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Detection of bacteremia with liquid media containing sodium polyanetholsulfonate.

Two liquid blood culture media, Tryptic soy broth (TSB) and Thiol broth, containing sodium polyanetholsulfonate were compared in 8,654 cultures. Pseudomonas and Corynebacterium (including Propionibacterium) were isolated significantly more frequently (P < 0.001) from TSB than from Thiol. Escherichia coli, Haemophilus, and Bacteroidaceae were isolated more frequently in TSB; however, the differences were not statistically significant. In no instance was Thiol superior to TSB in detecting bacteremia. In an additional 2,977 cultures, aerobic and anaerobic Vacutainer culture tubes with supplemented peptone broth were inoculated in parallel with TSB and Thiol. Significantly greater rates of detection (P < 0.01) in TSB or Thiol were noted with Pseudomonas, E. coli, Enterobacter, viridans, and group A streptococci, Bacteroidaceae, and staphylococci.

Aerobiosis↗

[Effect of DQ-2556, a new cephalosporin derivative, on fecal microflora].

A new cephalosporin derivative antibacterial agent, DQ-2556, was administered intravenously to 4 healthy adult male volunteers at 2 grams per dosage 2 times a day for 5 days, and degrees of effects of drug concentrations on the fecal microflora were investigated. 1. The total viable count remained unchanged during the study period due to the minimal change in the number of members of the family Bacteroidaceae, which were the most dominant organisms. In one of the volunteers, however, the total count was remarkably reduced immediately after the end of administration of the drug. 2. Among the obligate anaerobes, most of the organisms decreased except members of the family Bacteroidaceae. A marked increase of lecithinase-positive clostridia was noted in 2 volunteers. 3. The number of aerobes, including facultative anaerobes, members of the family Enterobacteriaceae were suppressed but bacilli and yeasts showed slight increases. In many cases, increases in enterococci were also observed. 4. DQ-2556 was not detected in the feces from any of the volunteers during the study period. 5. Neither Clostridium difficile nor its toxin D-1 was detected in any of the volunteers. 6. Diarrhoea or other side-effects were not noted in any of the volunteers.

Adult↗

Change in the bacterial flora of pouchitis.

BACKGROUND/AIMS: Pouchitis is the most frequent complication of ileal pouch-anal anastomosis for patients with ulcerative colitis. While the mechanism of pouchitis is still unknown, a role involving the bacterial flora is suspected. The aim of the present study is to investigate the association between pouchitis and change in the bacterial flora. METHODOLOGY: This study included 22 patients who underwent ileal pouch-anal anastomosis in our department to treat ulcerative colitis. The mean age was 46.2 years. The male-to-female ratio was 10:12. The mean interval between surgery and this study was 95.6 months. Collected feces was serially diluted 10-fold to 10-8 and each solution was cultured. We diagnosed pouchitis using the pouchitis disease activity index. RESULTS: Nine of the 22 patients were diagnosed as having pouchitis. In the obligate anaerobes, the numbers of Bacteroidaceae and Bifidobacterium were significantly lower in the pouchitis group (P<0.01 and P<0.001). In the facultative anaerobic bacteria, the number of Lactobacillus was significantly lower in the pouchitis group (P<0.05). CONCLUSIONS: The numbers of Bacteroidaceae, Bifidobacterium and Lactobacillus were significantly lower in the bacterial flora of the pouchitis group. Our findings suggest that there is an association between change in the bacterial flora and pouchitis.

Bacteria, Anaerobic↗

Use of nonprotein nitrogen in pigs: effects of dietary urea on the intestinal microflora.

The fecal flora of 2 pigs given an optimal protein diet (digestive crude protein, 13.3%) with 2% urea, 2 pigs given a low protein diet (digestive crude protein, 5.7%) with 2% urea, and 2 pigs given a low protein diet only were bacteriologically investigated before feeding of each diet and on feeding days 21 and 38. The addition of urea to a low protein diet was significantly decreased Lactobacillus (P less than 0.001), Bacteroidaceae (P less than 0.01), Peptococcaceae (P less than 0.05), and Megasphaera (P less than 0.01) in the feces. In addition, there was a marked suppression of anaerobic curved rods (mainly Selenomonas) to undetectable amounts in the feces. A significant reduction (P less than 0.01) of Bifidobacterium and a marked suppression of Megasphaera to undetectable amounts were observed in the feces of pigs given low protein diets without urea. Microbial groups in the ileum and colon of pigs on feeding day 38 were also analyzed. Effects of dietary urea on the intestinal flora included: (i) anaerobic curved rods and spiral rods were restricted mainly to the colon, (ii) smaller numbers of yeasts were present in the intestine, and (iii) Bacteroidaceae, Peptococcaceae, and Eubacterium in the colon were not changed. Many anaerobic isolates, including curved rods and spiral rods, in the colon of pigs may have an important role in urea use.

Animals↗

[Metronidazole sensitivity of anaerobes. A comparison with other chemotherapeutics (author's transl)].

In comparative in vitro studies, broad spectrum penicillins and cephalosporins showed a relatively weak action in the range of beta-lactamase-positive bacteroides species (B. fragilis, B. thetaiotamicron) bacteroidaceae activity being otherwise good. Clindamycin was more effective against bacteroides species than against bacteria of the fusobacterium-sphaerophorus group. Metronidazole, however, showed no gap in its efficacy against bacteroidaceae, peptococcaceae and clostridium spp. with the exception of Propionibacterium acnes. The substance is therefore suitable alone and in combination for the chemotherapy of anaerobic infections.20

Anaerobiosis↗

Susceptibility of Bacteroides fragilis, Bacteroides thetaiotaomicron, and Fusobacteria in Antibacterial agents.

The minimum inhibitory concentrations of 45 antimicrobial agents were established by the agar dilution method for 37 strains of Bacteroidaceae: Bacteroides fragilis (18 strains), B. thetaiotaomicron (8 strains), Fusobacterium nucleatum (6 strains), and Fusobacterium spp. (5 strains). The fusobacteria tested showed good susceptibility to beta-lactam-antibiotics, tetracyclines, lincomycins, and nitroimidazole compounds. Bacteria of the B. fragilis group were resistant to most of the penicillins and cephalosporins and could not be suppressed with certainty by nitroimidazoles and older tetracyclines. Doxycycline and minocycline, however, showed sufficient activity, and all our Bacteroides strains tested were sensitive to clindamycin. Especially the good in vitro susceptibility of all Bacteroidaceae strains investigated to cefoxitin may offer new therapeutic possibilities.

Anti-Bacterial Agents↗

Changes in microflora of the cloaca and oviduct of hens after intracloacal or intravaginal inoculation with Salmonella enteritidis.

Quantitative and qualitative microbiological examination was carried out on cloacal and oviductal contents pre- and postinfection with Salmonella enteritidis (SE) intracloacally or intravaginally. Before inoculation with SE, the means +/- standard deviation (SD) of total bacterial counts, anaerobic bacterial counts, and aerobic bacterial counts in the cloaca were log10 7.7 +/- 0.7, 7.4 +/- 0.2, and 6.9 +/- 0.8 colony-forming units (CFU)/g, respectively. The predominant bacteria were Bacteroidaceae, Lactobacillus, and Escherichia coli. Before inoculation with SE, the means +/- SD of total bacterial counts, anaerobic bacterial counts, and aerobic bacterial counts in the vagina were log10 5.7 +/- 1.4, 5.5 +/- 1.3, and 3.6 +/- 2.7 CFU/g, respectively. Bacteroidaceae and Lactobacillus were predominant. Following inoculation with SE, only the cloacal population of Lactobacillus in hens inoculated intracloacally was significantly increased compared to that before the inoculation. Other indigenous microflora were stable even after the inoculation. In the uterus, very few bacteria, Lactobacillus and Staphylococcus, were isolated. Five of 20 eggs (25%) from hens inoculated with SE intravaginally were positive for SE, whereas no SE was recovered from 22 eggs in hens inoculated with SE intracloacally. SE was recovered from the uterus after intravaginal inoculation with SE and from the vagina after intracloacal inoculation with SE. Contamination may ascend from the cloaca into the lower parts of the oviduct and subsequently contaminated eggs may occur.

Animals↗

Lipopolysacharides from anaerobic gram-negative rods.

The chemical composition and antigenic specificity of the cell wall lipopolysaccharides of the two main groups of Bacteroidaceae, Fusobacterium and Bacteroides species, are reviewed. Cell wall lipopolysaccharides from Fusobacterium species contain 2-keto-3-deoxy-octonate and heptose, but lipopolysaccharides isolated from Bacteroides species do not.

Bacteroidaceae↗

Serious infection with Leptotrichia buccalis. Report of a case and review of the literature.

Leptotrichia buccalis is an obligately anaerobic, gram-negative rod classified as a member of the family Bacteroidaceae. Organisms found in this family (Bacteroides and Fusobacteria) are frequent causes of severe infection in human beings. L. buccalis is a constituent of normal oral flora. Leptotrichia has rarely been recovered from mixed infections primarily of the head and neck. It has not previously been reported as a cause of serious infection in man. We report a case of cavitary pneumonia and septicemia caused by L. buccalis. The colonial morphology and gram stain of L. buccalis are characteristic. Definitive identification is made by gas-liquid chromatography. Leptotrichia have been shown to contain potent endotoxin. L. buccalis has the potential to be a virulent, pathogenic microorganism, especially in the compromised host.

Bacterial Infections↗

Current taxonomy of medically important nonsporing anaerobes.

This review deals mainly with the taxonomy of the genera and species in the family Bacteroidaceae. It has been proposed that the genus Bacteroides should be restricted to include the "Bacteroides fragilis group" and that the asaccharolytic black-pigmented Bacteroides species be transferred to a new genus, Porphyromonas. New Bacteroides, Fusobacterium, and Selenomonas species have been described. A high degree of heterogeneity is apparently present among Peptostreptococcus species. Mobiluncus is a novel genus including gram-variable curved rods isolated from the human vagina. Brachyspira has been proposed as the generic name for spirochetes isolated from patients with intestinal spirochetosis.

Bacteria, Anaerobic↗

Bacteremia due to Bacteroidacceae: a review of 92 cases.

The clinical courses of 92 patients who had bacteremia due to Bacteroidaceae were reviewed. The overall mortality rate was 21% (19 patients). There was no significant difference between mortality rates when patients were grouped by anticipated clinical course of underlying disease (nonfatal, ultimately fatal, and rapidly fatal) and when they were grouped by type of antibacterial therapy (appropriate, including clindamycin, chloramphenicol, lincomycin, and carbenicillin; or inappropriate, signifying no antibiotic treatment or treatment with antibiotics other than the four listed above). However, there was a correlation between recovery of the patient and portal of entry of the infectious agent; patients whose source of infection was the gastrointestinal tract had a mortality rate of 29% (17 of 58 patients), whereas there were no deaths among the 26 women whose presumed source of bacteremia was the genital tract. Increasing age appeared to be an important factor as well; 17 of 19 deaths occurred in patients who were older than 40 years. There was no significant difference between the mortality rate of patients who were treated with clindamycin (15%, eight of 52 patients) and that of patients who were treated with chloramphenicol (44%, four of nine).

Adult↗

Phylogenetic placement of Dialister pneumosintes (formerly Bacteroides pneumosintes) within the Sporomusa subbranch of the Clostridium subphylum of the gram-positive bacteria.

The nucleotide sequence of the 16S rRNA gene of the type strain of Dialister pneumosintes was determined. Phylogenetic analysis revealed that this species belongs to the Sporomusa branch of the Clostridium subphylum of the gram-positive bacteria and should therefore be excluded from the family Bacteroidaceae. Within this branch, which encompasses several other gram-negative taxa, such as Acidaminococcus, Pectinatus, Phascolarcobacterium, Quinella, Selenomonas, and Zymophilus, Dialister showed a specific, albeit distant, affinity with the genera Megasphaera and Veillonella.

Bacteroidaceae↗

Phylogenetic analysis of Prevotella nigrescens, Prevotella intermedia and Porphyromonas gingivalis clinical strains reveals a clear species clustering.

Prevotella nigrescens, Prevotella intermedia and Porphyromonas gingivalis are oral pathogens from the family Bacteroidaceae, regularly isolated from cases of gingivitis and periodontitis. In this study, the phylogenetic variability of these three bacterial species was investigated by means of 16S rRNA (rrs) gene sequence comparisons of a set of epidemiologically and geographically diverse isolates. For each of the three species, the rrs gene sequences of 11 clinical isolates as well as the corresponding type strains was determined. Comparison of all rrs sequences obtained with those of closely related species revealed a clear clustering of species, with only a little intraspecies variability but a clear difference in the rrs gene with respect to the next related taxon. The results indicate that the three species form stable, homogeneous genetic groups, which favours an rrs-based species identification of these oral pathogens. This is especially useful given the 7% sequence divergence between Prevotella intermedia and Prevotella nigrescens, since phenotypic distinction between the two Prevotella species is inconsistent or involves techniques not applicable in routine identification.

Bacteroidaceae Infections↗

Comparative evaluation of supplemented peptone broth with sodium polyanetholesulfonate and trypticase soy broth with sodium amylosulfate for detection of septicemia.

We compared the yield and speed of detection of clinically important microorganisms from 10,156 paired 5-ml samples of blood cultured in supplemented peptone broth (SPB) with 0.03% sodium polyanetholesulfonate (SPS) or Trypticase soy broth (TSB) with 0.5% sodium amylosulfate (SAS). The atmosphere of incubation (open venting units) and ratio of blood to broth (1:10) were the same for both samples. Only cultures with adequate blood samples (greater than or equal to 80% of stated volume) were compared statistically. Overall, SPB/SPS outperformed TSB/SAS. Bacteroidaceae and Eubacterium were found more often (P less than 0.05) and viridans streptococci were found sooner (P less than 10(-4)) in SPB/SPS than in TSB/SAS. Most importantly, staphylococci were found both more often (P less than 0.03) and sooner (P less than 10(-7)) in SPB/SPS than in TSB/SAS. In a separate experiment, SAS slowed the growth of a clinical strain of Staphylococcus aureus in TSB. Unless important advantages can be confirmed for SAS in controlled clinical trials, SAS cannot be recommended for routine use as an anticoagulant in blood culture media.

Amylopectin↗

Chitinolytic bacteria in the minke whale forestomach.

Minke whales consume large amounts of pelagic crustaceans. Digestion of the prey is initiated by indigenous bacteria in a rumen-like forestomach system. A major structural component of the crustacean exoskeleton is chitin, the beta-1,4-linked polymer of N-acetyl-D-glucosamine. The exoskeletons appear to dissolve completely in the non-glandular forestomach. Bacteria in the forestomach fluid of six krill-eating minke whales were enumerated and isolated using an anaerobic habitat-simulating culture medium. Median viable population densities ranged between 6.0 x 10(6) and 9.9 x 10(9) bacterial cells per mL forestomach fluid. Bacterial isolates (n = 44) cultured from the forestomach fluid of one minke whale mainly resembled strains of Eubacterium (25%), Streptococcus (18%), Clostridium (14%), and Bacteroides (11%). As much as 12% of the bacterial isolates were chitinolytic, while beta-N-acetylglucosaminidase activity was demonstrated in 54% of the isolates, and utilisation of N-acetyl-D-glucosamine was observed in 73%. The chitinolytic isolates resembled strains of Bacteroides, Bacteroidaceae, Clostridium, and Streptococcus. Scanning and transmission electron microscopy of partly digested krill from the minke whale forestomach revealed bacteria close to and inside the chitinous exoskeleton. The bacterial chitinase may act on the chitinous crustacean exoskeletons, thereby allowing other bacteria access to the nutritious soft inner tissues of the prey, and thus initiating its degradation and fermentation.

Animals↗

Changes in normal vaginal flora of African green monkeys (Cercopithecus aethiops) during the menstrual cycle.

Changes in normal vaginal flora of African green monkeys associated with the estrous cycle were examined by the swab method. Bacteroidaceae, corynebacteria, and streptococci were predominant throughout the whole estrous cycle, although individual differences were great. It was also clear that all bacterial species tended to decrease in the menstrual phase. Lactobacilli, the most predominant bacteria in the normal vagina of humans, were detected only in very low numbers in the African green monkey, suggesting that the normal vagina of the African green monkey has a different ecosystem from that of normal human vaginal flora.

Animals↗

Establishment of specific pathogen-free guinea-pig colonies using limited-flora guinea-pigs associated with conventional guinea-pig flora, and monitoring of their cecal flora.

Six groups of limited flora (LF) Hartley guinea-pigs were produced by inoculation of hysterectomy-derived GF guinea-pigs with various combinations of cecal bacteria of conventional (CV) guinea-pigs to determine the effective bacterial cocktails for the establishment of a specific pathogen free (SPF) colony. Bifidobacterium magnum (Bif) isolated from CV guinea-pigs was used for pretreatment. The mortality of LF guinea-pigs inoculated with only Bif was 75%, and that of those inoculated with Bif plus chloroform-treated cecal suspension (CHF) or Bif plus CHF plus 32 isolates from CV guinea-pigs was 40 to 66.7%. These three groups were in an unhealthy condition with mucoid enteritis-like diarrhea. However, the mortality of LF guinea-pigs inoculated with the anaerobic growth on EG plates injected with 10(-5) dilution of cecal contents (CF) or inoculated with Bif plus CF was 6.3 and 15%, respectively. These latter two groups of LF guinea-pigs were transferred to separate barrier rooms and some of the LF guinea-pigs were maintained in isolators as a source of intestinal flora for SPF guinea-pigs. The composition of cecal flora of LF guinea-pigs was stable for a long time, and bacteroidaceae and peptococcaceae were maintained as predominant components. The basic composition of the cecal flora of SPF guinea-pigs originated from LF guinea-pigs, which consists mainly of the anaerobic bacteria, was not changed over a long period, and the flora composition became similar to that in CV guinea-pigs. Guinea-pig-specific pathogens from the SPF colonies were not detected during experiments.

Animals↗

Establishment of specific pathogen-free (SPF) rat colonies using gnotobiotic techniques.

Gnotobiotic Wistar rats were produced using gnotobiotic techniques, which were established in the production of a SPF mouse colony, in order to establish a barrier-sustained colony. One strain of Escherichia coli, 28 strains of Bacteriodaceae (B-strains), three strains of Lactobacillus (L-strains) and a chloroform-treated fecal suspension (CHF, Clostridium mixture) were prepared from conventional Wistar rats as the microflora source. Two groups of limited-flora rats, E. coli plus B-strains and E. coli plus CHF, were produced. After confirmation that Clostridium difficile was not detected in the CHF-inoculated rats, two groups of limited-flora rats were transferred to an isolator and housed together in a cage. These rats were then orally inoculated with L-strains. The gnotobiotic rats showed colonization resistance to Pseudomonas aeruginosa, and the number of E. coli in the feces was 10(5) to 10(6)/g. The gnotobiotic rats were transferred to a barrier room as a source of intestinal flora for SPF colonies. In the SPF rats, basic cecal flora was mainly composed of Bacteroidaceae, clostridia, fusiform-shaped bacteria and lactobacilli, and did not change over a long period. Their flora became similar to that of conventional rats.

Animal Husbandry↗