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Intestinal bacteria antagonistic to Clostridium difficile in mice.

Overgrowth by Clostridium difficile has been reported in conventional mice injected intraperitoneally with ampicillin. In this study, we aimed to determine which types of indigenous intestinal bacteria were eliminated by ampicillin to allow overgrowth by C. difficile. C. difficile overgrowth was associated with a decrease in the numbers of lactobacilli, an increase in bacteroidaceae and a slight decrease in the frequency of isolation of fusiform-shaped bacteria (clostridia). C. difficile cytotoxin was detected in caeca from mice in which the numbers of C. difficile were greater than 10(5) per gram of faeces. Gnotobiotic mice were inoculated with various groups of intestinal anaerobes to determine which members of the indigenous flora would antagonize C. difficile. Gnotobiotic mice inoculated with three strains of lactobacilli, 37 strains of bacteroides or 46 strains of clostridia isolated from limited-flora mice were unable to eliminate C. difficile. C. difficile was eliminated, however, from the gastrointestinal tracts of gnotobiotic mice inoculated with whole faeces or chloroform-treated faeces from conventional mice or whole faeces from limited-flora mice containing only clostridia.

Ampicillin↗

Gastrointestinal flora of cotton rats.

The gastrointestinal (GI) flora of cotton rats was examined. No lactobacilli were detected in any part of the GI tract. Anaerobes, including Peptococcaceae, Bacteroidaceae, bifidobacteria and eubacteria, were the predominant bacteria in the stomach, small intestine, caecum and faeces. Aerobes and facultative anaerobes, including Enterobacteriaceae and streptococci, were detected at low numbers and very low frequency of occurrence in all parts of the GI tract. Sixty-one isolates of bifidobacteria were recovered from the stomach, small intestine, caecum and faeces of cotton rats. They were identified as Bifidobacterium animalis, B. pseudolongum biovar a and b. The study showed that the GI flora of cotton rats seem to be very different from the GI flora in other rodents.

Animals↗

A mixture of organisms affects cholesterol metabolism together with rat cecal flora.

The effects of a mixture of organisms on cecal fermentation and cholesterol metabolism in sham-operated and cecectomized rats were investigated. Male F344 rats, allocated into four groups: cecectomized rats fed a mixture of organisms (CEMO), cecectomized rats fed rice bran (CERB), sham-operated rats fed a mixture of organisms (SHMO), and sham-operated rats fed rice bran (SHRB) for 4 weeks. The diets had 0.5% cholesterol and 0.125% sodium cholate added. There were no significant differences in the body weight gain and food intake among the groups. The cecal pH in the SHMO group was significantly lower than that in the other groups. The total cholesterol and (VLDL + IDL + LDL)-cholesterol concentrations in serum were significantly lower in the SHMO group than that in the SHRB group, and the triacylglycerol concentration in the sham-operated rats tended to decrease compared to the cecectomized rats. The fecal cholesterol excretion in the CERB group was higher than that in the other groups, and that in the SHMO group was significantly higher than in the SHRB group. The acetic acid, propionic acid, n-butyric acid, and total short-chain fatty acid concentrations in the cecum contents were significantly higher in the SHMO group than those in the other groups. Streptococcus, Bifidobacterium, and Lactobacillus in the SHMO group tended to be higher than the other groups and Bacteroidaceae in the CEMO and CERB groups were significantly higher than that in the SHMO group. The results demonstrate that the mixture of organisms was fermented with the cecal contents and that the metabolites such as short-chain fatty acid lowered the serum total cholesterol and liver cholesterol concentrations in the rats fed a cholesterol-containing diet.

Animals↗

Effect of tea polyphenols on fecal flora and fecal metabolic products of pigs.

The effects of tea polyphenols on fecal flora and fecal metabolic activities were studied in eight 30-day-old pigs fed a basal diet (control) or the diet supplemented with 0.2% of tea polyphenols for 2 weeks. During tea polyphenol administration compared to before administration, the levels of lactobacilli were increased significantly (P < 0.05 and P < 0.01 on days 7 and 14 of administration, respectively), whereas the levels of total bacteria (P < 0.01) and bacteroidaceae (P < 0.01 and P < 0.001 on days 7 and 14 of administration, respectively) were decreased significantly. The detection rate of lecithinase positive clostridia including Clostridium perfrigens showed a tendency to decrease during administration. No detectable change occurred in the levels of other organisms throughout the experimental period. Fecal concentrations of ammonia were decreased significantly (P < 0.05 and P < 0.01 on days 7 and 14 of administration, respectively). Fecal phenol (P < 0.001), p-cresol (P < 0.05), and skatol (P < 0.01) were reduced significantly on day 14 of administration compared to before administration. The amounts of fecal short chain fatty acids, acetic acid (P < 0.05) and lactic acid (P < 0.05) were increased significantly on day 14 of administration. The pH values were decreased slightly during administration. The odour of the feces was reduced remarkably during tea polyphenol administration.

Animals↗

Establishment of specific pathogen-free rabbit colonies with limited-flora rabbits associated with conventional rabbit flora, and monitoring of their cecal flora.

In the present study we attempted to establish specific-pathogen-free (SPF) rabbit breeding colonies with two groups of limited-flora (LF) rabbits, both ex-germfree rabbits, and their offspring. Two groups of LF rabbits associated with cecal flora of conventional (CV) rabbits produced in a previous study [Exp. Animals, submitted], were transferred to individual barrier rooms and some of the LF rabbits were accommodated in isolators to maintain the basic flora for SPF rabbits. The composition of the cecal flora of LF rabbits was stable for a long period; bacteroides remained predominant and clostridia dominant. From the SPF rabbits, different types of bacteria, e.g., enterobacteriaceae and streptococci, which could not be isolated in the isolator were detected at a low population level at an early stage in the establishment of the SPF colonies, but the basic composition of the cecal flora was mainly bacteroidaceae and clostridia and did not change over a long period, and the floral composition became similar to that of CV rabbits. The fertility and weaning rates of the SPF rabbits were satisfactory for a SPF rabbit colony. In addition, these SPF colonies were free of more than one year rabbit-specific pathogens.

Animals↗

Bile acid metabolism, bacterial bowel flora and intestinal function following ileal pouch-anal anastomosis in dogs, with reference to the influence of administration of ursodeoxycholic acid.

The pathophysiology following a total colectomy with ileal pouch-anal anastomosis (IPAA) has not been sufficiently clarified yet. We investigated bile acid metabolism, bacterial bowel flora and transit of the alimentary tract after IPAA, with reference to administration of ursodeoxycholic acid (UDCA) in dogs undergoing IPAA. Ten adult beagle dogs underwent IPAA at one stage, and were observed for 12 months. UDCA (100 mg/day) was administered orally to five dogs, and the other five did not. In the UDCA(+) group, UDCA replaced other bile acids, especially cholic acid, accounting for 16.5% of gallbladder bile at 12 months after surgery. Both plasma levels and postprandial increase of total bile acids remained unchanged in the UDCA(+) group, but decreased in the UDCA(-) group at 12 months. Fecal excretion of bile acids tended to be smaller in the UDCA(+) group, and the ratio of secondary to primary bile acids was larger in the UDCA(-) group. Almost all the bile acids were in free form in stool, and UDCA constituted 19% in the UDCA(+) group. The transit time of the whole alimentary tract was elongated by administering UDCA, especially at an early period after IPAA. Although both anaerobic and aerobic bacteria decreased after IPAA, the latter decreased more in stool, resulting in an increase in the ratio of total anaerobes/total aerobes, especially in the UDCA(-) group. The decrease in Bacteroidaceae and Lactobacillus after IPAA was slightly smaller in the UDCA(+) group. Administration of UDCA following IPAA was efficient to induce rapid intestinal adaptation and also to keep the bile acid fraction in the ileal pouch less harmful.

Administration, Oral↗

Succinate accumulation in pig large intestine during antibiotic-associated diarrhea and the constitution of succinate-producing flora.

Succinate was the major organic acid detected in the hindgut content of pigs suffering from antibiotic-associated diarrhea. Antibiotic-associated diarrhea was induced by an oral dose of polymyxin B sulfate (3,000,000 units/day) or an intramuscular injection of enrofloxacin (0.6 g enrofloxacin/day). In the large intestine of enrofloxacin-treated pigs, Gram-negative facultative anaerobic rods phylogenetically related to Escherichia coli and Gram-positive facultative anaerobic non-spore-forming rods phylogenetically related to Lactobacilli were isolated as succinate producers. Succinate-producing Lactobacilli were only isolated as the succinate producer in polymyxin B sulfate-treated pigs. In contrast to antibiotic-associated diarrhea pigs, bacteria belonging to Bacteroidaceae, Fusobacteria, and Enterobacteriaceae were detected as succinate producers in a non-treated pig. In antibiotic-associated diarrhea conditions, antibiotic-resistant Enterobacteria, E. coli in particular, and Lactobacilli may contribute to an abnormal succinate accumulation and may affect water absorption in the hindgut that relates to an expression of antibiotic-associated diarrhea.

Animals↗

Does the hygroscopic property of the laminaria tent imply a risk for ascending infection in legal abortions? A microbiological study.

In 53 women admitted for first-trimester abortion, the cervical canal was dilated with laminaria tents prior to vacuum aspiration. Before insertion of the tents, endocervical specimens were taken for microbiological investigations, including Chlamydia trachomatis, and both aerobic and anaerobic microorganisms. The laminaria tents, widened by their hygroscopic properties, were removed just before the vacuum aspiration and sent for cultivation of aerobic and anaerobic microorganisms. The Gram-positive aerobic cocci, Staph. epidermidis, Strept. faecalis and Strept. agalactiae as well as Escherichia coli and Klebsiella/Enterobacter were the most frequent microorganism among the aerobic isolates from the endocervices as well as from the laminaria tents. These microorganisms were identified on the tents in greater numbers than in the cervices. Staph. aureus was isolated from three laminaria tents, but only one of these 3 women was proved to be primarily colonized endocervically. Among the anaerobes, Bacteroidaceae followed by peptococci and peptostreptococci were the predominant genera found in the cervix as well as on the removed laminaria tents. These three groups of anaerobic microorganisms were found markedly less frequently on the laminaria tents than in the endocervix. On the other hand, Strept. intermedius, another anaerobe of possible enteric origin, was recovered from 9 laminaria tents but not in the cervices. Since no cases of clinically verified endometritis/salpingitis were registered in the present study it is concluded that the risk of a pelvic inflammatory disease (PID) due to the laminaria tent is negligible as long as the laminaria treatment does not exceed 24 hours.

Abortion, Induced↗

Effects of transgalactosylated disaccharides on the human intestinal microflora and their metabolism.

The effects of transgalactosylated disaccharide (TD) intake on human fecal microflora and their metabolism were investigated in 12 Japanese males. TD is a mixture of sugars, galactosyl galactose, and galactosyl glucose, synthesized from lactose through the transgalactosylation reaction of Streptococcus thermophilus beta-galactosidase. Volunteers took 15 g of the test sugar daily for 6 days. The TD ingestion increased the number of bifidobacteria and lactobacilli, but decreased the number of Bacteroidaceae and Candida spp. in the feces. The ratio of bifidobacteria to total bacteria increased from 0.28 to 0.51. TD decreased the fecal concentrations of propionic acid, isobutyric acid, isovaleric acid, and valeric acid. This sugar also lowered the fecal pH, and the concentrations of fecal ammonia, p-cresol, and indole. Moreover, a positive correlation was found between the concentration of ammonia, and that of branched-chain fatty acids (isobutyric acid and isovaleric acid), p-cresol, and indole. All of these compounds are produced from amino acids through deamination by the intestinal bacteria. The depression of amino acid fermentation by intestinal bacteria may be involved in the reduction of fecal ammonia. These results suggest that a part of the transgalactosylated disaccharides passes into the colon, inducing changes in the colonic microflora composition, hastening carbohydrate fermentation, and depressing amino acid fermentation in the human gut.

Adult↗

Studies on the relationship between intestinal flora and cecal coccidiosis in chicken.

The bacteriological and histopathological changes in the ceca of young chickens after being infected with sporulated oocysts of Eimeria tenella were investigated. Lactobacilli and bifidobacteria showed remarkable decrease in number on the 5th day after infection, when shizont and gametocyate came to appear, and destruction of mucosa along with severe haemorrhaging was noticed. Other predominant bacteria like bacteroidaceae, catenabacteria and peptostreptococci showed only moderate and temporal decrease in number during the infection. Clostridium perfringens prolifereated after the 5th day post infection following the decline of lactobacilli and bifidobacteria. Proliferation of clostridia was so intense that the number was almost million times greater than that of the uninfected chicken at its peak on the 7th day after infection. Enterobacteria also showed a moderate but persistent increase in number. Most bacteria came to the normal population on the 10th day after infection. As for lactobacilli, bifidobacteria, clostridia and enterobacteria, the disturbances were still noticeable on the 17th day after injection. Antagonism between proliferated bacteria (clostridia and enterobacteria) and lactic acid bacteria (lactobacilli and bifidobacteria) whose intensity is suppose to be related to the cecal mucosa is suggested during cecal coccidiosis in chickens.

Actinomycetaceae↗

Enteral Nutrition Is Associated with a Distinct Gut Microbiome Composition and Fermentation Capacity Profile After Acute Colonic Injury in Rats.

Enteral nutrition (EN) is known to promote mucosal healing in inflammatory bowel disease, and multi-omics data suggest that the gut microbiome mediates its therapeutic effects. However, the impact of EN and its components on the gut community during recovery from acute epithelial injury remains incompletely understood. We used whole-genome metagenomic sequencing to investigate the effect of an EN formula based on extruded amaranth flour and pea protein on the gut microbiome in a dextran sulfate sodium (DSS) rat model of acute colonic injury. Three groups were compared, as follows: an unchallenged control (n = 9) with standard chow, a colonic injury (5% DSS; n = 9) group with standard chow, and a colonic injury (5% DSS; n = 9) group with EN. Injury was confirmed histologically (median MCHI score was 2, indicating epithelial damage without inflammation). DSS caused significant weight loss. Animals receiving EN regained baseline weight faster, by day 14, whereas animals on standard chow achieved recovery only by day 21. Differences in energy intake should be further investigated to validate the effect of EN on body weight recovery. At day 21, both injury groups demonstrated higher relative abundances of Bacteroidaceae and Erysipelotrichaceae, including the mucin-degrader Allobaculum mucilyticum, compared with the control group. Conversely, Lactobacillus abundance, notably Lactobacillus acidophilus, was higher in the EN group than in both other groups, as was the inferred capacity for lactate-producing fermentation. These findings suggest that EN is associated with a distinct microbial composition and inferred metabolic profile during the post-injury period, with lactobacilli as one of the potential mediators of its effects.

Animals↗

Changes in fecal microflora induced by intubation of mice with Bacillus subtilis (natto) spores are dependent upon dietary components.

We examined changes in mouse fecal microflora after various dietary components and Bacillus subtilis (natto) spores were delivered by intubation. The administration of intact spores of Bacillus subtilis (natto) did not affect fecal Enterobacteriaceae and Enterococcus spp. in all three diet groups; on the other hand, it did affect fecal Bacteroidaceae and Lactobacillus spp., depending upon the diets fed. The administration of autoclaved spores did not alter fecal microflora. In vitro cultures of Lactobacillus murinus obtained from mouse feces, together with Bacillus subtilis (natto) under aerobic conditions as a mixed culture, revealed that the growth of L. murinus was enhanced by the addition of intact spores of Bacillus subtilis (natto). This enhancement of growth was displayed only in media containing either sucrose, glucose, maltose, or fructose but not in media containing cornstarch, soluble starch, or microcrystalline cellulose. From these results it was evident that some metabolites of Bacillus subtilis (natto) produced during germination and (or) outgrowth of spores of this strain, requiring monosaccharides or oligosaccharides, participated in the enhancement of the growth of L. murinus.

Animals↗

Polyamine distribution profiles in newly validated genera and species within the Flavobacterium-Flexibacter-Cytophaga-Sphingobacterium complex.

Cellular polyamines of 58 strains belonging to the Flavobacterium-Flexibacter-Cytophaga-Sphingobacterium complex were analysed by HPLC. Homospermidine was found in all species of Flavobacterium, Chryseobacterium, Empedobacter, Myroides, Cellulophaga, Salegentibacter, Psychroserpens and Gelidibacter of the family Flavobacteriaceae. Flavobacterium ferrugineum located outside of this family also contained homospermidine. Cytophaga fermentans and C. xylanolytica belonging to the family Bacteroidaceae contained spermidine. Cytophaga marinoflava and C. latercula belonging to Flavobacteriaceae contained homospermidine. The Cytophaga hutchinsonii/C. aurantiaca group contained homospermidine which was the major polyamine in Flexibacter maritimus/ F. ovolyticus of the family Flavobacteriaceae. The Flexibacter sancti/F filiformis/ Cytophaga arvensicola group, F. elegans, F. ruber, F. canadensis, F. flexilis and F. tractuosus, were located separately in different six clusters, and contained homospermidine. The Flexibacter litoralis/F. polymorphus/F. aggregans group contained spermidine, which was detected in Flexibacter roseolus belonging to a divergent cluster. Sphingobacterium and Pedobacter species of the family Sphingobacteriaceae contained homospermidine. Polyamine profiles serve, as a phenotypic chemotaxonomic marker, for the classification of this complex.

Bacteria↗

Changes in intestinal microflora in patients with chronic severe hepatitis.

OBJECTIVE: To investigate changes in intestinal microflora in patients with chronic severe hepatitis (CSH), and their role in this life-threatening disease. METHODS: We classified nineteen patients with chronic severe hepatitis as the CSH group, thirty patients with chronic hepatitis (CH) as the CH group and thirty-one healthy volunteer as the control group. Fecal flora from all subjects were analyzed. Concentrations of plasma endotoxin, serum cytokines tumor necrosis factor alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) and liver function were assessed. RESULTS: The number of fecal bifidobacterium (P < 0.001, P < 0.05 respectively), as well as bacteroidaceae (P < 0.001, P < 0.01 respectively) were significantly deceased in patients with chronic severe hepatitis compared with the CH and control groups, while the number of enterobacteriaceae (P < 0.001, P < 0.05 respectively) and yeasts (P < 0.01, P < 0.05 respectively) were significantly increased. Levels of plasma endotoxin, serum TNF-alpha, IL-1 beta and total bilirubin (TBiL) were significantly increased in the CSH group. The concentration of endotoxin positively correlated with levels of both TNF-alpha, IL-1 beta and TBiL (P < 0.001, respectively). Levels of plasma endotoxin were positively correlated with the number of fecal enterobacteriaceae and negatively correlated with bifidobacterium (P < 0.05, P < 0.001, respectively). CONCLUSION: Intestinal flora in patients with chronic severe hepatitis were severely disturbed and gut microbiological colonization resistance was impaired. Changes in intestinal flora may have a pivotal role in both the elevation of plasma endotoxin and further hepatic lesions resulting in liver failure.

Adult↗

The effect of levofloxacin, an optically-active isomer of ofloxacin, on fecal microflora in human volunteers.

Following oral administration of levofloxacin (LVFX, (S)-(-)-Ofloxacin; formerly designated as DR-3355) at 200 mg per dose 3 times a day for 7 days to 6 healthy male volunteers, degrees of disturbance of the fecal microflora and fecal drug concentrations were examined. The total viable count remained unchanged during the study period due to the minimal change in the number of members of the family Bacteroidaceae, the most predominant organisms. Most of the aerobes including facultative anaerobes were suppressed by LVFX with only a slight increase in yeasts. In particular, the members of the family Enterobacteriaceae were reduced to below the detection limit on and after day 3 through the time of discontinuation of the drug in all subjects but one. Among the obligate anaerobes, peptostreptococci and bifidobacteria decreased or disappeared in some volunteers, but no significant changes were observed in other anaerobes. Neither Clostridium difficile nor its toxin D-1 was detected in any of the volunteers. No side effects attributable to the drug were observed. During administration, LVFX was detected in the feces at high concentrations which correlated well with the decrease of susceptible members of flora as well as to their detection rate.

Adult↗

[Preliminary study on intestinal flora in diarrhea type irritable bowel syndrome with pi-wei dampness-heat syndrome].

OBJECTIVE: To observe the changes of intestinal flora in diarrhea type irritable bowel syndrome with Pi-wei dampness-heat syndrome (IBS-PDS). METHODS: The seven kinds of common intestinal bacteria in feces, including enteri bacillus, enterococci, saccharomycete, bifid bacteria, lactobacillus, bacteroides and peptococcus were studied in 21 patients suffered from IBS-PDS, and compared with those in 22 patients with IBS with deficiency of Pi syndrome (DPS) and 25 healthy subjects as control. RESULTS: As compared with the healthy subjects, the levels of enteri bacillus and enterococci were significantly increased (P<0.01), the levels of bifid bacteria, Lactobacillus and Peptococcus were significantly decreased (P < 0.01), and saccharomycete and Bacteroides were insignificantly different in patients with PDS. As compared with patients with DPS, the levels of enteri bacillus, enterococci, bifid bacteria, Lactobacillus, Peptococcus and Bacteroidaceae were significantly increased except the level of saccharomycete. CONCLUSION: There may be alteration of intestinal flora in patients with IBS-PDS.

Adult↗

[Bacteriology of abdominal pus in 43 cases of acute appendicitis and appendiceal abscess at the Ivrea-Castellamonte Hospital, isolation of aerobic and anaerobic bacteria and drug sensitivity].

A retrospective study on the microbiology of abdominal pus from acute appendicitis or peritonitis was carried out by the authors. A total of 45 specimens were examined, 38 of them (84.4%) where found to be positive, of which 29 (76.3%) were polymicrobial and 9 (23.7%) were monomicrobial. The most represented species were Escherichia coli (28.4%), Bacteroides fragilis (7.8%), Streptococcus milleri (7.8%), Bacteroides oralis (3.9%) and Pseudomonas aeruginosa (3.9%). The polymicrobial associations more represented include Bacteroidaceae, Enterobacteriaceae and streptococcus genus. The most active drugs in vitro were found to be Piperacillin and Chloramphenicol, Cephalosporin (cefotaxime, ceftriaxone and cefoxitin) showed a good capability of being active against isolated microorganisms although they were less effective when used against anaerobic microorganisms then aerobic ones. On the contrary, Rifampicin, Metronidazole and Clindamycin were found to be quite effective against the anaerobes.

Abscess↗

[Community- and hospital-acquired bacteremia: a retrospective study in a regional hospital. III. Microbiological aspects].

A retrospective study was made of all blood cultures performed over a 40-month period at the Ivrea-Castellamonte Hospital (Turin, Italy). A total of 4386 vials from 619 patients were examined. There were 619 positive vials (14.1%) from 131 patients (21.2%) corresponding to 145 bacteremia episodes, including 129 monomicrobial (89%) and 16 polymicrobial (11%). Ten patients (1.6%) had more than one episode. There were 73 polluted vials (1.7%). A total of 165 microorganism were isolated: Gram-positive (52.7%) and Gram-negative (46%) bacteria, and mycetes (1.2%), anaerobic flora (9.7%). The predominant families were: Enterobacteriaceae (29.5%), Micrococcaceae (27.3%), Pseudomonadaceae (4.8%), Bacteroidaceae (4.8%) and Streptococcus "Genus" (18.8%). The species frequencies were: Escherichia coli (20%), Staphylococcus aureus (15.8%), Enterococcus (8.5%), Staphylococcus epidermidis (7.3%), Pseudomonas aeruginosa (4.8%), Proteus mirabilis (4.2%), Brucella spp. (2.4%), Bacteroides fragilis, Streptococcus bovis e Propionibacterium acnes (1.8%). These findings are compared with those published in the Italian and international literature. Stress is laid on periodical review of the isolations from samples of this kind as a useful aid towards the diagnosis and treatment of hospital infections, and in their monitoring and epidemiological evaluation.

Bacteremia↗