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In vitro activity of gemifloxacin (SB-265805) compared to eleven other antimicrobial agents against streptococcal isolates, excluding Streptococcus pneumoniae.

The purpose of the study presented here was to determine the in vitro activity of gemifloxacin compared with that of 11 other antimicrobial agents (5 of them quinolones) against 400 isolates of beta-haemolytic and viridans group streptococci. The minimum inhibitory concentration values for gemifloxacin against 90% of the streptococci tested were as follows: Lancefield groups A, C and G, 0.06 microg/ml; Lancefield group B, Streptococcus mitis, Streptococcus mutans and Streptococcus bovis, 0.125 microg/ml; and Streptococcus milleri, 0.03 microg/ml. Resistance to penicillin, ampicillin and erythromycin was found mainly in the Streptococcus mitis isolates; tetracycline showed variable results, and no vancomycin resistance was encountered. Higher rates of ciprofloxacin resistance were identified in the Streptococcus bovis, mitis and mutans isolates. In conclusion, gemifloxacin was the most active quinolone tested followed by trovafloxacin, sparfloxacin, grepafloxacin, ciprofloxacin and levofloxacin, especially against isolates resistant to beta-lactam agents, macrolides and tetracycline.

Anti-Bacterial Agents↗

Differentiation of mutans streptococci by intact cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

It is difficult to distinguish mutans streptococci on the species level, and even more so on the subspecies level. Intact cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) (ICM) was applied to reference strains of five of the species of the mutans group (Streptococcus criceti, Streptococcus downei, Streptococcus mutans, Streptococcus ratti, Streptococcus sobrinus), nonmutans streptococci (Streptococcus oralis, Streptococcus mitis, Streptococcus salivarius, and Streptococcus sanguinis), and 177 mutans streptococci isolated from saliva of 10 children. From the analysis of the reference strains, readily distinguishable ICM mass spectra were obtained for the different species. Based on multivariate statistical analysis, a correct and unambiguous assignment was made of the spectra of 159 isolated mutans streptococci to S. mutans and 16 isolates to S. sobrinus. Two isolates were sorted out and were identified by sequencing of their 16S rRNA genes as Streptococcus anginosus. In addition, ICM indicated a misclassification for some reference strains (AHT, V 100 and E 49) and re-classified AHT and E 49 as S. ratti and V 100 as S. sobrinus. This was confirmed by 16S rDNA sequencing. Based on a statistical similarity analysis of the spectra of reference strains and a quantitative assessment of the reproducibility of ICM, the isolates identified as either S. mutans or S. sobrinus were phenotyped on the subspecies level. In the population of the clinical isolates, 14 unambiguously different S. mutans and three different S. sobrinus phenotypes were detected. ICM proved to be a powerful tool for a differentiation of mutans streptococci down to the subspecies level.

Child↗

The acid-tolerant microbiota associated with plaque from initial caries and healthy tooth surfaces.

The intent of this study was to compare the inherent acid tolerance of bacteria in samples of dental plaque from tooth sites in subjects with and without initial caries. Plaque was collected from approximal surfaces showing early enamel caries and from healthy tooth surfaces in the same subjects, as well as from enamel surfaces of caries-free individuals. In addition to plating on blood agar, the plaque samples were plated directly on non-selective solid agar medium buffered to pH 7.0, 6.0, 5.5, 5.0, 4.5 and 4.0 to avoid any loss of adaptation to acid during primary isolation of plaque bacteria. The results showed that approximately 50% of the total cultivable plaque microbiota from caries, as well as healthy tooth sites, was able to grow at pH 5.5 and 1% at pH 5.0, pH values regarded as critical for the demineralization of tooth enamel. At pH 5.0, members of the genus Streptococcus were the dominant group, but mutans streptococci accounted for less than half of the streptococcal viable count. The other acid-tolerant streptococcal isolates included Streptococcus anginosus, Streptococcus constellatus, Streptococcus gordinii, Streptococcus intermedius, Streptococcus mitis, Streptococcus oralis, Streptococcus salivarius and SStreptococcus sanguis. Analysis of the results indicated that the mutans streptococci in dental plaque were highly variable with respect to acid tolerance, and that both caries and healthy sites harboured significant numbers of mutans streptococci that were not acid-tolerant.

Acids↗

The influences of hinge length and composition on the susceptibility of human IgA to cleavage by diverse bacterial IgA1 proteases.

The influences of IgA hinge length and composition on its susceptibility to cleavage by bacterial IgA1 proteases were examined using a panel of IgA hinge mutants. The IgA1 proteases of Streptococcus pneumoniae, Streptococcus sanguis strains SK4 and SK49, Neisseria meningitidis, Neisseria gonorrhoeae, and Haemophilus influenzae cleaved IgA2-IgA1 half hinge, an Ab featuring half of the IgA1 hinge incorporated into the equivalent site in IgA1 protease-resistant IgA2, whereas those of Streptococcus mitis, Streptococcus oralis, and S. sanguis strain SK1 did not. Hinge length reduction by removal of two of the four C-terminal proline residues rendered IgA2-IgA1 half hinge resistant to all streptococcal IgA1 metalloproteinases but it remained sensitive to cleavage by the serine-type IgA1 proteases of Neisseria and Haemophilus spp. The four C-terminal proline residues could be substituted by alanine residues or transferred to the N-terminal extremity of the hinge without affect on the susceptibility of the Ab to cleavage by serine-type IgA1 proteases. However, their removal rendered the Ab resistant to cleavage by all the IgA1 proteases. We conclude that the serine-type IgA1 proteases of Neisseria and Haemophilus require the Fab and Fc regions to be separated by at least ten (or in the case of N. gonorrhoeae type I protease, nine) amino acids between Val(222) and Cys(241) (IgA1 numbering) for efficient access and cleavage. By contrast, the streptococcal IgA1 metalloproteinases require 12 or more appropriate amino acids between the Fab and Fc to maintain a minimum critical distance between the scissile bond and the start of the Fc.

Amino Acid Sequence↗

Unusual asymptomatic giant sialolith of the submandibular gland: a clinical report.

This report presents an unusual case of asymptomatic sialolith of the submandibular gland. A 61-year-old man was referred to our department for multiple extractions. An ortopantomographic exam revealed the existence of a large radiopacity in the right premolar mandibular region. The patient was completely asymptomatic and no episodes of pain and swelling had occurred in the previous years. Ultrasonography and clinical examination confirmed the diagnosis of sialolithiasis of the submandibular duct. The calculus was removed trans-orally in local anaesthesia. The sialolith measured 22 mm and it was mainly constituted by phosphate, calcium and smaller amounts of magnesium. The bacteriological exam revealed the presence of Streptococcus Mitis, Streptococcus Salivarius and non-pathogenic Neisserie. Postoperative course was uneventful. Even a sialolith of significant dimensions may not be symptomatic. Nevertheless, the likelihood of future complications may constitute an indication for surgical removal of abnormal asymptomatic sialoliths.

Calcium Carbonate↗

Adherence of Veillonella species mediated by extracellular glucosyltransferase from Streptococcus salivarius.

The effect of extracellular products from Streptococcus salivarius on sucrose-dependent adherence to smooth surfaces by other oral bacteria was studied in vitro. Strains of Streptococcus mitis, Streptococcus pyogenes, and Veillonella parvula without innate ability to adhere to a steel wire were able to do so when incubated with sucrose and cell-free culture fluid from S. salivarius strains 9759, 25975, CNII, and MEPI. These culture fluids synthesized more adherent material and water-insoluble glucan than those from Streptococcus mutans C67-1 and seven other S. salivarius strains. Among the S. salivarius strains, glucosyltransferase (GT; dextransucrase, EC 2.4.1.5) activity varied more than 100-fold. Cells of Veillonella and S. mitis S3 that had been incubated in culture fluids from S. salivarius 25975 and 9759, respectively, and then washed adhered upon subsequent incubation with sucrose. This was due to adsorbed GT because (i) the adherence was sensitive to dextranase; (ii) it was observed only with the high-GT culture fluids; (iii) it was dependent on sucrose; and (iv) the washed Veillonella cells synthesized glucan, but not fructan, from sucrose. These results suggest that sucrose-dependent adherence of bacteria without such innate ability can be mediated by (i) entrapment in insoluble glucan synthesized by S. salivarius culture fluids, and (ii) prior adsorption of GT from S. salivarius culture fluids. The possibility that GT formed by high-yield strains of S. salivarius is distributed through the mouth by the action of salivary flow and contributes to sucrose-dependent adherence and plaque formation is considered.

Adsorption↗

Antibacterial temporary filling materials: the effect of adding various ratios of Ag-Zn-Zeolite.

The effect of a new type of antibacterial temporary filling material was evaluated. Ag-Zn-Zeolite (Bactekiller, Kanebo, Japan) and SiO2 filler were incorporated into urethane acrylate monomer paste in amounts of 5/55, 10/50, 20/40 and 30/30 wt%, respectively. The present study was designed to use a dye penetration test to measure direct inhibition of bacterial growth of four oral bacteria (Streptococcus mutans, Streptococcus mitis, Streptococcus salivarius, Streptococcus sanguis). The amounts of silver and zinc ions released from these materials were measured by atomic absorption spectrophotometry. The results indicated that the occurrence of marginal leakage was low in all of these materials. These materials exhibited prominent in-vitro antibacterial activity against S. mutans and S. mitis. The Ag-Zn-Zeolite in these materials was able to release very small but detectable amounts of Ag and Zn even 4 weeks after the immersion started. The larger the amounts of Ag-Zn-Zeolite that were incorporated, the greater the release of silver and zinc. However, it appears that increasing antibacterial activity is not promoted by the higher ratio of Ag-Zn-Zeolite.

Acrylic Resins↗

Emergence in human dental plaque and host distribution of amylase-binding streptococci.

Salivary amylase is known to bind specifically to several species of oral streptococci. To assess the importance of this interaction in bacterial colonization of the oral cavity, we determined the proportion and identity of amylase-binding bacteria (ABB) in dental plaque of humans and various salivary amylase-secreting and non-secreting mammalian species. The numbers of ABB in undisturbed plaque collected over time from tooth surfaces of six human volunteers or from 14 other mammalian species were determined by means of a replicating assay. The mean proportion of ABB cultured aerobically from human teeth at 2 h was 10.5% (SD 10), at 8 h 7.9% (8), at 24 h 13% (11), and at 48 h 12% (9). The mean proportion of anaerobically cultured ABB found at 2 h was 3% (SD 4), at 8 h 5% (5), at 24 h 12% (9), and at 48 h 16% (12). Amylase-binding bacteria cultured from these samples resembled Streptococcus mitis, Streptococcus gordonii, Streptococcus salivarius, Streptococcus crista, or unidentified streptococci. In addition, only animals exhibiting salivary amylase activity in their saliva harbored ABB (ranging from 2 to 31% of the total flora), with the exception of the pig, where no ABB were found to colonize, despite considerable amylase activity in saliva. Only strains resembling S. mitis and S. salivarius and unspeciated strains were isolated from these mammals. These results suggest that amylase-binding streptococci are the predominant ABB in human plaque, and their numbers generally increase as plaque develops. Since ABB colonized only the oral cavities of hosts demonstrating salivary amylase activity, the ability to bind amylase may play an important role in oral colonization by these bacteria.

Amylases↗

Preparation and antimicrobial activity of gelatin microparticles containing propolis against oral pathogens.

Gelatin microparticles containing propolis ethanolic extractive solution were prepared by spray-drying technique. Particles with regular morphology, mean diameter ranging of 2.27 microm to 2.48 microm, and good entrapment efficiency for propolis were obtained. The in vitro antimicrobial activity of microparticles was evaluated against microorganisms of oral importance (Enterococcus faecalis, Streptococcus salivarius, Streptococcus sanguinis, Streptococcus mitis, Streptococcus mutans, Streptococcus sobrinus, Candida albicans, and Lactobacillus casei). The utilized techniques were diffusion in agar and determination of minimum inhibitory concentration. The choice of the method to evaluate the antimicrobial activity of microparticles showed be very important. The microparticles displayed activity against all tested strains of similar way to the propolis, showing greater activity against the strains of E. salivarius, S. sanguinis, S. mitis, and C. albicans.

Anti-Infective Agents↗

Characterization of salivary alpha-amylase binding to Streptococcus sanguis.

The purpose of this study was to identify the major salivary components which interact with oral bacteria and to determine the mechanism(s) responsible for their binding to the bacterial surface. Strains of Streptococcus sanguis, Streptococcus mitis, Streptococcus mutans, and Actinomyces viscosus were incubated for 2 h in freshly collected human submandibular-sublingual saliva (HSMSL) or parotid saliva (HPS), and bound salivary components were eluted with 2% sodium dodecyl sulfate. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western transfer, alpha-amylase (EC 3.2.1.1) was the prominent salivary component eluted from S. sanguis. Studies with 125I-labeled HSMSL or 125I-labeled HPS also demonstrated a component with an electrophoretic mobility identical to that of alpha-amylase which bound to S. sanguis. Purified alpha-amylase from human parotid saliva was radiolabeled and found to bind to strains of S. sanguis genotypes 1 and 3 and S. mitis genotype 2, but not to strains of other species of oral bacteria. Binding of [125I]alpha-amylase to streptococci was saturable, calcium independent, and inhibitable by excess unlabeled alpha-amylases from a variety of sources, but not by secretory immunoglobulin A and the proline-rich glycoprotein from HPS. Reduced and alkylated alpha-amylase lost enzymatic and bacterial binding activities. Binding was inhibited by incubation with maltotriose, maltooligosaccharides, limit dextrins, and starch.

Adult↗

Purification and properties of sorbitol-6-phosphate dehydrogenase from oral streptococci.

The activity of sorbitol-6-phosphate (S6P) dehydrogenase (S6PDH) and the sorbitol transport system were studied in strains of the oral streptococci Streptococcus gordonii, Streptococcus mitis, Streptococcus sanguis and Streptococcus mutans. Genetically transformed (to ferment sorbitol) strains and their DNA donors were included. S6PDH was purified by anion exchange chromatography and gel filtration. The purity of the enzyme was confirmed by polyacrylamide gel electrophoresis. The purified enzyme from all the strains exhibited Michaelis-Menton saturation kinetics. The Km values for nicotinamide-adenine dinucleotide (NAD) and S6P ranged between 0.03 and 0.21 mM and 0.07 and 0.20 mM respectively. The relative molecular weights of the native enzyme were 229,000 for one donor-transformant pair (S. sanguis and S. gordonii), 107,000 for the other pair (S. mitis and S. gordonii) and 129,000 for S. mutans. The molecular weights of the S6PDH subunits ranged from 26,000 to 28,000. The pH optima (greater than 8.5) and the amino acid composition (15 amino acids examined) were similar for the S6PDH from the different strains. However, the chromatographic and electrophoretic patterns as well as the Km values for NAD and S6P were the same only between the S6PDHs from the strains within each donor-transformant pair. Purified S6PDH from S. mutans also exhibited low mannitol-1-phosphate dehydrogenase activity. Sorbitol-grown decryptified cells of all the strains phosphorylated sorbitol in the presence of phosphoenolpyruvate but not in the presence of adenosine triphosphate (ATP). ATP-mediated phosphorylation of glucose was observed with the same strains when grown on glucose. No evidence for a non-phosphotransferase transport system was found for sorbitol in any of the strains.(ABSTRACT TRUNCATED AT 250 WORDS)

Biological Transport, Active↗

Enamel demineralization by acid produced from endogenous substrate in oral streptococci.

Bovine enamel blocks were covered by a bacterial cell mass and held in the mouths of human volunteers by a palatal appliance, to study the effects of acid production from endogenous bacterial substrate on the pH of the mass and on enamel demineralization. Variables were cells with and without endogenous substrate, rinsing with 10 per cent glucose or sucrose solution, or no rinsing and the type of organism. Acid production from endogenous substrate by Streptococcus mutans strain IB-1600, due, at least in part, to intracellular glycogen degradation, caused a marked and prolonged pH drop and significant enamel demineralization. A similar but less marked effect was obtained with a strain of Streptococcus mitis, Streptococcus sanguis and Actinomyces viscosus. Thus acid production from endogenous sources by plaque bacteria may be of importance in the aetiology of dental caries.

Acids↗