双歧联菌株微胶囊制剂的改进

目的：提高微胶囊制剂的安全性。方法：采用ESM2为囊材,代替原用的ESM1。同时以95％乙醇代替丙酮作为囊材溶剂。结果：经实验确定了ESM2微囊化的最佳浓度为8％,并进一步确定了复合囊材中各种辅料的添加量以及微囊化操作条件。结论：在上述优化条件制备的双歧联菌株微胶囊具有良好的耐酸性和肠溶性。     

双歧杆菌NQ1501双层微胶囊制备及其特性研究

目的提高双歧杆菌NQ1501对环境的耐受性。方法采用流化床底喷工艺,以蔗糖、麦芽糊精、海藻糖和改性淀粉为内层壁材,聚丙烯酸树脂为外层壁材制备双歧杆菌NQ1501微胶囊,并对制得微囊进行评价。结果制得4种双歧杆菌NQ1501微胶囊均具有较好的耐酸和肠溶能力,25℃贮存6个月后对照菌粉存活率为4.8%,采用4种不同内壁材的微胶囊存活率分别为蔗糖28.6%,麦芽糊精45.7%,海藻糖38.9%,改性淀粉55.2%。结论采用该制备工艺制得双歧杆菌NQ1501微胶囊具备较好的耐酸性、肠溶性和常温保存稳定性。     

枯草芽胞杆菌微生态制剂的研制

采用液体发酵工艺,确定枯草芽胞杆菌的最适发酵条件为:发酵温度30℃,初始pH值7.2,并以1%海藻酸钠和3%明胶组成的混合胶体溶液为囊壁材料,以4%氯化钙作固化剂将枯草芽胞杆菌制成微胶囊剂,稳定性试验结果显示经微胶囊包埋的枯草芽胞杆菌制剂,室温下保存1个月,活菌存活率为98.8%,保存3个月,活菌存活率为50.6%,保存6个月,活菌存活率为15.7%,均高于未经微胶囊化的样品;在4℃冷藏下保存3个月,未经微胶囊化的样品活菌存活率仅为经微胶囊包埋制剂的66.2%。该微胶囊制剂提高了活菌存活率,延长了活菌常温保存期。     

一株广谱抑菌活性乳酸菌的筛选及特性研究

【目的】从贵州剑河采集的传统自然发酵豆酱中分离筛选具有广谱抑菌效果的乳酸菌,并进行肠道益生特性的研究。【方法】通过抑菌试验分离筛选得到菌株DJ-04,对其进行人工胃肠液耐受性、胆盐耐受性和渗透压耐受性的研究,并对其进行生理生化鉴定和16S r RNA鉴定。【结果】菌株DJ-04对大肠杆菌、沙门氏菌、金黄色葡萄球菌、志贺氏菌和铜绿假单胞菌的生长有很好的抑制作用;在p H值为2.5的人工胃液中处理3 h活菌数达到107 CFU/m L以上;在人工肠液中处理3 h活菌数达到108 CFU/m L以上,对人工胃肠液表现出良好的耐受性。能耐受一定浓度的牛胆盐,在质量浓度0.2 g/100 m L的牛胆盐环境中活菌数可达到107 CFU/m L;具有较高的渗透压耐受能力,在Na Cl质量浓度为10 g/100 m L的液体MRS中培养24 h后,活菌数仍在107 CFU/m L以上。经鉴定,DJ-04为植物乳杆菌。【结论】植物乳杆菌DJ-04具有良好的人工胃肠液耐受性以及耐胆盐和耐渗透压能力,具有肠道益生菌的潜能。     

新型高效微生态制品的研制

正交实验结果表明 ,4%淀粉水解液 ,10 %酵母提取液 ,2 0 %豆水解液 ,9%肝汤为双歧联菌株混合培养的最佳促生因子配方。经此优化培养基混合培养所得的双歧联菌株 ,制成冻干粉后 ,采用微囊化技术包裹成双歧杆菌微胶囊 ,其耐酸性强 ,肠溶性好 ,保存期长 ,由此可制得一系列新型高效微生态制品。     

低粘度辛烯基琥珀酸淀粉钠作微胶囊壁材包埋柠檬油

以蜡质玉米淀粉为原料在优化条件下制备具有乳化能力的低粘度辛烯基琥珀酸淀粉钠(SSOS)。用Brabender粘度仪测定淀粉糊的粘度及其稳定性,发现酶处理前SSOS糊的粘度较蜡质玉米淀粉提高,但稳定性变化不大。研究发现低粘度SSOS微胶囊包埋柠檬油的百分率在57.4%至87.6%(w/w)之间,并取决于SSOS的葡萄糖值。在壁材与芯材的质量比大于3比2,葡萄糖值为12.8时,微胶囊包埋的产率和效率达到最高值。微胶囊在40℃下放置贮存3周后,其微胶囊化效率仅降低3%。通过扫描电镜观察,发现微胶囊为球形的未粘连的颗粒,有些颗粒的表面并不平整。研究结果表明一定葡萄糖值的低粘度SSOS作为微胶囊壁材具有很好的包埋效果。     

抗肿瘤工程菌Escherichia coliNissle 1917发酵培养基优化及微胶囊制备

为了获得高活力抗肿瘤工程菌Escherichia coli Nissle 1917(EcNA)微胶囊制剂,对EcNA进行发酵培养基优化和微胶囊制剂的制备。首先利用单因素试验考察甘油、酵母提取物、蛋白胨及玉米浆对EcNA菌体浓度的影响,在以Box-Behnken设计试验的基础上,分别建立响应面模型和BP人工神经网络结合遗传算法模型优化发酵培养基组分,最后采用挤压法以海藻酸钠和壳聚糖为复合壁材,将EcNA包埋在微胶囊中。结果显示,最佳优化配方为甘油7 g/L,蛋白胨28.75 g/L,酵母提取物86.25 g/L,玉米浆10 g/L,K_2HPO_4 16.43g/L,KH_2PO_4 2.3 g/L,发酵液菌体浓度OD_(600)达到12.92,与未优化相比提高了3.86倍。通过正交试验得到最佳制备条件:海藻酸钠0.035 g/mL,壳聚糖0.004 g/mL,壁芯比2:2,氯化钙0.05 g/mL。结果表明,BP人工神经网络结合遗传算法在培养基优化中具有显著的优越性,制成的EcNA微胶囊有良好的耐酸性和肠溶性。     

不同类型益生菌制剂在胃肠道中耐受程度的对比

目的对市场销售的几种不同类型益生菌制剂进行胃、肠液耐受程度比较,评价各益生菌制剂活菌数量,为临床选择益生菌制剂提供参考。方法选择7种菌粉及某颗粒益生菌、国产某晶球、日本产某晶球、某胶囊益生菌、圣邦步步佳肠释益生菌共12种口服益生菌产品,在崩解时限仪中加入人工胃液进行崩解试验,在0.0 h、0.5 h、1.0 h、1.5 h和2.0 h时吸取上述样品并进行浓度梯度稀释,并在MRS固体培养基上培养后进行活菌计数。在上述5个时间点吸取部分益生菌-胃液混合物(针对益生菌粉末)或取出益生菌产品(针对晶球、肠溶胶囊、肠释益生菌)换人工肠液再进行40 min崩解,在MRS固体培养基上培养后进行活菌计数。结果粉末状益生菌制剂对人工胃液的耐受能力普遍较差。某颗粒益生菌、国产某晶球、日本产某晶球、某胶囊益生菌、圣邦步步佳肠释益生菌在人工胃液中不崩解。圣邦步步佳肠释益生菌在人工肠液中40 min完全崩解且活菌数峰值较高。结论圣邦步步佳肠释益生菌对人工胃、肠液耐受能力较强,且活菌数较高。     

生姜精油喷雾干燥法微胶囊化的研究

本文研究了以阿拉伯胶、麦芽糊精及大豆分离蛋白作为包埋生姜精油的壁材,采用三因子单形重心法优化确定壁材的最佳配方及工艺条件,采用喷雾干燥技术进行生姜精油的微胶囊包埋处理以获得固化的生姜精油微胶囊产品。     

正交法优化保加利亚乳杆菌NQ2508微囊制备工艺

目的制备保加利亚乳杆菌NQ2508双层微胶囊,并考察其包埋产率和耐胃酸能力。方法以保加利亚乳杆菌NQ2508改性淀粉微胶囊为研究对象,以聚丙烯酸树脂为包衣材料,采用流化床底喷工艺制备微胶囊,通过正交试验考察进风温度、雾化压力、进风风量、包衣增重4个因素对微胶囊化的影响。结果最佳工艺条件为进风温度50℃、雾化压力2.0bar、进风风量35m3/h、包衣增重30%,在该工艺条件下制得的微胶囊包埋产率为50.2%,微胶囊经人工胃酸处理后活菌存活率为39.2%。结论工艺优化后制得保加利亚乳杆菌NQ2508双层微胶囊的包埋产率和耐酸能力均较高。     

Lactobacillus cells in the rabbit digestive tract and the factors affecting their distribution

Lactobacillus fermentum was present in small numbers in the caecum and colon of only one of 16 rabbits. Studies aimed at elucidating the factors affecting the colonization of lactobacilli in rabbit gut were thus performed. These studies included investigation of the effect of gastric juice and bile salts on the viability of lactobacilli, assay of the survival rates of lactobacilli in the gastrointestinal tracts of the rabbits with or without ileum cannulation and measurement of the adhesive capability of lactobacilli to the rabbit intestinal epithelial cells. Results showed that, although some lactobacilli were resistant to the rather low pH levels of rabbit gastric juice, lack of adhesive capability may prevent them from colonizing in the intestinal tract. Therefore, if lactobacilli are to be used as probiotics for rabbit, factors that affect their viability and ability to colonize should be considered.     

蒙古族儿童源益生特性双歧杆菌的筛选及鉴定

【目的】双歧杆菌在人和动物胃肠道中发挥着重要的生理作用,然而双歧杆菌能否耐受胃酸、肠液及胆汁酸是影响活菌制剂益生效果的关键因素,本研究旨在从蒙古族儿童粪便中分离筛选出具有良好益生特性的双岐杆菌。【方法】本文采用双岐杆菌选择性培养基对样品进行分离纯化,并对菌株进行生理生化鉴定,以耐受人工胃肠液、耐受牛胆盐为手段对各菌株的益生特性进行评价,并且对B. animalisV9进行了16S rDNA分子生物学鉴定。【结果】本文从12位健康蒙古族儿童粪便中分离得到11株双歧杆菌,经传统生理生化试验鉴定为5株B. adolescentis：A1、H3、G4、A8、V10;3株B. longum：C6、C7、D11;1株B. pseudocatenlatum：B2;1株B. bifidum：G5;1株B. animalis：V9。B. animalis V9具有较强的耐酸性,在pH2.0的人工胃液中厌氧培养3h后存活率为92.4%,而其它10株双歧杆菌在此条件下的存活率均小于31.25%;B. animalis V9在pH2.0的人工胃液中厌氧培养3h后接入pH8.0的人工肠液中消化8h,存活率为99.7%,并且可以耐受0.3%的牛胆盐。进一步对V9菌株进行16S rDNA分子生物学鉴定,发现与Bifidobacterium animalis subsp. lactic BB12的同源性为99%。【结论】本研究结果显示B. animalis V9来源安全,并且具有良好耐酸、耐胆盐益生特性,有望在乳制品及保健类产品中得到广泛的应用。     

Effects of Orange Juice Formulation on Prebiotic Functionality Using an In Vitro Colonic Model System

A three-stage continuous fermentative colonic model system was used to monitor in vitro the effect of different orange juice formulations on prebiotic activity. Three different juices with and without Bimuno, a GOS mixture containing galactooligosaccharides (B-GOS) were assessed in terms of their ability to induce a bifidogenic microbiota. The recipe development was based on incorporating 2.75g B-GOS into a 250 ml serving of juice (65°Brix of concentrate juice). Alongside the production of B-GOS juice, a control juice – orange juice without any additional Bimuno and a positive control juice, containing all the components of Bimuno (glucose, galactose and lactose) in the same relative proportions with the exception of B-GOS were developed. Ion Exchange Chromotography analysis was used to test the maintenance of bimuno components after the production process. Data showed that sterilisation had no significant effect on concentration of B-GOS and simple sugars. The three juice formulations were digested under conditions resembling the gastric and small intestinal environments. Main bacterial groups of the faecal microbiota were evaluated throughout the colonic model study using 16S rRNA-based fluorescence in situ hybridization (FISH). Potential effects of supplementation of the juices on microbial metabolism were studied measuring short chain fatty acids (SCFAs) using gas chromatography. Furthermore, B-GOS juices showed positive modulations of the microbiota composition and metabolic activity. In particular, numbers of faecal bifidobacteria and lactobacilli were significantly higher when B-GOS juice was fermented compared to controls. Furthermore, fermentation of B-GOS juice resulted in an increase in Roseburia subcluster and concomitantly increased butyrate production, which is of potential benefit to the host. In conclusion, this study has shown B-GOS within orange juice can have a beneficial effect on the fecal microbiota.     

Microbiological characterization of wet wheat distillers' grain, with focus on isolation of lactobacilli with potential as probiotics

Wet wheat distillers' grain (WWDG), a residue from ethanol fermentation, was examined from a microbiological perspective. After storage, WWDG was characterized by a high content of lactobacilli, nondetectable levels of other bacteria, occasional occurrence of yeasts, and a pH of about 3.6 and contained a mixture of lactic acid, acetic acid, and ethanol. The composition of lactobacilli in WWDG was simple, including primarily the species Lactobacillus amylolyticus, Lactobacillus panis, and Lactobacillus pontis, as determined by 16S rRNA gene sequencing. Since the use of WWDG as pig feed has indicated a health-promoting function, some relevant characteristics of three strains of each of these species were examined together with basal physiological parameters, such as carbohydrate utilization and growth temperature. Seven of the strains were isolated from WWDG, and two strains from pig feces were included for comparison. It was clear that all three species could grow at temperatures of 45 to 50 degrees C, with L. amylolyticus being able to grow at temperatures as high as 54 degrees C. This finding could be the explanation for the simple microflora of WWDG, where a low pH together with a high temperature during storage would select for these organisms. Some strains of L. panis and L. pontis showed prolonged survival at pH 2.5 in synthetic stomach juice and good growth in the presence of porcine bile salt. In addition, members of all three species were able to bind to immobilized mucus material in vitro. Especially the isolates from pig feces but, interestingly, some isolates from WWDG as well possessed properties that might be of importance for colonization of the gastrointestinal tracts of pigs.     

Influence of oligosaccharides on the growth and tolerance capacity of lactobacilli to simulated stress environment

Aims:  Lactobacilli should resist stress environments in industry process and gastrointestinal tract before exerting their beneficial effects. To explore the possible stabilizers in probiotic products, prebiotic oligosaccharides were investigated.
Methods and Results:  We investigated the effect of four selected oligosaccharides on the survival of probiotic Lactobacillus plantarum and L. acidophilus to simulated stress conditions. It was found that the tolerance of lactobacilli to simulated artificial gastrointestinal juice, heat treatment and phenol solution was obviously enhanced in fructo-oligosaccharides (FOS) and xylo-oligosaccharides (XOS) group. In addition, chito-oligosaccharides (COS), manno-oligosaccharides (MOS) and glucose also had positive effect compared with control group (without sugar).
Conclusions:  Prebiotic oligosaccharides, especially XOS and FOS added in medium have protection function to lactobacilli in stress environments. The protection function of oligosaccharides may correlate with the bacteria growth, which was stimulated by these oligosaccharides.
Significance and Impact of the Study:  Prebiotic oligosaccharides may be used as stabilizers in probiotic products.     

The mutagenic and antimutagenic effects of Ecballium elaterium fruit juice in human peripheral lymphocytes

The aim of this study was to investigate the mutagenic and antimutagenic effects of Ecballium elaterium (EE) fruit juice, which has an anti-inflammatory effect, using in vitro human peripheral lymphocytes. To investigate the mutagenic effects of the EE fruit juice, human peripheral lymphocytes were treated with three doses (18, 36, and 72 μl/l) of fruit juice alone for 24 and 48 h. For investigating the antimutagenic effects of the EE fruit juice, the human lymphocytes were also treated with the mixture of the fruit juice and 0.25 μg/ml MMC. The EE fruit juice induced the percentage of total CA when used alone (especially the percentage of structural CA than the percentage of the numerical CA) and synergically induced the percentage of total CA when used as a mixture with MMC. The EE fruit juice did not affect the SCE frequency for 24 and 48 h treatment time. In contrast, EE and MMC as a mixture sinergically induced the SCE frequency at the highest concentration for 48 h treatment time only. EE alone did not decrease the RI while it decreased the MI in a dose-dependent manner. EE and MMC as a mixture have a higher cytotoxic effect than the cytotoxic effects of EE alone. As a result, it can be concluded that EE had no antimutagenic effect while EE had a mutagenic and a cytotoxic effect in human peripheral lymphocytes. This article was submitted by the authors in English.     

Studies on Pectolytic Enzymes of Molds

The clarification of apple juice has been studied using six pectolytic enzymes produced by Coniothyrium diplodiella, endo-PG (polygalacturonase) I, II and III, exo-PG and PE (pectinesterase) I and II. Each of these six enzymes had no effect on the clarification of apple juice when acted alone, whereas mixtures of any one of endo-PGs and PEs were all able to clarify the juice. Mixtures of exo-PG and either of PEs has no effect on the clarification. Clarifying activities of PG-PE mixtures were varied with the kind of endo-PG used in each mixture and not with the kind of PE. Clarifying activity of PG-PE mixture depended on either endo-PG or PE activities when the other was kept constant.

Crude enzyme from the mold and a mixture of the four PGs and PE in the ratio of the crude enzyme had essentially identical effect on apple juice as well as on artificial pectin and pectic acid.     

A MEDIUM FOR THE CULTIVATION OF LACTOBACILLI

SUMMARY: An improved growth medium for lactobacilli is described. It supports good growth of lactobacilli generally and also is particularly useful for a number of fastidious strains which grow only poorly in other general media. In addition, tomato juice, a highly variable material, is not required. In a slightly modified form, it can also be used as a basal medium for fermentation tests.     

Trophic relationships between Saccharomyces cerevisiae and Lactobacillus plantarum and their metabolism of glucose and citrate.

Glucose and citrate are two major carbon sources in fruits or fruit juices such as orange juice. Their metabolism and the microorganisms involved in their degradation were studied by inoculating with an aliquot of fermented orange juice a synthetic model medium containing glucose and citrate. At pH 3.6, their degradation led, first, to the formation of ethanol due to the activity of yeasts fermenting glucose and, eventually, to the formation of acetate resulting from the activity of lactobacilli. The yeast population always outcompeted the lactobacilli even when the fermented orange juice used as inoculum was mixed with fermented beet leaves containing a wider variety of lactic acid bacteria. The evolution of the medium remained similar between pH 3.3 and 5.0. At pH 3.0 or below, the fermentation of citrate was totally inhibited. Saccharomyces cerevisiae and Lactobacillus plantarum were identified as the only dominant microorganisms. The evolution of the model medium with the complex microbial community was successfully reconstituted with a defined coculture of S. cerevisiae and L. plantarum. The study of the fermentation of the defined model medium with a reconstituted microbial community allows us to better understand the behavior not only of fermented orange juice but also of many other fruit fermentations utilized for the production of alcoholic beverages.