五种益生菌在纤维固定化连续培养系统的共存定殖

模拟肠道的半固态生境,建立以玉米纤维为载体的固定化连续搅拌培养系统、并以非固定化连续搅拌培养系统为对照,稀释率为0.0417,分别连续培养12 d,双歧杆菌、酪酸菌、嗜酸乳杆菌、肠膜芽孢杆菌、粪肠球菌五种菌在两种系统中均可稳定共存,固定化系统固相五种菌数均高于非固定化系统,以双歧杆菌增幅最大,半固态的固定化连续培养较液态连续培养的菌相与肠道菌相较一致,能更好地模拟肠道微生态.扫描电镜观察五种菌在玉米纤维上固定形成了生物膜.     

不同剂量恩诺沙星对连续培养系统中肠道菌群的影响

目的 研究不同残留剂量恩诺沙星对连续培养系统中肠道菌群数量的影响.方法 建立4套连续培养系统,接种正常人粪便于发酵罐中,连续培养肠道菌群23 d;第23天发酵罐中分别泵入含恩诺沙星剂量为0、1.25、12.5和125 mg/L的培养基,连续加药7 d,第31天停止加药.加药前10 d和加药过程中每隔2 d从4个发酵罐中取样并进行活菌计数.结果 活菌计数结果表明,空白组菌群数量稳定,说明4套连续培养系统运行稳定;低剂量组(1.25 mg/L)恩诺沙星使双歧杆菌下降3个数量级,对其他菌基本无影响;中剂量组(12.5 mg/L)中除拟杆菌外数量均有所下降,厌氧总菌、乳酸菌和肠杆菌停药前数量恢复到加药前水平,需氧总菌、双歧杆菌和肠球菌数量下降后不能恢复原初水平;高剂量组(125 mg/L)中所计数细菌数量均发生变化,需氧总菌、乳酸菌、双歧杆菌、肠球菌和拟杆菌数量下降且停药前不能恢复原初水平,厌氧总菌和肠杆菌数量先降后升,最终计数结果比加药前数量增加.结论 中国现行规定恩诺沙星的日允许摄入量(ADI)可能对人体肠道菌群造成影响.     

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。     

大豆多糖对双歧杆菌及人肠道菌群生长的影响

目的研究大豆多糖对双歧杆菌及肠道菌群生长的影响。方法替换Bs培养基中的碳源,分为不加糖、加葡萄糖2％、加大豆多糖2％、加大豆多糖5％、加低聚果糖2％五组,加3种双歧杆菌（长双歧、青春双歧、两歧双歧）菌液1％,测其24h后的活菌数,比较大豆多糖对双歧杆菌生长的影响;替换Bs培养基中的碳源,分为不加糖、加葡萄糖2％、加大豆多糖2％,加低聚果糖2％四组,加人体粪便菌液1％,模拟人体肠道环境厌氧培养24h后,用选择性培养基测其肠杆菌、肠球菌、双歧杆菌、乳酸杆菌的活菌数,观察大豆多糖对人体肠道菌群的影响。结果大豆多糖添加量为5％时对长双歧的促进作用明显优于不加糖组（P〈0．05）;大豆多糖对人体肠道各菌群的生长促进作用与低聚果糖差异无显著性（P〉0．05）。结论大豆多糖对长双歧杆菌的体外促进作用较明显;以粪菌群发酵糖试验表明,大豆多糖对乳杆菌和双歧杆菌均有促进作用,和低聚果糖作用效果相比差异无显著性（P〉0．05）,具有益生元的特性。     

玉米紫色植株花色苷色素对小鼠肠道菌群的影响

目的了解玉米紫色植株花色苷色素对小鼠肠道菌群的影响。方法按中华人民共和国卫生部《保健食品检验与评价技术规范》中的动物实验调节肠道菌群功能的检验方法,观察玉米紫色植株色素对小鼠肠道中肠杆菌、乳杆菌、双歧杆菌数量的影响。结果灌服后小鼠肠道乳杆菌、双歧杆菌数量变化差异均无显著性,低剂量组肠扦菌数量明显低于灌服前（P〈0．05）,其他组肠杆菌数量变化差异无显著性。结论玉米紫色植株色素对小鼠肠道菌群可能有影响。     

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.     

长康双歧菌增殖胶囊对肠道菌群影响的研究

目的 研究以苏糖为主要功能组分的长康双歧菌增殖胶囊的特性及功效。方法 通过实验鼠及志愿者进行调节肠道菌群试验,观察该微生态制剂的作用。结果 小鼠肠道内双歧杆菌数量明显增加;受试人群肠道内肠杆菌、拟杆菌数量明显减少,肠球菌、乳酸杆菌、双歧杆菌数量明显增加,产气莱膜梭菌数量增加。结论 以水苏糖为主要功能组分的长康双歧菌增殖胶囊具有调节肠道菌群、改善肠道内环境的功能。     

断奶仔猪肠道菌群的研究

研究断奶仔猪在不同生理状况下的肠道菌群,结果表明：健康仔猪肠道内容物中,从十二指肠到结肠,所测定的８种细胞数量都随肠段后移而逐渐增加,４３日龄仔猪肠道内容物中以乳杆菌、肠球菌和梭菌占绝对优势,双歧杆菌次之;与健康仔猪相比,下痢仔猪肠道中肠球菌、梭菌和乳杆菌等优势菌群的数量明显低于对照组;饲喂了含肠球菌的活菌制剂的仔猪肠道中双歧杆菌和类杆菌的数量比对照组明显增加,但肠球菌数量却无显著改变。     

微生态制剂(双歧三联活菌胶囊)治疗肠易激综合征的临床观察

双歧三联活菌胶囊(贝飞达,Bifido)是中国预防医学科学院流研所海斯药业有限公司开发出的新一代微生态制剂,为长双歧杆菌、嗜酸乳杆菌、粪肠球菌经适当配合制成的活菌制剂,该3种菌皆分离自健康人体,易于在肠道定植,能直接补充肠道正常生理细菌,通过生物拮抗,抑制病原菌,调整肠道菌群失调,改善肠道功能和肝功能,增强免疫,预防肠道肿瘤等多种生理作用.我科用双歧三联活菌胶囊治疗腹泻型肠易激综合征(irritable bowel syndrome,IBS),旨在评价双歧在联活菌胶囊在IBS治疗中的疗效和安全性,探讨肠道感染和肠道菌群失调与IBS的发病关系.     

淡豆豉对人体肠道六种常住菌的调节作用

目的初探淡豆豉对人体肠道6种常住菌的调节作用。方法利用选择性培养基从人的粪便中分离培养肠杆菌、肠球菌、双歧杆菌、乳杆菌、产气荚膜梭菌和脆弱拟杆菌6种人体肠道常住菌。将高、中和低浓度淡豆豉药液加入选择性培养基中,以不加药液的培养基作为空白对照,根据各菌种培养条件分别于厌氧或好氧条件下培养,然后进行菌落计数。结果与空白对照相比,加药培养基脆弱拟杆菌增长20.00%以上;而其他5种菌的生长均被不同程度的抑制,其中对乳杆菌和肠球菌抑制作用最强,加药培养基上这2种菌均不生长;50.00%浓度药液对产气荚膜梭菌抑制率为32.81%;20.00%浓度药液对肠杆菌完全抑制,但随着药液浓度增大抑制作用减弱,50.00%和70.00%浓度药液对肠杆菌的抑制率分别为53.83%和22.11%;低、中和高浓度药液对双歧杆菌的抑制率分别为17.07%、17.83%和17.84%,差异无统计学意义。结论淡豆豉对人体6种肠道常住菌具有不同程度的调节作用。     

Specific ATP and specific oxygen uptake rate in immobilized cell aggregates: Experimental results and theoretical analysis using a structured model of immobilized cell growth

In this work the quality and activity of immobilized Beneckea natriegens have been measured using Specific ATP (SATP) [mg (ATP) g(-1) (dry biomass)] and Specific Oxygen Uptake Rate (SOUR) [mg O(2) g(-1) (dry biomass) h(-1)]. The cells were grown in a 3 L Three Phase Air Lift Bioreactor (TPALB) and were immobilized on diatomaceous earth (silica) support particles; sterile conditions were employed during the experiment, with n-propanol as the sole carbon source. Two sets of experiments were performed, one with varying dilution rate and the other with varying inlet substrate concentration. The average SATP and SOUR of the immobilized biomass was 3-4 times lower than the values obtained for suspended Beneckea natriegens growing at its maximum growth rate. The suspended biomass present in the TPALB was generated primarily through attrition from the outer layer of the biofilm, and maintained higher levels of SATP and SOUR than the immobilized biomass. This result indicates that the immobilized biomass quality/activity is higher at the external layer of the biofilm. A structured model in which biomass is divided into two compartments, active and inert, was used to describe the experimental results. This model predicts the biomass quality/activity and substrate concentration distributions in the biofilm. These distributions were integrated to give overall values of SATP and SOUR for the immobilized biomass which compared favorably with experimental data. The primary implication of the results is that the location of immobilized biomass in the biofilm affects its biocatalytic activity, and should be taken into account when modeling immobilized biomass bioreactors. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 660-673, 1997.     

Increased stress tolerance of Bifidobacterium longum and Lactococcus lactis produced during continuous mixed-strain immobilized-cell fermentation

AIMS: The effect of immobilization and long-term continuous culture was studied on probiotic and technological characteristics of lactic acid and probiotic bacteria. METHODS AND RESULTS: A continuous culture in a two-stage system was carried out for 17 days at different temperatures ranging from 32 to 37 degrees C, with a first reactor containing Bifidobacterium longum ATCC 15707 and Lactococcus lactis subsp. lactis biovar. diacetylactis MD immobilized separately in gel beads, and a second reactor operated with free cells released from the first reactor. The tolerance of free cells from both strains produced in the effluent medium of both reactors to hydrogen peroxide, simulated gastric and intestinal juices, antibiotics and nisin, and freeze-drying markedly increased with culture time and was generally higher after 6 days than that of stationary-phase cells produced during free-cell batch fermentations. The reversibility of the acquired tolerance of B. longum, but not L. diacetylactis, to antibiotics was shown during successive free-cell batch cultures. CONCLUSIONS: Free cells produced from continuous immobilized-cell culture exhibited altered physiology and increased tolerance to various chemical and physico-chemical stresses. SIGNIFICANCE AND IMPACT OF THE STUDY: Continuous culture with immobilized cells could be used to produce probiotic and lactic acid bacteria with enhanced technological and probiotic characteristics.     

Studies on formation,control and application of biofilm formed by food related microorganisms

Biofilms are sessile microbial aggregates on the interfaces, and they were usually considered as microbial contamination sources in medical care and various industries. We studied the control and application of biofilms formed by food-related microorganisms, and mechanism of the biofilm formation was also investigated. We studied the biofilm formation in mixed cultures using various combinations of two strains of food-related microorganisms. There were various microorganisms that showed decreased or increased biofilm formation in the mixed culture in comparison with that in a single culture. Biofilm formed by lactic acid bacteria and yeast isolated from traditional fermented food, Fukuyama pot vinegar, exhibited unique feature in that structure and formation mechanism, and expected to be used as an immobilized microorganism in fermentation production. Here our studies on the control and application of biofilms and the mechanisms of its formation were described.     

Growth model and metabolic activity of brewing yeast biofilm on the surface of spent grains: a biocatalyst for continuous beer fermentation

In the continuous systems, such as continuous beer fermentation, immobilized cells are kept inside the bioreactor for long periods of time. Thus an important factor in the design and performance of the immobilized yeast reactor is immobilized cell viability and physiology. Both the decreasing specific glucose consumption rate (q(im)) and intracellular redox potential of the cells immobilized to spent grains during continuous cultivation in bubble-column reactor implied alterations in cell physiology. It was hypothesized that the changes of the physiological state of the immobilized brewing yeast were due to the aging process to which the immobilized yeast are exposed in the continuous reactor. The amount of an actively growing fraction (X(im)act) of the total immobilized biomass (X(im)) was subsequently estimated at approximately X(im)act = 0.12 g(IB) g(C)(-1) (IB = dry immobilized biomass, C = dry carrier). A mathematical model of the immobilized yeast biofilm growth on the surface of spent grain particles based on cell deposition (cell-to-carrier adhesion and cell-to-cell attachment), immobilized cell growth, and immobilized biomass detachment (cell outgrowth, biofilm abrasion) was formulated. The concept of the active fraction of immobilized biomass (X(im)act) and the maximum attainable biomass load (X(im)max) was included into the model. Since the average biofilm thickness was estimated at ca. 10 microm, the limitation of the diffusion of substrates inside the yeast biofilm could be neglected. The model successfully predicted the dynamics of the immobilized cell growth, maximum biomass load, free cell growth, and glucose consumption under constant hydrodynamic conditions in a bubble-column reactor. Good agreement between model simulations and experimental data was achieved.     

漏斗多孔菌液体菌种培养基及其栽培条件

为实现漏斗多孔菌资源化利用,设计单因素试验,以菌丝生物量、菌球密度和菌球直径为指标,获得漏斗多孔菌Polyporus arcularius液体菌种培养基配方为马铃薯（去皮）200g、玉米粉20.0g、蛋白胨5.0g、KH₂PO₄ 3.0g、K₂HPO₄ 1.0g,MgSO₄·7H₂O 1.5g,初始pH 5.0,并优化培养条件为装液量250mL/500mL,接种量2%,培养温度32℃,转速170r/min,种龄6d;依上述条件制备液体菌种,以固体菌种为对照,进行出菇试验,调查农艺性状,试验结果表明液体菌种接种后,菌袋满袋时间提前7d,子实体原基产生时间提前8d,头潮菇鲜重提高22.5%;配方试验结果表明,以细木屑为主料的配方产量最高,头潮菇鲜重为270.6g/袋。     

Membrane process for biological treatment of contaminated gas streams

A hollow fiber membrane bioreactor was investigated for control of air emissions of biodegradable volatile organic compounds (VOCs). In the membrane bioreactor, gases containing VOCs pass through the lumen of microporous hydrophobic hollow fiber membranes. Soluble compounds diffuse through the membrane pores and partition into a VOC degrading biofilm. The hollow fiber membranes serve as a support for the microbial population and provide a large surface area for VOC and oxygen mass transfer. Experiments were performed to investigate the effects of toluene loading rate, gas residence time, and liquid phase turbulence on toluene removal in a laboratory-scale membrane bioreactor. Initial acclimation of the microbial culture to toluene occurred over a period of nine days, after which a 70% removal efficiency was achieved at an inlet toluene concentration of 200 ppm and a gas residence time of 1.8 s (elimination capacity of 20 g m-3 min-1). At higher toluene loading rates, a maximum elimination capacity of 42 g m-3 min-1 was observed. In the absence of a biofilm (abiotic operation), mass transfer rates were found to increase with increasing liquid recirculation rates. Abiotic mass transfer coefficients could be estimated using a correlation of dimensionless parameters developed for heat transfer. Liquid phase recirculation rate had no effect on toluene removal when the biofilm was present, however. Three models of the reactor were created: a numeric model, a first-order flat sheet model, and a zero-order flat sheet model. Only the numeric model fit the data well, although removal predicted as a function of gas residence time disagreed slightly with that observed. A modification in the model to account for membrane phase resistance resulted in an underprediction of removal. Sensitivity analysis of the numeric model indicated that removal was a strong function of the liquid phase biomass density and biofilm diffusion coefficient, with diffusion rates below 10(-9) m2 s-1 resulting in decreased removal rates.     

Biofilm model calibration and microbial diversity study using Monte Carlo simulations

Mathematical models are useful tools for studying and exploring biological conversion processes as well as microbial competition in biological treatment processes. A single‐species biofilm model was used to describe biofilm reactor operation at three different hydraulic retention times (HRT). The single‐species biofilm model was calibrated with sparse experimental data using the Monte Carlo filtering method. This calibrated single‐species biofilm model was then extended to a multi‐species model considering 10 different heterotrophic bacteria. The aim was to study microbial diversity in bulk phase biomass and biofilm, as well as the competition between suspended and attached biomass. At steady state and independently of the HRT, Monte Carlo simulations resulted only in one unique dominating bacterial species for suspended and attached biomass. The dominating bacterial species was determined by the highest specific substrate affinity (ratio of µ/K_S). At a short HRT of 20 min, the structure of the microbial community in the bulk liquid was determined by biomass detached from the biofilm. At a long HRT of 8 h, both biofilm detachment and microbial growth in the bulk liquid influenced the microbial community distribution. Biotechnol. Bioeng. 2013; 110: 1323–1332. © 2012 Wiley Periodicals, Inc.     

Effects of different sources of physically effective fiber on rumen microbial populations

Physically effective fiber is needed by dairy cattle to prevent ruminal acidosis. This study aimed to examine the effects of different sources of physically effective fiber on the populations of fibrolytic bacteria and methanogens. Five ruminally cannulated Holstein cows were each fed five diets differing in physically effective fiber sources over 15 weeks (21 days/period) in a Latin Square design: (1) 44.1% corn silage, (2) 34.0% corn silage plus 11.5% alfalfa hay, (3) 34.0% corn silage plus 5.1% wheat straw, (4) 36.1% corn silage plus 10.1% wheat straw, and (5) 34.0% corn silage plus 5.5% corn stover. The impact of the physically effective fiber sources on total bacteria and archaea were examined using denaturing gradient gel electrophoresis. Specific real-time PCR assays were used to quantify total bacteria, total archaea, the genus Butyrivibrio, Fibrobacter succinogenes, Ruminococcus albus, Ruminococcus flavefaciens and three uncultured rumen bacteria that were identified from adhering ruminal fractions in a previous study. No significant differences were observed among the different sources of physical effective fiber with respect to the microbial populations quantified. Any of the physically effective fiber sources may be fed to dairy cattle without negative impact on the ruminal microbial community.     

Two-dimensional model of biofilm detachment caused by internal stress from liquid flow

A two-dimensional model for biofilm growth and detachment was used to evaluate the effect of detachment on biofilm structures. The detachment process is considered to be due to internal stress created by moving liquid past the biofilm. This model generated a variety of realistic biofilm-formation patterns. It was possible to model in a unified way two different biofilm detachment processes, erosion (small-particle loss), and sloughing (large-biomass-particle removal). The distribution of the fraction from total biomass detached as a function of detached particle mass, gives indications about which of the two mechanisms is dominant. Model simulations indicate that erosion makes the biofilm surface smoother. Sloughing, in contrast, leads to an increased biofilm-surface roughness. Faster growing biofilms have a faster detachment rate than slow-growing biofilms, under similar hydrodynamic conditions and biofilm strength. This is in perfect accordance with the experimental evidence showing that detachment is dependent on both shear- and microbial-growth rates. High growth rates trigger instability in biofilm accumulation and abrupt biomass loss (sloughing). Massive sloughing can be avoided by high liquid shear, combined with low biomass growth rates. As the modeling results show, the causes for sloughing must be sought not only in the biofilm strength, but also in its shape. Several "mushroom-like" biofilm structures like those repeatedly reported in the literature occurred, due to a combined effect of nutrient depletion and breaking at the colony base. A rough carrier surface promotes biofilm development in hydrodynamic conditions in which the biofilm on a flat surface would not form. Although biofilm patches filled completely the cavity in which they started to grow, they were unable to spill over the carrier peaks and to fully colonize the substratum.