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PubMed1972

Carbon monoxide production from heme compounds by bacteria.

Carbon monoxide formation from heme compounds by bacteria was investigated to study microbial hemoprotein catabolism with reference to heme degradation by mammalian tissues. Hemolytic and nonhemolytic bacteria were incubated aerobically and anaerobically with the following substrates: erythrocytes, hemoglobin, myoglobin, cytochrome c, hematin, iron hematoporphyrin, copper hematoporphyrin, protoporphyrin, and bilirubin. After 18 hr at 37 C the evolved CO was measured by gas chromatography. None of the bacteria formed CO anaerobically. Under aerobic conditions both alpha-hemolytic Streptococcus mitis and hemolytic Bacillus cereus formed CO from all of the heme compounds tested, whereas nonhemolytic Streptococcus mitis did not evolve CO from any of the substrates. The hemolytic bacteria did not produce CO when the iron of heme was either replaced by copper or removed, as in copper hematoporphyrin and in protoporphyrin, respectively.

Aerobiosis↗
PubMed1994

The effect of indigenous bacterial populations on buccal epithelial cells on subsequent microbial adhesion in vitro.

Despite the numerous investigations on the adhesion of microorganisms to buccal epithelial cells, it is difficult to correlate different results, as disparate adherence values have been reported for a given organism. As one reason for this disparity may be the indigenous or natural bacterial populations on human buccal epithelial cells, the effect of the latter on subsequent microbial adherence in vitro was examined. There was a highly significant correlation between the degree of natural bacterial colonization on pooled buccal epithelial cells from 8 healthy donors and the adhesion of a single isolate each of Streptococcus mitis, Escherichia coli and Actinomyces naeslundii. However, no such relationship could be established for Candida albicans, Streptococcus milleri and another isolate of Streptococcus mitis. As in previous studies, variation in adherence values was found, both between samples from different donors, and from the same donor over time, but to a far lesser degree in pooled samples from different donors. These results imply that natural bacterial populations on buccal epithelial cells may affect the adhesion values derived from laboratory experimentation, and hence such data should be interpreted with caution.

Actinomyces↗
PubMed2005

Hematogenous infection of the human temporomandibular joint.

Objective This study was designed to discover the relationship between bacteremia and the presence of specific bacterial species in the synovial fluid of the human temporomandibular joint (TMJ). Study design Sixteen volunteers (female to male, 1:2.2; average age, 30.00 +/- 9.93 years) who received operations via intraoral incision participated in this study. Samples from the blood and TMJ synovial fluid of the patients were taken preoperatively and postoperatively and analyzed by PCR assays with specific primers for the clinically important bacteria Staphylococcus aureus , Streptococcus mitis , and beta-hemolytic Streptococcus . Results PCR-based assays revealed the occurrence of S aureus in 68.8% and 50% of the total blood and synovial fluid samples collected immediately after the operations, which statistically differed from the preoperative samples ( P < .05). However, the frequency of the other bacteria, S mitis and beta-hemolytic Streptococcus , in pre- and postoperative blood samples had no such a difference ( P > .05). The chi-square test showed a significant association between the presence of S aureus in the blood and in TMJ synovial fluid (chi 2 = 6.409, P < .05), and the probability of hematogenous infection of the TMJ was estimated as 55.5%. Conclusion Hence, the data obtained provided evidence that the invasion of S aureus into TMJ synovial fluid was due to the presence of these bacteria in the blood. To our knowledge, this is the first report of the bacteremia-related mechanism of S aureus invasion in the human TMJ.

Adult↗
PubMed1976

Metabolism of the reserve polysaccharide of Streptococcus mitior (mitis): is there a second alpha-1,4-glucan phosphorylase?

The alpha-1,4-glucan phosphorylase (alpha-1,4-glucan: orthophosphate glucosyltransferase; EC 2.4.1.1) associated with the particulate cell fraction of Streptococcus mitior strain S3 was compared with the soluble maltodextrin phosphorylase that had been previously isolated from the same organism (Walker et al., 1969). The particulate enzyme was more sensitive to the glycogen content of the cell than the soluble euzyme; its activity was highest when the cells were grown under conditions favoring high glycogen storage. Substrate specificities of the two high activity towards endogenous glycogen, whereas low-molecular-weight maltodextrins were the preferred substrates for the soluble phosphorylase. The purification of the particulate phosphorylase included incubation of the particulate fraction in 160 mM sodium phosphate-10 mM sodium citrate-0.1% (wt/vol) Triton X-100 buffer (pH 6.7) and ion-exchange chromatography on diethylamino-ethyl- Sephadex A-50. The purified enzyme was fully soluble. The value for the purification factor was variable and depended on (i) the substrate used and (ii) whether the synthetic or the degradative reaction was being measured. The solubilization resulted in considerable changes in the properties of the phosphorylase: the pH optimum for activity was raised from 6.0 to 7.0-7.5 and the substrate specificity was altered. Consequently, the purified enzyme bore greater similarity to the soluble maltodextrin phosphorylase. The reported results are best explained in terms of a single phosphorylase, the specificity which is determind by its binding state in the cell. The enzyme acts as a glycogen phosphorylase in the particulate state and as a maltodextrin phosphorylase when soluble. The equilibrium between the two forms is related to the glycogen content of the cells.

Glycogen↗
PubMed1992

In vitro antimicrobial susceptibility of viridans streptococci isolated from blood cultures.

The susceptibility of 211 viridans streptococci isolated from blood cultures to eight antimicrobial agents was determined. All the isolates were susceptible to cefotaxime, ceftriaxone, imipenem and vancomycin. Thirty eight percent of the isolates were resistant to penicillin (MICs greater than or equal to 0.25 micrograms/ml). Tetracycline resistance was found in 41% of the isolates and in 7% of these strains tetracycline resistance was combined with erythromycin resistance. Five Streptococcus mitis isolates exhibited increased (MIC 64 micrograms/ml and 128 micrograms/ml) or high-level (MIC greater than or equal to 500 micrograms/ml) resistance to gentamicin, kanamycin and tobramycin. Four of these isolates were also resistant to penicillin (MICs 16-32 micrograms/m). In vitro synergy was not demonstrated for combinations of penicillin and gentamicin against three Streptococcus mitis isolates with gentamicin MICs of 1000, 128 and 64 micrograms/ml. Results of this study indicate the importance of monitoring antibiotic resistance trends in viridans streptococci particularly with respect to penicillin and aminoglycoside resistance.

Drug Resistance, Microbial↗
PubMed2004

Mercuric resistance genes in gram-positive oral bacteria.

Mercury-resistant bacteria isolated from the oral cavities of children carried one of two types of merA gene that appear to have evolved from a common ancestor. Streptococcus oralis, Streptococcus mitis and a few other species had merA genes that were very similar to merA of Bacillus cereus strain RC607. Unlike the B. cereus RC607 merA gene, however, the streptococcal merA genes were not carried on Tn5084-like transposons. Instead, comparisons with microbial genomic sequences suggest the merA gene is located on a novel type II transposon. Coagulase-negative staphylococci and Streptococcus parasanguis had identical merA genes that represent a new merA variant.

Adolescent↗
PubMed1981

Immunosuppressive action of oral Streptococci.

The study on the immunosuppressive activity of the cytoplasmic component (SF) from Streptococcus mitis on the immune response of mice to the sheep red blood cells (SRBC) was carried out. The bacterial cell suspension was disrupted in a Braun shaker. The disrupted cell suspension was diluted and centrifuged at 10,000 x g for 30 minutes to remove the cell walls. The supernatant was centrifuged at 31,000 x g for 45 minutes and the final supernatant was dialyzed against distilled water, frozen and lyophilized. These products were employed as the cytoplasmic component (SF). The SF in the extracts of Streptococcus mitis suppresses the antibody formation in mice against SRBC. It is effective to inject it just one day before the inoculation with SRBC.

Animals↗
PubMed2006

Distribution and genetic diversity of the ABC transporter lipoproteins PiuA and PiaA within Streptococcus pneumoniae and related streptococci.

Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide. The existence of approximately 90 antigenically distinct capsular serotypes has greatly complicated the development of an effective pneumococcal vaccine. Virulence-associated proteins common and conserved among all capsular types now represent the best strategy to combat pneumococcal infections. PiuA and PiaA are the lipoprotein components of two pneumococcal iron ABC transporters and are required for full virulence in mouse models of infection. Here we describe a study of the distribution and genetic diversity of PiuA and PiaA within typical and atypical S. pneumoniae, Streptococcus oralis, and Streptococcus mitis strains. The genes encoding both PiuA and PiaA were present in all typical pneumococci tested, (covering 20 and 27 serotypes, respectively). The piuA gene was highly conserved within the typical pneumococci (0.3% nucleotide divergence), but was also present in "atypical" pneumococci and the closely related species S. mitis and S. oralis, showing up to 10.4% nucleotide divergence and 7.5% amino acid divergence from the typical pneumococcal alleles. Conversely, the piaA gene was found to be specific to typical pneumococci, 100% conserved, and absent from the oral streptococci, including isolates of S. mitis known to possess pneumolysin and autolysin. These are desirable qualities for a vaccine candidate and as a diagnostic tool for S. pneumoniae.

ATP-Binding Cassette Transporters↗
PubMed2006

Influence of biosurfactant on interactive forces between mutans Streptococci and enamel measured by atomic force microscopy.

Although interactive forces, influenced by environmental conditions, between oral bacteria and tooth surfaces are important for the development of plaque, they have never been estimated. It is hypothesized that interactive forces, as measured by atomic force microscopy, between enamel with or without a pellicle and two strains of mutans streptococci become less attractive by the application of a Streptococcus mitis BMS biosurfactant coating. Upon approach of each of the strains toward bare and pellicle-coated enamel, adsorbed biosurfactant increased the range of the repulsive forces. Upon retraction of the enamel surface, small adhesion forces (0.8-0.9 nN) were measured for bare enamel that almost disappeared after biosurfactant coating. The prevalence and magnitude of the adhesion forces also decreased upon pellicle-coating of the enamel, with a minor effect of adsorbed biosurfactant. These findings indicate that adsorbed S. mitis BMS biosurfactant changes the interactive forces between the mutans streptococci studied and enamel, explaining the effects of biosurfactant on adhesion.

Adsorption↗
PubMed2003

Real-time interaction of oral streptococci with human salivary components.

Oral streptococci are present in large numbers in dental plaque and several types interact with the enamel salivary pellicle to form a biofilm on tooth surfaces. The respective affinity of individual streptococci for salivary components has an influence on the etiologic properties of oral biofilm in the development of dental caries. We studied real-time biospecific interactions between oral streptococci and salivary components utilizing biosensor technology to analyze surface plasmon resonance. Streptococcus sanguis and Streptococcus mutans showed significant responses for binding to salivary components, in comparison with other bacteria. Further, the association rates (4.1 x 10-11/bacterium) and dissociation rate (5.7 +/- 0.9 x 10-3 Second(s)-1) were higher for S. sanguis than for S. mutans (2.4 x 10-11 and 2.9 +/- 0.8 x 10-3) and Streptococcus mitis (1.3 x 10-11 and 3.5 +/- 1.3 x 10-3). However, the association equilibrium constants (8.2 S/bacterium) for S. mutans was 2 times higher in than S. mitis (3.8) and slightly higher than S. sanguis (7.2). These findings may provide useful information regarding the mechanism of early biofilm formation by streptococci on the tooth surface.

Adult↗
PubMed2006

Sites in the CH3 domain of human IgA1 that influence sensitivity to bacterial IgA1 proteases.

The influence of regions, other than the hinge, on the susceptibility of human IgA1 to cleavage by diverse bacterial IgA1 proteases, was examined using IgA1 mutants bearing amino acid deletions, substitutions, and domain swaps. IgA1 lacking the tailpiece retained its susceptibility to cleavage by all of the IgA1 proteases. The domain swap molecule alpha1alpha2gamma3, in which the CH3 domain of IgA1 was exchanged for that of human IgG1, was resistant to cleavage with the type 1 and 2 serine IgA1 proteases of Neisseria meningitidis, Neisseria gonorrhoeae, and Haemophilus influenzae, but remained sensitive to cleavage with the metallo-IgA1 proteases of Streptococcus pneumoniae, Streptococcus oralis, Streptococcus sanguis, and Streptococcus mitis. Substitution of the IgA1 Calpha3 domain motif Pro440 -Phe443 into the corresponding position in the Cgamma3 domain of alpha1alpha2gamma3 resulted now in sensitivity to the type 2 IgA1 protease of N. meningitidis, indicating the possible requirement of these amino acids for sensitivity to this protease. For the H. influenzae type 2 protease, resistance of an IgA1 mutant in which the CH3 domain residues 399-409 were exchanged with those from IgG1, but sensitivity of mutant HuBovalpha3 in which the Calpha3 domain of bovine IgA replaces that of human IgA1, suggests that CH3 domain residues Glu403, Gln406, and Thr409 influence sensitivity to this enzyme. Hence, unlike the situation with the metallo-IgA1 proteases of Streptococcus spp., the sensitivity of human IgA1 to cleavage with the serine IgA1 proteases of Neisseria and Haemophilus involves their binding to different sites specifically in the CH3 domain.

Amino Acid Sequence↗
PubMedDate not supplied

Stress as a determinant of saliva-mediated adherence and coadherence of oral and nonoral microorganisms.

OBJECTIVE: The mucosal secretory proteins, such as the salivary proteins, play a key role in the acquisition and regulation of the mucosal microflora. Most notably, some microorganisms utilize the host's secretory proteins to adhere to the mucosa; a first step in colonization and infection. The secretory proteins also influence colonization by affecting the binding among microorganisms, a process denoted as coadherence. Previously we reported that acute stressors cause specific changes in saliva composition. The present study investigated to what extent these changes influence saliva-mediated microbial adherence and coadherence (ex vivo). METHODS: Thirty-two male undergraduates provided unstimulated saliva before and during a control condition and two stressors: A memory test and a surgery video presentation. We used saliva-coated microplates to test the adherence of bacteria for which the oral cavity is either a natural reservoir (eg, viridans streptococci) or a portal of entry (eg, Helicobacter pylori). We also tested the saliva-mediated co-adherence between Streptococcus gordonii and the yeast Candida albicans. Correlation analyses were performed to determine the relationships between changes in microbial adherence and the concentrations of potential salivary ligands, viz. cystatin S, the mucins MUC5B and MUC7, S-IgA, lactoferrin, alpha-amylase, and total salivary protein. RESULTS: During the memory test, saliva-mediated adhesion of Streptococcus sanguis, Streptococcus gordonii, and H. pylori increased, whereas the coadherence of C. albicans with S. gordonii decreased. During the surgical video presentation the saliva-mediated adherence of H. pylori, S. sanguis, and Streptococcus mitis increased. These changes were independent of salivary flow rate, but correlated with specific changes in salivary protein composition. CONCLUSION: The results show that even moderate stressors, by altering the activity of the mucosal secretory glands, may affect microbial colonization processes such as adherence and coadherence. This study hereby presents a mechanism by which stress may affect the mucosal microflora and susceptibility to infectious disease.

Adult↗
PubMed2005

Discrimination of Streptococcus pneumoniae from viridans group streptococci by genomic subtractive hybridization.

Two oligonucleotide primer sets for the discrimination of Streptococcus pneumoniae from "pneumococcus-like" oral streptococcal isolates by PCR were developed. Genomic subtractive hybridization was performed to search for differences between Streptococcus pneumoniae strain WU2 and the most closely related oral streptococcus, Streptococcus mitis strain 903. We identified 19 clones that contained S. pneumoniae-specific nucleotide fragments that were absent from the chromosomal DNA of typical laboratory strains of S. mitis and other oral bacteria. Subsequently, oligonucleotide PCR primers for the detection of S. pneumoniae were designed from the sequences of the subtracted DNA fragments, and the specificities of the 19 primer sets were evaluated by PCR using chromosomal DNAs extracted from four S. pneumoniae clinical isolates and from 20 atypical organisms classified as S. mitis or S. oralis, which harbored genes encoding the pneumococcal virulence factors autolysin (lytA) or pneumolysin (ply), as templates. Of the 19 primer sets, two (Spn9802 and Spn9828) did not amplify PCR products from any of the pneumococcus-like streptococcal strains that we examined. The genes containing the Spn9802 and Spn9828 sequences encoded proteins of unknown function that did not correspond to any previously described proteins in other bacteria. These new oligonucleotide primers may be very useful for early and correct diagnosis of S. pneumoniae infections.

Bacterial Proteins↗
PubMed2003

Echocardiographic presentations of endocarditis, and risk factors for rupture of a sinus of Valsalva in childhood.

BACKGROUND: In recent years, the diagnosis of infective endocarditis has been enhanced by the use of echocardiography. We sought, therefore, to review its effect on the management of endocarditis in children. METHODS: We reviewed all the patients presenting to our institution for evaluation for infective endocarditis from May 1994 to January 2002. The patients were stratified according to whether or not they had congenitally malformed hearts. RESULTS: Of the 90 referred patients identified, 46 (51%) had positive ultrasonic findings. Of these, we excluded 26 patients because of the presence of indwelling lines. The remaining 20 patients with features of endocarditis had a median age of 6.5 years, and a range from 0.14 to 8.5 years. There were 4 patients with normal hearts, and 16 with congenital cardiac malformations. We identified rupture of a sinus of Valsalva in four patients, with rupture into the left ventricle in two, and into the right ventricle and right atrium in one each. The mitral valve was involved in six patients, the aortic valve in another six, including all four with rupture of the sinus of Valsalva, both mitral and aortic valves in three, the pulmonary trunk in three patients, and the tricuspid valve and a Blalock-Taussig shunt in one patient each. Organisms isolated included Streptococcus mitis in 4 patients, Streptococcus pneumoniae in 2 patients, Streptococcus sanguis in 1, Staphylococcus aureus in 3, Staphylococcus epidermidis in 1, and Enteroccocus in 2. Cultures proved negative in 7 patients. Surgical intervention was needed in 12 patients, and one died (5%). Only the left-sided chambers were involved in those with normal hearts. Both patients infected with Streptococcus pneumoniae had rupture of a sinus of Valsalva. CONCLUSION: Involvement of the left-sided chambers is more likely in structurally normal hearts, and in cases with rupture of a sinus of Valsalva, in which case infection with Streptococcus pneumonia should be suspected.

Aortic Rupture↗
PubMedDate not supplied

The bacteriology of the supragingival plaque of child dental patients in Kuwait.

OBJECTIVE: To investigate the bacterial flora of the gingival plaques of pediatric dental patients in order to establish baseline data for future studies on the complex microbiota of the oral cavity of this group of child dental patients. SUBJECTS AND METHODS: Forty subjects, aged 3-12 years, were enrolled and investigated after informed consent was obtained. During the patients' first visit to the dental clinic supragingival plaque samples were collected from the gingival area of buccal and lingual tooth surfaces of all primary and permanent molar teeth using sterile curettes. Serially diluted samples were inoculated onto a set of selective and nonselective media and then incubated at the appropriate atmospheric conditions and temperatures. Representative colonies of isolates were identified by standard methods, including hemolytic characteristics on blood agar and profiling on API 20S and API 20A as appropriate. RESULTS: A total of 183 isolates (123 aerobes and 60 anaerobes) were recovered from the 40 patients with a mean count of 3 per sample of aerobes compared to 1.5 per sample for the anaerobes. Streptococcus sanguis was the predominant aerobic species (23.6%) isolated from 29 (72.5%) of the 40 patients, followed by Streptococcus mitis, 19.5 and 60%, Streptococcus salivarius, 17.1 and 52.5% and Streptococcus mutans, 17.1 and 52.5%. The frequency of isolation of anaerobic bacteria was Prevotella spp. 50% from 30 (75%) of the 40 patients, Fusobacterium spp., 18.3 and 27.5%, Bacteroides spp., Porphyromonas spp. and Peptostreptococcus spp., 6.7 and 10%, 6.7 and 10%, and 5 and 7.5%, respectively. Prevotella intermedia was the single most common species. CONCLUSION: The results show that there were more aerobic/facultative anaerobic bacteria than the obligate anaerobes in the dental plaque of the pediatric dental patients in Kuwait. The high prevalence of the pioneering streptococci as well as black-pigmented Prevotella spp. and Fusobacterium spp. indicates that the dental plaques of the children were in the developing stage, a precursor to the development of periodontal diseases.

Bacteria, Aerobic↗