昂立植物乳杆菌发酵及其稳定性能的研究

研究昂立植物乳杆菌(LP-Onlly)的发酵特性、耐药性及遗传稳定性.结果表明LP-Onlly发酵培养条件为:37 ℃培养16～18 h发酵结束,pH可达4.4左右;LP-Onlly对研究的33种抗生素敏感、7种耐受;LP-Onlly连续传代15代后,基因组DNA指纹图谱及代谢产物HPLC指纹图谱与原代保持一致.     

昂立植物乳杆菌耐酸、耐胆盐性能及相关表型、基因型分析

目的从益生菌功能性评价出发,探讨昂立植物乳杆菌(LP-Onlly)的耐酸、耐胆盐性能及其相关的表型和基因型信息.结果 LP-Onlly在耐酸、耐胆盐性能方面,优于其他实验室保存菌株,在pH 3、37 ℃培养3 h后,LP-Onlly活菌存活率达95%以上.在1.0%胆盐的MRS培养液中生长24 h后,LP-Onlly活菌存活率达95%以上.经表型及分子鉴定,LP-Onlly为植物乳杆菌,其16S rDNA序列保存于GenBank, 登录号为AY 590777,与实验室保存的其他植物乳杆菌的基因DNA指纹图谱相似率达71%.     

施普瑞螺旋藻对尾吊模拟失重大鼠肠道微生态失调的调整作用研究

目的：研究模拟失重条件下,施普瑞螺旋藻对大鼠肠道微生态失调的调整作用。方法：将基础饲料中加入5%的螺旋藻作为处理组饲喂大鼠,用选择性培养基分别对肠球菌、肠杆菌、类杆菌、乳杆菌以及双歧杆菌定量测定,扫描电镜观察大鼠盲肠上皮细胞的组织变化,结果：螺旋藻处理组过路菌群中肠杆菌和肠球菌数量变化并不明显;原籍菌群中双歧杆菌较对照组显著增多,类杆菌和乳杆菌的数量差异没有显著性,模拟失重条件下SD大鼠盲肠上皮有肿胀细胞出现,并且肠道上皮绒毛排列紊乱、稀疏、而螺旋藻处理组中只发现有少量的肿胀细胞,上皮绒毛致密,排列较整齐。结论：旋普瑞螺旋藻具有纠正在模拟失重条件下大鼠肠道微生态平衡失调的作用。     

模拟失重对大鼠肠道菌群影响的研究

目的：研究模拟失重条件下大鼠肠道菌群变化情况。方法：选择性培养基分别对肠球菌、肠杆菌、类杆菌、乳杆菌以及双歧杆菌进行定量测定,扫描电镜观察大鼠盲肠上皮细胞的组织变化。结果：过路菌群中肠杆菌和肠球菌数量增加显著;原籍菌群中双歧杆菌减少十分明显,乳杆菌的数量也有不同程度的减少,而类杆菌的数量有增加的趋势。SD大鼠盲肠出现了肿胀细胞,上皮细胞颈毛排列紊乱、稀疏。结论：模拟失重条件下大鼠肠道微生态出现平衡失调。     

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.     

婴儿肠道细菌及乳杆菌种群多样性研究

目的 对婴儿肠道细菌及乳杆菌组成进行分析。方法 以66例出生1～7d的婴儿为调查对象,用分子生物学的手段,借助肠道细菌DNA指纹图谱和16S rDNA PCR扩增克隆分类（RFLP）技术。结果 婴儿肠道菌群随出生天数增加而变化;乳杆菌特异性引物对PCR扩增结果表明,出生4h后乳杆菌已在婴儿肠道内定植,且定植时间与分娩方式无关;克隆多样性研究则显示婴儿肠道乳杆菌优势菌群为植物乳杆菌（74％）、嗜酸乳杆菌（14％）和约氏乳杆菌（12％）。结论 婴儿肠道中的优势乳杆菌主要为植物乳杆菌。     

益生菌对烧伤大鼠肠道膜菌群和sIgA的影响

目的探索益生菌对严重烧伤大鼠早期肠道菌群的调理作用,为防治肠源性感染寻找新途径.方法选用健康Wistar大鼠100只,体重180～220 g,雌雄各半,随机分为:益生菌治疗组(BR),烧伤对照组(BC)和正常对照组(NC),建立30% Ⅲ°烫伤肠源性感染的动物模型,按时分批活杀取材,检测盲肠膜菌群和肠黏膜sIgA的含量.结果 BC组盲肠中双歧杆菌数量与NC组比较,明显减少(P＜0.01),而BR组与NC组比较,差异无显著性(P＞0.05);BC组肠道中酵母菌和大肠埃希菌与NC比较明显增加,BR组差异无显著性;肠粘液sIgA水平与上述指标有类似变化.结论 30%Ⅲ°烫伤大鼠肠道内容物中,双歧杆菌明显下降,大肠埃希菌、酵母菌迅速过生长,导致肠道微生态失衡,应用BR治疗后,促进肠黏膜机械屏障功能的恢复,调理肠道微生态平衡,对肠黏膜起占位性保护作用,提高了肠道局部和全身益生菌免疫的功能.     

高尿酸血症鹌鹑肠道菌群结构分析

目的探讨高尿酸血症鹌鹑肠道菌群结构变化特点,为高尿酸血症病理机制研究提供参考。方法将迪法克鹌鹑随机分为两组:正常组和模型组,正常组喂饲正常饲料,模型组喂饲高嘌呤饲料; PCR-DGGE法构建两组鹌鹑盲肠内容物菌群DNA指纹图谱,对图谱进行数字化处理,聚类分析法比较两组鹌鹑肠道菌群结构整体差异,logistic回归法辨别差异条带,克隆和鉴定差异条带序列; HE染色观察盲肠组织结构变化。结果造模第28天,模型组鹌鹑与正常组比较,血尿酸显著升高、肠道菌群整体结构发生变化,DGGE指纹图谱上12条差异条带,序列鉴定归属于拟杆菌门、厚壁菌门、变形菌门;盲肠组织出现炎性变化。结论高嘌呤饮食诱导的高尿酸血症鹌鹑肠道菌群结构发生改变。     

昂立一号~优菌多颗粒人群试食研究

昂立一号○R优菌多颗粒是“昂立一号”家庭新成员,其人群试食研究结果表明,昂立一号○R优菌多颗粒在短时间(4 d)内就能显著改善食欲、胃肠道及代谢排便功能,随着服用时间的延长,改善作用越强;20 d服用后可极显著地提高人体肠道菌群中的益生菌(双歧杆菌和乳杆菌)数量,却极显著地抑制肠道有害菌产气荚膜梭菌数量,可调节人体肠道菌群平衡,腹泻便秘双重调节,促进肠道健康。     

燕麦麸皮对大鼠肠道菌群及糖脂代谢水平的影响

目的：研究富含燕麦麸皮饲料对正常大鼠的耐受性和安全性,及其对肠道功能、肠道菌群构成、血糖、血脂水平的影响。方法：SD大鼠分为对照组（基础饲料喂养）和干预组（含燕麦麸皮21.7%饲料喂养）,4w实验期间记录大鼠一般生活状况、进食量、体重、血糖、血脂水平、肠道转运时间等指标,第29d处死大鼠,记录盲肠总重、壁重及内容物重量,行肠球菌、肠杆菌、双歧杆菌及乳杆菌特异性培养及计数,测定盲肠内容物氨含量及pH值。结果：喂养4w后,与对照组相比,干预组大鼠盲肠内容物双歧杆菌数量明显上升,盲肠内容物pH值水平明显下降,其他各项指标在2组间未见显著性差异。结论：正常大鼠对干预组剂量的燕麦麸皮安全耐受,燕麦麸皮增加正常大鼠肠道双歧杆菌数量,降低肠道内pH值水平,未观察到其对肠道转运时间、盲肠结构及糖脂代谢水平等指标的影响。     

抗生素所致腹泻大鼠肠屏障功能损伤及乳酸杆菌保护机制的研究

目的探讨抗生素对所致腹泻大鼠肠道屏障功能、肠道菌群结构和肠道细菌移位的影响及乳酸杆菌制剂的保护机制。方法采用细菌培养法动态测定抗生素所致腹泻大鼠肠道菌群变化及肠系膜淋巴结、肝脏、脾脏和结肠组织的移位细菌量;应用光镜和电子显微镜观察肠黏膜组织超微结构变化。结果应用抗生素可致大鼠腹泻,肠道菌群失调,肠黏膜组织受损,发生肠道细菌移位。大肠埃希菌攻击可加重肠道菌群失调和肠黏膜损伤程度,促发细菌移位发生。乳酸杆菌可扶正肠菌群结构,修复损伤的肠黏膜,抑制肠细菌移位发生。结论阐明了抗生素、肠黏膜屏障功能、肠道菌群结构和肠道细菌移位间的互为因果,相互影响的关系。微生态制剂在维持机体微生态平衡、修复肠黏膜方面具有保护作用。     

大承气汤对MODS时肠道细菌微生态学影响的实验研究

目的探讨多器官功能不全综合征(MODS)大鼠肠道细菌微生态的变化及其与肠源性内毒素血症和细菌易位的关系,并观察大承气汤的影响。方法32只SD大鼠随机分成4组,对照组、模型组、大承气组和氨苄青霉素组。腹腔注射无菌酵母多糖A制备大鼠MODS模型。各组动物于造模后48 h无菌操作抽取外周静脉血和门静脉血进行内毒素含量测定;取肠系膜淋巴结进行细菌定量培养,取回肠和盲肠内容物进行肠腔内游离内毒素测定;取盲肠内容物进行肠道细菌微生态学分析。结果模型组外周血和门静脉血内毒素水平以及肠腔内游离内毒素含量均明显高于对照组(P<0.05);与对照组相比,模型组肠道菌群出现明显变化。肠球菌、肠杆菌数量明显增加,而双歧杆菌和乳酸杆菌数量出现显著下降,类杆菌数量亦出现明显下降(P<0.05)。模型组厌氧菌总数明显下降而需氧菌总数明显增加,同时厌氧菌总数/需氧菌总数的比值和B/E比值呈相应下降,发生倒置(P<0.05);正常对照组未发现肠道细菌向肠系膜淋巴结的易位,而模型组细菌易位阳性率是83.33%(P<0.05)。与模型组相比,大承气汤组上述各指标均出现明显变化(P<0.05);抗生素组作用不明显(P>0.05)。结论MODS时大鼠肠道细菌微生态出现明显变化,发生肠源性内毒素血症和细菌易位。大承气汤可以调整肠道菌群,恢复肠道微生态平衡,增加机体定植抗力,防治细菌易位和内毒素血症。     

维吾尔族正常黑胆质人群肠道菌群的分布情况分析

目的：了解维吾尔医学正常黑胆质人群肠道菌群分布情况、多样性并优势菌。方法：对健康人进行维吾尔医学体液分型并挑 取其中正常黑胆质人群,采集受检者粪便样品,提取总DNA,设计一对通用引物扩增16S rDNA 的V6～V8 可变区,扩增出来的 PCR产物稀释并进行变形梯度凝胶电泳DGGE,从DGGE 指纹图谱中选择条带,切胶回收、克隆、序列测定。结果：通过实验得到 了反映肠道菌群结构特征的DNA指纹图谱,从指纹图谱上选择一些特异性条带切下来回收,重新纯化扩增出来并测序,测出来 的基因序列在基因库进行比对检测相似性程度。最终用相似性程度大于95%以上的序列比对做出进化树了解菌群之间的亲缘 性。结论：正常黑胆质人群肠道菌群基因序列的亲缘性结果显示黑胆质人群肠道菌群具有丰富的多样性,其中肠道优势细菌乳酸 杆菌属GU269544.1 占优势。     

Autochthonous bacterial flora indicated by PCR-DGGE of 16S rRNA gene fragments from the alimentary tract of Costelytra zealandica (Coleoptera: Scarabaeidae)

Aims: To locate and identify putative autochthonous bacteria within the grass grub gut that may have a role in symbiosis. Methods and Results: Polymerase chain reaction-denaturing gradient gel electrophoresis (DGGE) fingerprinting was used to investigate bacterial diversity in the grass grub larval gut. The microbial community profiles from five geographically distinct populations were compared and the influence of feeding was analysed. Bacterial community in the midgut was highly variable between locations and was affected by feeding. The hindgut contained a more diverse but stable bacterial community that was less affected by external conditions. Forty-seven distinct DGGE bands, representing different bacterial genotypes, could be distinguished from all samples, with 34 different bands occurring in the hindgut. The 22 most common bands were isolated and DNA was sequenced. Sequence analysis revealed that most bacteria (16/22) were affiliated to the Clostridiales with the predominant bacteria affiliated to the genus Clostridium. The remaining bacteria were aligned to the Betaproteobacteria, Deltaproteobacteria and Bacteroidetes. Conclusions: The grass grub larva has an autochthonous microflora with predominance of Clostridium spp. in the hindgut. Significance and Impact of the Study: Occurrence of an autocthonous microflora in the grass grub hindgut suggests a symbiotic relationship which could help explain the ability of larval scarabs to feed on recalcitrant organic matter.     

ERIC-PCR指纹图谱技术分析糖尿病小鼠肠道细菌群落变化

目的通过比较1型糖尿病模型组和空白对照组雄性小鼠肠道菌群结构的变化,探索糖尿病造模与肠道菌群的关系。方法收集造模2周后空白对照组(n=5)、STZ造模成功组(n=5)和造模不成功组(n=3)ICR小鼠的新鲜粪便样品,提取粪便样品的总DNA,ERIC-PCR扩增形成DNA指纹图谱,借助多变量统计分析方法研究各组样品肠道菌群结构上的异同。结果ERIC-PCR指纹图谱结合偏最小二乘法(PLS-DA)分析表明造模成功组和造模不成功组小鼠的肠道菌群结构显著区别于空白对照组,而造模不成功组小鼠的肠道菌群结构与造模成功组仍有一定的区别。结论STZ诱导的1型糖尿病会造成小鼠的肠道菌群结构的变化,而部分小鼠造模失败可能与这些小鼠的肠道菌群结构有关。     

Application of denaturing gradient gel electrophoresis for analysing the gut microflora of Lumbricus rubellus Hoffmeister under different feeding conditions

The earthworm, Lumbricus rubellus, plays an essential role in soil ecosystems as it affects organic matter decomposition and nutrient cycling. By ingesting a mixture of organic and mineral material, a variety of bacteria and fungi are carried to the intestinal tract of the earthworm. To get a better understanding of the interactions between L. rubellus and the microorganisms ingested, this study tried to reveal if the diet affects the composition of the gut microflora of L. rubellus or if its intestinal tract hosts an indigenous, species-specific microbiota. A feeding experiment with L. rubellus was set up; individuals were collected in the field, transferred to a climate chamber and fed with food sources of different quality (dwarf shrub litter, grass litter or horse dung) for six weeks. DNA was extracted from the guts of the earthworms, as well as from the food sources and the surrounding soil, and further analysed by a molecular fingerprinting method, PCR-DGGE (Polymerase Chain Reaction -- Denaturing Gradient Gel Electrophoresis). We were able to demonstrate that the gut microbiota was strongly influenced by the food source ingested and was considerably different to that of the surrounding soil. Sequencing of dominant bands of the bacterial DGGE fingerprints revealed a strong occurrence of y-Proteobacteria in all gut samples, independent of the food source. A specific microflora in the intestinal tract of L. rubellus, robust against diet changes, could not be found.     

大蒜素对急性酒精性肝损伤小鼠 肠道菌群失调的预防作用研究

目的应用肠杆菌基因间重复共有序列基因扩增(ERIC-PCR)和聚合酶链式反应—变性梯度凝胶电泳(PCR-DGGE)研究大蒜素对急性酒精性肝损伤小鼠肠道菌群失调的预防作用。方法 SPF级昆明小鼠40只,分为正常对照组、急性酒精性肝损伤组、护肝片组、大蒜素高和低浓度组,每组8只。灌服相应药物30d,除正常对照组,其他组灌胃14 mL/kg红星二锅头,12h后收集鼠便,提取粪便细菌DNA,利用ERIC-PCR和PCR-DGGE电泳获得肠道菌群指纹图谱,主成分分析(PCA)和聚类分析(UPGMA)研究肠道菌群整体差异并鉴定优势条带序列。结果 ERIC-PCR表明急性酒精性肝损伤小鼠肠道中细菌条带较少,以300bp和500bp左右的条带为特征条带,PCR-DGGE显示肠球菌为优势菌型;大蒜素灌胃小鼠肠道菌群结构组成发生改变,优杆菌属为优势菌型。结论大蒜素预防给药可以扶持肠道中优杆菌属等益生菌生长,抑制肠球菌等病原菌的增殖,调节急性酒精性肝损伤伴有的肠道菌群失调。     

异育银鲫消化道微生物群落对恢复投喂的响应

以实验室养殖的异育银鲫（Carassius auratus gibelio）中科3号成鱼为对象, 利用PCR-DGGE技术比较研究了其经饥饿与恢复投喂后消化道微生物多样性的变化。实验共检测到23条谱带, 其中饥饿组、投喂组谱带数分别为18条和17条（共有谱带12条）, 投喂组大部分谱带亮度高于饥饿组。基于PCR-DGGE所得到的谱带进行的CA （Correspondence Analysis）分析和聚类都表明饥饿组与投喂组消化道微生物存在明显不同。结果表明, 在充分饥饿处理后, 恢复饲料投喂可导致消化道微生物种类减少及微生物平均密度的增加, 同时对消化道微生态系统内微生物种类组成有显著影响。研究为进一步阐述消化道微生物功能基因奠定了基础, 也为水产鱼类肠道菌群研究提供一些基础数据。       

Microbial manipulation of the rat dam changes bacterial colonization and alters properties of the gut in her offspring

The impact of an altered bacterial colonization on gut development has not been thoroughly studied, despite the increased risk of certain diseases with a disturbed microbiota after birth. This study was conducted to determine the effect of microbial manipulation, i.e., antibiotic treatment or Escherichia coli exposure, of the dam on bacterial colonization and gut development in the offspring. Pregnant rats were administered either broad-spectrum antibiotics 3 days before parturition or live nonpathogenic E. coli Culture Collection of University of G?teborg, Sweden type strain (CCUG 29300(T)) 1 wk before parturition and up to 14 days of lactation in the drinking water. Cecal bacterial levels, gut growth, intestinal permeability, digestive enzyme levels, and intestinal inflammation were studied in 2-wk-old rats. Pups from dams that were antibiotic-treated had higher densities of Enterobacteriaceae, which correlated with a decreased stomach growth and function, lower pancreatic protein levels, higher intestinal permeability, and increased plasma levels of the acute phase protein, haptoglobin, compared with pups from untreated mothers. Exposure of pregnant/lactating mothers to E. coli CCUG 29300(T), also resulting in increased Enterobacteriaceae levels, gave in the offspring similar results on the stomach and an increased small intestinal growth compared with the control pups. Furthermore, E. coli pups showed increased mucosal disaccharidase activities, increased liver, spleen, and adrenal weights, as well as increased plasma concentrations of haptoglobin. These findings indicate that disturbing the normal bacterial colonization after birth, by increasing the densities of cecal Enterobacteriaceae, appears to have lasting effects on the postnatal microflora, which affects gut growth and function.     

胆管结扎对大鼠肠道双歧杆菌组成的影响

目的分析胆管结扎对SD大鼠肠道双歧杆菌菌群结构组成的影响。方法采集手术前3 d和手术后2周5只模型组和5只对照组大鼠的粪便样品,提取粪便样品中微生物的混合DNA进行双歧杆菌类群特异性PCR-DGGE,结合主成分分析技术比较2组大鼠在手术前后肠道内双歧杆菌结构组成的变化。结果DGGE图谱及其主成分分析表明手术后对照组和模型组大鼠肠道双歧杆菌菌群的结构组成明显不同,主要表现在假长双歧杆菌的数量在模型组显著增加,而动物双歧杆菌却明显减少。结论胆管结扎导致SD大鼠肠道双歧杆菌结构发生异常变化。