Effects of Clostridium difficile, Lactobacillus casei, Bacillus subtilis and Lactobacillus sp. on anomalous lower bowel function on germfree mice

The aim of this work was to study the redressing effects which selected elements of the microbial flora have on intestinal and cardiovascular anomalies of germfree mice. In reference to the enlarged cecum, to the more positive redox potential in cecal contents and to the refractoriness of vascular smooth muscle to various agonist substances observed in germfree rodents, the following were determined after three weeks' microbial association: (i) the weight, (ii) dry percent, (iii) Eh and (iv) epinephrine inhibitory activity of ‘alpha pigment’ in cecal contents. The results indicated that none of the tested micro-organisms changed the weight and dry percent of cecal contents of the ex-germfree mice in comparison to germfree controls. L. casei caused a slight shift to less positive Eh values. Cl. difficile and L. casei as mono-associates reduced the activity of alpha pigment by approx. 80% in comparison to germfree controls. These findings suggest that in malfunctions which develop in gastrointestinal and cardiovascular aspects of rodents in the absence of the flora, specific microbial associates effect their remedial action by correcting specific details in various anomalous functions.     

4种常见淡水鱼肠道菌群的分析

为了解山东省东平湖库区常见淡水鱼肠道菌群的组成, 作者对该库区内的草鱼、鲢鱼、鲫鱼和鲤鱼4 种鲤科鱼肠道菌群中的需氧菌或兼性厌氧菌进行分离鉴定,并对每种鱼肠道中的菌群数量和种类组成进行分析。结果表明,在上述鱼肠道中均存在哈夫尼亚菌属、致病杆菌属、气单胞菌属、柠檬酸菌属、芽孢杆菌属和链球菌属,其中鲤鱼和鲫鱼肠道菌群的总量比较接近,两者共同含有菌属的数量差别不大。     

圈养大熊猫食性转换前期肠道细菌多样性

为了探讨大熊猫食性转换前期肠道菌群的多样性与差异性。对3只处于夏季食性转换前期大熊猫粪便细菌总DNA进行ERIC—PCR分析,并构建16SrDNA克隆文库,采用限制性内切酶HinfI、MsPI对其进行RFLP（restric—tionfragmentlengthpolymorphism）及测序分析。研究表明,食性转换前期大熊猫肠道细菌主要由硬壁菌门和变形菌门组成。其中7月份样品中以梭菌属和醋杆菌属为主;8月份以八迭球菌属和埃希氏菌属为主;9月份以链球菌属和埃希氏菌属为主。此外在3个月中均发现一定比例的未培养细菌。结果表明：随着食性转换前期的过渡,大熊猫肠道菌群发生了改变。由硬壁菌门中的梭菌属渐变到以球菌属为主,由变形菌门的醋杆菌属渐变到以肠杆菌为主。     

圈养老年大熊猫肠道菌群16SrDNA克隆文库的建立

为了研究老年大熊猫肠道菌群的构成,本试验对3只圈养老年大熊猫粪便细菌构建16S rDNA克隆文库,采用限制性内切酶Hinf Ⅰ、Msp Ⅰ对其进行限制性片段长度多态性(restriction fragment length polymorphism,RFLP)及测序分析.结果显示,老年大熊猫肠道细菌主要由变形菌门和厚壁菌门组成;变形菌门中又以大肠埃希氏菌属为主,其次为假单胞菌属、志贺氏菌属、气单胞菌属;而厚壁菌门中以链球菌属为主,其次为魏斯氏菌属、梭菌属;此外还发现一定比例的未培养细菌.本试验第一次建立了较丰富的老年大熊猫肠道菌群的克隆文库,为分析比较各年龄层大熊猫的肠道菌群结构的异同提供了参照,也为合理饲喂老年大熊猫,保障老年大熊猫的健康提供重要信息.     

阿拉善马鹿冬季肠道菌群组成的性别差异

肠道菌群对于动物消化及健康有着至关重要的作用,但在性别差异上的研究仍然有限。本研究通过高通量测序技术对贺兰山阿拉善马鹿(Cervus elaphus alashanicus)的20个粪便样本进行多样性分析,雌雄各半,并讨论性别因素对肠道菌群的影响。结果表明,阿拉善马鹿雌雄肠道菌群的多样性及结构均存在差异,且雌性肠道菌群的多样性更高。菌群构成的门水平上,雄性肠道菌群的优势菌门为厚壁菌门(Firmicutes 49.47%)和疣微菌门(Verrucomicrobia 14.21%),雌性肠道优势菌门为厚壁菌门(54.47%)和变形菌门(Proteobacteria 54.41%)。在属水平上,雌雄肠道菌群的优势菌属都为孢杆菌属(Sporobacter),含量为(雌性:6.79%,雄性:7.97%),拟杆菌属(Bacteroides),含量为(雌性:3.87%,雄性:4.42%),NMDS分析及LEfSe分析表明,性别因素对阿拉善马鹿雌雄肠道菌群的影响存在显著差异(P<0.05)。本研究表明性别因素对阿拉善马鹿冬季肠道菌群的多样性及构成存在影响,可为深入研究性别因素对阿拉善马鹿肠道菌群的影响提供依据。     

Effects of exocrine pancreatic insufficiency and replacement therapy on the bacterial flora of the duodenum in dogs

The influence of pancreatic secretions on the bacterial flora of the small intestine in 6 dogs was investigated by determining effects of exocrine pancreatic insufficiency on numbers and types of bacteria in duodenal juice, and by examining the subsequent response to dietary supplementation with bovine pancreatic extract. Exocrine pancreatic insufficiency was induced by ligation of pancreatic ducts and was confirmed by indirect assessment of exocrine pancreatic function. Duct ligation was followed by large increases (P less than 0.01) in total numbers of bacteria, reflecting increased numbers particularly of Lactobacillus spp and Streptococcus spp, in 3 dogs accompanied by obligate anaerobes. Total numbers of aerobes and anaerobes decreased markedly (P less than 0.05) after supplementation with bovine pancreatic extract to values that were not significantly different from those determined before duct ligation. Exocrine pancreatic insufficiency therefore resulted in small intestinal bacterial overgrowth that was reversed by pancreatic replacement therapy, indicating that pancreatic secretions can have an important influence on the small intestinal bacterial flora of dogs.     

Protection of chicks against Salmonella infantis infection induced by strict anaerobically cultured intestinal microflora

Summary

In two experiments newly hatched broilers were orally inoculated either with intestinal microfloras cultured under different conditions or with a suspension of intestinal homogenate and challenged with high doses (3 x 10⁵ c.f.u. per chick) of Salmonella infantis organisms.

Inocula were prepared from intestinal material of mature SPF WL hens under aerobic or strictly anaerobic conditions (< 5 ppm oxygen), and protected against atmospheric oxygen during storage. A very significant reduction of the number q/Salmonella infantis bacteria in caecal content was achieved one and two weeks after challenge in those groups of chicks which were inoculated with anaerobically cultured flora or with intestinal homogenate. In order to induce protection, obligate anaerobes had to be accompanied by facultative anaerobes. Amprolium or Monensin dit not interfere with protection. Protection induced by intestinal microflora depends on anaerobiosis during preparation, culturing, storage, and application of the protective bacteria. Starvation on the first day of life appeared to have a negative effect on protection.     

不同乳酸菌发酵乳对高血脂症大鼠肠道菌群的影响

试验旨在探究不同乳酸菌发酵乳对高血脂模型大鼠血脂与肠道菌群的调节作用。将体重为(120±10)g的 40 只 SD 雄性大鼠随机分为空白对照组(C 组)、高脂模型组(M 组)、L. rhamnosus LV108 发酵乳组(LV组)及混合乳酸菌(LV108 发酵乳∶grx08 发酵乳∶grx12 发酵乳=2∶1∶1)发酵乳组(H 组),每组 10 只,建模后干预(1 mL/100 g,1×108 CFU/mL)4 周,测定大鼠血脂及大鼠粪样细菌 16S rDNA 的 V3-V4 区,并对大鼠肠道微生物和血清血脂水平的相关性进行分析。结果表明:LV 组和 H 组大鼠血清中甘油三酯、总胆固醇及低密度脂蛋白胆固醇含量显著降低,体重增长被抑制;LV 组和 H 组中大鼠肠道菌群的丰度及多样性显著提高,且 H 组大鼠肠道菌群的丰度高于 LV 组(P<0.05);LV 组和 H 组大鼠粪便中 Lactobacillus spp.、S24-7 spp 和 Treponema spp.等菌属显著升高,Peptostreptococcus spp.、Clostridium spp.、Turicibacter spp、Escherichia spp.及 Desulfovibrio spp.等菌属显著降低。结果提示,L. rhamnosus LV108发酵乳及混合乳酸菌发酵乳均能改善大鼠血脂水平并恢复大鼠肠道菌群的多样性及菌群构成。     

肠道菌群对动物肠黏膜免疫的调控作用

动物肠道共生着数量庞大、结构复杂的菌群,而肠壁内存在着为数众多、功能强大的黏膜淋巴细胞。肠道菌群具有促进肠黏膜免疫系统生长与发育和调控肠黏膜屏障与免疫功能的双重作用。本文主要从动物肠道菌群的定植与特性、肠黏膜免疫细胞的功能以及肠道菌群对肠黏膜屏障与免疫功能的调控作用进行综述。     

不同饲养密度对笼养蛋鸡十二指肠肠道菌群的影响

本试验采用聚合酶链式反应(PCR)-变性梯度凝胶电泳(DGGE)方法探究不同饲养密度对笼养海兰灰蛋鸡十二指肠肠道菌群的影响。随机选取1 250只11周龄海兰灰蛋鸡,按照不同饲养密度随机分为5个组:A组900 cm2/只、B组675 cm2/只、C组540 cm2/只、D组450 cm2/只、E组380 cm2/只,每个组内50个重复。5个组均在相同的环境和饲养管理模式下进行饲养。分别于16、26、50周龄时,在每个组中随机选取5只鸡,取十二指肠内容物,用PCR-DGGE法进行肠道菌群分析。结果表明,在同一个饲养密度内,随着饲养周龄的增加笼养蛋鸡十二指肠肠道菌群结构相似系数逐渐减小;16周龄时高密度组与低密度组之间十二指肠肠道菌群结构相似性很高,即肠道菌群结构差异较小;26和50周龄时高密度组与低密度组之间十二指肠肠道菌群结构相似性很低,即肠道菌群结构差异很大。由此可见,随着饲养周龄的增加笼养蛋鸡十二指肠肠道菌群结构差异增大;2种有益菌(Lactobacillus gastricus和Lactobacillus alvi)在高饲养密度组(D组和E组)中消失,饲养密度高于450 cm2/只对笼养蛋鸡十二指肠肠道菌群结构具有不利的影响。     

微生态制剂对仔猪肠道微生态调控的研究与应用进展

微生态制剂是一种安全、无毒、无污染、无残留的生物类饲料添加剂,具有提高仔猪生产性能、减少腹泻、增强机体免疫力、调节肠道菌群数量和维持肠道微生态平衡等功能,对仔猪肠道微生态的调节起着重要的作用。本文从微生态制剂的分类、作用机理、对肠道的作用等阐述国内外有关微生态制剂在仔猪中的应用及前景。     

日粮中添加恩拉霉素对仔猪生产性能和肠道菌群的影响

试验研究日粮中添加恩拉霉素及停用后对断奶仔猪生长性能和肠道菌群结构的影响。采用单因素试验设计,将20头仔猪分为4个处理,每个处理5个重复;分别在基础饲粮中添加0、5、20 mg/kg和80 mg/kg恩拉霉素。试验分为3个阶段,加药期28 d,饲喂试验日粮;停药期7 d和LPS诱导应激期3 d,均饲喂基础日粮。结果表明:加药期与对照组相比,5、20mg/kg添加组平均日增重增加(P>0.05),80mg/kg添加组却显著降低(P<0.05),3个添加组直肠总菌数显著下降(P<0.05)。停药期,5、20 mg/kg添加组平均日增重与对照组相比有下降的趋势(P>0.05),20、80mg/kg添加组直肠总菌数和乳酸杆菌/大肠杆菌显著下降(P<0.05)。应激后,盲肠乙酸和丁酸含量显著下降(P<0.05)。     

肠道寄生虫与菌群互作关系研究进展

肠道菌群是动物机体的重要组成部分，与寄生虫和宿主的关系十分密切。寄生虫感染往往会对宿主产生不利影响，其致病作用与寄生虫种类、寄生部位及其与宿主的相互作用有关。寄生虫感染可导致肠道微生态的改变、失调和炎症性疾病；肠道菌群也影响寄生虫在宿主体内的定植、增殖和毒力。肠道寄生虫包括寄生蠕虫和原虫，近年来对肠道寄生虫与菌群相互作用研究发现，无论是原虫还是蠕虫感染均可导致肠道微生物组成和多样性的改变，对宿主产生致病作用，而蠕虫对炎症性肠病的潜在治疗作用一度颠覆了以往人们对于寄生虫的认知。肠道菌群可促进或抑制寄生虫对宿主的致病作用，且益生菌对肠道寄生虫感染有一定的预防或治疗作用。目前对寄生虫与菌群相互作用的研究仍然处于初期阶段，对其相互作用的机制尚不清楚。明确寄生虫与肠道菌群的相互影响及其作用机制，对深入理解寄生虫－肠道菌群－宿主的相互关系、研发有效的抗寄生虫微生态制剂具有重要意义。作者就近年来肠道蠕虫和原虫与菌群相互作用的研究进展进行了综述，并对该领域未来的研究方向进行展望，以期为预防和控制寄生虫病提供新的策略和理论依据。     

饲粮添加酶解蛋白肽对断奶仔猪生长性能、血清免疫指标和肠道菌群的影响

本试验旨在探究饲粮添加酶解蛋白肽对断奶仔猪生长性能、血清免疫指标和肠道菌群的影响,以确定断奶仔猪饲粮中酶解蛋白肽的适宜添加水平.试验选取150头21日龄体重为(7.01±0.17)kg的健康"杜×长×大"三元杂交断奶仔猪,随机分为5个组,每组5个重复,每个重复6头(公母各占1/2).各组断奶仔猪分别饲喂在基础饲粮中添加...     

术苦芩总多糖对湿热泄泻仔猪肠道菌群和免疫功能的影响

旨在探究术苦芩总多糖(ZKQPs)对湿热泄泻仔猪肠道菌群和免疫功能的影响。通过HPLC分析ZKQPs的单糖组成,体外抑菌试验研究ZKQPs体外对常见肠道致病菌的直接影响,同时建立仔猪腹泻模型,用白头翁散(PP)及3种剂量的ZKQPs (25、50和75 mg·kg^-1)治疗腹泻仔猪,治疗结束后,收集小肠各段组织、盲肠内容物,对菌落进行鉴定与计数;通过16S rDNA高通量测序分析肠道菌群的变化,测定小肠免疫相关细胞因子mRNA表达。结果显示:ZKQPs一级结构主要由鼠李糖、葡萄糖、半乳糖和木糖构成。在体外试验中,大肠杆菌、沙门菌、猪源产气荚膜梭菌肠道常见有害菌对ZKQPs敏感,并在一定范围内呈现浓度依赖性。动物试验中,ZKQPs有效促进了腹泻仔猪肠道乳酸杆菌的生长,对致病大肠杆菌生长有较强的抑制作用,其中,50 mg·kg^-1ZKQPs影响最为显著。16S rDNA高通量测序结果表明,ZKQPs调节了腹泻仔猪菌群Alpha多样性,对肠道菌群在纲和属水平上有显著影响,腹泻仔猪芽胞杆菌纲、乳酸杆菌属相对丰度显著下降,梭菌纲相对丰度显著上升;ZKQPs治疗后芽胞杆菌纲、乳酸杆菌属相对丰度均显著上升且高于正常值,梭菌纲相对丰度极显著下降,显著改善了腹泻仔猪肠道菌群结构。仔猪腹泻时,小肠各段Th1细胞因子IL-2 mRNA水平上升(P<0.01),Th2细胞因子IL-4、IL-5 mRNA水平均下降;ZKQPs治疗后,腹泻仔猪小肠各段IL-2 mRNA水平下降,IL-4、IL-5 mRNA水平普遍上升,且不同细胞因子mRNA水平在小肠不同位置变化有所差异,50和75 mg·kg^-1ZKQPs治疗后对其影响最为显著,综合分析50 mg·kg^-1ZKQPs治疗腹泻仔猪对细胞因子mRNA水平影响最大。综上表明,ZKQPs在体外对常见肠道致病菌生长具有良好的抑制作用,可以改变腹泻仔猪肠道微生物群的丰富度和多样性,提高乳酸杆菌属的相对丰度,并改善菌群结构,同时调节肠道免疫反应,有效增强了仔猪抗腹泻能力。     

The intestinal flora of the chicken in the period 2 to 6 weeks of age, with particular reference to the anaerobic bacteria

A study has been made of the intestinal flora of chickens aged between 2 and 6½ weeks. No major differences were found when rations containing 9 or 25% fish meal were compared or when two different sources of fish meal were used. Throughout the period investigated, it was confirmed that the lactobacilli are the only group of organisms generally present in the small intestine in numbers exceeding 104/g. Clostridia were regularly found at 102 to 104/g, but Clostridium welchii was isolated from only occasional samples.

In the caeca, the total number of bacteria present at 2, 3, 4 and 6½ weeks was about 1011/g. Of these it was generally possible to isolate more than 20% using an anaerobic roll‐tube technique and oxygen‐free CO2. Many different types of anaerobes were isolated including strictly anaerobic budding bacteria which had not previously been studied. During the growth of the bird the flora changed. Anaerobic streptococci (peptostreptococci) predominated at 2 weeks but gradually decreased so that by 6½ weeks these organisms formed only a small part of the flora. Many of the Gram‐negative non‐sporing anaerobes only appeared between the 4th and 6th weeks.

Throughout the 2 to 6½‐week period all the caecal samples examined contained 108 to 109 anaerobes/g which were able to attack uric acid and sometimes 1010/g. This property was shared by at least one group of peptostreptococci, four groups of Gram‐negative non‐sporing anaerobes, three groups of Gram‐variable or Gram‐positive anaerobic non‐sporing rods and several species of clostridia.     

肠道菌群与动物免疫的相互关系

肠道菌群形成了一个庞大而复杂的微生态系统,与动物免疫之间的关系极为密切.正常的肠道菌群具有屏障作用,与肠道黏膜的免疫作用、免疫器官的生长发育及免疫因子的激活都有关系.本文对肠道菌群的分布、肠道菌群对动物免疫的影响及抗生素、肠道菌群与动物免疫之间的相互作用等方面做了综述.     

果寡糖对蛋雏鸡生长性能和肠道菌群的影响

试验选择7日龄海兰褐蛋雏鸡192只,采用随机区组法设计,设3个处理,每处理4个重复,每重复16羽,A组为抗生素组:基础日粮+杆菌肽锌20 mg/kg+粘杆菌素4 mg/kg,B组为果寡糖组:基础日粮+0.1%果寡糖(FOS),对照组(CT):基础日粮。通过饲养试验和肠道菌群培养,研究果寡糖替代抗生素对蛋雏鸡生长性能和肠道菌群数量的影响。结果表明:果寡糖组(B组)和抗生素组(A组)的生长性能均显著优于对照组(CT组),且二者之间差异不显著;果寡糖比对照组和抗生素组显著增加了盲肠和回肠内容物中双歧杆菌和乳酸杆菌数量。     

Incorporation of nitrogen-15 from dietary [15N] diammonium citrate into amino acids of liver and muscle proteins in germfree and specific-pathogen-free neonatal pigs

Incorporation of 15N from dietary [15N]diammonium citrate ([15N]DAC) into amino acids isolated from hydrolyzed tissue proteins was investigated in the presence or absence of intestinal flora in neonatal pigs. The 15N was incorporated into all of the essential and nonessential amino acids from liver and muscles in one of two germfree pigs and two specific-pathogen-free pigs which were killed on the 5th day after the administration of [15N]DAC. But, a higher 15N concentration than natural abundance of 15N was not detected in histidine, lysine, and threonine from these tissues in another germfree pig killed the next day. Nitrogen transfer from DAC into all amino acids, including these three essential amino acids, may be possible in the specific-pathogen-free pig and even in the germfree pig.     

红芪多糖-1-1的分离纯化及其调节抗生素诱导小鼠肠道菌群失调的最佳剂量分析

旨在筛选红芪多糖-1-1（Radix Hedysari polysaccharide-1-1,RHPS-1-1）调节小鼠肠道菌群失调的最佳剂量。将红芪粗多糖分别用DEAE-52和Sephadex G-100进行分离纯化得单一多糖RHPS-1-1,采用高效阴离子交换色谱法及甲基化和红外光谱法鉴定其单糖组成与结构。将C57BL/6小鼠90只随机分为正常对照组、模型组、自愈组和不同剂量RHPS-1-1（12.5、25、50、100、200和400 mg·kg^-1）给药组,每组10只。通过连续14 d灌服抗生素鸡尾酒（氨苄西林、新霉素、甲硝唑、万古霉素）的方法建立小鼠肠道菌群失调模型,造模结束后,模型组小鼠处死采样,各给药组灌服不同剂量RHPS-1-1进行治疗,正常对照组与自愈组给予等量生理盐水,连续14 d。应用16S rDNA高通量测序法分析各组小鼠盲肠内容物的肠道菌群变化特征,并观测正常对照组、自愈组和25 mg·kg^-1 RHPS-1-1组主要脏器的器官指数和组织病理变化。结果表明,经纯化得到单一多糖RHPS-1-1,重均分子量为19.420 ku,主要由葡萄糖组成,主链连接方式为1,4-D-Glcp,并具有植物多糖的特征吸收峰。灌服抗生素鸡尾酒使小鼠出现了严重的肠道菌群失调,拟杆菌门、厚壁菌门和放线菌门等优势菌丰度极度减少,变形菌门相对增多,且不能自然恢复;经RHPS-1-1给药后肠道菌群的多样性和丰富度发生改变,25 mg·kg^-1 RHPS-1-1治疗组Chaol指数显著升高（P<0.05）,PCA和UPGMA聚类分析结果均显示,25 mg·kg^-1的RHPS-1-1给药组与正常对照组肠道菌群结构最接近,有害菌Muribaculaceae_unclassified相对丰度降低,有益菌Ruminococcaceae_UCG-014和Clostridiales_unclassified相对丰度增加。自愈组相比正常对照组仅肝脏指数显著减小（P<0.05）,经25 mg·kg^-1的RHPS-1-1治疗后显著升高（P<0.05）。但正常对照组、自愈组及25 mg·kg^-1的RHPS-1-1治疗组的心、肝、脾、肺、肾和脑均无明显病理变化。结果证明,所获RHPS-1-1是一种分子量约为19.420 ku的植物葡聚糖,治疗剂量为25 mg·kg^-1时,促进抗生素所致小鼠肠道菌群失调恢复的效果最佳,且可改善肝脏指数下降,本研究为红芪多糖调节肠道菌群的临床应用提供依据。