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[Viridans streptococci in human infections: identification and susceptibility to antibiotics].

A method for the speciation of viridans streptococci (devoided of group antigens) is described. The major identification criteria are based on the reaction of a series of biochemical tests such as acid production in lactose, inuline, raffinose, mannitol and sorbitol, hydrolysis of arginine, esculin and Na hippurate, and production of polysaccharides in 5% sucrose media. A total of 460 strains were isolated from human specimens and identified as follows: 118 Streptococcus mitis, 102 S. sanguis II, 75 S. Sanguis I, 87 S. milleri (Streptococcus MG-intermedius), 28 S. mutans, 25 S. salivarius, 14 S. morbillorium, 2 S. uberis and 9 unspeciated. Susceptibility to antibiotics was studied for 318 strains: 63% of them were susceptible to all drugs tested; 37% of the strains were resistant to one or several antibiotics as follows: 34% to tetracycline, 8.5% to macrolides and related drugs, 5.3% to streptomycin and/or kanamycin (MIC greater than 2,000 micrograms/ml), 5% to penicillin (MIC = 1-4 micrograms/ml) and 4% to chloramphenicol.

Anti-Bacterial Agents↗

PubMed1993

Phenotypic characterization of oral streptococci by classical methods.

The phenotypic characteristics of 215 strains of oral streptococci were compared on the basis of the results of sorbitol, raffinose and trehalose fermentation, ammonium from arginine production, aesculin hydrolysis, H2O2 production, susceptibility to 2 U bacitracin, and growth in 4% NaCl solution. Eleven different species were identified. Among the most noteworthy findings were the low number of H2O2-producing strains of Streptococcus sobrinus, the effective discrimination between Streptococcus sanguis and Streptococcus mitis on the basis of raffinose fermentation, and the phenotypic similarity between Streptococcus oralis and the former mutans group. To avoid the creation of new species, the term biotype is proposed to designate phenotypically distinct organisms which do not qualify as different species.

Dental Plaque↗

PubMed1979

Serologic studies of Streptococcus intermedius, Streptococcus constellatus, and Streptococcus morbillorum by crossed immunoelectrophoresis.

A reference antigen-antibody system for Streptococcus intermedius, Streptococcus constellatus, and Streptococcus morbillorum was established with crossed immunoelectrophoresis. A comparison of S. intermedius, S. constellatus, and S. morbillorum with crossed immunoelectrophoresis and crossed immunoelectrophoresis with intermediate gel indicated that S. intermedius and S. constellatus are closely related antigenically with as many as six common cytoplasmic antigens. S. morbillorum was antigenically more distinct; antiserum of one strain of S. morbillorum was monospecific, indicating that specific serogroups of S. morbillorum exist. Crossed immunoelectrophoresis and tandem crossed immunoelectrophoresis revealed that S. intermedius, S. constellatus, and S. morbillorum also share some common antigens with Streptococcus sanguis and Streptococcus mitis, but S. intermedius, S. constellatus, and S. morbillorum are antigenically distinct from Streptococcus mutans and Streptococcus bovis.

Antibodies, Bacterial↗

PubMed2001

High proportion of pharyngeal carriers of commensal streptococci resistant to erythromycin in Spanish adults.

The presence of erythromycin-resistant (ErR) commensal streptococci in the throat of 110 healthy subjects and 87 patients with pharyngitis was investigated. The resistance determinants were studied by PCR using the primers for mef and erm genes, followed by hybridization and sequencing analysis. Overall, 94.4% of the subjects carried one or more ErR strains in their pharynx. A total of 253 ErR strains was studied: 127 (50.2%) showed constitutive or inducible resistance to clindamycin (MLS(B) phenotype) and 126 (49.8%) were susceptible to clindamycin (M phenotype). In 50 subjects (25.4%) both phenotypes were detected. The ermB gene was predominant among the MLS(B) phenotype strains (97.6%). The mefA (mefA/mefE) gene was detected in 100% of the strains with the M phenotype. One Streptococcus oralis strain bearing the MLS(B) phenotype carried both mefA and ermB genes. The mefA gene from clinical isolates of Streptococcus mitis and S. oralis was transferred by conjugation to an erythromycin-susceptible Streptococcus pneumoniae strain.

Adolescent↗

PubMed1976

Determinants of the developing oral flora in normal newborns.

The ability of Streptococcus species to selectively adhere to the oral epithelial cells of newborns was studied in vitro. On day 1 of life, mucosal cells from normal infants demonstrated selective attraction for the natural distribution of streptococci that would soon colonize these surfaces. Streptococcus salivarius and Streptococcus mitis adhered well in vitro to scraped cells from cheek and tongue surfaces. Streptococcus mutans, on the other hand, exhibited feeble or no adherence to cheek or tongue cells. Adherence of Escherichia coli to oral epithelial cells was also studied. The ability of strains of E. coli to adhere to cheek and tongue cells correlated solely with the presence of cell surface substances, probably pili. These observations, made on infants at the critical moment of their developing flora, strengthen the hypothesis that the ability of bacteria to adhere to surfaces is an important determinant of their ecological place in the oral microflora.

Cells, Cultured↗

PubMed2005

Effect of mutations in the human immunoglobulin A1 (IgA1) hinge on its susceptibility to cleavage by diverse bacterial IgA1 proteases.

Components of the human immunoglobulin A1 (IgA1) hinge governing sensitivity to cleavage by bacterial IgA1 proteases were investigated. Recombinant antibodies with distinct hinge mutations were constructed from a hybrid comprised of human IgA2 bearing half of the human IgA1 hinge region. This hybrid antibody and all the mutant antibodies derived from it were resistant to cleavage by the IgA1 proteases from Streptococcus oralis and Streptococcus mitis biovar 1 strains but were cleaved to various degrees by those of Streptococcus pneumoniae, some Streptococcus sanguis strains, and the type 1 and 2 IgA1 proteases of Haemophilus influenzae, Neisseria meningitidis, and Neisseria gonorrhoeae. Remarkably, those proteases that cleave a Pro-Ser peptide bond in the wild-type IgA1 hinge were able to cleave mutant antibodies lacking a Pro-Ser peptide bond in the hinge, and those that cleave a Pro-Thr peptide bond in the wild-type IgA1 hinge were able to cleave mutant antibodies devoid of a Pro-Thr peptide bond in the hinge. Thus, the enzymes can cleave alternatives to their preferred postproline peptide bond when such a bond is unavailable. Peptide sequence analysis of a representative antibody digestion product confirmed this conclusion. The presence of a cleavable peptide bond near the CH2 end of the hinge appeared to result in greater cleavage than if the scissile bond was at the CH1 end of the hinge. Proline-to-serine substitution at residue 230 in a hinge containing potentially cleavable Pro-Ser and Pro-Thr peptide bonds increased the resistance of the antibody to cleavage by many IgA1 proteases.

Amino Acid Sequence↗

PubMed1989

Molecular surface characterization of oral streptococci by Fourier transform infrared spectroscopy.

In order to characterize the molecular composition of oral streptococci, infrared transmission spectroscopy on freeze-dried cells dissolved in KBr was used. All infrared spectra show similar absorption bands for the strains studied with the most important absorption bands located at 2930 cm-1 (CH), 1653 cm-1 (AmI), 1541 cm-1 (AmII) and two bands at 1236 cm-1 and 1082 cm-1, which were assigned to phosphate and sugar groups. However, calculation of absorption band ratios normalized with respect to the integrated intensity of the CH stretching region around 2930 cm-1, show significant differences between the strains. Both Streptococcus mitis strains possess high AmI/CH and AmII/CH absorption band ratios compared to the other strains. Streptococcus salivarius HBC12, a mutant strain devoid of all proteinaceous surface appendages, shows significantly lower AmI/CH and AmII/CH band ratios with respect to its parent strain S. salivarius HB. Two positive relationships could be established both between the AmII/CH absorption band ratio and the N/C elemental surface concentration ratio of the strains previously, determined from X-ray photoelectron spectroscopy (XPS) and also between AmI/CH and the fraction of carbon atoms at the surface involved in amide bonds, determined by XPS as well. From this comparison, it is concluded that transmission infrared spectroscopy can be employed as a technique to study the molecular surface composition of freeze-dried microorganisms.

Amides↗

PubMed1994

Bacterial interference in vitro. Comparison between a quantitative kinetic and a cocultivation blood agar test method.

The aim of the present study was to compare two methods for estimation of bacterial growth interference between various bacteria using a Bioscreen robot analyzer, allowing kinetic documentation, and a cocultivation test on blood agar plates. Six laboratory strains with different virulence and growth requirements were used: Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Fusobacterium nucleatum, Streptococcus mitis, Staphylococcus aureus, and Staphylococcus epidermidis. The interference activity was correlated with a reference system of Streptococcus sanguis (strain alpha 89) and Streptococcus pyogenes (group A streptococci, GAS serotypes T 9 and T 22). The methods used and results obtained were as follows: 1. Estimation of synergistic and antagonistic bacterial interferences using a Bioscreen robot analyzer. Suspensions of viable bacteria were added to microtiter plates with different concentrations of UV light-killed bacteria in liquid media. The Bioscreen analyzer monitored bacterial growth every 10 min for 24 h giving kinetic data during the growth period. Synergisms as well as antagonisms were demonstrated between the tested bacterial strains which have not earlier been reported in the literature. However, the antagonistic effect observed between the six strains was less than that induced by the S. sanguis strain on the two strains of S. pyogenes. 2. Cocultivation of bacterial strains on blood agar surface with precultivated or simultaneously stamped interfering bacteria indicated no detectable interference between the six tested bacterial strains, while the S. sanguis strain inhibited the growth of S. pyogenes strains as well as the hemolysis around the colonies. The Bioscreen method was found more sensitive for testing bacterial interference compared to the commonly used blood agar test.

Bacteria↗

PubMed1985

Quantitative analysis of cell walls of nutritionally variant streptococci grown under various growth conditions.

Strains of nutritionally variant streptococci are usually isolated from patients with subacute bacterial endocarditis. Only recently have these strains been subdivided into three serotypes; however, no group-specific antigen has been described. To understand the immunochemical basis for the serology of these microorganisms as well as set the groundwork for adherence studies, quantitative analysis of the cell walls of nutritionally variant streptococci was undertaken. The bacteria were grown in semisynthetic medium or pyridoxal-supplemented Todd-Hewitt broth and harvested during the exponential or stationary phase. Cell walls were isolated and analyzed for amino sugars, sugars, polyalcohols, amino acids, and phosphorus by gas chromatography, high-pressure liquid chromatography, or colorimetric assays. The peptidoglycans of the cell walls of the prototype strains from the three serotypes were representative of other streptococcal cell walls, including the presence of alanine as the possible cross-bridge. The composition of the peptidoglycan was similar for all three strains and included a decreased concentration of peptidoglycan in their cell walls during the stationary phase. Glucosamine, glucose, galactose, ribitol, and a small amount of rhamnose were found in each of the cell wall polysaccharides. Galactosamine was only found in serotype II and III cell walls and might be responsible for the previously described cross-reaction between these strains. The concentration of the other sugars and amino sugars varied in each of the cell wall preparations, depending on the growth conditions. Finally, all three strains expressed both ribitol and phosphorus in their cell walls, characteristic of the presence of a ribitol teichoic acid. Therefore the cell wall composition of the nutritionally variant streptococci varies depending on the growth conditions, and their composition appears similar to that of strains of Streptococcus mitis.

Carbohydrates↗

PubMed1989

Surface structures, co-aggregation and adherence phenomena of Streptococcus oralis and related species.

Seven strains of Streptococcus oralis were found to possess surface structures. Four strains possessed long fimbriae which ranged in length from 266-366 nm, while the remaining three strains possessed shorter peritrichously distributed fibrils which ranged in length from 80-197 nm. The fibrillar strains were morphologically similar to strains of Streptococcus sanguis I and II and Streptococcus mitis. No strain of S. oralis produced tufts of fibrils like certain strains of S. sanguis I. Strains of Streptococcus milleri produced peritrichously distributed fimbriae which were morphologically dissimilar to the fimbriae produced by S. oralis strains. S. oralis strains were moderately to highly hydrophobic, but hydrophobicity could not be related to adhesion parameters or type or length or density of surface structures. Furthermore, there appeared to be no correlation between type of surface structures and adhesion parameters. Two of the three fibrillar strains of S. oralis and the peritrichously fibrillar strains of S. sanguis, together with one strain of S. milleri, were able to co-aggregate with actinomycetes.

Actinomyces↗

PubMed1993

[Antibacterial and cytotoxic effects of N-butyl-2-cyanoacrylate used as a tissue adhesive].

In this study, the antibacterial and cytotoxic effects of a synthetic tissue adhesive called n-butyl-2-cyanoacrylate (Histoacryl) which provide rapid healing in incision in oral surgical procedures were investigated. The antibacterial effects of Histoacryl were tested by using standard reference strains of Streptococcus pyogenes, Streptococcus mitis, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus grown in Mueller-Hinton media (sheep blood supplemented Mueller-Hinton for streptococcus strains) by diffusion technique. The cytotoxic effects were also researched on continuous Vero and primary gingival fibroblast cell cultures by using agar overlay method. In these methods, the inhibition zones produced by Histoacryl were evaluated. As a result, Histoacryl was found to have antibacterial effect on the tested bacteria and produce cytotoxic and cytopathic effects on the tested cell cultures.

Animals↗

PubMed1999

Acid-induced acid tolerance and acidogenicity of non-mutans streptococci.

Acid tolerance and acidogenicity of non-mutans streptococci and their capacity of acid adaptation were studied. The cells of non-mutans streptococci (Streptococcus sanguis [Streptococcus sanguinis], Streptococcus gordonii, Streptococcus oralis and Streptococcus mitis) grown at pH 7.0 showed 0.0088% to 71% viability after acidification at pH 4.0 for 60 min, whereas the cells of mutans streptococci (Streptococcus mutans) were not killed by the acidification. Washed cells of non-mutans streptococci lowered pH to 4.04-4.33 in the presence of glucose, while mutans streptococci cells lowered pH to 3.70. When the growth pH was shifted to 5.5 for 30-90 min, the viability of non-mutans streptococci after the acidification at pH 4.0 for 60 min increased (0.25% to 91%) and the minimum pH values of the cells in the presence of glucose decreased (3.90 4.19). Along with the increase in acid tolerance and acidogenicity, non-mutans streptococci increased activities of H(+)-ATPase and arginine deiminase and amounts of stress proteins cross-reacting with 60 kDa and 70 kDa heat shock proteins (Hsp60 and Hsp70). These results indicate that non-mutans streptococci were capable of increasing their acid tolerance and acidogenicity in response to environmental acidification. Furthermore, it is suggested that the acid adaptation observed in non-mutans streptococci cells could involve the induction of H(+)-ATPase, arginine deiminase and stress protein syntheses. The strains of non-mutans streptococci, which are pioneer bacteria for dental plaque formation and predominant in plaque microbial flora, may play a significant role in shifting the dental plaque environment toward acidic and consequently promoting the colonization of more acid-tolerant and acidogenic bacteria such as mutans streptococci and lactobacilli.

Acids↗

PubMed2003

Distribution of selected bacterial species on intraoral surfaces.

BACKGROUND/AIM: To examine the proportions of 40 bacterial species in samples from 8 oral soft tissue surfaces and saliva in systemically healthy adult subjects and to compare these microbiotas with those of supra- and subgingival plaque. METHODS: Microbial samples were taken from 8 oral soft tissue surfaces of 225 systemically healthy subjects using a "buccal brush". Saliva was taken by expectoration. Forty-four of these subjects provided additional supra- and subgingival plaque samples. Samples were individually evaluated for their content of 40 bacterial species using checkerboard DNA-DNA hybridization. The percentage of total DNA probe count was determined for each species, at each sample location and averaged across subjects. The significance of differences among the proportions of the 40 test species at different sample locations was sought in the 225 and 44 subjects separately using the Quade test and adjusted for multiple comparisons. Cluster analysis was performed using the proportions of the 40 species at the different sample locations using the minimum similarity coefficient and an average unweighted linkage sort. The proportions of each species were averaged across subjects in the resulting cluster groups and the significance of differences was tested using the t-test and ANOVA. RESULTS: Microbial profiles differed markedly among sample locations in the 225 subjects, with 34 of 40 species differing significantly. Proportions of Veillonella parvula and Prevotella melaninogenica were higher in saliva and on the lateral and dorsal surfaces of the tongue, while Streptococcus mitis and S. oralis were in significantly lower proportions in saliva and on the tongue dorsum. Cluster analysis resulted in the formation of 2 clusters with >85% similarity. Cluster 1 comprised saliva, lateral and dorsal tongue surfaces, while Cluster 2 comprised the remaining soft tissue locations. V. parvula, P. melaninogenica, Eikenella corrodens, Neisseria mucosa, Actinomyces odontolyticus, Fusobacterium periodonticum, F. nucleatum ss vincentii and Porphyromonas gingivalis were in significantly higher proportions in Cluster 1 and S. mitis, S. oralis and S. noxia were significantly higher in Cluster 2. These findings were confirmed using data from the 44 subjects providing plaque samples. The microbial profiles of supra- and subgingival plaque differed from the other sample locations, particularly in the increased proportions of the Actinomyces species. Species of different genera exhibited different proportions on the various intraoral surfaces, but even within the genus Streptococcus, there were differences in colonization patterns. S. oralis, S. mitis and S. constellatus colonized the soft tissues and saliva in higher proportions than the samples from the teeth, while the other 4 streptococcal species examined colonized the dental surfaces in proportions comparable to the soft tissue locations and saliva. CONCLUSIONS: Proportions of bacterial species differed markedly on different intraoral surfaces. The microbiota of saliva was most similar to that of the dorsal and lateral surfaces of the tongue. The microbiotas of the soft tissues resembled each other more than the microbiotas that colonized the teeth both above and below the gingival margin.

Adult↗

PubMed2002

[Cerebral hemorrhage due to a ruptured mycotic aneurysm. Two cases].

INTRODUCTION: Cerebral hemorrhages due to rupture of mycotic aneurysms are rare but severe complications of infective endocarditis. We report two cases with a good outcome. EXEGESIS: The first patient presented with a parieto-occipital hematoma which occurred in the course of a relapsing infective endocarditis due to Streptococcus mitis. She fully recovered after neurosurgical treatment. In the second case, a right frontal hematoma revealed two mycotic aneurysms and an infective endocarditis due to Streptococcus gordonii. Motor weakness partially recovered after antibiotic therapy and angiography demonstrated complete resolution of aneurysms. CONCLUSION: Ruptured mycotic aneurysms are poor prognosis factors in infective endocarditis. Adapted antibiotic therapy is the first-intent treatment. Neurosurgery is indicated when hematomas are poorly tolerated and in cases requiring anticoagulant therapy.

Adult↗

PubMed1993

An analysis of the subgingival microflora in randomly selected subjects.

The aim of this study was to describe the presence of some microbial species in the subgingival microflora of a randomly selected subject sample. A further objective was to analyze the association between some microbiological characteristics and the periodontal conditions of the subjects. A total of 171 individuals aged 30-65 years were included. A pooled subgingival plaque sample was obtained from the mesial aspect of the 6 Ramfjord teeth by the use of the paper-point method. The samples were processed and analyzed according to standardized procedures. The periodontal condition of the subjects was examined clinically and included assessment of plaque, gingivitis, probing pocket depth and probing attachment level. The results showed that 81% of the subjects were positive for Campylobacter rectus, 80% for Fusobacterium nucleatum, 77% for Streptococcus sanguis, 72% for Streptococcus mitis, 66% for Eikenella corrodens, 58% for Prevotella intermedia, 32% for Capnocytophaga, 27% for Selenomonas, 25% for Actinobacillus actinomycetemcomitans, 14% for Porphyromonas gingivalis and 13% for Streptococcus mutans. Small differences in the periodontal conditions were observed between subjects harboring P. gingivalis, A. actinomytecomitans or > 5% P. intermedia and the rest of the sample. Similarly small differences in the clinical parameters were revealed when the subjects were grouped according to proportions of disease-associated and health-associated species in their subgingival samples.

Adult↗

PubMed1984

Characterization of a pH-dependent chromophore from nutritionally variant streptococci.

Strains of nutritionally variant streptococci and Streptococcus mitis produce a chromophore when they are heated at acid pH. No other strains of streptococci elaborated this chromophore. Furthermore, the nutritionally variant streptococci produced approximately twice the amount of chromophore as the S. mitis strains. The chromophore was localized in the cell wall of these streptococcal strains and appeared to be resistant to trypsin treatment. Hydrolysis apparently is required because elevated temperatures at pH 2 are necessary for demonstration of the chromophore. The chromophore has a maximal absorbance at 504 nm with a pKa of 3.6. The chromophore absorbance spectrum showed an isosbestic point at 400 nm. This is the first example of a pH-dependent chromophore to be found in streptococci and serves as a positive characteristic for the description of nutritionally variant streptococci as well as S. mitis.

Chromogenic Compounds↗

PubMed1987

Differential medium for mixed cultures of alpha-hemolytic streptococci from blood.

Alpha-hemolytic streptococci (AHS) were isolated from blood cultures from 100 patients, and species were identified by the Ruoff and Kunz scheme. When isolates were inoculated onto sheep blood agar, all 100 cultures appeared to be pure, with identifications based on colonial morphology and Gram stain. When isolates were subcultured onto mitis salivarius agar (MSA), mixtures of two species of AHS were detected in 10 cultures from patients (10%). These mixed cultures would have been reported as pure cultures of Streptococcus milleri (six cultures), Streptococcus salivarius (three cultures), Streptococcus sanguis I (one culture), with identifications based on biochemical profiles. Cultures on MSA demonstrated S. milleri (six cultures), Streptococcus mitis (five cultures), S. salivarius (three cultures), S. sanguis I (one culture), and S. sanguis II (five cultures). The inability to separate AHS species by colony morphology on sheep blood agar demands that a differential medium such as MSA be routinely used for subculture. Failure to use such a medium may account for some of the confusing biochemical profiles associated with AHS species identification.

Blood↗

PubMed1999

Identification of oral mitis group streptococci by arbitrarily primed polymerase chain reaction.

"Mitis group" streptococci are commensal but may play some role in dental caries, septicemia or endocarditis. Rapid genotypic identification would aid studies of dental plaque ecology, or diagnostic use. AP-PCR with 58 unpaired arbitrary primers was used to characterize 7 Streptococcus gordonii, 11 Streptococcus sanguis, 2 Streptococcus crista, 5 Streptococcus parasanguis, 18 Streptococcus oralis, and 36 Streptococcus mitis (22 biovar 1 and 14 biovar 2). S. parasanguis 16S rRNA variable region primer RR2 produced species-specific bands with all S. gordonii and S. sanguis. Human V beta 1 T-cell receptor primer 434 yielded concordant genotypic identification of all phenotypically defined S. crista and S. parasanguis, 83% of S. oralis, and 74% of S. mitis biovar 1. Amplicon patterns for S. mitis biovar 2 were heterogeneous. Findings suggest that primers RR2 and 434 in succession will allow rapid identification of genotypic groups corresponding closely to mitis group species established by phenotype.

Bacterial Typing Techniques↗