Bettina Spitzmüller

Plasmid-Mediated Quinolone Resistance in Isolates Obtained in German Intensive Care Units

ABSTRACT Screening of 703 isolates of Enterobacteriaceae, obtained from 34 German intensive care units (ICUs), revealed qnr-positive, integron-containing isolates of Enterobacter sp. and Citrobacter freundii from four patients in 2 German ICUs. This is one of the first reports of qnr-positive strains obtained from patients in Europe.

Intrinsic enzymatic crosslinking and maturation of the in situ pellicle

AIM The acquired enamel pellicle is a proteinaceous layer formed on all solid substrata exposed to the oral cavity. It has been supposed that the pellicle undergoes maturation after protein adsorption. The aim of the present study was to investigate enzyme activities with an impact on intrinsic maturation processes in in situ formed pellicles. METHODS Bovine enamel specimens were exposed to the oral cavity in six subjects to allow in situ pellicle formation over 3, 30 and 120 min. The slabs were fixed on the buccal and palatal surfaces of individual splints fixed with silicone impression material. After rinsing with deionised water, the pellicle samples were tested fluorimetrically for transglutaminase, protease and elastase activity. Phosphatase activities were tested photometrically. Separate samples were used for each of the enzymes tested. RESULTS Transglutaminase was detected in in situ pellicle (16.7+/-21.2 mU/cm(2)) as was alkaline phosphatase activity (0.87+/-0.99 mU/cm(2)). For both enzymes, there was no correlation of enzyme activities with time or localisation of pellicle formation. Acidic phosphatase- and protease-activities were not detectable. Only traces of elastase activity were found in 57% of the samples. CONCLUSION Transglutaminase and phosphatase activity are detectable within in situ pellicle. Enzymatic crosslinking and dephosphorylation appear more important for intrinsic maturation of the acquired enamel pellicle than proteolysis.

Food-borne Enterococci Integrate Into Oral Biofilm: An In Vivo Study

INTRODUCTION Enterococci, particularly Enterococcus faecalis, are still a primary concern in endodontic infections. To date, enterococci have been considered to be only transiently present in the oral cavity. The aim of this study was to examine whether different enterococci from food are able to reside in oral biofilm. METHODS Six healthy volunteers wore dental splints loaded with enamel slabs. After 3 days, the volunteers consumed cheese containing enterococci. The fate of the enterococci was analyzed by culture technique and 16S rRNA gene sequencing. All isolates were characterized genotypically by macrorestriction analysis (SmaI) and pulsed-field gel electrophoresis. E. faecalis was also analyzed by using fluorescent in situ hybridization (FISH). RESULTS E. faecalis, E. faecium, E. avium, and E. durans were detected in the initial biofilm after 2 hours, as well as in the 5-day-old oral biofilm. E. faecalis, E. faecium, and E. avium isolated from the initial biofilm and from the 5-day-old biofilm, as well as those isolated from cheese, showed genetic homogeneity. E. faecium and E. avium had integrated into a pre-existing 3-day-old biofilm. No genetic similarity between E. durans strains isolated from cheese and those from the initial and 5-day-old oral biofilm was detected. E. faecalis was also detected in the oral biofilm by using FISH. CONCLUSIONS Food-borne enterococci, particularly E. faecalis, might not only be transient but could also survive in the oral biofilm and become a source for endodontic infections. Moreover, genotypic analysis is required to study the source of oral enterococci.

Effects of Cistus-tea on bacterial colonization and enzyme activities of the in situ pellicle

OBJECTIVES Polyphenols are expected to have antibacterial properties. Cistus is a tea rich in polyphenols. The aim of the present in situ study was to investigate the effect of Cistus-tea on the pellicle and on the initial oral biofilm. METHODS For in situ pellicle formation and initial biofilm formation, bovine enamel slabs were fixed on maxillary splints and carried by four subjects at buccal sites for up to 2 h. Bacteria present in 120-min pellicles were determined with DAPI-staining and fluorescence in situ hybridization with and without a 10 min rinse with Cistus-tea performed 1 min after incorporation of the slabs. In addition, amylase, lysozyme, glucosyltransferase and peroxidase activities immobilised in the pellicle layer were measured before and after rinsing for 10 min with Cistus-tea. RESULTS The amount of bacteria detected in the 120-min biofilm was reduced significantly, if a 10 min rinse with Cistus-tea was performed one min after insertion of the enamel slabs. DAPI-staining yielded 13.2+/-3.5 for controls and 6.5+/-1.1 x 10(4) bacteria/cm(2), if a rinse with Cistus-tea was applied. Lysozyme, amylase and glucosyltransferase activities immobilised in the pellicle were not affected following a rinse with Cistus-tea. However, peroxidase activity was reduced significantly. CONCLUSIONS Cistus-tea may be used to reduce the initial bacterial adhesion in the oral cavity.

Electron microscopic detection and activity of glucosyltransferase B, C, and D in the in situ formed pellicle.

OBJECTIVE Glucosyltransferases (GTFs) represent a virulence factor of mutans streptococci. The aim of the present in situ study was to investigate the distribution of different GTF-isoforms in the pellicle. DESIGN Bovine enamel slabs were fixed on buccal and palatal sites of individual splints worn by five subjects for 30 and 120 min to allow pellicle formation. Pellicle specimens were processed for transmission electron microscopy (TEM) and field emission in-lens scanning electron microscopy (FEI-SEM). Gold-immunolabelling was used for detection of GTF-isoforms B, C and D. Furthermore, glucosyltransferase activity of 3-, 30- and 120-min pellicles was tested via determination of fructose release. RESULTS All isoforms of the enzyme were found to be randomly distributed within all layers of the pellicle. In cross-sections (TEM), GTF D was the most abundant isoform. More labelled molecules were detected on buccal sites compared with palatal surfaces, the number of molecules detected increased with time. The amount of GTF B, C and D found on the pellicle surface by FEI-SEM showed no correlation with pellicle formation time or localisation in the oral cavity. Overall, GTF D was detected more frequently on the surface than GTF B and C. All pellicles tested showed GTF-activity. CONCLUSION The study shows for the first time the presence of the GTF-isoforms B, C and D within all layers of the in situ formed pellicle. This emphasises the impact of streptococcal products on the composition of the pellicle and illustrates a mechanism used by bacteria to colonize dental surfaces.

Bacterial and Candida albicans Adhesion on Different Root Canal Filling Materials and Sealers

INTRODUCTION Microbial adhesion and subsequent biofilm formation on endodontic root canal filling materials and sealers lead to survival of microorganisms in treated root canals and subsequently to endodontic treatment failures. The present study focused on initial microbial adhesion to different endodontic filling materials. METHODS The following endodontic biomaterials were tested: AH-Plus, Tubli Seal, gutta-percha, Real Seal SE, EndoREZ, Apexit Plus, GuttaFlow, and dentin. Samples of each material were prepared. Bovine dentin samples were used as a control. The initial adhesions of salivary bacteria as well as the subsequent single species were quantified by determination of colony-forming units (CFUs) and visualized by scanning electron microscopy and confocal microscopy (CLSM): Enterococcus faecalis, Streptococcus mutans, Streptococcus sanguis, Candida albicans, and Prevotella nigrescens. RESULTS Initially adherent microorganisms could be detected and microscopically visualized on each of the materials tested. Considering the values of the CFUs and the covering grade as detected by CLSM, there were significant differences among the materials. Fewer bacteria tended to adhere to Apexit Plus, whereas Real Seal SE and the widely used gutta-percha showed the highest number of adherent bacteria. This tendency was not detected for C. albicans. CONCLUSIONS Endodontic microorganisms have a high affinity to root canal filling materials and sealers, especially to gutta-percha. Because of this high level of bacterial adhesion, subsequent biofilm formation on these materials could be suggested as leading to the persistence of microorganisms in root canals.

Antimicrobial Effects of Dental Luting Glass Ionomer Cements on Streptococcus mutans

Objective. To reduce secondary caries, glass ionomer luting cements are often used for cementing of indirect restorations. This is because of their well-known antimicrobial potential through the release of fluoride ions. The aim of this in vitro study was to investigate the antimicrobial effect of five dental luting cements which were based on glass ionomer cement technology. Methods. Five different glass ionomer based luting cements were tested for their antimicrobial effects on Streptococcus mutans in two different experimental setups: (i) determination of colony-forming units (CFUs) in a plate-counting assay; (ii) live/dead staining (LDS) and fluorescence microscopy. All experiments were conducted with or without prior treatment of the materials using sterilized human saliva. Antimicrobial effects were evaluated for adherent and planktonic bacteria. Bovine enamel slabs (BES) were used as negative control. BES covered with 0.2% chlorhexidine (CHX) served as positive control. Results. Each of the tested materials significantly reduced the number of initially adhered CFUs; this reduction was even more pronounced after prior incubation in saliva. Antimicrobial effects on adherent bacteria were confirmed by live-dead staining. Conclusion. All five luting cements showed an antimicrobial potential which was increased by prior incubation with human saliva, suggesting an enhanced effect in vivo.