehalose (1) and 4-beta-galactosyl-trehalose (2) with a 9:1 ratio (w/w). The beta-GTO mixture showed a highly enhanced hygroscopicity as compared to those of trehalose and other sugars used. At 72 h of incubation under 90% relative humidity and room temperature, it had a large increase in weight due to its moisture absorption, which was five times larger than that of trehalose, 1.9 times larger than that of sucrose, and 1.5 times larger than that of maltotriose. It was very effective in the growth promotion of Bifidobacteria, such as Bifidobacterium longum and Bifidobacterium bifidum, which was better than the growth promotion in the cases of trehalose and galactooligosaccharide. It also showed a highly anticariogenic property; it had only 10% cell proliferation of Streptococcus sobrinus for that of the sucrose control and 60% inhibition of insoluble glucan synthesis. Its effectiveness of inhibition was two and 1.5 times better than that of trehalose and one and two times than xylitol, respectively, against cell growth and glucan synthesis. Conclusively, the functionality of the beta-GTO in terms of hygroscopicity, bifidogenicity, and anticariogenicity was considerably improved as compared to that of trehalose. It is thus suggested that the beta-GTO might be applied as an effective humectant and prebiotic substitute with enhanced noncariogenicity in food applications.     

Production of conjugated linoleic acid (CLA) by Bifidobacterium breve LMC520 and its compatibility with CLA-producing rumen bacteria

This study was performed to characterize the ability of an active Bifidobacterium strain to produce conjugated linoleic acid (CLA) and to test its possible utilization as a probiotic compatible to the ruminal condition. Bifidobacterium breve LMC520 can actively convert linoleic acid (LA) to cis-9,trans-11-CLA, which is a major isomer derived from microbial conversion. LMC520 showed reasonable tolerance under acidic conditions (pH 2.5 with 1% pepsin) and in the presence of oxgall (0-3%). The growth and CLA production of LMC520 were tested under ruminal conditions and compared with those of Butyrivibrio fibrisolvens A38, which is a major CLA producer in the rumen as an intermediate in the biohydrogenation (BH) process. LMC520 converted 15% of LA to CLA under ruminal conditions, which was 2 times higher activity than that of A38, and there was no decline in CLA level during prolonged incubation of 48 h. The BH activity of LMC520 was comparable to that of A38. When LMC520 was cocultured with A38, even with slight decrease of CLA due to high BH activity by A38, but the level of CLA was maintained by the high CLA-producing activity of LMC520. This comparative study shows the potential of this strain to be applied as a functional probiotic not only for humans but also for ruminants as well as to increase CLA production.     

Andean yacon root (Smallanthus sonchifolius Poepp. Endl) fructooligosaccharides as a potential novel source of prebiotics

The ability of three known probiotic strains (two lactobacilli and one bifidobacterium) to ferment fructooligosaccharides (FOS) from yacon roots (Smallanthus sonchifolius Poepp. Endl) was compared to commercial FOS in this study. Results indicate that Lactobacillus acidophilus NRRL-1910, Lactobacillus plantarum NRRL B-4496, and Bifidobacterium bifidum ATCC 15696 were able to ferment yacon root FOS. FOS consumption apparently depended on the degree of polymerization and the initial FOS composition. L. plantarum NRRL B-4496 and L. acidophilus NRRL B-1910 completely utilized 1-kestose molecules, while B. bifidum was able to utilize 1-kestose molecules as well as molecules with a higher degree of polymerization.     

In vitro fermentation of lactulose-derived oligosaccharides by mixed fecal microbiota

Fermentation properties of oligosaccharides derived from lactulose (OsLu) and lactose (GOS) have been assessed in pH-controlled anaerobic batch cultures using lactulose and Vivinal-GOS as reference carbohydrates. Changes in gut bacterial populations and their metabolic activities were monitored over 24 h by fluorescent in situ hybridization (FISH) and by measurement of short-chain fatty acid (SCFA) production. Lactulose-derived oligosaccharides were selectively fermented by Bifidobacterium and lactic acid bacterial populations producing higher SCFA concentrations compared to GOS. The highest total SCFA production was from Vivinal-GOS > lactulose > OsLu > GOS. Longer incubation periods produced a selective fermentation of OsLu when they were used as a carbon source reaching the highest selective index scores. The new oligosaccharides may constitute a good alternative to lactulose, and they could belong to a new generation of prebiotics to be used as a functional ingredient for improving the composition of gut microflora.