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PubMed1994

Characterization of biofilm and encrustation on ureteric stents in vivo.

OBJECTIVE: To examine the relationship between encrustation and microbial biofilm formation on indwelling ureteric stents. PATIENTS AND METHODS: Ureteric stents from 40 patients were examined for the presence of a microbial biofilm and encrustations. Bacteria in stent biofilms were isolated and identified. RESULTS: A profuse biofilm (> 10(4) c.f.u. cm-3) was identified on 11 (28%) stents. Enterococcus faecalis was the most common biofilm organism identified and Proteus spp. were not present. Encrustation was seen in 23 (58%) of stents and was not associated with the level of urinary calcium. The major risk factor for stent encrustation was the presence of urolithiasis. Importantly, there was no causative link between stent biofilm formation and encrustation. Both biofilm formation and encrustation increased with the duration of stenting. CONCLUSION: The results indicate that polyurethane is readily encrusted and colonized by bacteria in vivo despite antibiotic prophylaxis. Newer materials must be sought if effective long-term stenting is to be achieved.

Adult↗
PubMed1987

Effect of vancomycin hydrochloride on Staphylococcus epidermidis biofilm associated with silicone elastomer.

Peritonitis is a major complication of continuous ambulatory peritoneal dialysis. Relapsing peritonitis after the cessation of antimicrobial therapy is frequently reported and often involves Staphylococcus epidermidis. To investigate the potential role of catheter-associated biofilm in the pathogenesis of relapsing peritonitis, we describe an in vitro model permitting the development of an S. epidermidis biofilm on silicone elastomer biomaterial. This model has been used to investigate the ability of vancomycin hydrochloride to kill biofilm-encased organisms by using an antibiotic regimen typical of peritonitis therapy. No significant differences were seen between vancomycin-exposed and control groups in biofilm viable and total cell counts after 10 days. Vancomycin-exposed silicone-associated biofilm populations decreased by only 0.5 log10 CFU/cm2 over the study period. MICs and MBCs for the original S. epidermidis suspension were 3.125 and 6.25 micrograms/ml, respectively. For biofilm homogenate suspensions, MICs were 3.125 micrograms/ml, but MBCs were greater than 400 micrograms/ml. These data indicate that the biofilm organisms associated with an indwelling peritoneal catheter may display a form of tolerance, thereby suggesting one possible mechanism behind relapsing peritonitis.

Humans↗
PubMed1988

Inactivation of biofilm bacteria.

The current project was developed to examine inactivation of biofilm bacteria and to characterize the interaction of biocides with pipe surfaces. Unattached bacteria were quite susceptible to the variety of disinfectants tested. Viable bacterial counts were reduced 99% by exposure to 0.08 mg of hypochlorous acid (pH 7.0) per liter (1 to 2 degrees C) for 1 min. For monochloramine, 94 mg/liter was required to kill 99% of the bacteria within 1 min. These results were consistent with those found by other investigators. Biofilm bacteria grown on the surfaces of granular activated carbon particles, metal coupons, or glass microscope slides were 150 to more than 3,000 times more resistant to hypochlorous acid (free chlorine, pH 7.0) than were unattached cells. In contrast, resistance of biofilm bacteria to monochloramine disinfection ranged from 2- to 100-fold more than that of unattached cells. The results suggested that, relative to inactivation of unattached bacteria, monochloramine was better able to penetrate and kill biofilm bacteria than free chlorine. For free chlorine, the data indicated that transport of the disinfectant into the biofilm was a major rate-limiting factor. Because of this phenomenon, increasing the level of free chlorine did not increase disinfection efficiency. Experiments where equal weights of disinfectants were used suggested that the greater penetrating power of monochloramine compensated for its limited disinfection activity. These studies showed that monochloramine was as effective as free chlorine for inactivation of biofilm bacteria. The research provides important insights into strategies for control of biofilm bacteria.

Bacteria↗
PubMed1994

Determination of diffusion coefficients in biofilms by confocal laser microscopy.

Microbial exopolymer may hinder the diffusion of nutrients, antibiotics, and other materials to the cell surface. Studies of diffusion in biofilms have been limited to indirect measurements. This study demonstrated the use of fluorescein and size-fractionated fluor-conjugated dextrans in conjunction with scanning confocal laser microscopy to directly monitor and determine diffusion coefficients within biofilms. The monitoring approaches were simple and, when combined with computerized image collection, allowed assembly of a data set suitable for calculation of one-dimensional diffusion coefficients for biofilm regions. With these techniques, it was shown that regional variability in the mobility of the dextrans occurred within mixed-species biofilms. Some regions exhibited rapid diffusion of all test molecules, while adjacent regions were only penetrated by the lower-molecular-weight compounds. The effective diffusion coefficients (D(e)) determined in a mixed-species biofilm were a function of the molecular radius of the probe (i.e., fluorescein, D(e) = 7.7 x 10 cm s; 4,000 molecular weight, D(e) = 3.1 x 10 cm s; and 2,000,000 molecular weight, D(e) = 0.7 x 10 cm s). These results demonstrated that diffusion in the biofilm was hindered relative to diffusion in the bulk solution. The study indicated that in situ monitoring by scanning laser microscopy is a useful approach for determining the mobility of fluorescently labeled molecules in biofilms, allowing image acquisition, appropriate scales of study, both xy and xz monitoring, and calculation of D(e) values.

Journal Article↗
PubMed1997

Methods for observing microbial biofilms directly on leaf surfaces and recovering them for isolation of culturable microorganisms.

Epifluorescence microscopy, scanning electron microscopy, and confocal laser scanning microscopy were used to observe microbial biofilms directly on leaf surfaces. Biofilms were observed on leaves of all species sampled (spinach, lettuce, Chinese cabbage, celery, leeks, basil, parsley, and broad-leaved endive), although the epifluorescent images were clearest when pale green tissue or cuticle pieces were used. With these techniques, biofilms were observed that were about 20 (mu)m in depth and up to 1 mm in length and that contained copious exopolymeric matrices, diverse morphotypes of microorganisms, and debris. The epifluorescence techniques described here can be used to rapidly determine the abundance and localization of biofilms on leaves. An additional technique was developed to recover individual biofilms or portions of single biofilms from leaves and to disintegrate them for isolation of the culturable microorganisms they contained. Nineteen biofilms from broad-leaved endive, spinach, parsley, and olive leaves were thus isolated and characterized to illustrate the applications of this technique.

Journal Article↗
PubMed1998

Structural and functional dynamics of sulfate-reducing populations in bacterial biofilms

We describe the combined application of microsensors and molecular techniques to investigate the development of sulfate reduction and of sulfate-reducing bacterial populations in an aerobic bacterial biofilm. Microsensor measurements for oxygen showed that anaerobic zones developed in the biofilm within 1 week and that oxygen was depleted in the top 200 to 400 &mgr;m during all stages of biofilm development. Sulfate reduction was first detected after 6 weeks of growth, although favorable conditions for growth of sulfate-reducing bacteria (SRB) were present from the first week. In situ hybridization with a 16S rRNA probe for SRB revealed that sulfate reducers were present in high numbers (approximately 10(8) SRB/ml) in all stages of development, both in the oxic and anoxic zones of the biofilm. Denaturing gradient gel electrophoresis (DGGE) showed that the genetic diversity of the microbial community increased during the development of the biofilm. Hybridization analysis of the DGGE profiles with taxon-specific oligonucleotide probes showed that Desulfobulbus and Desulfovibrio were the main sulfate-reducing bacteria in all biofilm samples as well as in the bulk activated sludge. However, different Desulfobulbus and Desulfovibrio species were found in the 6th and 8th weeks of incubation, respectively, coinciding with the development of sulfate reduction. Our data indicate that not all SRB detected by molecular analysis were sulfidogenically active in the biofilm.

Journal Article↗
PubMed2005

Biofilm reactors for industrial bioconversion processes: employing potential of enhanced reaction rates.

This article describes the use of biofilm reactors for the production of various chemicals by fermentation and wastewater treatment. Biofilm formation is a natural process where microbial cells attach to the support (adsorbent) or form flocs/aggregates (also called granules) without use of chemicals and form thick layers of cells known as "biofilms." As a result of biofilm formation, cell densities in the reactor increase and cell concentrations as high as 74 gL(-1) can be achieved. The reactor configurations can be as simple as a batch reactor, continuous stirred tank reactor (CSTR), packed bed reactor (PBR), fluidized bed reactor (FBR), airlift reactor (ALR), upflow anaerobic sludge blanket (UASB) reactor, or any other suitable configuration. In UASB granular biofilm particles are used. This article demonstrates that reactor productivities in these reactors have been superior to any other reactor types. This article describes production of ethanol, butanol, lactic acid, acetic acid/vinegar, succinic acid, and fumaric acid in addition to wastewater treatment in the biofilm reactors. As the title suggests, biofilm reactors have high potential to be employed in biotechnology/bioconversion industry for viable economic reasons. In this article, various reactor types have been compared for the above bioconversion processes.

Journal Article↗
PubMed2006

Effect of temperature and pipe material on biofilm formation and survival of Escherichia coil in used drinking water pipes: a laboratory-based study.

Segments of used drinking water pipes of galvanised steel (GS), cross-linked polyethylene (PEX), copper pipes (Cu) or new medium-density polyethylene (PE) were investigated for the formation of biofilm and survival of E. coli in biofilm and in the water phase. Pipes were filled with water and incubated at 15 degrees C or 35 degrees C under static conditions. Biofilm formation was followed during 32, 40 and 56 (58) d. The most dense biofilm was formed on GS, reaching approximately 4.7 x 10(5) CFU/cm2 measured as heterotrophic plate count (HPC), and at the other materials the density reached 3 x 10(3) CFU/cm2 on PE and PEX and 5 x 10(1) and 5 x 10(2) CFU/cm2 on Cu pipes after 58d at 15 degrees C. Biofilm HPC values were higher at 35 degrees C than at 15 degrees C, with only slightly higher values on the metals, but 100-fold higher on PE and PEX. Adenosine triphosphate (ATP) measurements confirmed the general trends observed by HPC. Higher temperature was seen to be an important factor reducing E. coli survival in the water phase in drinking water pipes. At 15 degrees C E. coli survived more than 4 d in GS and Cu pipes and 8 d in PE pipes, but was not detected after 48 h at 35 degrees C. The E. coli survived longer at both temperatures in the glass control bottles than in the drinking water pipes. Despite the obvious biofilm formation, E. coli was not detected in the biofilm at any of the investigated surfaces.

Colony Count, Microbial↗
PubMed2003

Influence of ethanol concentration on biofilm bacterial composition from a denitrifying submerged filter used for contaminated groundwater.

The influence of the ethanol concentration on the composition and activity of a developed biofilm in a denitrifying submerged unidirectional filter was studied. Process yields (represented as inorganic nitrogen removal), total platable bacteria, denitrifying bacteria, nitrate- and sulphate-reducing bacteria and denitrifying activity (N2O production) were compared at different ethanol concentrations (0 to 46.74 mg l(-1)). The biofilm exhibited a diverse bacterial composition and higher microbial development at the entrance of the unidirectional biofilter. The number of cells per gram of dry weight of biofilm was increased when the ethanol concentration increased, with the exception of nitrate reducers, for which the number of cells decreased per gram of biofilm in relation to height. Five different species of denitrifying bacteria were isolated from the biofilm, all of which were gram-negative rods. All of the species manifested an increase in denitrifying activity when the ethanol concentration was increased. In this sense, the number of denitrifying bacteria in the biofilm was positively correlated with the ethanol concentration. Both nitrate- and sulphate-reducing bacteria were present in the biofilm in the lower and higher part of the column. Nitrate-nitrogen removal in the submerged filter showed a high correlation with the influent ethanol concentration.

Journal Article↗
PubMed2021

Affordable oral health care: dental biofilm disruption using chloroplast made enzymes with chewing gum delivery.

Current approaches for oral health care rely on procedures that are unaffordable to impoverished populations, whereas aerosolized droplets in the dental clinic and poor oral hygiene may contribute to spread of several infectious diseases including COVID-19, requiring new solutions for dental biofilm/plaque treatment at home. Plant cells have been used to produce monoclonal antibodies or antimicrobial peptides for topical applications to decrease colonization of pathogenic microbes on dental surface. Therefore, we investigated an affordable method for dental biofilm disruption by expressing lipase, dextranase or mutanase in plant cells via the chloroplast genome. Antibiotic resistance gene used to engineer foreign genes into the chloroplast genome were subsequently removed using direct repeats flanking the aadA gene and enzymes were successfully expressed in marker-free lettuce transplastomic lines. Equivalent enzyme units of plant-derived lipase performed better than purified commercial enzymes against biofilms, specifically targeting fungal hyphae formation. Combination of lipase with dextranase and mutanase suppressed biofilm development by degrading the biofilm matrix, with concomitant reduction of bacterial and fungal accumulation. In chewing gum tablets formulated with freeze-dried plant cells, expressed protein was stable up to 3 years at ambient temperature and was efficiently released in a time-dependent manner using a mechanical chewing simulator device. Development of edible plant cells expressing enzymes eliminates the need for purification and cold-chain transportation, providing a potential translatable therapeutic approach. Biofilm disruption through plant enzymes and chewing gum-based delivery offers an effective and affordable dental biofilm control at home particularly for populations with minimal oral care access.

Biofilms↗
PubMed2025

A novel regulation on the developmental checkpoint protein Sda that controls sporulation and biofilm formation in Bacillus subtilis.

UNLABELLED: Biofilm formation by Bacillus subtilis is triggered by an unusually simple environmental sensing mechanism. Certain serine codons, the four TCN codons (N for A, T, C, or G), in the gene for the biofilm repressor SinR caused lowered SinR translation and subsequent biofilm induction during transition from exponential to stationary growth. Global ribosome profiling showed that ribosomes pause when translating the four UCN (U for T on the mRNA) serine codons on mRNA, but not the two AGC/AGU serine codons. We proposed a serine codon hierarchy (AGC/AGT vs TCN) in that genes enriched in the TCN serine codons may experience reduced translation efficiency when serine is limited. In this study, we designed an algorithm to score all protein-coding genes in B. subtilis NCIB3610 based on the serine codon hierarchy. We generated a short list of 50 genes that could be subject to regulation by this novel mechanism. We further investigated one such gene from the list, sda, which encodes a developmental checkpoint protein regulating both sporulation and biofilm formation. We showed that synonymously switching the TCN serine codons to AGC in sda led to delayed biofilm formation and sporulation. This engineered strain also outgrew strains with other synonymously substituted sda alleles (TCN) in competition assays for biofilm formation and sporulation. Finally, we showed that the AGC serine codon substitutions in sda elevated the Sda protein levels. This serine codon hierarchy-based novel signaling mechanism could be exploited by bacteria in adapting to stationary phase and regulating important biological processes. IMPORTANCE: Genome-wide ribosome profiling in Bacillus subtilis shows that under serine limitation, ribosomes pause on the four TCN (N for A, C, G, and T), but not AGC/AGT serine codons, during translation at a global scale. This serine codon hierarchy (AGC/T vs TCN) differentially influences the translation efficiency of genes enriched in certain serine codons. In this study, we designed an algorithm to score all 4,000+ genes in the B. subtilis genome and generated a list of 50 genes that could be subject to this novel serine codon hierarchy-mediated regulation. We further investigated one such gene, sda, encoding a developmental checkpoint protein. We show that sda and cell developments controlled by Sda are also regulated by this novel mechanism.

Bacillus subtilis↗
PubMed1998

Oscillation characteristics of biofilm streamers in turbulent flowing water as related to drag and pressure drop.

Mixed population biofilms consisting of Pseudomonas aeruginosa, P. fluorescens, and Klebsiella pneumoniae were grown in a flow cell under turbulent conditions with a water flow velocity of 18 cm/s (Reynolds number, Re, =1192). After 7 days the biofilms were patchy and consisted of cell clusters and streamers (filamentous structures attached to the downstream edge of the clusters) separated by interstitial channels. The cell clusters ranged in size from 25 to 750 microm in diameter. The largest clusters were approximately 85 microm thick. The streamers, which were up to 3 mm long, oscillated laterally in the flow. The motion of the streamers was recorded at various flow velocities up to 50.5 cm/s (Re 3351) using confocal scanning laser microscopy. The resulting time traces were evaluated by image analysis and fast Fourier transform analysis (FFT). The amplitude of the motion increased with flow velocity in a sigmoidal shaped curve, reaching a plateau at an average fluid flow velocity of approximately 25 cm/s (Re 1656). The motion of the streamers was possibly limited by the flexibility of the biofilm material. FFT indicated that the frequency of oscillation was directly proportional to the average flow velocity (u(ave)) below 9.5 cm/s (Re 629). At u(ave) greater than 9.5 cm/s, oscillation frequencies were above our measurable frequency range (0.12-6.7 Hz). The oscillation frequency was related to the flow velocity by the Strouhal relationship, suggesting that the oscillations were possibly caused by vortex shedding from the upstream biofilm clusters. A loss coefficient (k) was used to assess the influence of biofilm accumulation on pressure drop. The k across the flow cell colonized with biofilm was 2.2 times greater than the k across a clean flow cell.

Biofilms↗
PubMed2002

Sensitivity analysis of a biofilm model describing a one-stage completely autotrophic nitrogen removal (CANON) process.

A mathematical model for nitrification and anaerobic ammonium oxidation (ANAMMOX) processes in a single biofilm reactor (CANON) was developed. This model describes completely autotrophic conversion of ammonium to dinitrogen gas. Aerobic ammonium and nitrite oxidation were modeled together with ANAMMOX. The sensitivity of kinetic constants and biofilm and process parameters to the process performance was evaluated, and the total effluent concentrations were, in general, found to be insensitive to affinity constants. Increasing the amount of biomass by either increasing biofilm thickness and density or decreasing porosity had no significant influence on the total effluent concentrations, provided that a minimum total biomass was present in the reactor. The ANAMMOX process always occurred in the depth of the biofilm provided that the oxygen concentration was limiting. The optimal dissolved oxygen concentration level at which the maximum nitrogen removal occurred is related to a certain ammonium surface load on the biofilm. An ammonium surface load of 2 g N/m2. d, associated with a dissolved oxygen concentration level of 1.3 g O2/m3 in the bulk liquid and with a minimum biofilm depth of 1 mm seems a proper design condition for the one-stage ammonium removal process. Under this condition, the ammonium removal efficiency is 94% (82% for the total nitrogen removal efficiency) (30 degrees C). Better ammonium removal could be achieved with an increase in the dissolved oxygen concentration level, but this would strongly limit the ANAMMOX process and decrease total nitrogen removal. It can be concluded that a one-stage process is probably not optimal if a good nitrogen effluent is required. A two-stage process like the combined SHARON and ANAMMOX process would be advised for complete nitrogen removal.

Anaerobiosis↗
PubMed2002

Effect of flow regime on the architecture of a Pseudomonas fluorescens biofilm.

A comparison of the effects of laminar versus turbulent flow regime on the characteristics of a single-species biofilm is presented. The study was carried out by growing Pseudomonas fluorescens biofilms in a flow cell and studying the different layers of the biological matrix with a confocal laser-scanning microscope. The following conclusions were obtained: i) a higher concentration of cells was found in the upper layers of the microbial films than in their inner layers, regardless of the flow regime; ii) the fraction of cells in the overall biofilm mass decreased with time as the film grew; and iii) under laminar flow the total number of cells was higher than in biofilms formed under turbulent flow, but the latter had a higher number of cells per unit volume. Such conclusions, together with the fact that the biofilms were more dense and stable when formed in contact with turbulent flows, favor the design of more compact and efficient biofilm reactors operating in turbulent conditions.

Bacterial Adhesion↗
PubMed2006

Community structures and activities of nitrifying and denitrifying bacteria in industrial wastewater-treating biofilms.

The bacterial community structure, in situ spatial distributions and activities of nitrifying and denitrifying bacteria in biofilms treating industrial wastewater were investigated by combination of the 16S rRNA gene clone analysis, fluorescence in situ hybridization (FISH) and microelectrodes. These results were compared with the nitrogen removal capacity of the industrial wastewater treatment plant (IWTP). Both nitrification and denitrification occurred in the primary denitrification (PD) tank and denitrification occurred in the secondary denitrification (SD) tank. In contrast, nitrification and denitrification rates were very low in the nitrification (N) tank. 16S rRNA gene clone sequence analysis revealed that the bacteria affiliated with Alphaproteobacteria, followed by Betaproteobacteria, were numerically important microbial groups in three tanks. The many clones affiliated with Alphaproteobacteria were closely related to the denitrifying bacteria (e.g., Hyphomicrobium spp., Rhodopseudomonas palustris, and Rhodobacter spp.). In addition, Methylophilus leisingeri affiliated with Betaproteobacteria, which favorably utilized methanol, was detected only in the SD-tank to which methanol was added. Nitrosomonas europaea and Nitrosomonas marina were detected as the ammonia-oxidizing bacteria affiliated with Betaproteobacteria throughout this plant, although the dominant species of them was different among three tanks. Nitrifying bacteria were mainly detected in the upper parts of the PD-biofilm whereas their populations were low in the upper parts of the N-biofilm. The presence of denitrifying bacteria affiliated with Hyphomicrobium spp. in SD- and N-biofilms was verified by FISH analysis. Microelectrode measurements showed that the nitrifying bacteria present in the N- and PD-biofilms were active and the bacteria present in the SD-biofilm could denitrify.

Bacteria↗
PubMed2004

Confocal imaging of biofilm formation process using fluoroprobed Escherichia coli and fluoro-stained exopolysaccharide.

We developed a novel method of evaluating biofilm architecture on a synthetic material using green fluorescent protein-expressing Escherichia coli and red fluorescence staining of exopolysaccharides. Confocal laser scanning microscopy observation revealed the time course of the change in the in situ three-dimensional structural features of biofilm on a polyurethane film without structural destruction: initially adhered cells are grown to form cellular aggregates and secrete exopolysaccharides. These cells were spottily distributed on the surface at an early incubation time but fused to form a vertically grown biofilm with incubation time. Fluorescence intensity, which is a measure of the number of cells, determined using a fluorometer and biofilm thickness determined from confocal laser scanning microscopy vertical images were found to be effective for quantification of time-dependent growth of biofilms. The curli (surface-located fibers specifically binding to fibronectin and laminin)-producing Escherichia coli strain, YMel, significantly proliferated on fibronectin-coated polyurethane, whereas the curli-deficient isogenic mutant, YMel-1, did not. The understanding of biofilm architecture in molecular and morphological events and new fluorescence microscopic techniques may help in the logical surface design of biomaterials with a high antibacterial potential.

Bacterial Adhesion↗
PubMed2005

Simulation of biofilm formation at different assimilable organic carbon concentrations under lower flow velocity condition.

A simulation model was established in this study to evaluate the impacts of assimilable organic carbon concentration on the growth of biofilm and bulk bacteria under lower flow velocity conditions. The experimental results showed that the growth trends of heterotrophic plate count bacteria of biofilm and bulk bacteria were coincident with the pattern of logistic growth model. The effects of the assimilable organic carbon level on the (specific) growth rates of biofilm and bulk bacteria were quite significant, and the higher the assimilable organic carbon level was, the higher the (specific) growth rate was. There was a growth and decline relationship between the maximum quantities of biofilm and bulk water bacteria at steady state. The maximum growth rate of biofilm bacteria exponentially increased with the increase of assimilable organic carbon level. The length of time that the maximum growth rate occurred was inversely decreased with the increase of assimilable organic carbon level. Some results of this study support the inference that the increase of bulk bacteria results mainly from the release or detachment of biofilm bacteria, not from the growth itself.

Bacterial Physiological Phenomena↗
PubMed2003

Photodestruction of human dental plaque bacteria: enhancement of the photodynamic effect by photomechanical waves in an oral biofilm model.

BACKGROUND AND OBJECTIVES: Periodontal disease results from the accumulation of subgingival bacterial biofilms on tooth surfaces. There is reduced susceptibility of these biofilms to antimicrobials for reasons that are not known. The goals of this study were to investigate the photodynamic effects of a conjugate between the photosensitizer (PS) chlorin(e6) (c(e6)) and a poly-L-lysine (pL) with five lysine residues on human dental plaque bacteria as well as on biofilms of the oral species Actinomyces naeslundii after their exposure to photomechanical waves (PW) generated by a laser in the presence of the conjugate. STUDY DESIGN/MATERIALS AND METHODS: Subgingival plaque samples from 12 patients with chronic destructive periodontitis were divided in 3 groups that were incubated for 5 minutes with 5 microM c(e6) equivalent from the pL-c(e6) conjugate in the presence of fresh medium (group I), PBS (group II), and 80% PBS/20% ethylenediaminetetra-acetic acid (EDTA) (group III) and were exposed to red light. Also, biofilms of A. naeslundii (formed on bovine enamel surfaces) were exposed to PW in the presence of 5 microM c(e6) equivalent from the pL-c(e6) conjugate and were then irradiated with red light. The penetration depth of the conjugate was measured by confocal scanning laser microscopy (CSLM). In both cases, after illumination serial dilutions were prepared and aliquots were spread over the surfaces of blood agar plates. Survival fractions were calculated by counting bacterial colonies. RESULTS: The PS/light combination achieved almost 90% killing of human dental plaque species. In biofilms of A. naeslundii, CSLM revealed that PW were sufficient to induce a 50% increase in the penetration depth of the pL-c(e6) conjugate into the biofilm. This enabled its destruction (99% killing) after photodynamic therapy (PDT). CONCLUSIONS: PW-assisted photodestruction of dental plaque may be a potentially powerful tool for treatment of chronic destructive periodontal disease.

Actinomyces↗