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PubMed1995

Role of anaerobic bacteria in chronic otitis media and cholesteatoma.

Otitis media (OM), a common infection in children, can cause significant morbidity. Selection of the most appropriate treatment regimen directed against the pathogens responsible for the OM can minimize complications. The most frequently isolated bacteria from chronic OM are Staphylococcus aureus, Pseudomonas aeruginosa and anaerobic bacteria. The predominant anaerobes are Peptostreptococcus spp., pigmented Prevotella and Porphyromonas spp., Bacteroides spp. and Fusobacterium spp. Many of the organisms causing OM can produce beta-lactamase, which can contribute to the failure of penicillins therapy. The appropriate surgical and medical therapy for chronic OM is reviewed.

Bacteria, Anaerobic↗
PubMed1996

Immunopharmacological activities of the nontoxic monophosphoryl lipid A of Porphyromonas gingivalis.

The low endotoxic lipid A derived from Porphyromonas gingivalis 381, 1-phospho beta(1-6)-linked glucosamine disaccharide with 3-hydroxy-15-methylhexadecanoyl and 3-hexadecanoyloxy-15-methylhexadecanoyl groups at the 2- and 2'-positions, respectively, induced mitogenic responses in LPS low responder C3H/HeJ as well as LPS responder C3H/HeN mouse splenocytes. The mitogenic activities of P. gingivalis lipid A in splenocytes of LPS responder mice were comparable to that of the synthetic Escherichia coli-type lipid A (compound 506). The addition of polymyxin B resulted in the inhibition of mitogenic activity. P. gingivalis lipid A also stimulated strongly nonspecific host resistance against Pseudomonas aeruginosa infection in BALB/c mice, and specific immune response in guinea pigs against infection with P. gingivalis. Furthermore, P. gingivalis lipid A demonstrated antitumour activity against MH134 hepatoma in C3H/HeN mice. These life-preserving with tumour regression properties were comparable to those of monophosphoryl lipid A derived from Salmonella minnesota Re 595. Thus, P. gingivalis lipid A appears to have beneficial properties as an immunopharmacological agent.

Adjuvants, Immunologic↗
PubMed1987

Organisms isolated from severe odontogenic soft tissue infections: their sensitivities to cefotetan and seven other antibiotics, and implications for therapy and prophylaxis.

Cefotetan is a new cephamycin antibiotic characterised by excellent B-lactamase stability and anti-anaerobe activity, coupled with a long half life of 3-4 h which permits twice daily dosage. A clinical trial of cefotetan in the treatment of severe oro-facial infections is presented, together with a detailed analysis of the causative organisms and their sensitivities to eight antibiotics. 50/50 patients achieved clinical cure with a treatment regime comprising cefotetan therapy and incision and drainage, with patients being transferred to oral cephradine for the final phase of therapy. Side effects were minimal and there were no instances of relapse. Significant resistance among alpha-haemolytic streptococci and bacteroides organisms to penicillin was observed. The streptococci were resistant due to mechanisms other than beta-lactamase production. In the light of these findings and the reports of other workers it is suggested that penicillin V may no longer be the most appropriate drug for endocarditis prophylaxis, despite the most recent recommendations of the American Heart Association. Furthermore, if penicillin V is used for this purpose, a penicillin free interval of 6-8 weeks may be inadequate before this drug is used. Cefotetan is not suitable for prophylaxis against endocarditis.

Acute Disease↗
PubMed1987

Selective suppression of the anaerobic oropharyngeal microflora with local metronidazole.

Anaerobic microorganisms of the normal oropharyngeal flora have been shown to be the main pathogens in orofacial infections of odontogenic origin. Reduction of the density of anaerobes in the oral cavity as a prophylactic measure before oral surgery may be a rational way to reduce the frequency of post-operative infections. This report describes a local antibiotic regimen that can reduce anaerobic pathogens in the oral cavity. Over a period of 7 days, 10 healthy individuals applied locally 1.5-2 g of an antibiotic preparation consisting of 0.5% metronidazole in 99.5% Orabase paste three times daily. The paste had antibacterial activity against obligate anaerobic micro-organisms such as bacteroides, fusobacteria and leptotrichia, known pathogens in orofacial infections. Fusobacteria and leptotrichia were eliminated in all subjects. Bacteroides species were eliminated in five subjects and significantly decreased in three subjects. The aerobic microflora was not affected. No new colonisation of the oropharynx was observed during the treatment period. The pre-treatment numbers of different micro-organisms were re-established within 2-9 days after the withdrawal of metronidazole treatment.

Adult↗
PubMedDate not supplied

Clinical significance of dental root canal microflora.

OBJECTIVES: Previous work by this group has shown that a significant association exists between pain and the presence of either Prevotella or Peptostreptococcus spp. in dental root canals. The aim of this study was to examine a more extensive series of canals microbiologically, to determine whether any other particular endodontic symptoms or clinical signs showed specific associations with individual bacterial species. METHODS: Seventy root canals were examined microbiologically and clinical data collected to investigate in detail such associations. RESULTS: Of the canals studied, 37 were associated with pain, 49 with tenderness to percussion, 23 with swelling, six with purulent exudate and 57 presented with wet root canals. Anaerobes were isolated from 70.3% of painful canals and from 29.7% of pain-free canals. Significant associations were found between (a) pain and either Prevotella spp. or peptostreptococci, both with P < 0.01; (b) tenderness to percussion and Prevotella spp. (P < 0.01) or anaerobes (P < 0.05); (c) swelling and Eubacterium spp. (P < 0.01), or with Prevotella spp. or Pstr. micros, both with P < 0.05; (d) purulent exudate and any one of F. necrophorum (P < 0.01), Prev. loescheii, Streptoccoccus constellatus or Bacteroides spp. (each P < 0.05); (e) wet canal and facultative anaerobes (P < 0.01), and any one of the genera of Eubacterium, Peptostreptococcus, Prevotella or Propionibacterium (each P < 0.05). CONCLUSION: It was concluded that several different endodontic clinical signs and symptoms are significantly associated with specific bacterial species.

Adult↗
PubMed1991

Hemin-binding property of Porphyromonas gingivalis outer membranes.

The present study indicates that the outer membranes of Porphyromonas gingivalis have a high affinity for hemin, an essential growth factor for this suspected periodontal pathogen. The hemin-binding property appears to be mediated by the lipopolysaccharides, particularly the lipid A region. It is suggested that this binding mechanism may be important for the growth and establishment of P. gingivalis in environments with a low hemin content such as periodontal pockets.

Bacteroidaceae↗
PubMed1991

Propionigenium modestum: a separate line of descent within the eubacteria.

The complete nucleotide sequence of 16S rRNA from Propionigenium modestum was determined and compared with 380 16S rRNA sequences from representatives of all eu- and archaebacterial phyla known so far. The phylogenetic analysis of this data set indicated P. modestum to represent a new separated line of descent within the radiation of eubacterial phyla moderately related to cyanobacteria and Gram-positive bacteria with low DNA GC content.

Bacteroidaceae↗
PubMed1995

Interaction of Porphyromonas gingivalis with transferrin.

In this study, we characterized the binding of transferrin to Porphyromonas gingivalis using a classical receptor-binding assay, and examined the relationship between the binding and availability of transferrin for the growth of P. gingivalis. The binding of 125I-labeled human transferrin to P. gingivalis occurred rapidly, reversibly and specifically. Scatchard analysis yielded a Kd of 1.37 +/- 0.16 microM and an apparent number of 1.13 +/- 0.26 x 10(5) receptors per cell. The binding of transferrin was much increased when organisms were grown in iron-limited conditions. Among the species of black-pigmented anaerobic.rods, those strains of P. gingivalis which had high transferrin-binding activity exhibited unrestricted growth following the addition of transferrin to the hemin-free culture medium. On the other hand, the presence of transferrin in the culture medium did not support unrestricted growth of organisms that had low transferrin-binding activity. These results suggest that the binding of transferrin to P. gingivalis cells may be a preliminary step in iron acquisition, which allows them to survive in the healthy periodontal environment.

Animals↗
PubMed1995

Partial purification and characterisation of Bfi57I and Bfi89I, restriction endonucleases from different strains of Butyrivibrio fibrisolvens.

Two class-II restriction endonucleases (ENases), Bfi57I and Bfi89I, were partially purified from Butyrivibrio fibrisolvens OB157 and OB189, respectively. Bfi57I (isoschizomer Sau3AI) had the DNA recognition/cleavage sequence 5'-/GATC-3'; it is not inhibited by Dam methylation, but is partially inhibited by M.BamHI methylation. Bfi89I (isoschizomer EaeI) had the recognition/cleavage sequence 5'-Y/GGCCR-3'; unlike the EaeI isoschizomer it is not fully inhibited by M.HaeIII methylation.

Bacteroidaceae↗
PubMed1995

Isolation and characterization of a new restriction endonuclease, Sru30DI, from Selenomonas ruminantium.

The restriction endonuclease (ENase) Sru30DI, an isoschizomer of StuI, which recognizes the sequence 5'-AGG/CCT-3', was purified from a natural isolate of Selenomonas ruminatinum. The ENase was isolated from cell extracts using single-step purification by phosphocellulose column chromatography. Activity of Sru30DI is inhibited by overlapping Dcm methylation. The ENase is extremely stable at 37 degrees C and is active over a wide range of pH, temperature and salt concentrations.

Animals↗
PubMed1983

Discrepancies between in vitro activity of and in vivo response to antimicrobial agents.

The correlation between in vitro antimicrobial activity and the in vivo response to antimicrobial therapy is affected by multiple host factors, the site and nature of the infection, and the pharmacokinetics of the antimicrobial and its penetration into areas of infection. In certain instances, discrepancies are also caused by methodologically-related variables of in vitro susceptibility tests. Examples of discrepancies between in vitro and in vivo response to antimicrobial are discussed.

Anti-Bacterial Agents↗
PubMed1994

Anaerobic susceptibility testing. Slight differences in inoculum size can make a difference in minimum inhibitory concentrations.

In the present study, we compared the actual inoculum density from inoculated broth microdilution wells to the targeted inoculum size (10(5) CFU/well) when the inoculum was prepared using a McFarland nephelometer to achieve the standard density. Three target inoculum sizes (10(5), 5 x 10(5), and 10(6) CFU/well) were used to compare the effect of slight inoculum size increases of both ATCC and clinical strains of anaerobes on MICs of various antimicrobials. Actual colony counts of Bacteroides fragilis, Bacteroides thetaiotaomicron, Eubacterium lentum, and Veillonella parvula ranged from 0.7 x 10(5) to 1.4 x 10(5) CFU/well. As the inoculum size rose above the desired 10(5) CFU/well level, the MICs of certain antimicrobials became elevated. Ceftizoxime, cefotaxime, and ceftriaxone MICs rose 4- to 16-fold with as little as 0.5 log10 increase in inoculum size. Other increases were also noted with E. lentum and Clostridium perfringens, but were primarily between the low and high inoculum sizes. Results with cefoxitin, cefotetan, mezlocillin, and imipenem did not show an appreciable increased inoculum effect. This study demonstrates that the variation in organism size among anaerobes (both between species and within species) does make a difference in actual inoculum size and certain anaerobes may require special adjustment to ensure proper MIC results from susceptibility testing.

Anti-Bacterial Agents↗
PubMedDate not supplied

Phenotypic characterization of human and animal biotypes within the species Porphyromonas gingivalis.

Ninety-nine strains of Gram-negative black-pigmented anaerobic rods, grown on Todd-Hewitt blood agar plates, were identified and characterized according to a typing scheme including UV fluorescence, catalase, trypsin-like and haemagglutinating activities, biochemical tests with the ATB 32A kit, and gas-liquid chromatography. To determine the taxonomic position of the Porphyromonas gingivalis biotypes, 68 strains (31 of human origin and 37 of animal origin) were compared to 31 strains of closely related species or of uncertain generic status. Most animal strains were isolated in our laboratory by subculturing samples from the oral cavity of five mammalian species (bear, cat, coyote, dog and wolf). Those strains differed from human P. gingivalis strains in that they were positive for catalase, beta-galactosidase and glutamyl-glutamic acid arylamidase; from Bacteroides macacae by more rapid pigmentation, positive haemagglutination, failure to produce propionic acid, and negative alpha-galactosidase; and from Bacteroides salivosus by more rapid pigmentation, positive haemagglutination and failure to produce propionic acid. These data demonstrate that phenotypic heterogeneity within the taxon P. gingivalis can be resolved into two biotypes, each corresponding to a human source or an animal source.

Animals↗
PubMedDate not supplied

Randomly amplified polymorphic DNA analysis confirms the biotyping scheme of Porphyromonas gingivalis.

The application of the arbitrarily primed PCR (AP-PCR) procedure to generate randomly amplified polymorphic DNA (RAPD) fingerprints for the study of the taxon Porphyromonas gingivalis was investigated. Nine human strains and seven animal strains of P. gingivalis as well as eighteen strains other than P. gingivalis were analysed. Four nanomer primers of random sequence were evaluated for their ability to distinguish genetic diversity. Three primers generated RAPD fingerprints that allowed the sixteen strains to be differentiated; two of the primers yielded species-specific markers, and two of the primers permitted biotype distinction. Cluster analysis of the RAPD fingerprints revealed two major phenetic groups that matched the human and animal biotypes. Our results indicate that AP-PCR (i) can generate strain-specific fingerprints, (ii) confirms genetic heterogeneity and the biotype grouping of the P. gingivalis taxon, and (iii) enables identification of potential genetic markers at the species, biotype and subtype levels and is thus a promising tool for bacterial systematics. Our results also underline the potential of AP-PCR for epidemiological studies of periodontal pathogens.

Animals↗
PubMed2006

An ortho dimer of butylated hydroxyanisole inhibits nuclear factor kappa B activation and gene expression of inflammatory cytokines in macrophages stimulated by Porphyromonas gingivalis fimbriae.

Butylated hydroxyanisole, BHA, is widely used as a potent antioxidant, but its adverse effects such as carcinogenesis and proinflammatory activity have been reported, which are possibly due to the prooxidant property of this compound. We recently demonstrated that the dimer of 2-methoxyphenols exhibits cyclooxygenase-2 inhibition, because of lessening of its prooxidant property caused by the dimerization. In the present study, toward our goal of developing a chemopreventive agent for chronic periodontal diseases, we examined whether 2-BHA (2-tert-butyl-4-methoxyphenol) and its synthetic ortho dimer, bis-BHA (3,3'-di-tert-butyl-5,5'-dimethoxy-1,1'-biphenyl-2,2'-diol) could inhibit the Porphyromonas gingivalis fimbria-stimulated inflammatory reaction. The fimbria-induced expression of interleukin-1beta and neutrophil chemoattractant KC genes in RAW264.7 murine macrophages was strongly inhibited by bis-BHA. In contrast, 2-BHA showed only slight inhibition. bis-BHA also significantly inhibited the fimbria-stimulated phosphorylation-dependent degradation of the alpha inhibitor of nuclear factor-kappaB and the transcriptional activity of this factor in the cells. These findings suggest that bis-BHA possesses anti-inflammatory activity against chronic periodontal diseases.

Animals↗
PubMed2004

Experimental intrauterine infection with Prevotella bivia in New Zealand White rabbits.

OBJECTIVE: The purpose of this study was to develop a model of chronic intrauterine and fetal infection with Prevotella bivia, an anaerobe of the lower genital tract that is associated often with bacterial vaginosis. STUDY DESIGN: Thirty timed pregnant New Zealand White rabbits on gestational day 21 were inoculated with P bivia or saline solution in a planned ratio of 4:1 (24 P bivia: 6 saline solution). Rabbits were inoculated 6 cm transcervically with 10(5) to 10(8) colony-forming units/uterine horn of P bivia or with saline solution. Necropsy was scheduled on days 4, 6, or 7 after inoculation. Cultures were collected from blood, uterus, amniotic fluid and fetal brain, lung, and heart. Tissues from placenta, uterus, fetal brain, and lung were evaluated with the histologic inflammation score, with a range of 0 to 13. Amniotic fluid was assayed for tumor necrosis factor-alpha by bioassay. Animals with contamination by other organisms were excluded. Categoric data were evaluated with the use of the Fisher exact test, and continuous data were evaluated with the use of the Wilcoxon rank sum. RESULTS: After the exclusion of 8 animals because of contamination with other organisms, 22 animals were evaluated. Of 3 rabbits with an inoculum of 10(8) P bivia colony-forming units/horn, 2 animals (67%) had fever within 24 hours. These results were not compatible with chronic, subclinical infection. Therefore, 14 does had inocula of 10(5-6) P bivia colony-forming units/horn, with necropsy planned at day 4 (n=5 animals), day 6 (n=3 animals), and day 7 (n=6 animals), and 5 animals were inoculated with saline solution. Animals that had been inoculated with P bivia were significantly more likely to have a positive culture than were those animals that were inoculated with saline solution (64% vs 0%; P<.04). Preterm delivery without fever occurred in 21% of does (3/14 does) that were inoculated with P bivia overall and in 33% of the does (3/9 does) that were followed for 6 to 7 days. No saline-solution inoculated animal had preterm birth. There was an increase in amniotic fluid tumor necrosis factor-alpha levels over time in the P bivia group (P=.12). Histologic inflammation scores were not significantly different between P bivia and saline solution groups. CONCLUSION: Inoculation with P bivia at 10(5-6) colony-forming units/horn leads to chronic intrauterine and fetal infection that are accompanied by preterm birth in up to 33% of cases. This model may serve to explore the mechanism of preterm birth that is induced by chronic infection with genital tract anaerobes.

Animals↗