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Anaerobic infections in childhood.

Anaerobic bacteria are part of the normal flora of mucous membranes and outnumber aerobic bacteria in the oral cavity and gastrointestinal tract. Anaerobes can be isolated from pediatric patients with various infections when appropriate techniques for transportation and cultivation of samples are employed. Frequently anaerobes are isolated in combination with other facultative or aerobic bacteria. The genera or groups of anaerobes most frequently isolated from pyogenic infections in children are (in order of decreasing frequency) OFFteroides, Clostridium, gram-positive cocci, Fusobacterium, gram-positive rods (Eubacterium, Lactobacillus, Propionibacterium, Actinomyces, and Bifidobacterium), and gram-negative cocci (Veillonella and Acidaminococcus). Clostridium perfringens causes bacteremia and wound infections. Clostridium botulinum can produce a paralytic toxin that causes a lethal illness in adults and a paralytic syndrome in infants. Clostridium difficile can cause antibiotic-associated colitis or diarrhea. Bacteroides fragilis is most frequently involved in intraabdominal infections, infections of the female genital tract, subcutaneous abscesses, and bacteremia. Bacteroides melaninogenicus and Bacteroides oralis are the predominent anaerobes in orofacial infections and aspiration pneumonia. Fusobacterium species are pathogens in aspiration pneumonia, brain abscesses, and orofacial infections. Anaerobic gram-positive cocci can be recovered from all types of infections but predominate in respiratory tract and intra-abdominal infections. Recognition of the pathogenic qualities of the various anaerobic organisms can assist in their prompt identification and in the initiation of appropriate therapy.

Actinomyces↗

Mechanisms of beta-lactam resistance in anaerobic bacteria.

The known mechanisms of beta-lactam resistance in anaerobic bacteria involve production of beta-lactamases, alteration of penicillin-binding proteins, and blocking of the penetration of beta-lactams through the outer membranes. The most important factor in beta-lactam resistance is the production of beta-lactamase. beta-Lactamases in various Bacteroides, Fusobacterium, and Clostridium species have been described. beta-Lactam resistance in Bacteroides fragilis is most commonly mediated by the production of beta-lactamase, primarily of the cephalosporinase type. Studies have also shown that B. fragilis can produce a penicillinase that inactivates piperacillin and carbenicillin. Enzymes that inactivate cefoxitin and imipenem have also been found in B. fragilis. The nonfragilis Bacteroides species produce beta-lactamases mainly of the penicillinase type. Recently a penicillinase from Fusobacterium nucleatum has been characterized. Among the clostridia, Clostridium butyricum, clostridium clostridiiforme and Clostridium ramosum have been shown to produce penicillinases.

Anti-Bacterial Agents↗

Susceptibility of anaerobic bacteria to metronidazole: relative resistance of non-spore-forming gram-positive baccilli.

Susceptibility of 358 clinical isolates of obligate anaerobes to metronidazole was determined by an agar-dilution technique. Only 66% of all isolates were inhibited by 6.25 mug/ml, whereas 30% required larger than or equal to 50 mug/ml. Considerable variation in susceptibility was observed among different genera and species of bacteria. Fusobacterium was most senstitive, followed by Clostridium, Bacteroides and Peptococcus, Peptostreptococcus, Veillonella and Acidaminococcus, and non-spore-forming gram-positive bacilli. Bacteroides fragilis was more sensitive than other species of Bacteroides. Similarly, Clostridium perfringens was more susceptible than other species of Clostriduim. While metronidazole appears to be a promising antimicrobial agent for infections caused by Fusobacterium, Clostrididium, and B.fragilis, therapy for infections with other anaerobic bacteria should be guided by in vitro tests of sensitiivity.

Anaerobiosis↗

Isolation and identification of bacteria from subgingival plaque with low susceptibility to minocycline.

Twenty subgingival plaque samples from patients with chronic periodontitis were screened for the presence of three periodontal 'pathogens'--Porphyromonas gingivalis, Fusobacterium nucleatum and Eikenella corrodens. Nineteen of the samples were found to contain at least one of the three organisms and six samples contained all three. The subgingival plaque samples were cultured in the presence of various concentrations of minocycline, and those organisms which appeared to be least susceptible to the antibiotic were isolated and identified. A total of 40 isolates were obtained and these consisted of 18 different species, only one of which, Fusobacterium nucleatum, is generally recognized as being associated with chronic periodontitis. The minimum inhibitory concentrations of minocycline for many of the organisms isolated were greater than the concentration of attainable in gingival crevicular fluid following routine parenteral administration of the antibiotic.

Adult↗

Review of the in vitro activity of gemifloxacin against gram-positive and gram-negative anaerobic pathogens.

Published reports on the in vitro activity of gemifloxacin mesylate (SB 265805), a new fluoronaphthyridone, against anaerobic pathogens are reviewed here. The studies used a variety of media, inocula and antimicrobial agents. Using a proposed breakpoint of 0.5 mg/L, these studies showed that gemifloxacin had generally higher potency against Gram-positive anaerobes (Clostridium perfringens, all Peptostreptococcus spp.) and fusobacteria (Fusobacterium nucleatum, Fusobacterium necrophorum) and moderate but variable potency against Gram-negative anaerobes. Bacteroides stercoris, Bacteroides tectum and many Bacteroides fragilis isolates were inhibited by concentrations of < or =0.5 mg/L, while the other species of the B. fragilis group required higher concentrations for inhibition. Species variability was evident: Porphyromonas asaccharolytica, Porphyromonas canoris, Porphyromonas gingivalis, Porphyromonas macaccae, Prevotella heparinolytica and Prevotella intermedia were susceptible to 0.5 mg/L of gemifloxacin while most other Porphyromonas and Prevotella spp. were not. These data suggest that gemifloxacin may have a clinical role in the treatment of certain dental, head and neck and pleuropulmonary infections in which Gram-positive anaerobes, fusobacteria and some Prevotella and Porphyromonas spp. may predominate.

Anti-Infective Agents↗

Lemierre syndrome.

Lemierre syndrome is characterized by pharyngitis followed by Fusobacterium necrobacillosis sepsis complicated by internal jugular vein thrombosis and infectious metastatic abscesses. It has been considered to be a rare disease until the last decade when a larger number of cases have been reported. We discuss a case of Lemierre syndrome in a 17-year-old girl and review the pathophysiology of Fusobacterium necrobacillosis and diagnosis and treatment of Lemierre syndrome.

Abscess↗

The isolation and identification of Bacteroides spp. from the normal human gingival flora.

Gram-negative anaerobic bacilli were isolated on an enriched selective medium from specimens of subgingival plaque from 20 normal, healthy adults without periodontal disease. a heavy growth of Bacteroides and Fusobacterium spp. was obtained from all specimens and 10 representative colonies from each subject were selected for identification by conventional bacteriological methods. Results were obtained in these tests with 188 isolates; some strains of the B. melaninogenicus/oralis group would not grow in pure culture on solid media and showed satellitism in mixed culture, and some strains of fusobacteria did not survive repeated subculture. Most isolates (68%) belonged to the B. melaninogenicus/oralis group. The commonest species identified in this group were B. oralis (41%), B. melaninogenicus ss. melaninogenicus (26%) and ss. intermedius (17%). B. oralis was isolated from all subjects and at least one strain of B. melaninogenicus was isolated from 18 out of 20 subjects. Only eight strains of the B. fragilis group were detected and they probably represented transient carriage. Fusobacterium spp. and L. buccalis were isolated from 15 out of 20 subjects and accounted for 21% of the isolates studied, but most could not be identified specifically by the set of tests used. Asaccharolytic species of Bacteroides were isolated in small numbers from seven subjects.

Adult↗

The identification of Bacteroides ureolyticus from patients with non-gonococcal urethritis by conventional biochemical tests and by DNA and protein analyses.

Twenty-seven strains of gram-negative anaerobic bacteria isolated from patients with non-gonococcal urethritis (NGU-associated strains) were compared with reference strains of Bacteroides ureolyticus and representatives of other species of Bacteroides and Fusobacterium. Conventional biochemical tests, electrophoretic (PAGE) protein profiles and DNA base compositions of 29.5 +/- 1.2 mol% G + C indicated striking similarities between the NGU-associated strains and B. ureolyticus. Three of the reference strains previously assigned to that species were atypical in some respects as was one of the strains from urethritis and one strain, NCTC 10939, was incorrectly classified as B. ureolyticus. Casein hydrolysis by B. ureolyticus was the principal test difference between the two sets of strains. DNA-DNA hybridisation results of greater than or equal to 72% at optimum temperature conditions provided further evidence of the close relationship between B. ureolyticus and the NGU-associated strains and we conclude that these organisms should be identified as B. ureolyticus. There was insufficient evidence for recognising subspecies or biotypes within this revised concept of B. ureolyticus although several types (PAGE-type I, 17 strains; type II, 6 strains; type III, 3 strains) were detected which might provide the basis for future studies of hospital strains. The DNA base composition and hybridisation data indicated that B. ureolyticus was not related to other species of Bacteroides or Fusobacterium.

Bacterial Proteins↗

A longitudinal study of the cultivable subgingival anaerobic bacteria isolated from sheep during the development of broken mouth periodontitis.

In a longitudinal bacteriological study of the cultivable subgingival anaerobic flora isolated from developing broken mouth periodontitis in sheep, samples were taken from five sheep on each of three farms on seven occasions over a period of 2.5 years. Ten different bacterial genera were isolated regularly but with fluctuating frequencies. Bacteroides and Fusobacterium organisms accounted for nearly 70% of the isolates. The Bacteroides and Fusobacterium isolates studied in detail from one farm were identified to species level. The fusobacteria comprised F. nucleatum-like organisms (68.6%). F. necrophorum (29.6%) and F. naviforme (1.8%). The Bacteroides spp. were divided into 11 main groups and included black-pigmented species similar to B. asaccharolyticus and B. gingivalis. On the farm studied in detail, the sheep could be allocated to two groups according to progression of periodontal disease. Most of the B. gingivalis-like isolates were from sheep with actively progressing disease, indicating that this organism may play a role in periodontal destruction in sheep.

Animals↗

Identification of fusobacteria in a routine diagnostic laboratory.

A scheme for differentiating Fusobacterium spp. and Leptotrichia spp. from Bacteroides spp. was devised after examining 114 strains of fusobacteria and asaccharolytic bacteroides (17 reference strains and 97 clinical isolates). Sensitivity to a 300 micrograms/ml plate of phosphomycin and an acid reaction on a lysine plate were found to be reliable for differentiating Fusobacterium spp. and L. buccalis from Bacteroides. Using a short set of simple cultural and biochemical tests, isolates could be identified as F. necrophorum, F. necrogenes, F. nucleatum, F. varium or L. buccalis. These tests were: indole, lecithinase, phosphatase, DNase and gas production, aesculin and casein hydrolysis, greening of casein/methylene blue agar, nitrite reduction, bile tolerance and haemolysis on horse blood agar.

Anti-Bacterial Agents↗

Comparison of populations of human faecal bacteria before and after in vitro incubation with plant cell wall substrates.

Human faecal slurries were incubated anaerobically with larchwood xylan, oat spelt xylan, wheat bran, apple cell walls or sugar beet pulp as sole carbon sources. The populations which developed during incubation were different from the inoculum, the most marked changes being an increase in the number of Bacteroides species and a decrease in the number of Fusobacterium species for all carbon sources tested. With a water-soluble preparation of larchwood xylan the population was dominated by species able to ferment this substrate, in contrast to the population which developed with the insoluble substrates. The ability to use one plant cell wall substrate appeared to be related to the ability to use others. Strains capable of using plant cell wall substrates included Bacteroides spp., Clostridium clostridiiforme, Bifidobacterium longum, Fusobacterium spp. and Escherichia coli. When incubation with two contrasting substrates (bran and larchwood xylan) was replicated, the populations which developed were reproducibly different from the inoculum and from each other.

Bacteroides↗

Microbiology of infected atopic dermatitis.

BACKGROUND: Bacterial infections occur frequently in lesions of atopic dermatitis (AD). The objectives of the study were to establish the aerobic and anaerobic microbiology of secondarily infected AD. METHODS: A retrospective review was carried out of clinical and microbiology laboratory records and of data obtained from patients with secondarily infected AD lesions, whose specimens of infected sites were processed for the presence of aerobic and anaerobic bacteria. RESULTS: Bacterial growth was noted in 41 specimens. Aerobic or facultative anaerobic bacteria only were present in 15 patients (36%), anaerobic bacteria only in eight (20%), and mixed anaerobic-aerobic flora was present in 18 (44%). Seventy-two isolates were recovered (1.8 per specimen), 34 aerobic or facultative bacteria, 35 strict anaerobes, and three Candida sp. The predominant aerobic and facultative bacteria were Staphylococcus aureus (12 isolates), Group A beta hemolytic streptococcus in five isolates, and Escherichia coli in four. The predominant anaerobes were Peptostreptococcus spp. (13 isolates), pigmented Prevotella and Porphyromonas spp. in eight, and Fusobacterium spp. in four isolates. Single bacterial isolates were recovered in 14 patients (34%), seven of which were S. aureus. Twenty-one of the organisms isolated from 16 patients (39%) produced the enzyme beta-lactamase. Organisms that resided in the mucous membranes close to the lesions predominated in these infections. Enteric gram-negative rods and Bacteroides fragilis group predominated in lesions on legs and buttocks. Group A beta-hemolytic streptococci, pigmented Prevotella, and Porphyromonas and Fusobacterium spp. were most frequently recovered in lesions of the finger, scalp, face, and neck. CONCLUSIONS: The polymicrobial etiology of secondarily infected AD lesions and the association of bacterial flora with the anatomic site of the lesions are demonstrated.

Adolescent↗

Effects of periodontopathic bacteria on IL-1 and IL-1 inhibitor production by human polymorphonuclear neutrophils.

The effect of heat-killed periodontopathic bacteria on the production of interleukin-1 (IL-1) and an IL-1 inhibitor by human polymorphonuclear neutrophils (PMN's) was examined. Peripheral blood was obtained from 18 healthy volunteers and the PMN's were separated using dextran sedimentation and Ficoll-Paque density gradient centrifugation. The PMN's (5 x 10(5) cells/well) were cultured in serum-free media with or without heat-killed periodontopathic bacteria. Four gram-negative periodontopathic bacteria were used; Bacteroides gingivalis FDC 381, Bacteroides forsythus FDC 338, Actinobacillus actinomycetemcomitans Y4 and Fusobacterium nucleatum FDC 263. The non-oral Fusobacterium mortiferum ATCC 25557 was used as a control organism. IL-1 activity was assayed using thymocyte proliferation. The non-oral organism F. mortiferum stimulated IL-1 production by PMN's, in contrast none of the periodontopathic bacteria stimulated IL-1 release although the bacteria themselves had an IL-1 enhancing effect. Following fractionation of the periodontopathic bacteria stimulated PMN supernatants, an IL-1 inhibitory fraction was identified. These results may illustrate a further mechanism by which periodontopathic bacteria may evade the protective effect of PMN's and may also suggest a regulatory role for PMN's in chronic inflammatory periodontal disease.

Actinomyces↗

Microbiology of early supragingival plaque development after delmopinol treatment.

The aim of this double blind, cross-over, microbiological study was to compare the effect of topical application of the plaque control agent 0.5% delmopinol HCl with placebo on early supragingival plaque formation. Six subjects underwent 7 periods (0.5, 1, 2, 8 and 24 h and 3 and 7 days) of placebo and delmopinol application, respectively. At the start of each study period the teeth were professionally cleaned and 2 ml of placebo and delmopinol 0.5%, respectively, were applied on all teeth (twice daily for periods lasting 24 h or more). At the end of each period, supragingival plaque samples of one upper and one lower buccal tooth surface were collected separately and cultured on anaerobically incubated Brucella blood agar, on aerobically incubated blood agar and on selective media for the enumeration of Streptococcus spp., Haemophilus spp., Actinomyces spp., Veillonella spp., Neisseria spp. and Fusobacterium spp. The total anaerobic cultivable microflora after delmopinol use was 10-100 times lower than after placebo use. Compared with placebo, the proportion of cultivable aerobes (61.3%), Streptococcus spp. (104.8%) and Haemophilus spp. (82.3%) increased and the proportion of Actinomyces spp. (86.1%), Veillonella spp. (60.5%), Neisseria spp. (96.9%) and Fusobacterium spp. (60.6%) decreased after 7 days. Short-term application of 0.5% delmopinol HCl on supragingival dental plaque regrowth resulted in a reduction of the number of cultivable microorganisms in the plaque and produced a shift in the cultivable plaque composition.

Actinomyces↗

The oral gram-negative anaerobic microflora in young children: longitudinal changes from edentulous to dentate mouth.

Eruption of primary teeth has a great influence on the oral environment by providing suitable niches for bacterial colonization. The composition of oral gram-negative anaerobic microflora was investigated in 21 young children (mean age 32 months) with primary dentition. The bacterial findings of samples were compared with those of the same children collected at their edentulous infant period (mean age 3 months). During the primary period, 2 samples were collected from each child: a sample with dental floss from gingival margin of 2 teeth and stimulated saliva pooled with a mucosal swab sample. Both samples were cultured aerobically and anaerobically using nonselective and selective media. Prevotella melaninogenica, nonpigmented Prevotella spp., Fusobacterium nucleatum group and Capnocytophaga spp. were found in all children at the older age, whereas they occurred in edentulous mouth in 76%, 62%, 67% and 19%, respectively. The occurrence of Prevotella loescheii increased from 14% to 90%, Prevotella intermedia from 10% to 67%, Leptotrichia spp. from 24 to 71%, Campylobacter (Wolinella) spp. from 5 to 43% and Eikenella corrodens from 5 to 57%. Only the occurrence of Bacteroides gracilis and Veillonella spp. remained at about the same level. Species not isolated from the edentulous mouth, such as Prevotella denticola, Fusobacterium spp. other than the F. nucleatum group and Selenomonas spp. were now detected in 71%, 71% and 43% of the children. The stability of the colonizing P. melaninogenica strain(s) in the oral cavity was determined by using ribotyping; 1-2 isolates per child from the edentulous infant period of 9 children and 3-15 isolates per child from their primary dentition period were analyzed.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Typing Techniques↗

Cell envelope and cell wall immunization of Macaca fascicularis: effect on the progression of ligature-induced periodontitis.

The nonhuman primate, Macaca fascicularis, was used to study the role of immunization with selected members of the periodontopathic microbiota in the longitudinal progression of ligature-induced periodontitis. Animals were immunized with cell envelope antigens prepared from Porphyromonas gingivalis and Prevotella intermedia, and a mixture prepared from Fusobacterium nucleatum, Campylobacter rectus, and Actinomyces viscosus. Serum immunoglobulin G (IgG), IgM and IgA isotype antibodies increased significantly in all immunization groups and were specific for each of the immunogens. P. gingivalis and P. intermedia immunization resulted in a stabilization of the proportions of these species throughout most of the experiment. The high P. gingivalis antibody titer resulted in low P. gingivalis numbers being recovered. P. gingivalis immunization, while lowering recoverable viable P. gingivalis, resulted in significantly increased levels of Prevotella loescheii, Prevotella buccae, Bacteroides macacae and Prevotella melaninogenica compared with preligation and preimmunization levels. Actinobacillus actinomycetemcomitans, Capnocytophaga spp. and Eikenella spp. remained at preligation levels postimmunization. Campylobacter spp. increased significantly during the course of the experiment in all groups, whereas the levels of Fusobacterium spp. decreased. Plaque indices and bleeding on probing showed significant increases in all groups following ligation, with the placebo group showing the greatest increase. Pocket depth measurements revealed that , whereas the placebo animals showed an approximate 5% increase, the P. gingivalis- and P. intermedia-immunized groups showed nearly a 20% increase in pocket depth. Attachment level measurements showed significantly greater attachment loss in the P. gingivalis- and P. intermedia-immunized groups, and the F. nucleatum + C. rectus + A. viscosus immunization appeared to prevent significant changes in pocket depth/attachment level loss. Radiographic measurement of bone loss by computer-assisted densitometric image analysis revealed that the placebo group lost bone throughout the experiment. P. gingivalis- and P. intermedia-immunized groups showed an exacerbated loss of bone density and the group immunized with F. nucleatum + C. rectus + A. viscosus exhibited significantly lower amounts of bone loss when analyzed by computer-assisted densitometric image analysis, compared with the other immunized groups. Although immunization with P. gingivalis and P. intermedia cell envelope antigens had an effect on their emergence in the complex microbiota of the developing periodontal pocket, this immunization also resulted in greater bone loss than immunization with F. nucleatum + C. rectus + A. viscosus, suggesting that, whereas selective members of the putative periodontopathic microbiota may play a direct role in periodontal tissue destruction, the complexity of the subgingival microbiota dictates that considerable scrutiny is required to select useful immunogens that can elicit functional protection from periodontal tissue destruction induced by oral microorganisms that already colonize or infect the host.

Actinomyces viscosus↗

Profiling of bacterial flora in crevices around titanium orthodontic anchor plates.

OBJECTIVES: The aims of this study were to characterize the microflora in crevices around titanium orthodontic anchor plates using anaerobic culture and molecular biological techniques for bacterial identification, and to compare the microbial composition between crevices around anchor plates and gingival crevices. MATERIAL AND METHODS: Samples from crevices around titanium anchor plates and healthy gingival crevices of 17 subjects (aged 20-29) were cultured anaerobically, and isolated bacteria were identified by 16S rRNA sequencing. RESULTS: The average logarithm colony-forming units/ml were 6.84, 7.51 and 8.88 in healthy anchor plate crevices, inflamed anchor plate crevices and healthy gingival crevices, respectively, indicating that the bacterial density of anchor plate crevices was lower than that of healthy gingival crevices. Of 184 strains isolated from healthy anchor plate crevices of seven subjects, 108 (59%) were anaerobic bacteria, while 73 (40%) were facultative bacteria. Predominant isolates were Gram-negative rods, such as Campylobacter (12%), Fusobacterium (10%) and Selenomonas (10%), and Gram-positive facultative bacteria, such as Actinomyces (17%) and Streptococcus (8.2%). Of 133 strains isolated from inflamed anchor plate crevices of three subjects, 110 (83%) were anaerobic bacteria, while predominant isolates were Gram-negative rods, such as Prevotella (47%), Fusobacterium (33%) and Campylobacter (16%). On the other hand, of 146 strains isolated from healthy gingival crevices of seven subjects, 98 (67%) were facultative bacteria, while 45 (31%) were anaerobic bacteria. Predominant isolates were Gram-positive facultative bacteria, such as Actinomyces (37%) and Streptococcus (20%). CONCLUSIONS: These results suggest that the environment in crevices around titanium orthodontic anchor plates is anaerobic and supportive of anaerobic growth of bacteria, which may trigger inflammation in the tissue around the plates.

Actinomyces↗

The microbiology of HIV-associated periodontal lesions.

2 intraoral lesions associated with human immunodeficiency virus (HIV) infection have recently been described: an atypical gingivitis and a rapidly progressive periodontitis. The microbiota associated with these gingival and periodontal lesions was investigated. Subgingival plaque samples were taken from 45 HIV-seropositive homosexual men and from 44 HIV-seronegative control subjects. Each sampled site was clinically and radiographically classified as HIV-associated gingivitis, HIV-associated periodontitis, healthy in an HIV-seropositive subject, or healthy, conventional gingivitis or classical periodontitis in a control subject. Plaque samples were examined by indirect immunofluorescence with polyclonal antisera to detect Bacteroides gingivalis, B. intermedius, Fusobacterium nucleatum, and Actinobacillus actinomycetemcomitans. Anaerobic culturing was used to detect black-pigmented Bacteroides species, Fusobacterium species, and A. actinomycetemcomitans to confirm the immunofluorescence findings. We detected B. gingivalis, B. intermedius, F. nucleatum, and A. actinomycetemcomitans in significantly more HIV-periodontitis sites (80, 65, 59 and 61% of sites, respectively) and HIV-gingivitis sites (61, 70, 52 and 52%, respectively) than in HIV-seropositive healthy and control sites (p less than 0.05). The results indicate that the microbiota found in HIV-periodontitis is similar to that of classical periodontitis. In contrast, however, the microbiota associated with HIV-gingivitis is strikingly different from that of conventional gingivitis. The similarity in the prevalence of periodontopathic organisms in both HIV-gingivitis and HIV-periodontitis suggests that the HIV-gingivitis lesion may be a precursor to the tissue destruction observed in HIV-periodontitis. Hence, early detection and treatment of the HIV-gingivitis lesion may prevent the rapid and extensive breakdown of periodontal tissues associated with HIV-periodontitis.

Actinobacillus↗