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Biomedical subjects

N E Fiehn

Publications and source records attributed to N E Fiehn.

At least 19 recordsLinked to original sources

The effect of Sterilex Ultra for disinfection of dental unit waterlines.

AIM: To evaluate the effect of a disinfectant agent based on hydrogen peroxide (Sterilex Ultra, Sterilex Corporation, Maryland, USA) on the microbiological water quality in dental unit waterlines (DUWL). SETTING: Six older dental units were disinfected with Sterilex Ultra and another six units with sodium hypochlorite. The results were compared to six non-disinfected units examined during a period without patient treatment. OUTCOME MEASURE: The number of colony forming units of bacteria (cfu)/ml of water was determined according to European Standards on water quality. DESIGN: The water quality was initially determined during a six week period, and subsequently in more detail for two weeks. Finally, the effect of prolonged administration of Sterilex Ultra was investigated for six weeks. RESULTS: Instillation of Sterilex Ultra according to the recommendations of the manufacturer initially reduced the number of bacteria in DUWL to <10(2) cfu/ml. However, following daily, as well as prolonged administration of Sterilex Ultra a gradual recolonisation was observed resulting in bacterial numbers >10(4) cfu/ml in a number of units. Major differences between the number of cfu/ml in individual units were observed. CONCLUSION: Neither daily nor extended administration of Sterilex Ultra was capable of maintaining an acceptable water quality in these older dental units.

Bacteria↗

The effect of drying dental unit waterline biofilms on the bacterial load of dental unit water.

AIM: To evaluate drying of the dental unit waterlines (DUWL) as a new method of controlling the bacterial biofilm therein and thereby to reduce the number of living bacteria in dental unit water. SETTING: 18 dental units were incorporated into the study. Six units constituted the experimental units, which were emptied for stagnant water in the DUWL every night; six units were chlorinated every night (positive controls), and six units were left untreated (negative controls). OUTCOME MEASURE: Water samples from the ultrasonic scaler were examined microbiologically according to the guidelines from the Danish Standard Association and the number of colony forming units (cfu) per ml of water was determined. RESULT: Drying of DUWL did not reduce the number of cfu per ml in dental unit water below the levels found in DUWL left untreated. CONCLUSION: The drying of the waterlines for about 16 hours per day during a 19-day period did not result in reduced counts of bacteria in water samples from the experimental units.

Bacteria↗

Disinfection of dental impressions and occlusal records by ultraviolet radiation.

As chemical disinfection of dental impressions may cause adverse effects on materials and the dental personnel this study examined disinfection by ultraviolet radiation. Alginate, addition silicone rubber and red wax contaminated by Streptococcus salivarius, Fusobacterium nucleatum and five other bacteria in different suspension media were radiated for up to 18 min, and the number of colony forming units was compared to non-radiated controls. The effect of ultraviolet radiation differed among bacterial species and depended on the organic content in the suspension. Generally, the bacterial reduction after ultraviolet radiation was below 4 log steps and thus insufficient for disinfection of dental impressions.

Alginates↗

Effect of steam sterilization inside the turbine chambers of dental turbines.

OBJECTIVE: It has been demonstrated that contamination of the insides of high-speed dental turbines occurs and that bacteria as well as viruses may remain infectious when expelled from such turbines during subsequent use. Consequently, it has been widely recommended that a high-speed turbine be sterilized after each patient. The purpose of this study was to evaluate the effect of steam autoclaving on a high-speed dental turbine with a contaminated turbine chamber. STUDY DESIGN: Streptococcus salivarius and endospores of Bacillus stearothermophilus were used as test organisms to determine the effectiveness of 4 different small non-vacuum autoclaves and one vacuum autoclave. RESULTS: The study demonstrated different efficiencies among the small non-vacuum autoclaves, the best showing close to a 6 log reduction of the test organisms inside the turbine chamber. When cleaning and lubrication of the high-speed dental turbine was carried out before autoclaving, this level of reduction was observed for all the examined non-vacuum autoclaves. CONCLUSIONS: It is concluded that cleaning before sterilization is essential for safe use of high-speed dental turbines and that small non-vacuum autoclaves should be carefully evaluated before being used for the reprocessing of hollow instruments such as high-speed turbines.

Dental High-Speed Equipment↗

Bacterial binding to extracellular matrix proteins -- in vitro adhesion.

A binding assay was developed and used to study the binding of oral streptococcus to immobilized human fibronectin, laminin, vitronectin, fibrinogen, heparin, and collagen IV. The protein binding was dependent on the broth used for bacterial growth. The binding after growth in brain heart infusion broth, trypticase soy broth, Todd-Hewitt broth, and Dulbecco's modified Eagle's medium was examined. Most of the strains were able to bind to immobilized fibronectin and laminin, and to a minor extent vitronectin. Binding was not observed on immobilized fibrinogen, collagen IV, or heparin. Measured surface hydrophobicity correlated well with the bacterial binding strength to the proteins. Streptococcal incubation with putative inhibitors indicates multiple binding mechanisms of a lectin-like and protein nature, possibly involving protein receptors.

Animals↗

The basic science teaching experience in the Nordic countries.

The article concerns the dental education at 10 different faculties in Denmark, Iceland, Norway, Sweden, and Finland. The information is based upon a questionnaire to the faculties concerning the definition, the structure and succession of the basic science subjects/courses/themes/topics in the dental curriculum, the time span of the basic science teaching, and the methods of teaching. Furthermore, there is information about integration among the basic science subjects and the coordination/integration between the basic science teaching and the clinical teaching. Finally, the teachers' educational background and the cowork with the medical education is elucidated. The main findings are: the basic science teaching is structured in different ways in the Nordic countries. In Reykjavik, Bergen, and Aarhus this teaching is subject specific, while in Copenhagen it is mainly subject specific, as it has few integrated courses. In Gothenburg, Stockholm, and Umeå, the basic science teaching consists of a mixture of integrated courses and subject specific teaching. In Helsinki, Oslo and Malmö the basic sciences are theme and topic based, and for the two last-mentioned institutions, integrated with the clinic, except at the faculty in Malmö. Here the basic science teaching is totally integrated in the clinical teaching throughout the study. Usually, the basic science teaching is placed mainly in the first part of the curriculum; however, in Umeå, Copenhagen, and Oslo, some integration takes place.

Curriculum↗

Epidemiology of Pseudomonas aeruginosa in cystic fibrosis and the possible role of contamination by dental equipment.

Cystic fibrosis (CF) patients often suffer from Pseudomonas aeruginosa lung infection yet the source of this organism is not known. In order to determine whether CF patients might be contaminated with P. aeruginosa from dental equipment, a total of 103 water samples from 25 dental sessions in Frederiksberg Municipal Oral Health Care Service were examined. Three samples (2.9%) were positive for P. aeruginosa. Three hundred and twenty-seven water samples from 82 dental sessions from various other Municipal Oral Health Services in Denmark, attended by CF patients, were also examined. Eighteen of 327 samples (5.5%) from nine sessions (11%) were positive for P. aeruginosa. In one case, genotypically identical (RFLP, pulsed-field gel electrophoresis) P. aeruginosa strains were found both in water from the dental equipment and in the CF patients sputum. This indicates a small risk for acquiring P. aeruginosa from dental sessions, which is however equal to the yearly 'natural background' incidence (1-2%) of acquisition of P. aeruginosa in our CF centre.

Case-Control Studies↗

Evaluation of a new device for sterilizing dental high-speed handpieces.

Dental high-speed turbines and handpieces can take up and expel microorganisms during operation and thus need regular sterilization. This study established a method for validating devices used to sterilize high-speed turbines and handpieces. The air and water channels and turbine chambers were contaminated with suspensions of Streptococcus salivarius or endospores of Bacillus stearothermophilus. The effect of flushing and/or autoclaving performed by a new device combining both procedures was evaluated by counting the number of viable bacteria recovered from these devices. Further, the effect on clinically used handpieces was evaluated. In an initial experiment, the device partially reduced S. salivarius, and the endospores survived. In a second experiment, a 5 to 6 log reduction of S. salivarius in air and water channels was obtained. No growth was observed in clinically used high-speed handpieces, and both S. salivarius and endospores were eliminated from the turbine chambers. Thus, the method of validation proved capable of discriminating between different levels of bacterial reduction.

Air↗

Development of resistance to metronidazole and minocycline in vitro.

By local delivery of antibiotics to periodontal pockets, very high initial concentrations are often quickly succeeded by subinhibitory concentrations, which may facilitate development of bacterial resistance. The purpose of the present study was to investigate possible development of resistance in suspected periodontal pathogens after exposure to subinhibitory concentrations of metronidazole and minocycline. The minimal inhibitory concentration (MIC) of 18 reference strains and 12 clinical isolates was determined by a broth dilution method. Subsequently, all strains with MIC < 8 micrograms/ml were exposed to serial passage on plates containing subinhibitory and gradually increasing concentrations of antibiotics, until growth was inhibited. Initially, most strains were inhibited at < or = 0.250 microgram/ml of minocycline and < or = 0.5 microgram/ml of metronidazole, though A. actinomycetemcomitans was resistant to metronidazole. After growth at subinhibitory concentrations, 8 strains survived 1-2 x and 11 stains survived 8-32 x their initial MIC of metronidazole, growing at up to 8 micrograms/ml. All A. actinomycetemcomitans survived 8-64 x their initial MIC of minocycline, growing at > or = 2 micrograms/ml, while all other strains were inhibited at < or = 0.250 microgram/ml, corresponding to a 1-8 x increase in their initial MIC. Thus, development of resistance was observed for periodontal bacteria growing at up to 64 x their initial MIC, but the final level of resistance was moderate.

Aggregatibacter actinomycetemcomitans↗

Microbiology of ligature-induced marginal inflammation around osseointegrated implants and ankylosed teeth in cynomolgus monkeys (Macaca fascicularis).

The microbiota associated with ligature-induced marginal inflammation around osseointegrated dental implants, ankylosed teeth, and normal control teeth was investigated in 8 cynomolgus monkeys (Macaca fascicularis). Submucosal/subgingival plaque was sampled with paper points on the day of literature placement and after 7 weeks. The samples were evaluated by phase-contrast microscopy and by cultivation on enriched non-selective and various solid media. The submucosal/subgingival flora was changed 7 weeks after ligation. The total number of cultivable bacteria and the proportions of motile rods, anaerobic Gram-negative rods, black-pigmented rods, Porphyromonas gingivalis, and Prevotella intermedia increased significantly around implants, ankylosed teeth, and normal control teeth. Except for a significantly higher proportion of anaerobic Gram-positive cocci around implants compared to ankylosed teeth and normal control teeth at the end of the study, no significant microbiological differences were observed between implants, ankylosed teeth, and normal control teeth neither at baseline nor at the end of the study. Consequently, the microbiota associated with marginal inflammation around implants, ankylosed teeth, and normal control teeth appears to be rather similar in cynomolgus monkeys.

Animals↗

Resistance of Streptococcus sanguis biofilms to antimicrobial agents.

Bacteria living in biofilms as dental plaque on tooth surfaces are generally more resistant to antimicrobial agents than bacteria in batch culture normally used for in vitro susceptibility testing. In order to compare the resistance of free-living and surface-grown oral bacteria, the MIC of Streptococcus sanguis 804 and ATCC 10556 to amoxicillin, doxycycline and chlorhexidine was determined by a broth dilution method. Subsequently, S. sanguis biofilms established in an in vitro flow model were perfused with the antimicrobial agents for 48 h at concentrations equal to and up to 500 times the MIC, and biofilm cell number was determined during this period. The antibiotics at the MIC did not affect the cell number of S. sanguis biofilms compared to the starting point, and only after 48 h at 500 times the MIC were the biofilm bacteria eliminated. At intermediate concentrations biofilm cell number gradually decreased. Chlorhexidine also gradually reduced biofilm cell number, but was inhibitory at concentrations closer to the MIC than was the case for the antibiotics. Thus S. sanguis in biofilms survived up to 500 times the MIC found in batch culture for up to 48 h.

Amoxicillin↗

Ribotyping on small-sized spirochetes isolated from subgingival plaque.

In the present study DNA restriction patterns and corresponding ribotypes of 17 subgingival small-sized spirochetes (1:2:1 and 2:4:2 isolates), 2 Treponema socranskii strains and two Treponema denticola strains were examined. Purified chromosomal DNA was digested by BamHI, HindIII, PstI and ClaI. The DNA fragments were separated in a horizontal slab of 0.7% agarose containing ethidium bromide and transferred by nylon membranes. Hybridization was carried out with digoxigenin-labelled copy DNA of 16S and 23S ribosomal RNA from Escherichia coli. Depending on the restriction endonuclease used, up to 4 distinct bands were observed for the 2:4:2 isolates and the T. denticola strains. For each of the endonucleases used, identical band patterns were always observed for this group of isolates, and these patterns differed persistently from the T. denticola strains. For the 1:2:1 strains, up to 11 distinct bands were observed after digestion with HindIII, whereas a maximum of 6 bands were observed when PstI or ClaI was used. By using ClaI, the examined 1:2:1 isolates were separated into 8 groups, whereas PstI and HindIII separated these isolates into 5 groups. The ribotyping showed that the tested 1:2:1 spirochetes were more heterogeneous than the 2:4:2 spirochetes examined.

Bacterial Typing Techniques↗

Identity of 1:2:1 and 2:4:2 subgingival spirochetes by DNA hybridization.

A group of 1:2:1 and 2:4:2 subgingival spirochetes, well characterized by transmission electron microscopy, biochemical tests, cellular fatty acid and carbohydrate analyses, and ribotyping, was recently suggested to represent new treponemal species. The present study used DNA hybridization to examine this possibility. When DNA of a representative strain (no. 16) of the 8 1:2:1 spirochetes examined was labeled by iodination, it showed, after S1 nuclease treatment, from 58 to 104% (average 76%) homology with DNA from the 1:2:1 spirochetes, 94% homology with DNA from the type strain of Treponema socranskii and of T. socranskii subsp. socranskii, i.e., ATCC 35536T, and 62% homology with DNA from T. socranskii subsp. buccale, strain ATCC 35534T. Similarly treated DNA from a representative strain (no. 3) of 8 2:4:2 spirochetes exhibited from 90 to 105% (average 97%) homology with DNA from the 2:4:2 spirochetes, and 85% and 87% homology, respectively, with DNA from Treponema denticola strains ATCC 33520 and FDC T1. There was a negligible degree of homology between the 1:2:1 and 2:4:2 spirochetes. Thus, all the 2:4:2 spirochetes belonged to T. denticola. 1:2:2 strains with DNA homology levels >70% (5 strains) belonged to T. socranskii or T. socranskii subsp. socranskii, while those with homology levels from 58 to 63% (3 strains) most likely belonged to other subspecies of T. socranskii.

Bacterial Typing Techniques↗

Development of a flow method for susceptibility testing of oral biofilms in vitro.

Bacteria in biofilms are known to be more resistant than bacteria in batch cultures to antimicrobial agents. The purpose of the present study was to develop a flow method for formation of oral biofilms permitting susceptibility testing of plaque bacteria. A brain heart infusion (BHI) Streptococcus sanguis 804 culture was pumped through a modified Robbins Device (MRD) with 25 exchangeable silicone disks at 40 ml/h. After 24-48 h disks were removed and biofilm cells dispersed by vortex mixing and low-output ultrasonication. Colony forming units (cfu)/cm2 were determined after aerobic incubation on blood agar plates. Optimal biofilm formation was found after growth for 48 h at 37 degrees C in BHI + 1% sucrose, using saliva-coated silicone disks in inverted MRDs, yielding on average 4.4 x 10(5) cfu/cm2. Similar results were obtained for S. sanguis ATCC 10556 and five clinical isolates. Testing the susceptibility of S. sanguis to chlorhexidine gluconate showed increased resistance of biofilms compared to batch culture. Thus an appropriate biofilm model for susceptibility testing of oral microorganisms has been established.

Biofilms↗

Restriction fragment length polymorphism of rRNA genes for molecular typing of members of the family Legionellaceae.

Typing of Legionella pneumophila remains important in the investigation of outbreaks of Legionnaires' disease and in the control of organisms contaminating hospital water. We found that the discriminatory power of a nonradioactive ribotyping method could be improved by combining results obtained with four restriction enzymes (HindIII, NciI, ClaI, and PstI). Fifty-eight clinical and environmental L. pneumophila strains including geographically unrelated as well as epidemiologically connected isolates were investigated. Epidemiologically related strains had the same ribotypes independent of the combinations of enzymes used. Some strains belonging to the same serogroup were assigned to different ribotypes, and some ribotypes contained members of different serogroups, indicating, as others have found, that serogroup and genotype are not always related. The discriminatory power of the method was estimated by calculating an index of discrimination (ID) for individual enzymes and combinations thereof. The combined result with all four enzymes was highly discriminatory (ID = 0.97), but results for three enzymes also yielded ID values acceptable for epidemiological purposes. In addition, the testing of 27 type strains and 6 clinical isolates representing Legionella species other than L. pneumophila indicated that ribotyping might be of value for species identification within this genus, as previously suggested.

Bacterial Typing Techniques↗

Decontamination of dental equipment. A validation of three devices designed for cleaning, disinfecting, and lubricating of dental high-speed turbines and handpieces.

This study deals with the decontamination of dental turbines and handpieces (DTH) aiming at eliminating the cross-infection risk in dental practice. Three types of equipment designed for cleaning/lubricating DTH were validated. The effect of the three devices was determined for four different types of DTH, which were artificially contaminated with Streptococcus salivarius ATCC 13419 in a controlled manner. Sampling was carried out by collecting one ml water flushed through the DTH connected to a dental unit. The study resulted in a model for testing the decontamination potential of a cleaning/lubrication device for DTH. One of the devices could not reduce the contamination, while two others were able to reduce the bioburden with at least 3.9 logarithmic steps. However none of the devices tested could constantly eliminate the contamination, for which reason an additional heat treatment of the DTH is recommended.

Alcohols↗

Ribotyping for differentiating Flavobacterium meningosepticum isolates from clinical and environmental sources.

On the basis of DNA-DNA hybridization data, two main genomic relatedness groups (I and II) have been reported for a geographically varied collection of 52 strains of Flavobacterium meningosepticum. Herein, we have shown that genomic group II can be further divided into four subgroups (II:1 to II:4). To examine the taxonomic relevance of the ribosomal patterns of the 52 F. meningosepticum strains, the patterns were compared with existing DNA-DNA hybridization data with restriction enzymes PstI and HindIII. Ribotyping of the 52 F. meningosepticum strains showed banding patterns that could identify them correctly to one of the five genomic groups or subgroups. To assess the value of ribotyping for the interpretation of epidemiological data, the discriminatory power of the method was investigated for the 52 F. meningosepticum strains. With one to four restriction enzymes (PstI, HindIII, ClaI, EcoRI), a discriminatory index of 0.95 to 0.97 was found. The value of ribotyping in an epidemiological setting was assessed for three clinical isolates of F. meningosepticum from an outbreak of meningitis and bacteremia in the neonatal intensive care unit, Rigshospitalet, Copenhagen, Denmark. The three clinical isolates were shown to belong to the same ribotype, characteristic of genomic subgroup II:1. This ribotyping method will prove to be a useful tool for epidemiological studies concerning F. meningosepticum in the future.

Bacteremia↗