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PubMed2005

Control of oral biofilm formation by an antimicrobial decapeptide.

Oral biofilms are mixed-species microbial communities, and their uncontrolled outgrowth can express as oral diseases. Antimicrobial peptides represent alternative classes of antimicrobials that exhibit selectivity for prokaryotes. We wanted to test the effect of a synthetic decapeptide antimicrobial, KSL, on the development of oral biofilms formed by isolated human salivary bacteria. We used differential interference contrast microscopy, coupled with a dual-flow cell system, to determine the effect of KSL on oral biofilm development. We used reductions of viable counts and confocal microscopy to assess the bactericidal activity of KSL on mature oral biofilms. KSL effectively blocked biofilm development. A significant effect on the viability of mature biofilms was observed when KSL was used in the presence of a surface-active agent, or after biofilms were mechanically disrupted. This study shows that KSL may be a useful adjunct for conventional oral hygiene to prevent plaque-mediated dental diseases.

Anti-Bacterial Agents↗
PubMed2007

Effects of plant lactones on the production of biofilm of Pseudomonas aeruginosa.

Sixteen plant sesquiterpene lactones, thirteen from four species of the Family Asteraceae, and three from a species of Hepaticae, as well as seven annonaceous acetogenins isolated from the seeds of the tropical tree Annona cherimolia (Family Annonaceae), were evaluated for their ability to inhibit or stimulate the production of biofilm by a strain of Pseudomonas aeruginosa. The tested compounds carry a gamma-lactone moiety in their structures. This structural feature is similar to the lactone moiety present in N-acyl homoserine lactones, compounds that play the important role of "quorum sensors" in the mechanisms of biofilm formation observed in many gram-negative bacteria. A special assay was employed to evaluate the influence of the tested plant compounds to inhibit or stimulate the production of biofilm in a P. aeruginosa wild strain. Most of the tested compounds affected the biofilm formation mechanism. Six sesquiterpene lactones isolated from Acanthospermum hispidum and one from Enydra anagallis as well as an acetogenin from Annona cherimolia strongly inhibited (69-77%) the biofilm formation when incorporated to a bacterial culture at a concentration of 2.5 microg/ml. However, one of the acetogenins, squamocin, stimulated the biofilm formation even at a concentration of 0.25 microg/ml. The study of substances affecting the biofilm formation can lead to the design of new strategies to control P. aeruginosa infections.

Biofilms↗
PubMed2005

Comparison of two cleansing pastes for the removal of biofilm from dentures and palatal lesions in patients with atrophic chronic candidiasis.

The efficacy of 2 oral hygiene products, an experimental toothpaste specific for complete denture cleansing and a regular standard toothpaste, was compared in terms of denture biofilm removal and cure of palatal lesions in patients with atrophic chronic candidiasis. The degree of correlation between presence of biofilm and mucosa erythema was also evaluated. Twenty-four complete denture wearers (45-80 years old) were divided into 2 groups: experimental paste and standard toothpaste (Sorriso-Kolynos, Brazil). Both groups received soft toothbrushes. The internal surfaces of upper dentures were stained using 1% sodium fluorescein and photographed at a 45 masculine angle at 0, 15, 30 and 60 days. The slides were scanned and the areas of interest (denture total area and biofilm area) were measured (Image Tool software). The degree of erythema was evaluated on slides according to the Prosthesis Tissue Index. There was a significant reduction (1%) in the degree of biofilm (ANOVA/Tukey) between the two initial visits (0 and 15 days) and the two final visits (30 and 60 days), and in the average erythema scores (Kruskal-Wallis) between 0 and 60 days, in both groups. The Mann-Whitney test showed a significant difference (1%) between pastes in terms of biofilm degree, but no difference was found for the erythema score. Correlation values between biofilm and erythema degree were 0.3801 (experimental paste) and (0.3678 (standard toothpaste). We may therefore conclude that the experimental product was efficient for the removal of denture plaque biofilm.

Aged↗
PubMedDate not supplied

[Relation between biofilm, caries activity and gingivitis in HIV + children].

The utilization of medicines to treat HIV-infected children has been promoting a decrease in the prevalence of soft-tissue oral lesions, as years pass by. In contrast, it has been observed that the experience of caries and gingivitis is constant in this population, mostly because of the chronic influence of some factors involved in the HIV-infection process, such as the chronic utilization of sweetened liquid medicines and carbohydrate-enriched diet, as well as frequent episodes of hospitalization. So, the purpose of this study was to evaluate if the quality and quantity of biofilm are important factors in the activity of dental caries and gingivitis, also in this special group. After examination of the biofilm (biofilm index - Ribeiro23, 2000), the activity of caries and gingivitis was assessed in 56 children, aging from 0 to 14 years, who were patients with definitive diagnosis of HIV infection. It was observed that only 7 subjects (12.5%) did not present with clinically visible biofilm, and 33 (58.9%) presented with gingivitis, with the average of 4.44 bleeding sites. As to dental caries, 73.2% of the patients presented with active carious lesions. A strong correlation was verified between Biofilm Index, gingival status and active carious lesions (Spearman's correlation test, r s = +0.57 and r s = +0.49, respectively). It was concluded that, also in HIV-infected children, the quality and quantity of biofilm over the dental surfaces are important etiologic factors related to the activity of caries and gingivitis. Biofilm should, thus, be controlled in order to reestablish the oral health of HIV-infected children.

Adolescent↗
PubMed2006

Comparison between two visible biofilm indices in the primary dentition.

The aim of this study was to compare two visible biofilm indices in the primary dentition. The sample consisted of 90 children of both sexes, aged up to 4 years old outpatients of the University Hospital of the Rio de Janeiro State University. A single examiner, aided by an assistant, performed the children's dental examination for biofilm assessment. A simplified visible biofilm index (BF1), which classifies biofilm as absent, thin or thick, in anterior and/or posterior teeth, and provides a score for the patient and not for each tooth, was compared to a conventional visible biofilm index, the visible plaque index, (BF2), which classifies biofilm as absent or present and provides scores for three surfaces of each tooth. A statistically significant association and a strong positive correlation between BF1 and BF2 was found (Kruskal-Wallis p < 0.001/rs = 0.81 p < 0.001). The time required to BF1 evaluation was approximately one third of the time required to BF2 evaluation (t test p < 0.001). These results suggest that the use of a simplified visible biofilm index is feasible in the primary dentition as it showed similar findings when compared to a conventional one, besides being more practical and quicker.

Biofilms↗
PubMed2003

Formation and decontamination of biofilms in dental unit waterlines.

BACKGROUND: Biofilms are a natural occurrence in aquatic environments, including community drinking water systems. The interior of small-diameter tubings in dental unit waterlines (DUWL) are also sites of biofilm formation. In the lumen of the tubings, the flow is minimal, and the water becomes stagnant when the units are not in use. Molecules precipitate from the water onto the interior wall and promote the adherence of planktonic microorganisms from the water. Once they become sessile, the microorganisms change their phenotype. After adherence, there is a so-called surface-associated lag time, and the organisms then enter a growth phase and produce exopolysaccharides that coat the organisms in a slime layer. Within the biofilm, the microorganisms can signal one another, transfer nutrients, and exchange genetic material. The insoluble exopolysaccharides shield the microorganisms from displacement and from penetration by predator organisms, antibiotics, and disinfectants. The external surface layer of microorganisms is faster growing and may detach as "swarmer" cells. Detachment of microorganisms from dental unit biofilm flushed into the oral cavity could theoretically infect the patient. Splatter and aerosols from dental procedures may possibly infect health care personnel. METHODS: This study compared three DUWL cleaners (an alkaline peroxide product, a freshly mixed chlorine dioxide product, and a buffer-stabilized chlorine dioxide product) in 16 dental units with self-contained water systems, 6 months after installation in a periodontal teaching clinic. One unit treated by flushing and drying served as a control. Units were sampled daily for 10 days with heterotrophic plate count (HPC) sampler plates. The plates were incubated for 7 days at room temperature, and colonies were counted at 10.5x magnification. Samples of internal water tubing before and after the use of waterline cleaners were processed and examined by scanning electron microscopy. RESULTS: The estimated mean HPC was derived from original and replicate independent counts of two investigators of undiluted and diluted samples, reported as colony forming units (CFU)/ml. Shock treatments with the alkaline peroxide product (n = 5) reduced the HPC from baseline, but in the ratio of daily counts to control, there was a large variance and a trend to return of high counts as days passed. The mean daily HPC was significantly better than the control for only 3 of the 9 days of treatment and exceeded the goal of 200 on 3 days. Freshly mixed chlorine dioxide (n = 4) and the buffer-stabilized chlorine dioxide (n = 5) both reduced HPC to near 0 on all days. Their ratios of daily estimated means to that of the control were significantly (P < 0.001) better at all times. In comparing treatments, the freshly mixed chlorine dioxide was better (P < 0.001) than the alkaline peroxide on 8 of 9 days. The buffered chlorine dioxide treatment was better than the alkaline peroxide at all times. The two chlorine dioxide treatments each had so many HPC counts of 0 that a meaningful statistical difference between them was not calculated. Scanning electron microscopy of plastic waterline tubing samples taken before and after treatments showed reductions in biofilm coverage, but the differences were not statistically significant. CONCLUSIONS: Chlorine dioxide waterline cleaners are effective in decontaminating DUWL biofilm. Chlorine dioxide has advantages over other chlorine products. Controlling DUWL biofilm may have beneficial effects on nosocomial infections.

Alkalies↗
PubMed2005

Biofilm formation by the periodontopathic bacteria Treponema denticola and Porphyromonas gingivalis.

BACKGROUND: Periodontitis develops as a result of the interaction of the host with subgingival plaque bacteria. Both Porphyromonas gingivalis and Treponema denticola are frequently associated together in these oral biofilms. METHODS: The molecular basis for in vitro biofilm formation was investigated for P. gingivalis 381, T. denticola 35405, and mixtures of the two organisms using microtiter plate assays. In addition, the biofilms were examined following confocal laser scanning microscopy. RESULTS: P. gingivalis 381, but not T. denticola strains, formed biofilms in vitro. This property was dependent, in part, on the strain 381 fimA, ppk, and usp genes. Microarray and Northern blot analyses suggested that the expression of the ppk gene was required for maximal expression of the uspA gene. P. gingivalis 381 formed synergistic biofilms when incubated with T. denticola strains. This process was dependent upon the strain 381 rgpB and fimA genes as well as the T. denticola flgE and cfpA genes. CONCLUSIONS: P. gingivalis 381 formed synergistic biofilms with T. denticola 35405. These results may be relevant to the previous observations that the two organisms are frequently observed together in subgingival plaque with the spirochetes localized to the exterior of the oral biofilms. It is suggested that other such synergistic effects may also occur between other plaque bacteria.

Bacterial Proteins↗
PubMed2006

Survival of Campylobacter jejuni in potable water biofilms: a comparative study with different detection methods.

Campylobacteria are important foodborne pathogens. C. jejuni bacteria have caused several drinking water-related epidemics in Finland. Normally, C. jejuni is not able to multiply in drinking water or in biofilms although it may survive in biofilms. The survival of C. jejuni in biofilms was studied using the Propella biofilm reactor. The number of bacteria was analysed with traditional culture methods and with fluorescence in situ hybridisation (FISH). By culture methods C. jejuni was detectable for only 1 d after spiking whereas bacteria were found from biofilms for at least 1 week after spiking and from outlet water of the reactor for 3 weeks when using FISH. These results suggested that C. jejuni may survive in biofilms and culture methods probably seriously underestimate the real number in water and in biofilms.

Biofilms↗
PubMed1998

Structure and functional characteristics of bacterial biofilms in fluid processing operations.

Bacterial biofilms create a number of serious problems for industrial fluid processing operations. Mechanical blockages, impedance of heat transfer processes, and biodeterioration of the components of metallic and polymeric systems result in billions of dollars in losses each year. Product spoilage and possible risks to public health are also consequences of biofilm-mediated contamination. Fundamentally, these biofouling activities can be described in terms of the physicochemical properties that are associated with bacterial metabolism and biofilm development. Treatment of biofouling is also complicated by the unique structural attributes of biofilms: extracellular polymeric substances create diffusional barriers to antimicrobial agents, protecting labile cellular targets from both oxidizing and nonoxidizing compounds. The mechanisms associated with the initial events of bacterial adhesion to engineered surfaces and subsequent fouling of biofilm formation are poorly understood. However, studies of bacterial biofilm architecture have been greatly facilitated by the application of confocal laser microscopy, scanning or transmission electron microscopy, and Fourier transform infrared spectroscopy. This paper reviews the genesis of biofilm formation and describes the influence of structure on biofouling activities in industrial fluid handling systems.

Animals↗
PubMed2002

Biofilms: microbial life on surfaces.

Microorganisms attach to surfaces and develop biofilms. Biofilm-associated cells can be differentiated from their suspended counterparts by generation of an extracellular polymeric substance (EPS) matrix, reduced growth rates, and the up- and down- regulation of specific genes. Attachment is a complex process regulated by diverse characteristics of the growth medium, substratum, and cell surface. An established biofilm structure comprises microbial cells and EPS, has a defined architecture, and provides an optimal environment for the exchange of genetic material between cells. Cells may also communicate via quorum sensing, which may in turn affect biofilm processes such as detachment. Biofilms have great importance for public health because of their role in certain infectious diseases and importance in a variety of device-related infections. A greater understanding of biofilm processes should lead to novel, effective control strategies for biofilm control and a resulting improvement in patient management.

Bacteria↗
PubMed2000

Naturally occurring biofilms on alfalfa and other types of sprouts.

Scanning electron microscopy was used to examine the cotyledons, hypocotyls, and roots of alfalfa, broccoli, clover, and sunflower sprouts purchased from retail outlets as well as alfalfa sprouts grown in the laboratory using a tray system equipped with automatic irrigation. Biofilms were observed on all plant parts of the four types of commercially grown sprouts. Biofilms were also commonly observed on alfalfa sprouts grown in the laboratory by 2 days of growth. Rod-shaped bacteria of various sizes were predominant on all sprouts examined both as free-living cells and as components of biofilms. Occasionally, cocci-shaped bacteria as well as yeast cells were also present in biofilms. The microbes contained in the biofilms appeared to be attached to each other and to the plant surface by a matrix, most likely composed of bacterial exopolysaccharides. Biofilms were most abundant and of the largest dimensions on cotyledons, sometimes covering close to the entire cotyledon surface (approximately 2 mm in length). Naturally occurring biofilms on sprouts may afford protected colonization sites for human pathogens such as Salmonella and Escherichia coli O157:H7, making their eradication with antimicrobial compounds difficult.

Biofilms↗
PubMed2005

Formation of biofilms by Listeria monocytogenes under various growth conditions.

Eight strains of Listeria monocytogenes (7644, 19112, 15313, Scott A, LCDC, 10403S, SLCC, and 1370) produce biofilms when grown on polyvinyl chloride microtiter well plates. The growth medium (tryptic soy broth [TSB] or modified Welshimer's broth [MWB] at 32 degrees C) influenced the amount of biofilm formed; maximum biofilms were formed in MWB by six strains and in TSB by the remaining two strains. This result suggests that the growth medium is critical in development of L. monocytogenes biofilm. This organism also produced biofilms on stainless steel chips. Biofilm formation on these chips was observed following growth in TSB at 4, 20, and 37 degrees C. After 20 h of incubation at 20 or 37 degrees C, the cell density was approximately 10(6) CFU per chip, and after 4 days incubation at 4 degrees C, the cell density was 10(5) CFU per chip. L. monocytogenes strain Scott A biofilm formation on stainless steel chips was visualized using scanning electron microscopy, which revealed dense aggregates of cells held together by meshlike webbing.

Bacterial Adhesion↗
PubMed2005

Effect of temperature, pH, and water activity on biofilm formation by Salmonella enterica enteritidis PT4 on stainless steel surfaces as indicated by the bead vortexing method and conductance measurements.

An assay was developed in an effort to elucidate the effect of important environmental parameters (temperature, pH, and water activity [aw]) on Salmonella Enteritidis biofilm formation on stainless steel surfaces. To achieve this, a modified microbiological technique used for biofilm studying (the bead vortexing method) and a rapid method based on conductivity measurements were used. The ability of the microorganism to generate biofilm on the stainless surfaces was studied at three temperatures (5, 20, and 37 degrees C), four pH values (4.5, 5.5, 6.5, and 7.4), and four aw values (0.5, 1.5, 5.5, and 10.5% NaCl). Results obtained by the bead vortexing method show that maximum numbers of adherent bacteria per square centimeter (106 CFU/cm2) were attained in 6 days at 20 degrees C. Biofilm formation after 7 days of incubation at 20 degrees C was found to be independent of the pH value. In addition, the high concentration of sodium chloride (10.5% NaCl, aw = 0.94) clearly inhibited the adherence of cells to the coupons. Conductance measurements were used as a supplementary tool to measure indirectly the attachment and biofilm formation of bacterial cells on stainless steel surfaces via their metabolic activity (i.e., changes in the conductance of the growth medium due to microbial growth or metabolism). Results obtained by conductance measurements corresponded well to those of the bead vortexing method. Furthermore, we were able to detect cells that remained attached on the metal surfaces even after vortexing via their metabolic activity. The results, except for demonstrating environmental-dependent Salmonella Enteritidis biofilm formation, indicated that traditional vortexing with beads did not remove completely biofilm cells from stainless steel; hence, conductance measurements seem to provide a more sensitive test capable to detect down to one single viable organism.

Bacterial Adhesion↗
PubMed2005

Enhancing the bactericidal effect of electrolyzed water on Listeria monocytogenes biofilms formed on stainless steel.

Biofilms are potential sources of contamination to food in processing plants, because they frequently survive sanitizer treatments during cleaning. The objective of this research was to investigate the combined use of alkaline and acidic electrolyzed (EO) water in the inactivation of Listeria monocytogenes biofilms on stainless steel surfaces. Biofilms were grown on rectangular stainless steel (type 304, no. 4 finish) coupons (2 by 5 cm) in a 1:10 dilution of tryptic soy broth that contained a five-strain mixture of L. monocytogenes for 48 h at 25 degrees C. The coupons with biofilms were then treated with acidic EO water or alkaline EO water or with alkaline EO water followed by acidic EO water produced at 14 and 20 A for 30, 60, and 120 s. Alkaline EO water alone did not produce significant reductions in L. monocytogenes biofilms when compared with the control. Treatment with acidic EO water only for 30 to 120 s, on the other hand, reduced the viable bacterial populations in the biofilms by 4.3 to 5.2 log CFU per coupon, whereas the combined treatment of alkaline EO water followed by acidic EO water produced an additional 0.3- to 1.2-log CFU per coupon reduction. The population of L. monocytogenes reduced by treatments with acidic EO water increased significantly with increasing time of exposure. However, no significant differences occurred between treatments with EO water produced at 14 and 20 A. Results suggest that alkaline and acidic EO water can be used together to achieve a better inactivation of biofilms than when applied individually.

Biofilms↗
PubMed2001

Single stage biological nitrogen removal by nitritation and anaerobic ammonium oxidation in biofilm systems.

In full scale wastewater treatment plants with at times considerable deficits in the nitrogen balances, it could hitherto not be sufficiently explained which reactions are the cause of the nitrogen losses and which micro-organisms participate in the process. The single stage conversion of ammonium into gaseous end-products--which is henceforth referred to as deammonification--occurs particularly frequently in biofilm systems. In the meantime, one has succeeded to establish the deammonification processes in a continuous flow moving-bed pilot plant. In batch tests with the biofilm covered carriers, it was possible for the first time to examine the nitrogen conversion at the intact biofilm. Depending on the dissolved oxygen (DO) concentration, two autotrophic nitrogen converting reactions in the biofilm could be proven: one nitritation process under aerobic conditions and one anaerobic ammonium oxidation. With the anaerobic ammonium oxidation, ammonium as electron donor was converted with nitrite as electron acceptor. The end-product of this reaction was N2. Ammonium and nitrite did react in a stoichiometrical ratio of 1:1.37, a ratio which has in the very same dimension been described for the ANAMMOX-process (1:1.31 +/- 0.06). Via the oxygen concentration in the surrounding medium, it was possible to control the ratio of nitritation and anaerobic ammonium oxidation in the nitrogen conversion of the biofilm. Both processes were evenly balanced at a DO concentration of 0.7 mg/l, so that it was possible to achieve a direct, almost complete elimination of ammonium without addition of nitrite. One part of the provided ammonium did participate in the nitritation, the other in the anaerobic ammonium oxidation. Through the aerobic ammonium oxidation into nitrite within the outer oxygen supplied layers of the biofilm, the reaction partner was produced for the anaerobic ammonium oxidation within the inner layers of the biofilm.

Bacteria, Anaerobic↗
PubMed1999

[Influence of roxithromycin combined with fleroxacin on bacterial biofilm induced by Pseudomonas aeruginosa].

OBJECTIVE: To investigate the influence of roxithromycin (RXM) on the production of glycocalyx (GLX) and the synergic effect of RXM and fleroxacin (FLRX) on P. Aeruginosa biofilms. METHODS: Bacterial biofilm (BF) was established and was influenced by RXM and FLRX with different concentrations. The samples were detected with scanning electron microscope and by a rapid method after BF was stained with AgNO3 solution. The synergism of antibacterial activities of RXM and FLRX to P. Aeruginosa was studied by computer image analysis and MTT method. RESULTS: GLX production was reduced significantly by both 1/16 MIC and 1/4 MIC RXM (P < 0.01). 1/16 MIC and 1/4 MIC RXM showed no bactericidal activities to P. Aeruginosa in biofilms, while viable counts in biofilms almost had no difference. However, RXM could enhance the bactericidal activity of FLRX to P. Aeruginosa in biofilms. When FLRX of different concentrations was combined with 1/16 MIC and 1/4 MIC RXM respectively, the difference of the stained BF was counted up by computer image analysis system with significance of P < 0.05 and the viable counts were reduced significantly (P < 0.05). The results was in accordance with SEM pictures. CONCLUSION: RXM has no direct bactericidal activities to P. Aeruginosa in biofilms, but it could inhibit GLX production. It can be considered that RXM could enhance antibacterial activities of FLRX by enhancing the permeability of FLRX into biofilms.

Anti-Bacterial Agents↗
PubMed2002

Development of photosynthetic biofilms affected by dissolved and sorbed copper in a eutrophic river.

Photosynthetic biofilms are capable of immobilizing important concentrations of metals, therefore reducing bioavailability to organisms. But also metal pollution is believed to produce changes in the microalgal species composition of biofilms. We investigated the changes undergone by natural photosynthetic biofilms from the River Meuse, The Netherlands, under chronic copper (Cu) exposure. The suspended particles in the river water had only a minor effect on reduction of sorption and toxicity of Cu to algae. Biofilms accumulated Cu proportionally to the added concentration, also at the highest concentration used (9 microM Cu). The physiognomy of the biofilms was affected through the growth of the chain-forming diatom Melosira varians, changing from long filaments to short tufts, although species composition was not affected by the Cu exposure. The Cu decreased phosphate uptake and algal biomass measured as chl a, which degraded exponentially in time. Photosynthetic activity was always less sensitive than algal biomass; the photon yield decreased linearly in time. The protective and insulating role of the biofilm, supported by ongoing autotrophic activity, was indicated as essential in resisting metal toxicity. We discuss the hypothesis that the toxic effects of Cu progress almost independently of the species composition, counteracting ongoing growth, and conclude that autotrophic biofilms act as vertical heterogeneous units. Effective feedback mechanisms and density dependence explain several discrepancies observed earlier.

Adsorption↗
PubMed2002

An integrated approach to analyse biofilms of a full scale wastewater treatment plant.

A rotating biological contactor (RBC) system operating in a full-scale wastewater treatment plant has been described by several approaches accounting for performance, composition and structure of biofilms in three stages through biological wastewater treatment (RBC1, RBC 2, RBC 3). RBC biofilms were effective in removing the BOD loading from 13 g BOD5 d(-1) m(-2) in RBC 1 to 6 g BOD5 d(-1) m(-2) in RBC 3. Analysis of biofilm composition showed: i) the volatile solids were similar in the three RBCs (0.6 g m(-2) VS per g m(-2) of TS); ii) the protozoan and metazoan biocenosis was mainly made up of ciliated protozoa, which were most abundant in RBC 2 (1.84 x 10(6) ciliates g(-1) VS). Relationship between ciliate species and physical-chemical profile of the system by cluster analysis indicated that the species Acineria uncinata, Amphileptus punctatus, Cinetochilum margaritaceum and Holosticha mancoidea were associated with the best RBC performance; iii) the exopolymeric matrix of the three RBC biofilms was mainly constituted by proteins, although humic substances, polysaccharides, uronic acids and DNA were also found. Analysis of biofilm structure by confocal microscopy indicated changes in biofilm organisation with depth. Results have been brought together and a graphic representation of the composition and architecture of RBC biofilms is presented.

Animals↗