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In vitro activity of DU-6859a against anaerobic bacteria.

The activity of a new quinolone agent, DU-6859a, against 330 strains of anaerobic bacteria was determined by using the National Committee for Clinical Laboratory Standards-approved Wadsworth brucella laked blood agar method; the activity of DU-6859a was compared with those of amoxicillin-clavulanate (2:1), chloramphenicol, ciprofloxacin, clindamycin, fleroxacin, imipenem, lomefloxacin, metronidazole, sparfloxacin, and temafloxacin. DU-6859a and chloramphenicol inhibited all of the isolates at concentrations of 1 and 16 micrograms/ml, respectively; amoxicillin-clavulanate, imipenem, and metronidazole inhibited > or = 94% of the isolates at their respective breakpoints (8, 8, and 16 micrograms/ml). MICs of DU-6859a at which 90% of the strains were susceptible were 1 to 5 twofold dilutions lower than those of the other quinolones for every group of organisms. MICs of DU-6859a at which 90% of the strains were susceptible (total numbers of strains tested are in parentheses) were < or = 0.25 micrograms/ml for Bacteroides fragilis (57), other B. fragilis group species (84), Bilophila wadsworthia (15), Clostridium species (27) (including C. difficile, C. perfringens, and C. ramosum), Fusobacterium nucleatum (16), Fusobacterium mortiferum-F. varium group species (10), Peptostreptococcus species (20), non-spore-forming gram-positive rods (20), and Prevotella species (25).

Anti-Infective Agents↗

Effect of CO2 on susceptibilities of anaerobes to erythromycin, azithromycin, clarithromycin, and roxithromycin.

The Oxyrase agar dilution method (Oxyrase, Inc., Mansfield, Ohio), which provides an anaerobic environment without added CO2, was compared with the reference agar dilution method recommended by the National Committee for Clinical Laboratory Standards (anaerobic chamber with 10% CO2) to test the susceptibilities of 302 gram-negative and gram-positive anaerobes to erythromycin, azithromycin, clarithromycin, and roxithromycin. For erythromycin, the overall MIC for 50% of isolates tested (MIC50) was 0.5 micrograms/ml and the MIC90 was 8.0 micrograms/ml by the Oxyrase method, whereas they were 4.0 and 64.0 micrograms/ml, respectively, under standard anaerobic conditions with CO2. At a breakpoint of 4.0 micrograms/ml, 88% of strains were susceptible to erythromycin by the Oxyrase method, whereas 63% were susceptible in the chamber. The corresponding MIC50s and MIC90s of azithromycin, clarithromycin, and roxithromycin by the Oxyrase method were 0.5 and 8.0, 0.25 and 4.0, and 0.5 and 16.0 micrograms/ml, respectively, whereas in the chamber they were 4.0 and > 64.0, 2.0 and 64.0, and 2.0 and 64.0 micrograms/ml, respectively. At a breakpoint of 8.0 micrograms/ml for these three drugs, 89, 92, and 85% of the isolates, respectively, were susceptible by the Oxyrase method, whereas 67%, 72, and 68% of the isolates, respectively, were susceptible in the chamber. Most strains resistant to all four compounds by both methods were Bacteroides distasonis, Fusobacterium mortiferum, Fusobacterium varium and non-Clostridium perfringens Clostridium species. Results of the study may lead to a reappraisal of the role played by macrolides and azalides in the treatment of anaerobic infections.

Azithromycin↗

Antianaerobic activity of the ketolide RU 64004 compared to activities of four macrolides, five beta-lactams, clindamycin, and metronidazole.

Agar dilution methodology (with added Oxyrase in the case of the macrolide group to allow incubation without added CO2) was used to compare the activity of RU 64004, a new ketolide, with the activities of erythromycin, azithromycin, clarithromycin, roxithromycin, clindamycin, amoxicillin with and without clavulanate, piperacillin with and without tazobactam, metronidazole, and imipenem against 379 anaerobes. Overall, RU 64004 yielded an MIC at which 50% of the isolates are inhibited (MIC50) of 1.0 microg/ml and an MIC90 of 16.0 microg/ml. In comparison, MIC50s and MIC90s of erythromycin, azithromycin, clarithromycin, and roxithromycin were 2.0 to 8.0 and >64.0 microg/ml, respectively. MICs of macrolides, including RU 64004, were higher for Bacteroides ovatus, Fusobacterium varium, Fusobacterium mortiferum, and Clostridium difficile than for the other species. RU 64004 was more active against gram-positive rods and cocci, Prevotella and Porphyromonas spp., and fusobacteria other than F. mortiferum and F. varium than against the Bacteroides fragilis group. Overall MIC50s and MIC90s (in micrograms per milliliter), respectively, of other compounds were as follows: clindamycin, 1.0 and 16.0; amoxicillin, 4.0 and 64.0; amoxicillin-clavulanate, 0.5 and 4.0; piperacillin, 8.0 and >64.0; piperacillin-tazobactam, 1.0 and 16.0; metronidazole, 1.0 and 4.0; and imipenem, 0.25 and 1.0.

Anti-Bacterial Agents↗

In vitro activity of Bay 12-8039, a new 8-methoxyquinolone, compared to the activities of 11 other oral antimicrobial agents against 390 aerobic and anaerobic bacteria isolated from human and animal bite wound skin and soft tissue infections in humans.

The in vitro activity of Bay 12-8039, a new oral 8-methoxyquinolone, was compared to the activities of 11 other oral antimicrobial agents (ciprofloxacin, levofloxacin, ofloxacin, sparfloxacin, azithromycin, clarithromycin, amoxicillin clavulanate, penicillin, cefuroxime, cefpodoxime, and doxycycline) against 250 aerobic and 140 anaerobic bacteria recently isolated from animal and human bite wound infections. Bay 12-8039 was active against all aerobic isolates, both gram-positive and gram-negative isolates, at < or = 1.0 microg/ml (MICs at which 90% of isolates are inhibited [MIC90s < or = 0.25 microg/ml) and was active against most anaerobes at < or = 0.5 microg/ml; the exceptions were Fusobacterium nucleatum and other Fusobacterium species (MIC90s, > or = 4.0 microg/ml) and one strain of Prevotella loeschii (MICs, 2.0 microg/ml). In comparison, the other quinolones tested had similar in vitro activities against the aerobic strains but were less active against the anaerobes, including peptostreptococci, Porphyromonas species, and Prevotella species. The fusobacteria were relatively resistant to all the antimicrobial agents tested except penicillin G (one penicillinase-producing strain of F. nucleatum was found) and amoxicillin clavulanate.

Administration, Oral↗

Comparative antianaerobic activities of the ketolides HMR 3647 (RU 66647) and HMR 3004 (RU 64004).

HMR 3647 (RU 66647) and HMR 3004 (RU 64004), two ketolides, had MICs at which 50% of the strains are inhibited (MIC50s) of 0.06 to 0.125 microg/ml and MIC90s of 16.0 microg/ml against 352 anaerobes. MIC50s and MIC90s of erythromycin, azithromycin, clarithromycin, and roxithromycin were 0.5 to 2.0 microg/ml and 32.0 to >64.0 microg/ml, respectively. HMR 3647 and HMR 3004 were more active against non-Bacteroides fragilis-group anaerobes (other than Fusobacterium mortiferum, Fusobacterium varium, and Clostridium difficile).

Anti-Bacterial Agents↗

Activities of telithromycin (HMR 3647, RU 66647) compared to those of erythromycin, azithromycin, clarithromycin, roxithromycin, and other antimicrobial agents against unusual anaerobes.

The comparative activity of telithromycin (HMR 3647) against 419 human anaerobic isolates was determined by the agar dilution method. At concentrations of </=0.5 microgram/ml, telithromycin was active against Actinomyces israelii, Actinomyces odontolyticus, Bacteroides tectum, Bacteroides ureolyticus, Bacteroides gracilis (now Campylobacter gracilis), Porphyromonas spp. (including Porphyromonas gingivalis and Porphyromonas macacae), Prevotella intermedia, Prevotella heparinolytica, and almost all Peptostreptococcus species. Clostridia showed species and strain variability, often with a biphasic pattern. Fusobacterium species, except Fusobacterium russii, were relatively resistant.

Anti-Bacterial Agents↗

Comparative in vitro activities of ABT-773 against aerobic and anaerobic pathogens isolated from skin and soft-tissue animal and human bite wound infections.

We studied the comparative in vitro activities of ABT-773, a new ketolide, against 268 aerobic and 148 anaerobic recent isolates from clinical bites using an agar dilution method and inocula of 10(4) CFU/spot for aerobes and 10(5) CFU for anaerobes. The following are the MIC ranges and MICs at which 90% of isolates are inhibited (MIC(90)s) of ABT-773 for various isolates, respectively: Pasteurella multocida and Pasteurella septica, 0.125 to 2 and 1 microg/ml; other Pasteurella species, 0.125 to 1 and 0.5 microg/ml; Corynebacterium spp., 0.015 to 0.06 and 0.015 microg/ml; Staphylococcus aureus, 0.03 to 0.06 and 0.06 microg/ml; coagulase-negative staphylococci, 0.015 to >32 and 32 microg/ml; streptococci, 0.015 to 0.03 and 0.03 microg/ml; Eikenella corrodens, 0.25 to 1 and 1 microg/ml; and Bergeyella zoohelcum, 0.03 to 0.25 and 0.06 microg/ml. For anaerobes the MIC ranges and MIC(90)s of ABT-773 were as follows, respectively: Prevotella heparinolytica, 0. 06 to 0.125 and 0.125 microg/ml; Prevotella spp., 0.015 to 0.125 and 0.06 microg/ml; Porphyromonas spp., 0.015 to 0.03 and 0.015 microg/ml; Fusobacterium nucleatum, 0.5 to 8 and 8 microg/ml; other Fusobacterium spp., 0.015 to 8 and 0.5 microg/ml; Bacteroides tectum, 0.015 to 0.5 and 0.06 microg/ml; and Peptostreptococcus spp., 0.015 to 0.25 and 0.03 microg/ml. ABT-773 was more active than all macrolides tested against S. aureus, E. corrodens, and anaerobes, but all compounds were poorly active against F. nucleatum. The activity of ABT-773 was within 1 dilution of that of azithromycin against Pasteurella spp., and ABT-773 was four- to eightfold more active than clarithromycin against Pasteurella spp. ABT-773 may offer a therapeutic alternative for bite wound infections.

Animals↗

In vitro activities of the des-fluoro(6) Quinolone BMS-284756 against aerobic and anaerobic pathogens isolated from skin and soft tissue animal and human bite wound infections.

BMS-284756, a new des-fluoro(6) quinolone, was very active against 240 aerobic and 180 anaerobic isolates from bite victims. It inhibited 403 of 420 (96%) isolates, including those of Moraxella spp., CDC group EF-4, and Eikenella corrodens at < or = 2 microg/ml and those of all Pasteurella spp. and Bergeyella zoohelcum at < or = 0.015 microg/ml. Fusobacterium russii and 6 of 11 Fusobacterium nucleatum isolates of animal bite origin were resistant, but isolates of human bite origin were susceptible, which suggests that they were of a different subspecies.

Animals↗

Antimicrobial susceptibilities of anaerobic bacteria: recent clinical isolates.

Minimal inhibitory concentrations of clindamycin, minocycline, metronidazole, penicillin, and carbenicillin were determined by agar dilution against 150 recent clinical isolates of anaerobic bacteria. Ninety-nine percent of Bacteroides fragilis and all B. melaninogenicus, Clostridium perfringens, and Fusobacterium were inhibited by clindamycin at 3.1 mug/ml. Only 58% of other clostridial species were inhibited by this concentration of clindamycin. Minocycline at 3.1 mug/ml inhibited 72% of C. perfringens, 81% of other Clostridium species, and 66, 75, and 100% of B. fragilis, B. melaninogenicus, and Fusobacterium, respectively. Metronidazole at 12.5 mug/ml inhibited all bacteria tested. B. fragilis was resistant to both penicillin and carbenicillin at 6.2 mug/ml. Concentrations of 25 mug/ml for penicillin and 100 mug/ml for carbenicillin were needed to inhibit more than 90% of B. fragilis. Organisms other than B. fragilis were moderately or extremely susceptible to the penicillins.

Anti-Bacterial Agents↗

Bacteria isolated from the duodenum, ileum, and cecum of young chicks.

Facultatively anaerobic and strictly anaerobic bacteria colonizing the intestinal tracts of 14-day-old chicks fed a corn-based diet were enumerated, isolated, and identified. Colony counts from anaerobic roll tubes (rumen fluid medium) or aerobic plates (brain heart infusion agar) recovered from homogenates of the duodenum, upper and lower ileum, and cecum varied appreciably among samples from individual birds. Anaerobic and aerobic counts from the duodenum and ileum were similar. Anaerobic counts were highest from the cecum (0.7 X 10(11) to 1.6 X 10(11)/g of dry tissue) and exceeded aerobic plate counts by a factor of at least 10(2). Facultatively anaerobic groups (Streptococcus, Staphylococcus, Lactobacillus, and Escherichia coli) comprised the predominant flora of the duodenum and ileum, although large numbers of anaerobes (9 to 39% of the small intestine isolates), represented by species of Eubacterium, Propionibacterium, Clostridium, Gemmiger, and Fusobacterium, were also recovered. Strict anaerobes (anaerobic gram-positive cocci, Eubacterium, Clostridium Gemmiger, Fusobacterium, and Bacteriodes) made up nearly the entire microbial population of the cecum. Scanning electron microscopy of the intestinal epithelia of chicks revealed populations of microbes on the duodenal, ileal, and cecal mucosal surfaces.

Anaerobiosis↗

Bacteriology of experimental gingivitis in children.

Children are more resistant to gingivitis than are adults. To determine possible differences in their periodontal floras, an experimental gingivitis study, identical in design to one reported earlier with young adults, was conducted with four 4- to 6-year-old children. The incidence of sites that developed gingival index scores of 2 in children was less than one-third of the incidence observed in adults. The composition of the flora of each child was statistically significantly different from that of any other child or adult. The floras of the children as a group were statistically significantly different from those of the adults. Children had 3-fold greater proportions of Leptotrichia species, 2.5-fold greater proportions of Capnocytophaga species, 2.3-fold greater proportions of Selenomonas species, 2-fold greater proportions of bacterial species that require formate and fumarate, and 1.5-fold greater proportions of Bacteroides species. Adults had greater proportions of Fusobacterium, Eubacterium, and Lactobacillus species. Fusobacterium nucleatum, Actinomyces WVa 963, Selenomonas D04, and Treponema socranskii were predominant species that correlated with increasing gingival index scores in both children and adults.

Age Factors↗

Microbial ecology of plaque in rats with naturally occurring gingivitis.

The microbial ecology of adherent plaque was investigated in relation to the pathological findings of gingivitis in plaque-susceptible rats. Plaque developed in the gingiva of the lower incisor in plaque-susceptible rats, but not in plaque-resistant rats, after they were fed a commercial powder diet. With increase in plaque volume, the total counts of bacteria increased 10(9) to 10(11)/g. In the first 3 months, Bacteroides species increased and became the predominant population. Streptococcus species also increased at the same time. After 9 months, Fusobacterium species and oral Treponema species were recognized in increasing numbers. The anaerobic bacteria increased in proportion with the progression of plaque development. Bacteroides intermedius, Fusobacterium nucleatum, Streptococcus salivarius, and other species were isolated. Acute gingivitis was observed within 3 months, and subacute-chronic gingivitis was observed between 2 and 12 months. These findings suggest that proportional changes in the gingival plaque flora may uniquely contribute to the development of gingival inflammation in this experimental model.

Animals↗

Structural analysis of lipopolysaccharides from Eikenella corrodens by use of murine monoclonal antibodies.

The structure of lipopolysaccharides (LPSs) of Eikenella corrodens was analyzed with prepared murine monoclonal antibodies. A common core epitope was found in three of seven LPS preparations from E. corrodens strains and Fusobacterium nucleatum ATCC 25586. Four E. corrodens LPSs were found to possess an O-side-chain epitope which cross-reacted with LPSs from Fusobacterium necrophorum ATCC 25286 and Capnocytophaga ochracea M-12. Lipid A of E. corrodens LPS shared an epitope common among LPSs from various gram-negative rods.

Animals↗

Fibrinogenolytic and fibrinolytic activity in oral microorganisms.

Samples were taken from blood accumulated in dental alveoli after surgical removal of mandibular third molars, from subgingival plaque of teeth with advanced periodontal destructions, from teeth with infected necrotic pulps, and from subjects suffering from angular cheilitis. Of the microorganisms subcultured from these samples, 116 strains were assayed for enzymes degrading fibrinogen and fibrin. Enzymes degrading fibrinogen were assayed with the thin-layer enzyme assay cultivation technique. This assay involves the cultivation of microorganisms on culture agars applied over fibrinogen-coated polystyrene surfaces. Enzymes degrading fibrin were assayed with both a plate assay and a tube assay, in which fibrin was mixed with a microbial culture medium. Microorganisms degrading fibrinogen or fibrin or both were isolated from all sampling sites. Activity was mainly detected in strains of Actinomyces, Bacteroides, Fusobacterium, Peptococcus, Propionibacterium, and Staphylococcus aureus. Most Fusobacterium strains degraded fibrinogen only. Enzymes degrading fibrinogen as well as enzymes degrading fibrin via activation of plasminogen were revealed in strains of Clostridium, S. aureus, and Streptococcus pyogenes. It was generally found that fibrinogen was degraded by more strains than was fibrin, which indicates that different proteases may be involved.

Bacteria↗

Evaluation of the RapID-ANA system for identification of anaerobic bacteria of veterinary origin.

This study evaluated the ability of the RapID-ANA system (Innovative Diagnostic Systems, Inc., Atlanta, Ga.) to accurately identify a spectrum of freshly isolated veterinary anaerobes. A total of 183 isolates were tested and included 7 Actinomyces spp., 53 Bacteroides spp., 32 Clostridium spp., 2 Eubacterium spp., 65 Fusobacterium spp., 1 Peptococcus spp., 22 Peptostreptococcus spp., and 1 Propionibacterium spp. All isolates were initially identified by conventional biochemical testing and gas-liquid chromatography of short-chain fatty acid metabolites. Additional tests were performed as required by the RapID-ANA system. Of these isolates, 81.4% were correctly identified to the genus level, including 59.6% to the species level, 14.2% were incorrectly identified at the genus level, and 4.4% were not identified. Initially, 20.2% of the strains were not identified because the microcodes were not in the code book. The majority of the incorrect identifications were caused by the misidentification of Fusobacterium spp. as Bacteroides spp. Errors also occurred when veterinary anaerobes not included in the data base were assigned an identification from the existing data base. The RapID-ANA system appears to be a promising new method for rapid identification of veterinary anaerobes; however, further evaluation with an extended data base is needed before the system can accurately identify all clinically significant anaerobes.

Animals↗

Comparison of four commercial brucella agar media for growth of anaerobic organisms.

Four different commercial brucella blood agar plating media (Anaerobe Systems, BBL Microbiology Systems, Remel, and Scott Laboratories) were compared for the abilities to recover anaerobic organisms from clinical specimens and to support the growth of American Type Culture Collection anaerobic stock cultures. Following 24 h of incubation in an anaerobe chamber, Anaerobe Systems prereduced, anaerobically sterilized brucella plates yielded 63% of the total clinical anaerobe isolates, the Scott medium yielded 51%, the Remel medium yielded 42%, and the BBL medium yielded 37%. Poor growth of Peptostreptococcus magnus, P. anaerobius, Fusobacterium necrophorum, F. nucleatum, and pigmented Bacteroides spp. was observed on brucella media obtained from BBL, Remel, and Scott. Data obtained with stock anaerobic cultures showed that Anaerobe Systems plates yielded good growth and produced a larger colony size with all of the strains tested in 1 day, whereas poor growth of Peptostreptococcus spp., B. melaninogenicus, and Fusobacterium spp. was noted on brucella media from BBL, Remel, and Scott.

Agar↗

Bacterial vaginosis and vaginal microorganisms in idiopathic premature labor and association with pregnancy outcome.

The vaginal microflora of 49 women in idiopathic preterm labor was compared with that of 38 term controls to determine whether the presence of bacterial vaginosis (BV) and/or specific microorganisms would influence the rate of preterm delivery. Demographic factors, pregnancy outcome, and reproductive history were also studied. BV, as defined by the presence of clue cells in a vaginal wet mount and characteristic microbial findings in a stained vaginal smear and vaginal culture, was more common in women with preterm labor and delivery than in controls (P < 0.01). The condition, diagnosed in 41% of women who had both preterm labor and delivery (n = 22) and in 11% each of women who had preterm labor but term delivery (n = 27) and controls, was associated with a 2.1-fold risk (95% confidence intervals, 1.2 to 3.7) for preterm birth prior to 37 weeks of gestation. BV was associated with low birth weight. Of 49 women with preterm labor, 67% (8 of 12) of women with BV were delivered of low-birth-weight neonates (< 2,500 g) compared with 22% (8 of 37) of women without the condition (P < 0.0005). The presence of hydrogen peroxide-producing facultative Lactobacillus spp. was strongly negatively associated with both preterm delivery and BV. BV-associated microorganisms, i.e., Mobiluncus, Prevotella, and Peptostreptococcus species, Porphyromonas asaccharolytica, Fusobacterium nucleatum, Mycoplasma hominis, and high numbers of Gardnerella vaginalis were significantly associated with preterm delivery; all species also strongly associated with BV (P = 0.0001 for each comparison). Mobiluncus curtisii and Fusobacterium nucleatum were recovered exclusively from women with preterm delivery. Our study clearly indicates that BV and its associated organisms are correlated with idiopathic premature delivery.

Adult↗

Rapid characterization of periodontal bacterial isolates by using fluorogenic substrate tests.

Eighty-nine species of subgingival bacteria, represented by 121 reference strains and 892 patient isolates, including gram-negative, gram-positive, aerobic, facultatively anaerobic, microaerophilic, and anaerobic species, were characterized with a panel of fluorogenic, 4-methylumbelliferyl-linked substrate tests. Identifications of all patient isolates were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell proteins relative to reference strains. Characteristic profiles of positive fluorogenic reactions differentiated most of the species, including five Porphyromonas species, six pigmenting and five nonpigmenting Prevotella species, Bacteroides forsythus, three Capnocytophaga species, six Actinomyces species, four Propionibacterium species, and eight Streptococcus species. Two mannoside isomers differentiated Actinomyces israelii and Actinomyces gerencseriae. In addition to Porphyromonas gingivalis, B. forsythus, and Capnocytophaga species, Fusobacterium alocis, Actinomyces odontolyticus, Actinomyces meyeri, and Bifidobacterium dentium were all positive for so-called trypsin-like activity. Fusobacterium nucleatum, Eikenella corrodens, Actinobacillus actinomycetemcomitans, and Campylobacter species were nonreactive with the carbohydrate-based substrates tested. Fluorogenic substrate tests provided a sensitive and simple method for biochemical characterization that could presumptively identify to species level most subgingival isolates within 4 h. The method was ideal for rapidly obtaining presumptive identifications of isolates prior to confirming identifications by definitive methods, such as SDS-PAGE.

Bacteria↗