In vitro enterococcus faecalis biofilm formation on five adhesive systems

Objective: To determine the E. faecalis biofilm formation on the surface of five adhesive systems (AS) and its relationship with roughness. Study Design: The formation of E. faecalis biofilms was tested on the surface of four dual-cure AS: AdheSE DC, Clearfil DC Bond, Futurabond DC and Excite DSC and one light-cure antimicrobial AS, Clearfil Protect Bond, after 24 hours of incubation, using the MBEC high-throughput device. Results: E. faecalis biofilms grew on all the adhesives. The least growth of biofilm was on Excite DSC, Clearfil Protect Bond, and the control. Futurabond DC resulted in the greatest roughness and biofilm amount. There was a close relationship between the quantity of biofilm and roughness, except for Clearfil Protect Bond, which showed little biofilm but high roughness. Conclusion: None of the tested AS prevented E. faecalis biofilm formation, although the least quantity was found on the surface of Clearfil Protect Bond.

Effectiveness of Tooth Wipes in Removing Babies' Dental Biofilm

Purpose: To assess the effectiveness of tooth wipes in removing dental biofilm from babies' anterior teeth, as well as to evaluate the babies' behaviour and the guardians' preference concerning hygiene methods. Materials and Methods: In this random blind cross-over study, 50 high caries risk babies, from 8 to 15 months old, were divided into two groups: babies with oral hygiene performed by caregivers (n = 25) or by their mothers (n = 25). The caregivers and mothers removed biofilm using three methods of oral hygiene (tooth wipes, toothbrushes and gauze), one in each experimental phase. Professional cleaning was done before each phase, which had 2 days of biofilm accumulation and 1 experimental day, when caregivers and mothers used one method to remove biofilnn. Examiners blinded to the study design assessed the biofilm index at baseline, prior to and following biofilm removal using each method. The babies' behaviour and the mothers'/caregivers' preference were assessed. Results: The tooth wipes, toothbrushes and gauze significantly reduced the amount of biofilm (P < 0.001). The mothers' group removed more biofilm than the caregivers' group, using toothbrushes or tooth wipes (P < 0.05). Babies in the mothers' group had better behaviour using tooth wipes than toothbrushes (P < 0.05). Mothers and caregivers preferred to use tooth wipes. Conclusions: Tooth wipes are effective in removing biofilrn from babies' anterior teeth and are the method best accepted by mothers, caregivers and babies.

Antimicrobial effect of endodontic solutions used as final irrigants on a dentine biofilm model

Aim To evaluate the residual biovolume of live bacterial cells, the mean biofilm thickness and the substratum coverage found in mixed biofilms treated with different endodontic irrigant solutions. Methodology Twenty-five bovine dentine specimens were infected intraorally using a removable orthodontic device. Five samples were used for each irrigant solution: 2% chlorhexidine, 1% sodium hypochlorite (NaOCl), 10% citric acid, 17% EDTA and distilled water. The solutions were used for 5 min. The samples were stained using the Live/Dead technique and evaluated using a confocal microscope. Differences in the amount of total biovolume (mu m3), number of surviving cells (mu m3), mean biofilm thickness (mu m) and substratum coverage (%) of the treated biofilms were determined using nonparametric statistical tests (P < 0.05). Results Similar values of biovolume total, biovolume of live subpopulations and substratum coverage were found in 2% chlorhexidine, 10% citric acid, 17% EDTA and distilled water-treated biofilms (P > 0.05). The lower values of the studied parameters were found in 1% NaOCl-treated dentine (P < 0.05) with the exception of the mean biofilm height criteria that did not reveal significant differences amongst the irrigant solutions (P > 0.05). Conclusions One per cent sodium hypochlorite was the only irrigant that had a significant effect on biofilm viability and architecture.

Toward the Optimization of an e-Tongue System Using Information Visualization: A Case Study with Perylene Tetracarboxylic Derivative Films in the Sensing Units

The wide variety of molecular architectures used in sensors and biosensors and the large amount of data generated with some principles of detection have motivated the use of computational methods, such as information visualization techniques, not only to handle the data but also to optimize sensing performance. In this study, we combine projection techniques with micro-Raman scattering and atomic force microscopy (AFM) to address critical issues related to practical applications of electronic tongues (e-tongues) based on impedance spectroscopy. Experimentally, we used sensing units made with thin films of a perylene derivative (AzoPTCD acronym), coating Pt interdigitated electrodes, to detect CuCl(2) (Cu(2+)), methylene blue (MB), and saccharose in aqueous solutions, which were selected due to their distinct molecular sizes and ionic character in solution. The AzoPTCD films were deposited from monolayers to 120 nm via Langmuir-Blodgett (LB) and physical vapor deposition (PVD) techniques. Because the main aspects investigated were how the interdigitated electrodes are coated by thin films (architecture on e-tongue) and the film thickness, we decided to employ the same material for all sensing units. The capacitance data were projected into a 2D plot using the force scheme method, from which we could infer that at low analyte concentrations the electrical response of the units was determined by the film thickness. Concentrations at 10 mu M or higher could be distinguished with thinner films tens of nanometers at most-which could withstand the impedance measurements, and without causing significant changes in the Raman signal for the AzoPTCD film-forming molecules. The sensitivity to the analytes appears to be related to adsorption on the film surface, as inferred from Raman spectroscopy data using MB as analyte and from the multidimensional projections. The analysis of the results presented may serve as a new route to select materials and molecular architectures for novel sensors and biosensors, in addition to suggesting ways to unravel the mechanisms behind the high sensitivity obtained in various sensors.

Comparative Analysis of the Closed Quotient for Lip and Tongue Trills in Relation to the Sustained Vowel /epsilon/

Various types of trill exercises have been used for a long time as a tool in the treatment and preparation of the voice. Although they are reported to produce vocal benefits in most subjects, their physiology has not yet been studied in depth. The aim of this study was to compare the mean and standard deviation of the closed quotient in exercises of lip and tongue trills with the sustained vowel /epsilon/ in opera singers. Ten professional classical (operatic) singers, reportedly in perfect laryngeal health, served as subjects for this study and underwent electroglottography. During the examination, the subjects were instructed to deliver the sustained vowel /epsilon/ and lip and tongue trills in a same preestablished frequency and intensity. The mean values and standard deviation of the closed quotient were obtained using the software developed for this purpose. The comparison of the results was intrasubjects; maximum intensities were compared only among them and so were minimum intensities. The means of closed quotient were statistically significant only in the strong intensities, and the lip trill was different from the tongue trill and the sustained vowel /epsilon/. The standard deviation of the closed quotient distinguished the sustained vowel /epsilon/ from the lip and tongue trills in the two intensities. We concluded that there is oscillation of the closed quotient during the exercises of tongue and lip trills, and the closed quotient is higher during the performance of exercises of the lip trill, when compared with the two other utterances, only in the strong intensities.

Performance of an electronic tongue during monitoring 2-rnethylisoborneol and geosmin in water samples

2-Methylisoborneol (MIB) and geosmin (GSM) are sub products from algae decomposition and, depending on their concentration, can be toxic: otherwise, they give unpleasant taste and odor to water. For water treatment companies it is important to constantly monitor their presence in the distributed water and avoid further costumer complaints. Lower-cost and easy-to-read instrumentation would be very promising in this regard. In this study, we evaluate the potentiality of an electronic tongue (ET) system based on non-specific polymeric sensors and impedance measurements in monitoring MIB and GSM in water samples. Principal component analysis (PCA) applied to the generated data matrix indicated that this ET was capable to perform with remarkable reproducibility the discrimination of these two contaminants in either distilled or tap water, in concentrations as low as 25 ng L-1. Nonetheless, this analysis methodology was rather qualitative and laborious, and the outputs it provided were greatly subjective. Also, data analysis based on PCA severely restricts automation of the measuring system or its use by non-specialized operators. To circumvent these drawbacks, a fuzzy controller was designed to quantitatively perform sample classification while providing outputs in simpler data charts. For instance, the ET along with the referred fuzzy controller performed with a 100% hit rate the quantification of MIB and GSM samples in distilled and tap water. The hit rate could be read directly from the plot. The lower cost of these polymeric sensors allied to the especial features of the fuzzy controller (easiness on programming and numerical outputs) provided initial requirements for developing an automated ET system to monitor odorant species in water production and distribution. (C) 2012 Elsevier B.V. All rights reserved.

Effect of Chlorhexidine on Denture Biofilm Accumulation

Purpose: Adequate denture hygiene can prevent and treat infection in edentulous patients, who are frequently elderly and have difficulty brushing their teeth. This study evaluated the efficacy of complete denture biofilm removal using a chlorhexidine solution in two concentrations: 0.12% and 2.0%. Materials and Methods: Sixty complete denture wearers participated in a trial for 21 days after receiving brushing instructions. They were distributed into three groups, according to the tested solution and regimen (n = 20): (G1) Control (daily overnight soaking in water); (G2) daily immersion at home in 0.12% chlorhexidine for 20 minutes after dinner; and (G3) a single immersion in 2.0% chlorhexidine for 5 minutes at the end of the experimental period, performed by a professional. Biofilm coverage area (%) was quantified on the internal surface of maxillary dentures at baseline and after 21 days. Afterward, the differences between initial and posttreatment results were compared by means of the Kruskal-Wallis test (a = 0.05). Results: Median values for biofilm coverage area after treatment were: (G1) 36.0%; (G2) 5.3%; and (G3) 1.4%. Differences were significant (KW = 35.25; p < 0.001), although G2 and G3 presented similar efficacy in terms of biofilm removal. Conclusions: Both chlorhexidine-based treatments had a similar ability to remove denture biofilm. Immersion in 0.12% or 2.0% chlorhexidine solutions can be used as an auxiliary method for cleaning complete dentures.

Abundance of biofilm on intertidal rocky shores: Can trampling by humans be a negative influence?

Trampling by human visitors to rocky shores is a known stressor on macroorganisms. However, the effects of trampling on rocky intertidal biofilm, a complex association of microorganisms of ecological importance in coastal communities, have not been quantified. We evaluated the impact of trampling frequency and intensity on total biomass of epilithic microalgae on intertidal rocky shores in the southeast of Brazil. There was a trend of increase in the variability of biomass of biofilm in function of intensity of trampling, but no significant effects emerged among trampling treatments. The low influence of trampling on biofilm might be a result of the small dimensions of the organisms coupled with their natural resilience and roughness of the substrate; the former preventing the removal of biofilm layers by shoes and facilitating their quick recovery. Our results provide insights for management and conservation of coastal ecosystems revealing a weaker impact of trampling on biofilm than that reported on macroorganisms. (C) 2012 Elsevier Ltd. All rights reserved.

Macroscopic and microscopic analysis of the tongue of the common opossum (Didelphis marsupialis)

We performed a macroscopic and microscopic study of the tongues of common opossums, Didelphis marsupialis, from South America. We studied two males and two females. We collected morphometric data on the tongue with precision calipers. For the light microscopy and scanning electron microscopy analyses, we fixed tissue fragments in 10% formaldehyde and 2.5% glutaraldehyde, respectively. The opossum tongues averaged 5.87 +/- 0.20 cm in length, 3.27 +/- 0.15 cm in width at the lingual body, and 3.82 +/- 0.15 cm in width at the root. The mean thickness of the lingual body was 1.8 +/- 0.1 cm, and the thickness of the root was 3.82 +/- 0.15 cm. Sharp filiform papillae were scattered across the entire tongue; conical filiform papillae occurred on the lingual body and tongue tip; fungiform papillae were scattered among the filiform papillae on the lingual body and tongue tip; and there were three vallate papillae at the root of the tongue. We found two strands of papillary projections in the tongue root. Despite the low variability observed in the lingual papillae, the morphological data obtained in this study may be related to the opossum's diverse food habits and the extensive geographic distribution of the species throughout America. Microsc. Res. Tech. 2012. (c) 2012 Wiley Periodicals, Inc.

Comparative analysis of Enterococcus Faecalis biofilm formation on different substrates

Introduction: The aim of this study was to compare Enterococcus faecalis biofilm formation on different substrates. Methods: Cell culture plates containing growth medium and E. faecalis (ATCC 29212) were used to grow biofilm on bovine dentin, gutta-percha, hydroxyapatite, or bovine bone. Substrates were incubated at 37 C for 14 or 21 days, and the medium was changed every 48 hours. After the growth induction periods, specimens (n = 5 per group and per induction period) were stained by using Live/Dead, and the images were analyzed under a confocal microscope. The total biovolume (mm3), live bacteria biovolume (mm3), and substrate coverage (%) were quantified by using the BioImage_L software. Results obtained were analyzed by nonparametric tests (P = .05). Results: Biofilm formation was observed in all groups. Gutta-percha had the lowest total biovolume at 14 days (P < .05) and hydroxyapatite the highest at 21 days (P < .05). No significant difference was observed in green biovolume at 14 days. At 21 days, however, hydroxyapatite had the highest volume (P < .05). The percentages of coverage were similar among all substrates at 21 days (P > .05), but at 14 days, bovine bone presented the highest coverage (P < .05). Conclusions: E. faecalis was capable of forming biofilm on all substrates during both growth periods; hydroxyapatite presented the highest rates of biofilm formation. The type of substrate influenced the biofilm characteristics, according to the parameters evaluated

Antimicrobial activity of triantibiotic paste, 2% chlorhexidine gel, and calcium hydroxide on an intraoral-infected dentin biofilm model

Introduction: The purpose of this study was to evaluate the antimicrobial activity of calcium hydroxide, 2% chlorhexidine gel, and triantibiotic paste (ie, metronidazole, minocycline, and ciprofloxacin) by using an intraorally infected dentin biofilm model. Methods: Forty bovine dentin specimens were infected intraorally using a removable orthodontic device in order to induce the biofilm colonization of the dentin. Then, the samples were treated with the medications for 7 days. Saline solution was used as the control. Two evaluations were performed: immediately after the elimination of the medication and after incubation in brain-heart infusion medium for 24 hours. The Live/Dead technique (Invitrogen, Eugene, OR) and a confocal microscope were used to obtain the percentage of live cells. Nonparametric statistical tests were performed to show differences in the percentage of live cells among the groups (P < .05). Results: Calcium hydroxide and 2% chlorhexidine gel did not show statistical differences in the immediate evaluation. However, after application of the brain-heart infusion medium for 24 hours, 2% gel chlorhexidine showed a statistically lesser percentage of live cells in comparison with calcium hydroxide. The triantibiotic paste significantly showed a lower percentage of live cells in comparison with the 2% chlorhexidine gel and calcium hydroxide groups in the immediate and secondary (after 24 hours) evaluations. Conclusions: The triantibiotic paste was most effective at killing the bacteria in the biofilms on the intraorally infected dentin model in comparison with 2% chlorhexidine gel and calcium hydroxide

Analysis of the biofilm proteome of Xylella fastidiosa

Abstract Background Xylella fastidiosa is limited to the xylem of the plant host and the foregut of insect vectors (sharpshooters). The mechanism of pathogenicity of this bacterium differs from other plant pathogens, since it does not present typical genes that confer specific interactions between plant and pathogens (avr and/or hrp). The bacterium is injected directly into the xylem vessels where it adheres and colonizes. The whole process leads to the formation of biofilms, which are considered the main mechanism of pathogenicity. Cells in biofilms are metabolically and phenotypically different from their planktonic condition. The mature biofilm stage (phase of higher cell density) presents high virulence and resistance to toxic substances such as antibiotics and detergents. Here we performed proteomic analysis of proteins expressed exclusively in the mature biofilm of X. fastidiosa strain 9a5c, in comparison to planktonic growth condition. Results We found a total of 456 proteins expressed in the biofilm condition, which correspond to approximately 10% of total protein in the genome. The biofilm showed 37% (or 144 proteins) different protein than we found in the planktonic growth condition. The large difference in protein pattern in the biofilm condition may be responsible for the physiological changes of the cells in the biofilm of X. fastidiosa. Mass spectrometry was used to identify these proteins, while real-time quantitative polymerase chain reaction monitored expression of genes encoding them. Most of proteins expressed in the mature biofilm growth were associated with metabolism, adhesion, pathogenicity and stress conditions. Even though the biofilm cells in this work were not submitted to any stress condition, some stress related proteins were expressed only in the biofilm condition, suggesting that the biofilm cells would constitutively express proteins in different adverse environments. Conclusions We observed overexpression of proteins related to quorum sensing, proving the existence of communication between cells, and thus the development of structuring the biofilm (mature biofilm) leading to obstruction of vessels and development of disease. This paper reports a first proteomic analysis of mature biofilm of X. fastidiosa, opening new perspectives for understanding the biochemistry of mature biofilm growth in a plant pathogen.

Papillary atrophy of the tongue and nutritional status of hospitalized alcoholics

BACKGROUND: Atrophy of the papillae, mucosa, and dorsum of the tongue are considered classical signs of nutritional deficiencies. OBJECTIVE: To assess the nutritional status of hospitalized alcoholics with or without papillary atrophy of the tongue. METHODS: This study was performed with 21 hospitalized alcoholics divided into Atrophic Glossitis Group (n=13) and Normal Tongue Group (n=8). Healthy, non-alcoholic volunteers composed the Control Group (n=8). Anthropometry and bioelectric impedance were performed, and serum vitamins A, E, and B12 were determined. RESULTS: There were no statistical differences in relation to age (46.7±8.7 vs. 46.8±15.8 years) or gender (92.3% vs. 87.5% male), respectively. Control Group volunteers were also paired in relation to age (47.5±3.1 years) and male predominance (62.5%). In relation to hospitalized alcoholics without atrophic lesions of the tongue and Control Group, patients with papillary atrophy showed lower BMI (18.6 ± 2,5 vs 23.8 ± 3.5 vs 26.7 ± 3,6 kg/m² ) and body fat content 7.6 ± 3.5 vs 13.3 ± 6.5 vs 19.5 ± 4,9 kg). When compared with the Control Group, alcoholic patients with or without papillary atrophy of the tongue showed lower values of red blood cells (10.8 ± 2.2 vs 11.8 ± 2.2 vs 14.5 ± 1,6g/dL) and albumin (3.6 ± 0.9 vs 3.6 ± 0.8 vs 4.4 ± 0.2g/dL). The seric levels of vitamins A, E, and B12 were similar amongst the groups. CONCLUSION: Hospitalized alcoholics with papillary atrophy of the tongue had lower BMI and fat body stores than controls, without associated hypovitaminosis.

Characterization of immobilized biomass by amplified rDNA restriction analysis (ARDRA) in an anaerobic sequencing-batch biofilm reactor (ASBBR) for the treatment of industrial wastewater

The performance of an anaerobic sequencing-batch biofilm reactor (ASBBR- laboratory scale- 14L )containing biomass immobilized on coal was evaluated for the removal of elevated concentrations of sulfate (between 200 and 3,000 mg SO4-2·L-1) from industrial wastewater effluents. The ASBBR was shown to be efficient for removal of organic material (between 90% and 45%) and sulfate (between 95% and 85%). The microbiota adhering to the support medium was analyzed by amplified ribosomal DNA restriction analysis (ARDRA). The ARDRA profiles for the Bacteria and Archaea domains proved to be sensitive for the determination of microbial diversity and were consistent with the physical-chemical monitoring analysis of the reactor. At 3,000 mg SO4-2·L-1, there was a reduction in the microbial diversity of both domains and also in the removal efficiencies of organic material and sulfate.