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Mosaic genes and mosaic chromosomes: intra- and interspecies genomic variation of Streptococcus pneumoniae.

Streptococcus pneumoniae remains a major causative agent of serious human diseases. The worldwide increase of antibiotic resistant strains revealed the importance of horizontal gene transfer in this pathogen, a scenario that results in the modulation of the species-specific gene pool. We investigated genomic variation in 20 S. pneumoniae isolates representing major antibiotic-resistant clones and 10 different capsular serotypes. Variation was scored as decreased hybridization signals visualized on a high-density oligonucleotide array representing 1,968 genes of the type 4 reference strain KNR.7/87. Up to 10% of the genes appeared altered between individual isolates and the reference strain; variability within clones was below 2.1%. Ten gene clusters covering 160 kb account for half of the variable genes. Most of them are associated with transposases and are assumed to be part of a flexible gene pool within the bacterial population; other variable loci include mosaic genes encoding antibiotic resistance determinants and gene clusters related to bacteriocin production. Genomic comparison between S. pneumoniae and commensal Streptococcus mitis and Streptococcus oralis strains indicates distinct antigenic profiles and suggests a smooth transition between these species, supporting the validity of the microarray system as an epidemiological and diagnostic tool.

Alleles↗

Streptococcus salivarius fimbriae are composed of a glycoprotein containing a repeated motif assembled into a filamentous nondissociable structure.

Streptococcus salivarius, a gram-positive bacterium found in the human oral cavity, expresses flexible peritrichous fimbriae. In this paper, we report purification and partial characterization of S. salivarius fimbriae. Fimbriae were extracted by shearing the cell surface of hyperfimbriated mutant A37 (a spontaneous mutant of S. salivarius ATCC 25975) with glass beads. Preliminary experiments showed that S. salivarius fimbriae did not dissociate when they were incubated at 100 degrees C in the presence of sodium dodecyl sulfate. This characteristic was used to separate them from other cell surface components by successive gel filtration chromatography procedures. Fimbriae with molecular masses ranging from 20 x 10(6) to 40 x 10(6) Da were purified. Examination of purified fimbriae by electron microscopy revealed the presence of filamentous structures up to 1 microm long and 3 to 4 nm in diameter. Biochemical studies of purified fimbriae and an amino acid sequence analysis of a fimbrial internal peptide revealed that S. salivarius fimbriae were composed of a glycoprotein assembled into a filamentous structure resistant to dissociation. The internal amino acid sequence was composed of a repeated motif of two amino acids alternating with two modified residues: A/X/T-E-Q-M/phi, where X represents a modified amino acid residue and phi represents a blank cycle. Immunolocalization experiments also revealed that the fimbriae were associated with a wheat germ agglutinin-reactive carbohydrate. Immunolabeling experiments with antifimbria polyclonal antibodies showed that antigenically related fimbria-like structures were expressed in two other human oral streptococcal species, Streptococcus mitis and Streptococcus constellatus.

Amino Acids↗

Use of restriction fragment polymorphism analysis of rRNA genes to assign species to unknown clinical isolates of oral viridans streptococci.

This study evaluated restriction fragment length polymorphisms of rRNA genes (ribotyping) for genotypic identification of 53 oral isolates classified as "Streptococcus sanguis" by colony morphology. Isolates were from 8-h buccal plaque on lower first permanent molars of 20 subjects. DNA was digested with AatII and hybridized with digoxygenin-labeled cDNA of Escherichia coli 16S and 23S rRNA. Strains were ribotyped again with AlwNI or PvuII on the basis of the presence or absence of a 2,290-bp AatII band. Band patterns were compared with reference ribotypes for Streptococcus gordonii, Streptococcus sanguis, Streptococcus crista, Streptococcus oralis, Streptococcus mitis, and Streptococcus parasanguis strains. Forty-eight isolates could be assigned to a species (22 S. sanguis, 14 S. oralis, 12 S. gordonii). Multiple species were seen in 14 subjects; multiple strains of the same species occurred in 11 subjects. Our findings suggest that ribotyping can be used for genotypic identification of S. sanguis, S. oralis, and S. gordonii isolates.

Bacterial Typing Techniques↗

Pacemaker-related endocarditis. Report of 7 cases and review of the literature.

We report on 7 patients with pacemaker endocarditis diagnosed during the workup of long-standing fever. Persistent positive blood cultures and echocardiography led to the diagnosis in 6 patients whereas autopsy was diagnostic in another. Causative microorganisms were Staphylococcus epidermidis (3), Staphylococcus lugdunensis (1), Pseudomonas aeruginosa (1), Streptococcus bovis (1), and Streptococcus mitis-Streptococcus sanguis (1). Pulmonary embolism was present in nearly 50% of the cases, a figure clearly higher than previously reported. In all but 1 case the initial medical approach was not successful, and thus the pacing system was finally removed. None of the cases relapsed after the removal. We have reviewed the literature regarding pacemaker endocarditis, particularly with respect to treatment.

Aged↗

A simplified diagnostic system for cultural detection and enumeration of Streptococcus mutans.

A simple dip-slide test (Cariescreen SM) based on MSB selective agar was devised for detection and quantitation of Streptococcus mutans in oral samples. For this test, a bacitracin tablet is dissolved in a vial containing buffered saline diluent. Paraffin-stimulated saliva is collected in this diluent vial. A dip slide which incorporates a modified MSB agar (minus bacitracin) is immersed briefly in the diluted saliva. After addition of a CO2-generating tablet, the screw-cap dip slide is closed tightly in the vial and incubated for two days at 37 degrees C and one day at room temperature. S. mutans populations in saliva are estimated by comparison with a colony density chart. Growth of reference strains of S. mutans was equivalent on Cariescreen SM dip slides and on MSB agar plates. Reference strains of Streptococcus sanguis, Streptococcus salivarius, Streptococcus mitis, and Streptococcus milleri did not grow on Cariescreen SM dip slides. Aliquots of saliva from 50 schoolchildren and 51 adults were tested by the dip-slide method and by conventional plating methods in MSB agar. Very good correlation (0.93) between the two methods was obtained. This simplified S. mutans detection system is suitable for use by clinical personnel in dental clinics or other non-laboratory settings for identification of subjects potentially at risk for caries.

Bacitracin↗

Adhesion in vitro of oral streptococci to porcelain, composite resin cement and human enamel.

We investigated the initial adhesion in vitro of oral streptococci to porcelain inlays. Four strains of streptococci, Streptococcus mitis NCTC12261, Streptococcus oralis ATCC9811, Streptococcus sanguis ATCC10556 and Streptococcus sobrinus OMZ176, were used in this study. The disc specimens were made of porcelain, composite resin cement and human enamel. These specimens, with or without a saliva-coat, were immersed in a suspension of each streptococci strain at 37 degrees C for 1 hr, and the numbers of cells adhering to specimens were counted after staining. The saliva-coat significantly decreased the numbers of adhering cells in all strains tested (t-test, p < 0.05). The adhesion to the porcelain and resin cements could be explained by a thermodynamic approach, although the adhesion to enamel could not be explained. These results indicated the possibility that the mechanism is different in initial adhered strains of streptococci between enamel and dental restorative materials.

Bacterial Adhesion↗

Molecular analysis of colonized bacteria in a human newborn infant gut.

The complex ecosystem of intestinal microflora is estimated to harbor approximately 400 different microbial species, mostly bacteria. However, studies on bacterial colonization have mostly been based on culturing methods, which only detect a small fraction of the whole microbiotic ecosystem of the gut. To clarify the initial acquisition and subsequent colonization of bacteria in an infant within the few days after birth, phylogenetic analysis was performed using 16S rDNA sequences from the DNA isolated from feces on the 1st, 3rd, and 6th day. 16S rDNA libraries were constructed with the amplicons of PCR conditions at 30 cycles and 50 degrees c annealing temperature. Nine independent libraries were produced by the application of three sets of primers (set A, set B, and set C) combined with three fecal samples for day 1, day 3, and day 6 of life. Approximately 220 clones (76.7%) of all 325 isolated clones were characterized as known species, while other 105 clones (32.3%) were characterized as unknown species. The library clone with set A universal primers amplifying 350 bp displayed increased diversity by days. Thus, set A primers were better suited for this type of molecular ecological analysis. On the first day of the life of the infant, Enterobacter, Lactococcus lactis, Leuconostoc citreum, and Streptococcus mitis were present. The largest taxonomic group was L. lactis. On the third day of the life of the infant, Enterobacter, Enterococcus faecalis, Escherichia coli, S. mitis, and Streptococcus salivarius were present. On the sixth day of the life of the infant, Citrobacter, Clostridium difficile, Enterobacter sp., Enterobacter cloacae, and E. coli were present. The largest taxonomic group was E. coli. These results showed that microbiotic diversity changes very rapidly in the few days after birth, and the acquisition of unculturable bacteria expanded rapidly after the third day.

Bacteria↗

Partial characterisation of the inhibitory substances produced by Streptococcus oralis and related species.

The production of inhibitory substances was sought by deferred antagonism. Streptococcus oralis, Streptococcus mitis and Streptococcus sanguis produced bactericidal concentrations of hydrogen peroxide, but no detectable bacteriocin. Actinomycetes and Neisseria pharyngis were particularly susceptible to the lethal action of hydrogen peroxide. Streptococcus milleri produced bactericidal concentrations of hydrogen peroxide and bacteriocin-like substances. S. oralis strains were inhibited by the bacteriocin-like products of strains of Streptococcus bovis, Streptococcus faecalis, Streptococcus mutans and Streptococcus salivarius. Inhibition was due neither to complete bacteriophage nor to fermentation end-products. Extracellular polysaccharides formed during growth could not protect sensitive strains from the effect of hydrogen peroxide or bacteriocin-like substances.

Actinomycetales↗

Spontaneous bacterial peritonitis caused by a viridans Streptococcus or Neisseria perflava.

Eight patients had nine episodes of presumed spontaneous bacterial peritonitis (SBP) caused by commensal bacteria that usually inhabit the oropharynx, including Neisseria perflava and the viridans streptococci Streptococcus milleri, Streptococcus mitis, and Streptococcus sanguis. Nonpurulent ascites (ascitic fluid WBC count, less than 1,000/cu mm) was present in four episodes, possibly caused by either bacterial colonization or early peritonitis; purulent ascites was present in five episodes. None died of their infection, and three episodes remitted without treatment. Poor condition of gums and teeth may play a pathogenic role in SBP caused by these bacteria.

Adult↗

Vitamin B6-dependent Streptococcus mitior (mitis) isolated from patients with systemic infections.

Strains of nutritional-variant viridans Streptococcus were isolated from two patients with bacterial endocarditis and from one patient with a pancreatic abscess. All three strains grew as satellite colonies of other bacteria but did not grow as pure cultures in media lacking sufficient thiol compounds. For one-half maximal growth these organisms required the addition of one of the active forms of the B6 vitamin, pyridoxal with HCl (0.16-1.45 mug/ml) or pyridoxamine dihydrochloride (0.67-2.0 mug/ml), or addition of l-cysteine (0.235-0.425 mg/ml) to routine bacteriological media. The active forms of vitamin B6 are essential coenzymes in the synthesis of l-cysteine and of other thiol compounds. Use of media supplemented with 0.001 percent pyridoxal with HCL led to identification of the strains as Streptococcus mitior (mitis). Incorporation of pyridoxal with HCl, pyridoxamine dihydrochloride, or l-cysteine into media is recommended for the isolation and identification of vitamin B6-dependent viridans streptococci.

Abscess↗

Effect of penicillin resistance on presentation and outcome of nonenterococcal streptococcal infective endocarditis.

BACKGROUND AND AIMS: Most cases of infective endocarditis are caused by nonenterococcal streptococci. The emergence of strains resistant to penicillin is increasingly recognized worldwide. This study sought to assess the effect of penicillin resistance on presentation and outcome of streptococcal endocarditis. METHODS: A retrospective study was conducted in a single tertiary care hospital. RESULTS: Between August 1996 and December 2004, patients infected with nonenterococcal streptococcal endocarditis and known minimal inhibitory concentrations (MICs) to penicillin were included in this study. A total of 62 cases were identified: 48 (77%) cases of the streptococcal endocarditis were caused by viridans streptococci. The most common species groups identified were Streptococcus mitis in 12 (19%) cases, Streptococcus oralis in 9 (15%) cases, Streptococcus sanguis in 7 (11%) cases and Streptococcus mutans in 7 (11%) cases. There were 36 male and 26 female patients with a median age of 46 years (range: 1-85). Twenty-two patients (35%) had peripheral embolization and 10 patients (16%) died in hospital. Twenty-eight (45%) of the 62 patients were infected with streptococcal strains of MICs > or =0.125 microg/ml to penicillin. Infection with streptococcal strains of MICs > or =0.125 microg/ml to penicillin was associated with a lower incidence of all-site embolization, but was not associated with higher mortality or increased frequency of surgery. CONCLUSIONS: High-level penicillin resistance of the streptococci responsible for endocarditis was increasingly common and medical therapy with vancomycin had a good response. Penicillin resistance was associated with a decreased risk of embolization, but was not associated with higher mortality.

Adolescent↗

Proportional distribution and relative adherence of Streptococcus miteor (mitis) on various surfaces in the human oral cavity.

A group of streptococci possessing the characteristics of Streptococcus miteor (S. mitis) was found to predominate on nonkeratinized human oral mucosa. These organisms averaged from 76 to 89% of the total flora cultivable on anaerobically incubated blood agar plates from cheek, lip, and ventral tongue surfaces. They averaged 34, 40, and 18% of the streptococci in dental plaque, in saliva, and on the tongue dorsum, respectively. Their ability to adhere to oral surfaces was studied by introducing mixtures of streptomycin-resistant strains of S. miteor, S. salivarius, and S. mutans into the mouths of volunteers. Samples of oral surfaces taken 1 hr later indicated S. miteor adhered far better than the other streptococci to buccal mucosa and to teeth, but S. salivarius showed a higher affinity to the tongue dorsum. Glucose-grown cells of S. mutans adhered feebly to all oral surfaces studied and were rapidly cleared from the mouth. Cells of S. miteor and S. salivarius present naturally in saliva adhered to cleaned teeth comparable to in vitro cultivated strains. Electron microscopy of cells of S. miteor attached to buccal epithelial cells obtained from germfree rats indicated that the organisms possessed a fibrillar "fuzzy" coat which appeared to mediate their attachment to the epithelial cell membrane. This "fuzzy" coat was removed by treatment with trypsin, and it appears to be similar to that previously observed on cells of S. pyogenes and S. salivarius.

Cell Adhesion↗

Molecular characterization of disease-associated streptococci of the mitis group that are optochin susceptible.

Eight optochin-susceptible (Opt(s)) alpha-hemolytic (viridans) streptococcus isolates were characterized at the molecular level. These isolates showed phenotypic characteristics typical of both viridans streptococci and Streptococcus pneumoniae. Comparison of the sequence of housekeeping genes from these isolates with those of S. pneumoniae, Streptococcus mitis, Streptococcus oralis, and Streptococcus pseudopneumoniae suggested that the Opt(s) isolates corresponded to streptococci of the mitis group. Besides, the Opt(s) streptococci were negative by a Gen-Probe AccuProbe pneumococcus test and hybridized with specific pneumococcal probes (lytA and ply) but also with ant, a gene not present in most S. pneumoniae strains. Moreover, the isolates were insoluble in 1% sodium deoxycholate but completely dissolved in 0.1% deoxycholate. Sequence analysis of the lytA gene revealed that the Opt(s) streptococci carried lytA alleles characteristic of those present in nonpneumococcal streptococci of the mitis group. The determination of the partial nucleotide sequence embracing the atp operon encoding the F(o)F(1) H(+)-ATPase indicated that the optochin susceptibility of the isolates was due to the acquisition of atpC, atpA, and part of atpB from S. pneumoniae by horizontal gene transfer.

Alleles↗

Salivary IgA antibody to oral streptococcal antigens in predentate infants.

Salivary IgA antibodies to oral streptococci which colonize the infant oral cavity soon after birth were analyzed in 53 whole salivas of 17 infants who were between three and 27 weeks old. Antibody activities to Streptococcus mitis cells were detected by enzyme linked immunosorbent assay in 78% of the whole salivas by the twelfth week of age. This antibody activity was associated with polymeric IgA as determined after gel filtration of salivas on Superose 6, followed by ELISA. Western blot analyses were used to detect IgA antibodies to Streptococcus mitis and Streptococcus salivarius culture supernatants. Forty one, and 91% of saliva samples contained IgA antibody which reacted in Western blot analyses with S. salivarius and S. mitis culture supernatants, respectively. The youngest infant to show reactive IgA antibody with either oral streptococcal antigen preparation was five week old. Salivary IgA antibody to either bacterial culture supernatant was detected in Western blot only after the isolation of the respective streptococcal species from the oral cavity of these young infants. Some heterogeneity was observed among patterns developed with salivas from different infants. These results suggest that salivary IgA antibody responses may be induced by oral colonization (S. mitis, S. salivarius) by the end of the first month of life.

Age Factors↗

An evaluation of the bactericidal effect of the Nd:YAP laser.

The aim of this in vitro study was to compare the efficacy of a classical irrigant with that of a laser in disinfecting a contaminated root canal. Thirty canals of extracted single-root teeth were prepared with files to size #20. The teeth were sterilized with Germispad (Spad, France) for 30 min and then inoculated with Streptococcus mitis ATCC 33399. By randomization, the teeth were divided into six groups of five teeth each. In the first group, teeth were neither inoculated nor prepared. This was the sterility control group (1). In the second group (2) teeth were inoculated without any preparation: as positive controls. The third group was inoculated and then hand-instrumented with files to size #30 with 5.25% NaOCl as irrigant. This was the hand instrumentation group. The other groups were prepared with hand instrumentation with files to size #30, using sterile water as an irrigant, and the canal was then lased with different frequencies as follows: group 4, frequency of 5 Hz and power of 260 mJ; group 5, frequency of 10 Hz and power of 310 mJ; and group 6, frequency of 30 Hz and power of 300 mJ. After experimentation, the residual colonies were counted. The results indicated that (i) the treatment with NaOCl and manual instrumentation effectively inhibited the growth of Streptococcus mitis ATCC 33399; and (ii) the antibacterial effect of the Nd:YAP laser depended on the frequency. Only a frequency of 30 Hz of the Nd:YAP laser inhibited the growth of Streptococcus mitis ATCC 333999.

Dental Disinfectants↗

Update on novel, validly published, and included bacterial taxa derived from human clinical specimens and taxonomic revisions published in 2025.

This review summarizes novel taxon designations ascribed to prokaryotes derived from human primary clinical material during calendar year 2025, as well as proposed revisions to existing taxonomy. Major activity took place in the Streptococcus genus, as more than one dozen novel species were validly and effectively published, with two of these later classified as synonyms of Streptococcus thalassemiae. Moreover, whole genome sequencing and phylogenetic investigation of Streptococcus mitis group organisms resulted in a proposal to designate five species-level Streptococcus spp. taxa as Streptococcus mitis and an additional taxon as Streptococcus oralis subsp. dentisani. More than one dozen taxa were newly included in order Enterobacterales in 2025. Select novel taxa within genera Providencia and Enterobacter commonly possessed genotypes that conferred resistance to carbapenem and/or higher-generation cephem agents. Stenotrophomonas muris sp. nov. and Terrisporobacter muris sp. nov., initially characterized in gnotobiotic murine systems within the past 4 years, had clinical significance in human infection that was demonstrated in primary literature. The vast majority of the more than 70 novel obligate anaerobic taxa were derived from microbiome studies and had little ascribed clinical significance. Four novel obligate anaerobic Gram-negative taxa were shown to be of greater abundance in persons with Parkinson's disease than in those without. Updates to taxa previously published in the Journal of Clinical Microbiology compendia reveal that several could serve as reservoirs for multiple antimicrobial resistance determinants. One example is the non-glucose fermentative Gram-negative bacillus Pseudomonas juntendi.

human clinical specimens↗

In-vitro adherence of oral streptococci in the presence of sucrose and its relationship to cariogenicity in the rat.

Streptococcus sanguis I gave a significantly greater percentage coverage (cell-pellicle attachment) of saliva-coated glass in the presence of sucrose than did Strep. sanguis II (p less than 0.025), and both these gave greater percentage coverages than the other species tested, between which no significant differences were noted. There was a large number of significant differences in clump size (cell-cell attachment) between species-pairs. Among the mutans streptococci, there were significant differences in the percentage coverage between Streptococcus rattus/Streptococcus mutans (p less than 0.05) and Strep. rattus/Streptococcus sobrinus (p less than 0.01), and in clump size between all species-pairs with the exception of Streptococcus cricetus or Strep. sobrinus and Strep. mutans. The rank order of species in relation to fissure caries was mutans streptococci greater than Streptococcus salivarius greater than Streptococcus milleri greater than Strep. sanguis greater than Streptococcus faecalis greater than Streptococcus mitis greater than Streptococcus lactis. There was a significant correlation between percentage coverage in vitro and fissure caries in vivo for strains of Strep. sanguis (p less than 0.05) and pooled strains of Strep. mitis and Strep. sanguis (p less than 0.01). On comparing data for adherence in the presence or the absence of sucrose, the sugar had no effect on the percentage coverage of the seven species tested, but significantly increased the clump size of the mutans streptococci (p less than 0.01) and Strep. sanguis (p less than 0.05).

Animals↗