Variation of mucin distribution in the rat intestine, caecum and colon: effect of the bacterial flora.

The influence of the intestinal microflora on mucin types was studied in the small intestine, caecum and colon of conventional (CV) rats as compared to germ-free (GF) rats. A colorimetric method was used on purified water-soluble mucin extracted from mucosal scrapings and contents. Variations occurred between the three anatomical sites both in the mucosas and intestinal contents of GF rats. In CV rats, the presence of the bacterial flora led to different effects depending on the intestinal site: in the small intestinal mucosa, neutral and sulphomucins values were higher whereas sialomucin was much lower. Conversely, sialomucin was higher in the caecal and colonic mucosas and contents whereas sulphated mucins were decreased significantly in caecal contents and caecal and colonic mucosas. These variations in the contents may reflect the bacterial mucolytic activity and the effect of bacterial metabolites on the mucosa.     

Effect of antibiotics on intestinal microflora and production of metabolites

Methodical approaches to detection of relation between intestinal microflora and its metabolites are described. The microbial origin of certain compounds can be asserted by a decrease in their production after exposure to antibacterial drugs or the absence of their production in microbe-free animals. The authors consider that parallel investigation of intestinal microflora and its metabolites after exposure to various agents e.g. narrow spectrum antibiotics or specific substrates is the most accurate methodical approach to detection of their interrelations. Data on the effect of four drugs i.e. kanamycin, metronidazole, cefotaxime and bactrim on production of 10 bacterial metabolites: p-cresol, phenol, indican, acetic, propionic, butyric, isobutyric, valeric, isovaleric and caproic acids in rats are presented. Correlation between the metabolites and the intestinal microflora composition was revealed. It is concluded that detection of microorganisms responsible for production of definite metabolites requires at the maximum: (1) exposure to drugs of different spectra, (2) detection of changes in intestinal microflora by biotope++ and (3) investigation of mucosa microflora which more exactly characterizes metabolism of definite biotops.     

Morphological and physiological characteristics of bacteria inhabiting the intestinal mucosa of pike (<Emphasis Type="Italic">Esox lucius</Emphasis> L.)

Predominance of gram-negative bacteria belong ing to the class Gammaproteobacteria on the intestinal mucosa of pike was determined. The morphophysiological features of the isolated microorganisms suggest that they belong to symbiotic intestinal microflora. The morphological characteristics of the intestinal symbiotic microflora of pike include the formation of the capsule, pseudovacuoles, and spheroplastic forms of cells. The bacteria that were found can produce hydrolytic enzymes and possess persistence properties. The morphophysiological features that were found contribute to the adaptation of bacteria to environmental conditions and indicate that the intestinal microflora offish is autochthonous.     

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。     

鸡肠道菌群总DNA三种提取方法的比较

目的比较不同方法提取鸡肠道菌群总DNA的差异,为分子方法分析肠道菌群组成提供质量较高的DNA模板。方法采用反复冻融法、酶裂解法和试剂盒法（E.N.Z.A Stool DNA Kit）来提取鸡肠道菌群的总DNA,并根据DNA浓度及纯度、16S DNA扩增产物和ERIC-PCR产物所反映的片段多态性4个指标,对这3种方法提取的DNA质量进行比较。结果3种方法均能提取DNA,所得DNA都可以用于16S DNA的扩增,但后2种方法所得DNA的ERIC-PCR结果能反映出更高的菌群多样性。结论试剂盒法和酶裂解法所提取的DNA质量好,适合用于肠道菌群的分子生态研究。     

Effect of the lavage of the digestive tract on microflora in patients with polyps in the large intestine

The microbial status of the intestine and the influence of lavage with polyethylene glycol and balanced electrolyte solution (PEG + E), used in the process of the preparation of patients to polypectomy, on this status were evaluated. The study of microflora was made before oral lavage after, and 48-72 hours later its completion. For control, a group of healthy volunteers, also subjected to oral lavage with PEG + E, was used. The lavage of the digestive tract with PEG + E led to a sharp change in the microbial status in both groups. Some microorganisms, previously absent in the intestine, were found after lavage. The processes of the restoration of intestinal microflora after lavage in healthy volunteers and in patients with polyps had certain differences. In healthy volunteers intestinal microflora was completely restored, and even improved, 48-72 hours after lavage with PEG + E, while at the expiration of this time intestinal microflora in the patients with polyps could be characterized as dysbiotic.     

The normal faecal microflora does not affect the adhesion of probiotic bacteria in vitro

Adhesion of probiotic microorganisms to the intestinal mucosa is considered important for many of the reported health effects. The influence of the endogenous microflora on the adhesion of four probiotic lactobacilli to immobilised intestinal mucus was investigated. It was observed that pre-treatment of the immobilised mucus with faecal extract slightly increased the adhesion of Lactobacillus GG. Pre-treatment of the immobilised mucus with faecal bacteria did not affect the adhesion of the tested strains. These results suggest that the normal microflora may not greatly affect the initial adhesion of the probiotic bacteria. This validates the results of earlier reports where the influence of the normal microflora was not taken into account.     

Use of a semicontinuous culture system as a model for determining the role of human intestinal microflora in the metabolism of xenobiotics

The use of semicontinuous culture system for determining the role of the human intestinal microflora int he biotransformation of xenobiotics is discussed. This model system, which stimulates the lumen of the large intestine, has been used to investigate human intestinal microflora metabolism of compounds representative of three different classes of chemicals; a benzidine-based azo dye, Direct Black 38, a nitropolycyclic aromatic hydrocarbon, 1-nitropyrene and a substituted pyrimidine, 5-fluorocytosine. Metabolites of each of the test compounds were identified, and the kinetics of production and biological activity of the metabolites determined. Metabolic adaptation was observed with 1-nitropyrene and 5-fluorocytosine incubations. This microbial culture system could be quite useful, especially in concert with other in vitro models and animal studies, for determining the pharmacological and toxicological role of the human intestinal microflora in the transformation of xenobiotics. Particular emphasis on the application of this method for toxicological studies in elucidating the role of the intestinal microflora in the etiology of cancer is described.     

黑麂肝、小肠和大肠的组织学结构及Ghrelin的分布

研究了成年雌性黑麂(Muntiacus crinifrons)的肝、小肠和大肠的组织学结构及Ghrelin的分布。采用H.E染色法观察组织学结构,免疫组化PV-9000两步法并结合DAB显色技术确定Ghrelin的分布。结果表明,黑麂的肝组织分为被膜、肝小叶、肝中央静脉、门管区等结构。被膜为浆膜结构,肝小叶不明显。肝细胞以中央静脉为中心,呈放射状排列。肝血窦的形状不规则。肠黏膜上皮为单层柱状上皮,小肠、盲肠和结肠的黏膜肌层很薄,管壁皱襞与肠绒毛等形态在消化道各部也存在差异。免疫组化结果显示肝细胞中有Ghrelin阳性细胞的表达;在肠道,免疫阳性细胞在十二指肠、空肠、回肠、盲肠和直肠的黏膜、黏膜下层和肌层均有分布,尤其在肠绒毛上皮和黏膜下层分布较多。黑麂肝、小肠和大肠结构与哺乳动物基本相似,但无十二指肠腺;Ghrelin阳性细胞在肝、小肠和大肠均有分布,这表明Ghrelin可能对消化有一定的调节作用。     

Investigation of piceid metabolites in rat by liquid chromatography tandem mass spectrometry

There is considerable evidence that stilbenes provide health benefits. Trans-piceid is one of the major stilbenoid compounds in red wine and other plants. The purpose of this study is to investigate the metabolism of piceid in rats, including its conversion product by intestinal microflora in vitro and urinary metabolites. A HPLC-MS/MS method with electrospray ionization (ESI), negative ion mode and collision induced dissociation (CID), was used to elucidate the structures of the major metabolites of piceid. Three metabolites resveratrol, dihydropiceid and dihydroresveratrol were detected after incubating with gut microbiota for 5h. Four urinary metabolites of piceid were identified as resveratrol, dihydroresveratrol monosulfate, piceid monosulfate and piceid monoglucuronide.     

Hydrolytic activity of symbiotic microflora enzymes in pike (<Emphasis Type="Italic">Esox lucius</Emphasis> L.) intestines

This study has revealed the existence of microflora which is, to different degrees, associated with the intestinal mucosa of pike. A total of 82 bacterial strains have been isolated. These microorganisms produce enzymes that hydrolyze the major food substrates (proteins and carbohydrates). These enzymes are produced by the pool of various microorganisms living in the intestines, as well as by separate strains isolated in pure cultures. These strains produce enzymes with different levels of activity. Most of the isolated strains (68%) produce proteases. The calculated values of the C/P factor (the ratio of carbohydrase to protease activity) indirectly testify to the autochthonous nature of the microflora associated with the intestinal mucosa of pike. Presumably, the contribution that the microflora enzymes make to the enzymatic activity of the pike intestine is substantial, but it is difficult to estimate now.     

Bacteriology of the oral cavity of BALB/c mice

To be used as a model in dental research, an animal must fulfil experimental needs and information on the composition and variation of its oral flora must be available. Only limited data are available on the indigenous oral bacterial flora of BALB/c mice. In this work, a total of 671 isolates from different sites (saliva, tongue, teeth, and mucosa) of the oral cavity of BALB/c mice were identified. Only 18 different species were isolated, which indicates the relative simplicity of the flora. The predominant species of the total cultivable flora were "Lactobacillus murinus" (38%), Staphylococcus aureus (37%), Streptococcus faecalis (8%), Staphylococcus sciuri (4%), and Escherichia coli (3%). The other species each represent less than 2% of the flora. "Lactobacillus murinus" is found in greater proportion on mucosa than in the other sites, Staph. aureus predominates in saliva, and Strep. faecalis was found in greater proportion in tooth samples. Statistical analyses, using the minimum percentage of similarity, indicate that there is some variation among the microflora of different mice but that this difference is smaller for mice from the same lot. These results set the basis for the study of the variations of the indigenous oral microflora of BALB/c mice under different conditions.     

Morphological and ultrastructural characteristics of symbiotic bacteria colonizing the surface of the helminth Triaenophorus nodulosus and the intestine of pike Esox lucius

The microflora inhabiting the intestinal mucosa of the pike and the tegument surface of the intestinal parasite Triaenophorus nodulosus is investigated. By means of electron microscopy, the ultrastructural features of bacteria and pattern of their interaction with the colonized surfaces are estimated. Specific distribution of microorganisms belonging to different morphotypes and subdivided into subpopulations on the colonized surfaces is found. Predominance of gram-negative bacteria in all studied microbiocenoses and abundance of nanobacteria in the “superficial” subpopulations are observed. The bacteria of “a deep population” associated with the basal parts of microtrichiae and microvilli are described. It is concluded that the helminthes, the fish parasites possess the normal microflora characterized by specific composition, by ultrastructural features of cells, and by patterns of interaction with the helminth surface. The obtained data characterize the mechanisms of symbiotic interaction of prokaryotes and eukaryotes in the system parasite-host-symbiotic microflora.     

Reconstitution of the gastrointestinal microflora of lactobacillus-free mice

A colony of mice that do not harbor lactobacilli in their digestive tracts but whose intestinal microflora is otherwise functionally similar to that of conventional animals was derived. Methods used to reconstitute the intestinal microflora of the mice included inoculation of the animals with cultures of specific microbes, noncultivable microbes attached to epithelial cells, and cecal contents from conventional mice treated with chloramphenicol. Twenty-six microflora-associated characteristics were monitored by using relatively simple tests to determine the microflora status of the mice.     

Microflora of the upper respiratory tract in young children under normal conditions and in respiratory tract diseases. Microbial count of the upper respiratory tract in healthy children and in children with pneumonia

The results of the inoculation of material taken from the anterior section of the nasal cavity and from the pharyngeal mucosa of 50 healthy young children and 298 acute pneumonia patients were analyzed. 23 microbial species were isolated. In the samples taken from the anterior section of the nasal cavity, monocultures were detected in 86 samples and 54 variants of associations including 2-4 species, in 139 samples. In the samples taken from the pharynx, monocultures were detected in 59 samples and 180 variants of associations including 2-6 species, in 282 samples. Differences in the contamination of the nasal cavity and the pharynx in healthy children and in pneumonia patients were revealed. These differences were manifested in the structure of the microflora (monocultures, associations, their composition), the assortment of microbial species and their concentration. In young children with pneumonia the microflora of the upper respiratory tract was found to reflect the severity of acute pneumonia and the intensity of the pathological process in the lungs (uncomplicated, pyodestructive pneumonia, pyodestructive pneumonia with fatal termination, acute purulent pleurisy).     

Modulatory Effects of <Emphasis Type="Italic">Lactobacillus salivarius</Emphasis> on Intestinal Mucosal Immunity of Piglets

Recent studies have demonstrated that lactobacilli or their cell components can improve certain immune function in animals. The aim of this study is to investigate the effects of porcine lactobacilli on the intestinal mucosal immunity of piglets. Neonatal piglets were used as a model and were orally administrated with Lactobacillus salivarius B1 isolated from the duodenal mucosa of a healthy piglet. The feces of the piglets were collected on days 7, 14, and 21 for intestinal microflora analysis. On day 28, the piglets were sacrificed, and their intestinal mucosa samples were immediately collected to investigate the changes in intestinal morphological and immunocompetent cells. Finally, the expression of cytokines and TLRs was detected in the different intestinal segments. The results indicate that L. salivarius B1 can partially ameliorate the microflora of the feces and increase the number of intestinal immunocompetent cells, as the intraepithelial lymphocyte (P < 0.05), and the IgA-producing cells (P < 0.01) in the lactobacilli-treated group were all increased compared with those in the control group. Enhanced expression of the cytokine IL-6 gene was also observed in the ileum (P < 0.05). Moreover, L. salivarius B1 can also upregulate the expression of TLR2 in the intestinal tract at the gene and protein levels (P < 0.05). The results demonstrate that L. salivarius B1 is beneficial for the maturation of the intestinal mucosal immune system and elicited local immunomodulatory activities. In addition, the modulatory effects of L. salivarius B1 on mucosal immunity mainly depend on its extracellular components.     

Reconstitution of the gastrointestinal microflora of lactobacillus-free mice.

A colony of mice that do not harbor lactobacilli in their digestive tracts but whose intestinal microflora is otherwise functionally similar to that of conventional animals was derived. Methods used to reconstitute the intestinal microflora of the mice included inoculation of the animals with cultures of specific microbes, noncultivable microbes attached to epithelial cells, and cecal contents from conventional mice treated with chloramphenicol. Twenty-six microflora-associated characteristics were monitored by using relatively simple tests to determine the microflora status of the mice.     

Dietary influence of kefir on microbial activities in the mouse bowel

AIMS: In this work the microflora present in kefir, a fermented milk product, was studied together with the effect of kefir administration on different groups of indigenous bacteria of mouse bowel. METHODS AND RESULTS: Kefir microflora was composed of lactic acid bacteria, acetic acid bacteria and yeasts. Yeast population was composed of Saccharomyces cerevisiae, S. unisporus, Candida kefir, Kluyveromyces marxianus and K. lactis. The streptococci levels in kefir treated mice increased by 10-fold and the levels of sulfite-reducing clostridia decreased by 100-fold. The number of lactic acid bacteria increased significantly. CONCLUSIONS: The administration of kefir significantly increased the lactic acid bacteria counts in the mucosa of the bowel. Ingestion of kefir specifically lowered microbial populations of Enterobacteriaceae and clostridia. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first long-term study about the effects of the kefir administration on the intestinal microflora of mice.     

Micro-ecological aspects of studies on the effect of antibacterial drugs on intestinal microflora

Possible characterization of intestinal microflora as an integral system after exposure to antibacterial drugs was studied. Microflora of the contents and mucosa of the jejunum and large intestine in control rats and in rats exposed to metronidazole was studied and numerical indicators characterizing ratios of dominating and accompanying microbial groups in the intestine biotope++ were developed. With this purpose the proportion of the microbial groups in the total quantity of the microbes of a microbiocenosis was determined by the data on microflora quantitative composition. On the basis of detected wide species variety of microorganisms potentially dominating by their biotope numerical limits of the norm were determined only for the microbial groups of the accompanying microflora. The total proportion of the accompanying microbial populations under the normal conditions and the detected measure of deviation (reverse, partial) from the ratio of the dominating and accompanying microorganisms peculiar of the given biotope++ in separate subjects promoted estimation of microbiocenoses of definite biotope of the intestine as a whole.     

多器官功能不全综合征时肠道细菌微生态学改变的实验研究

为研究多器官功能不全综合征（ＭＯＤＳ）时肠道细菌微生态的变化以及Ｇ＾－杆菌消长与肠道和血液中内毒素水平的关系,本实验选用ＳＤ大鼠,应用无菌Ｚｙｍｏｓａｎ腹腔注射制备大鼠ＭＯＤＳ模型,并对ＭＯＤＳ大鼠肠道菌群进行定量分析,对门静脉和外周静脉血中的内毒以及肠道游离内毒素含量进行定量测定。结果发现,ＭＯＤＳ状态下肠道细菌微生态发生明显变化,表现为肠杆菌和肠球菌等肠道内需氧菌的数量明显增多,双歧杆菌和乳酸