Agreement between transthoracic echocardiography and esophageal Doppler on aortic flow variables in anesthetized mechanically ventilated dogs.

The use of transthoracic echocardiography (TTE) and esophageal Doppler (ED) for the measurement of hemodynamic variables in anesthetized dogs was studied. Fourteen mixed-breed dogs, without cardiac disease, undergoing general anesthesia for diagnostic or therapeutic procedures were included in this prospective preliminary study. Dogs were premedicated with dexmedetomidine (3 µg/kg) and methadone (0.3 mg/kg), intramuscularly. General anesthesia was induced with propofol intravenously titrated to effect and maintained with isoflurane in oxygen. Animals were positioned in dorsal recumbency. Transthoracic echocardiography was performed using a 5-2 MHz cardiac ultrasound probe placed in the subcostal window, whereas esophageal Doppler was performed using a CardioQ probe (MP50; Deltex Medical, Chichester, UK). Once an appropriate view of the aortic flow was obtained, the variables peak velocity (PV) and velocity-time integral (VTI) were measured. Agreement between methods was evaluated using the Bland-Altman method with single observation per individual. The bias and the limits of agreement (LOA) between the two methods were determined. Mean (± SD) PV was 99.46 cm/s (± 42.73 cm/s) and 110.29 cm/s (± 35.86 cm/s), and VTI was 13.24 cm (± 4.33 cm) and 13.05 cm (± 4.47 cm), for TTE and ED, respectively. Mean differences and LOA were 10.83 cm/s (range: -20.50 to 42.16 cm/s) and -0.19 cm (range: -3.32 to 2.95 cm) for PV and VTI, respectively. No statistically significant differences were determined in the variables measured between TTE and ED in anesthetized dogs without cardiac disease, positioned in dorsal recumbency. This could be of clinical relevance when an evaluation of the intraoperative hemodynamic status of anesthetized dogs is desired.

Concordance entre l'échocardiographie transthoracique et le Doppler oesophagien sur les variables du débit aortique chez des chiens anesthésiés ventilés mécaniquement. L'utilisation de l'échocardiographie transthoracique (TTE) et du Doppler oesophagien (ED) pour la mesure des variables hémodynamiques chez les chiens anesthésiés a été étudiée. Quatorze chiens de race mixte, sans maladie cardiaque, subissant une anesthésie générale pour des procédures diagnostiques ou thérapeutiques ont été inclus dans cette étude préliminaire prospective. Les chiens ont reçu une prémédication avec de la dexmédétomidine (3 µg/kg) et de la méthadone (0,3 mg/kg), par voie intramusculaire. L'anesthésie générale a été induite avec du propofol intraveineux titré à effet et maintenue avec de l'isoflurane dans de l'oxygène. Les animaux ont été placés en décubitus dorsal. L'échocardiographie transthoracique a été réalisée à l'aide d'une sonde à ultrasons cardiaque 5-2 MHz placée dans la fenêtre sous-costale, tandis que le Doppler oesophagien a été réalisé à l'aide d'une sonde CardioQ (MP50; Deltex Medical, Chichester, Royaume-Uni). Une fois qu'une vue appropriée de l'aorte était obtenue, les variables vitesse maximale (PV) et intégrale vitesse-temps (VTI) étaient mesurées. La concordance entre les méthodes a été évaluée à l'aide de la méthode de Bland-Altman avec une seule observation par individu. Le biais et les limites d'accord (LOA) entre les deux méthodes ont été déterminés. La PV moyenne (± SD) était de 99,46 cm/s (± 42,73 cm/s) et 110,29 cm/s (± 35,86 cm/s), et la VTI était de 13,24 cm (± 4,33 cm) et 13,05 cm (± 4,47 cm), pour TTE et ED, respectivement. Les différences moyennes et la LOA étaient de 10,83 cm/s (intervalle : −20,50 à 42,16 cm/s) et de −0,19 cm (intervalle : −3,32 à 2,95 cm) pour PV et VTI, respectivement. Aucune différence statistiquement significative n'a été déterminée dans les variables mesurées entre TTE et ED chez des chiens anesthésiés sans maladie cardiaque, positionnés en décubitus dorsal. Cela pourrait être cliniquement pertinent lorsqu'une évaluation de l'état hémodynamique peropératoire des chiens anesthésiés est souhaitée.(Traduit par Dr Serge Messier).

Mechanism of Hg(II) Immobilization in Sediments by Sulfate-Cement Amendment.

Reactive amendments such as Portland and super-sulfate cements offer a promising technology for immobilizing metalloid contaminants such as mercury (Hg) in soils and sediments through sequestration in less bioavailable solid forms. Tidal marsh sediments were reacted with dissolved Hg(II) in synthetic seawater and fresh water solutions, treated with Portland cement and FeSO4 amendment, and aged for up to 90 days. Reacted solids were analyzed with bulk sequential extraction methods and characterized by powder X-ray diffraction (XRD), electron microscopy, and synchrotron X-ray absorption spectroscopy at the Hg LIII- and S K-edge. In amended sediments, XRD, SEM and sulfur K-edge XANES indicated formation of gypsum in seawater experiments or ettringite-type (Ca6Al2(SO4)3(OH)12.26H2O) phases in fresh water experiments, depending on the final solution pH (seawater ∼8.5; freshwater ∼10.5). Analysis of Hg EXAFS spectra showed Cl and Hg ligands in the first- and second-coordination shells at distances characteristic of a polynuclear chloromercury(II) salt, perhaps as a nanoparticulate phase, in both seawater and fresh water experiments. In addition to the chloromercury species, a smaller fraction (∼20-25%) of Hg was bonded to O atoms in fresh water sample spectra, suggesting the presence of a minor sorbed Hg fraction. In the absence of amendment treatment, Hg sorption and resistance to extraction can be accounted for by relatively strong binding by reduced S species present in the marsh sediment detected by S XANES. Thermodynamic calculations predict stable aqueous Hg-Cl species at seawater final pH, but higher final pH in fresh water favors aqueous Hg-hydroxide species. The difference in Hg coordination between aqueous and solid phases suggests that the initial Hg-Cl coordination was stabilized in the cement hydration products and did not re-equilibrate with the bulk solution with aging. Collectively, results suggest physical encapsulation of Hg as a polynuclear chloromercury(II) salt as the primary immobilization mechanism.

Membrane bioreactor wastewater treatment plants reveal diverse yeast and protist communities of potential significance in biofouling.

The yeast community was studied in a municipal full-scale membrane bioreactor wastewater treatment plant (MBR-WWTP). The unexpectedly high diversity of yeasts indicated that the activated sludge formed a suitable environment for them to proliferate, with cellular concentrations of 2.2 ± 0.8 × 10(3) CFU ml(-1). Sixteen species of seven genera were present in the biological reactor, with Ascomycetes being the most prevalent group (93%). Most isolates were able to grow in a synthetic wastewater medium, adhere to polyethylene surfaces, and develop biofilms of variable complexity. The relationship between yeast populations and the protists in the MBR-WWTP was also studied, revealing that some protist species preyed on and ingested yeasts. These results suggest that yeast populations may play a role in the food web of a WWTP and, to some extent, contribute to membrane biofouling in MBR systems.

Beyond Harmful: Exploring Biofilm Formation by Enterococci Isolated from Portuguese Traditional Cheeses.

This study investigated the biofilm-forming capabilities of Enterococcus isolates from Portuguese traditional cheeses with protected designation of origin (PDO) status, specifically Azeitão and Nisa. Given the absence of added starter cultures in the cheesemaking process, the characteristics of these cheeses are intrinsically linked to the autochthonous microbiota present in the raw materials and the production environment. Our findings demonstrate that all isolates possess biofilm production abilities, which are crucial for their colonization and persistence within cheese factories, thereby maintaining factory-specific microbial heritage. Through an integrated analysis utilizing principal component analysis (PCA), a direct correlation between biofilm formation and cell viability was established. Notably, these results underscore the adaptive capacity of enterococci to survive environmental fluctuations and their role in the unique characteristics of Portuguese traditional cheeses. Overall, this research enhances our understanding of the microbial dynamics in cheese production and highlights the importance of enterococci in preserving cheese quality and heritage.

Antimicrobial and Antibiofilm Potential of Thymus vulgaris and Cymbopogon flexuosus Essential Oils against Pure and Mixed Cultures of Foodborne Bacteria.

The spread of pathogenic and food spoilage microorganisms through the food chain still faces major mitigation challenges, despite modern advances. Although multiple cleaning and disinfection procedures are available for microbial load reduction in food-related settings, microbes can still remain on surfaces, equipment, or machinery, especially if they have the ability to form biofilms. The present study assessed the biofilm-forming properties of pure and mixed cultures of foodborne and spoilage bacteria (Listeria monocytogenes, Enterococcus faecalis, Aeromonas hydrophila, Brochothrix thermosphacta), using polystyrene and stainless steel contact surfaces. Subsequently, the antimicrobial and antibiofilm properties of Thymus vulgaris and Cymbopogon flexuosus essential oils-EOs-were evaluated against these bacteria. Moreover, in silico prediction of the absorption and toxicity values of the EOs' major constituents was also performed, perceiving the putative application in food-related settings. Overall, biofilm formation was observed for all microbes under study, at different temperatures and both contact surfaces. In polystyrene, at 25 °C, when comparing pure with mixed cultures, the combination Listeria-Aeromonas achieved the highest biofilm biomass. Moreover, at 4 °C, increased biofilm formation was detected in stainless steel. Regarding thyme, this EO showed promising antimicrobial features (especially against A. hydrophila, with a MIC of 0.60 µg/µL) and antibiofilm abilities (MBEC of 110.79 µg/µL against L. monocytogenes, a major concern in food settings). As for lemongrass EO, the highest antimicrobial activity, with a MIC of 0.49 µg/µL, was also observed against L. monocytogenes. Overall, despite promising results, the in situ effectiveness of these essential oils, alone or in combination with other antimicrobial compounds, should be further explored.

Immobilization of Hg(II) by coprecipitation in sulfate-cement systems.

Uptake and molecular speciation of dissolved Hg during formation of Al- or Fe-ettringite-type and high-pH phases were investigated in coprecipitation and sorption experiments of sulfate-cement treatments used for soil and sediment remediation. Ettringite and minor gypsum were identified by XRD as primary phases in Al systems, whereas gypsum and ferrihydrite were the main products in Hg-Fe precipitates. Characterization of Hg-Al solids by bulk Hg EXAFS, electron microprobe, and microfocused-XRF mapping indicated coordination of Hg by Cl ligands, multiple Hg and Cl backscattering atoms, and concentration of Hg as small particles. Thermodynamic predictions agreed with experimental observations for bulk phases, but Hg speciation indicated lack of equilibration with the final solution. Results suggest physical encapsulation of Hg as a polynuclear chloromercury(II) salt in ettringite as the primary immobilization mechanism. In Hg-Fe solids, structural characterization indicated Hg coordination by O atoms only and Fe backscattering atoms that is consistent with inner-sphere complexation of Hg(OH)(2)(0) coprecipitated with ferrihydrite. Precipitation of ferrihydrite removed Hg from solution, but the resulting solid was sufficiently hydrated to allow equilibration of sorbed Hg species with the aqueous solution. Electron microprobe XRF characterization of sorption samples with low Hg concentration reacted with cement and FeSO(4) amendment indicated correlation of Hg and Fe, supporting the interpretation of Hg removal by precipitation of an Fe(III) oxide phase.

Reactive Transport Modeling of Subaqueous Sediment Caps and Implications for the Long-Term Fate of Arsenic, Mercury, and Methylmercury.

A 1-D biogeochemical reactive transport model with a full set of equilibrium and kinetic biogeochemical reactions was developed to simulate the fate and transport of arsenic and mercury in subaqueous sediment caps. Model simulations (50 years) were performed for freshwater and estuarine scenarios with an anaerobic porewater and either a diffusion-only or a diffusion plus 0.1-m/year upward advective flux through the cap. A biological habitat layer in the top 0.15 m of the cap was simulated with the addition of organic carbon. For arsenic, the generation of sulfate-reducing conditions limits the formation of iron oxide phases available for adsorption. As a result, subaqueous sediment caps may be relatively ineffective for mitigating contaminant arsenic migration when influent concentrations are high and sorption capacity is insufficient. For mercury, sulfate reduction promotes the precipitation of metacinnabar (HgS) below the habitat layer, and associated fluxes across the sediment-water interface are low. As such, cap thickness is a key design parameter that can be adjusted to control the depth below the sediment-water interface at which mercury sulfide precipitates. The highest dissolved methylmercury concentrations occur in the habitat layer in estuarine environments under conditions of advecting porewater, but the highest sediment concentrations are predicted to occur in freshwater environments due to sorption on sediment organic matter. Site-specific reactive transport simulations are a powerful tool for identifying the major controls on sediment- and porewater-contaminant arsenic and mercury concentrations that result from coupling between physical conditions and biologically mediated chemical reactions.

Analysis of the usefulness of biological parameters for the control of activated sludge wastewater treatment plants in an interlaboratory study context.

The quality of the sludge in Wastewater Treatment Plants (WWTPs) depends on the suitable colonization of the flocs by microorganisms. Due to the functional importance of these biological constituents, several biological or biological-related parameters have been commonly used for the control of depuration efficiency. According to national and international water regulation recommendations, interlaboratory studies have a great relevance to determine which parameters are more reliable for their extensive application in routine control. However, these studies are also very useful to demonstrate consistency in results from multiple laboratories and to develop reliable and reproducible methodologies which might be necessary for protocol validation and also for accreditation issues to meet regulatory environmental requirements. The main purpose of this work was to assess the results obtained in consecutive interlaboratory assays in order to determine the concordance degree in the application of biological parameters by participating laboratories. Following the international recommendations about these studies, a common working protocol was proposed. Statistical tests indicated that Sludge Index and several routine physical-chemical analyses [V30, Mixed Liquor Suspended Solids (MLSS), Mixed Liquor Volatile Solids (MLVS) and Sludge Volumetric Index (SVI)] show low variability and therefore are suitable tools for laboratory control. Shannon Index and Sludge Biotic Index also presented low variability although a more precise protocol would be necessary, in particular the methodology to count small flagellates. The abundance and identification of protist species showed low concordance among laboratories and three factors were responsible for the low reliability of data: population density, size and morphological distinguishable characters of the specimens.

Haloarchaea have a high genomic diversity for the biosynthesis of carotenoids of biotechnological interest.

Haloarchaea are mostly components of the microbial biomass of saline aquatic environments, where they can be a dietary source of heterotrophic metazoans or contribute to flamingo's plumage coloration. The diversity of secondary metabolites (SMs) produced by haloarchaea, which might play multiple ecological roles and have diverse biotechnological applications has been largely understudied. Herein, 67 haloarchaeal complete genomes were analyzed and 182 SMs biosynthetic gene clusters (BGCs) identified that encode the production of terpenes (including carotenoids), RiPPs and siderophores. Terpene BGCs were further analysed and it was concluded that all haloarchaea might produce squalene and bacterioruberin, which one a strong antioxidant. Most of them have other carotenoid BGCs that include a putative ß-carotene ketolase that was not characterized so far in haloarchaea, but may be involved with canthaxanthin's biosynthesis. The production of bacterioruberin by Haloferax mediterranei ATCC 33500 was found to be not related to its antimicrobial activity.

Niche differentiation drives microbial community assembly and succession in full-scale activated sludge bioreactors.

Network models and community phylogenetic analyses are applied to assess the composition, structure, and ecological assembly mechanisms of microbial communities. Here we combine both approaches to investigate the temporal dynamics of network properties in individual samples of two activated sludge systems at different adaptation stages. At initial assembly stages, we observed microbial communities adapting to activated sludge, with an increase in network modularity and co-exclusion proportion, and a decrease in network clustering, here interpreted as a consequence of niche specialization. The selective pressure of deterministic factors at wastewater treatment plants produces this trend and maintains the structure of highly functional and specialized communities responding to seasonal environmental changes.

Arsenic speciation in cooked food and its bioaccessible fraction using X-ray absorption spectroscopy.

Thermal processing or the digestion process can alter the forms of arsenic (As) present in food. Identification of As species is necessary to accurately determine the risk associated with food consumption. X-ray absorption near-edge structure (XANES) was used to investigate As species in rice, asparagus, and garlic boiled in water containing As(V), and in their bioaccessible fractions (solubilized As after gastrointestinal digestion). The XANES analysis revealed the presence of As(III) (11871.5 eV) or As(III)-S [As(III)-Cys, 11869.6 eV] solution in the cooked foods and in their bioaccessible fractions. The percentage of trivalent species (12-55%) followed the order asparagus ≫ rice ≈ garlic. In the asparagus and garlic samples, part of the As(V) (tetrahedral form) [11875 eV] that had been added appeared in the form of an octahedral As(V) compound [As(V)-glycerol, 11876 eV]. All these changes could considerably modify the risk associated with ingestion of As-contaminated food.

A new species of the genus Metacystis (Ciliophora, Prostomatida, Metacystidae) from a wastewater treatment plant.

Unusual prostomatid specimens were found in the biological reactor of a wastewater treatment plant in a health resort in Valencia, Spain. These ciliates were attached to flocs unlike other free-swimming prostomatid ciliates described to date in the mixed liquor of activated sludge plants. The morphological study of this species led to a typically different combination of characteristics: elongated cell shape, 20-30 somatic kineties, 2 perioral kineties, and 1 circumoral kinety, 1 large vacuole protruding at the terminal end, a lorica tapered toward the aperture with a smooth neck, and 11-16 annular ridges. These characteristics place this representative as a new species of the genus Metacystis-Metacystis galiani n. sp. This species became the dominant population within the biological reactor when high values of conductivity (4,244 mS/cm) and temperature (26.8 degrees C) were recorded.

Effect of immediate and delayed post space preparation on coronal bacterial microleakage in teeth obturated with a methacrylate-based sealer with and without accelerator.

PURPOSE: To investigate the sealing properties of root fillings with resin-coated gutta-percha cones and a methacrylate-based resin endodontic sealer with and without an accelerator component in root canals subjected to immediate or delayed post space preparation. METHODS: Forty-eight extracted human teeth with single straight root canals were treated endodontically. Specimens were then assigned to four groups of 10 teeth each (n=10). After autoclaving, the following operative procedures were carried out under strict aseptic conditions. In Group 1 the root canals were filled with resin-coated gutta-percha cones and a methacrylate based resin endodontic sealer (EndoREZ). Post space preparations were performed 2 minutes after the sealer had set. In Group 2 the root canals were filled as in Group 1 but with the addition of a chemical accelerator. The post space preparations were also performed 2 minutes after the sealer had set. Groups 3 and 4 were filled as in Groups 1 and 2 respectively, however the post space preparations were done 7 days after the root canal filling was completed. One positive and one negative control tooth per group was added. All specimens were subjected to a coronal bacterial leakage of E. faecalis during a 60-day period using a dual chamber microbial leakage model. Data was analyzed among groups with the Kaplan-Meier survival analysis while significant pairwise differences were analyzed with the log-rank test (P<0.05). RESULTS: No significant differences (P>0.05) in bacterial leakage were observed between Groups 1 and 2 and between Groups 3 and 4. However, Groups 1 and 2 differed significantly from Groups 3 and 4 (P<0.05).

Sealing properties of a calcium aluminate luting agent.

PURPOSE: To determine the self-sealing properties of Ceramir C&B, a new calcium aluminate-based luting material and to compare it to two commercially available cements, using a bacterial model system. METHODS: 30 premolars were prepared in a standardized fashion with a total angle of convergence of approximately 10-12 degrees for complete cast restorations. All margins were placed in dentin. Gold copings were fabricated in a Type III alloy using standardized laboratory methods. Thirty teeth were randomly assigned to three groups of 10: Group 1: Ceramir C&B (CM), a calcium aluminate/glass ionomer hybrid (formerly XeraCem), Group 2: Ketac-Cem (KC), a glass-ionomer cement (control) and Group 3: Rely X Luting Plus (RX), a resin-modified glass-ionomer cement. After cementation the samples were allowed to bench set for 10 minutes, after which they were subjected to thermal cycling (x2000, between 5 and 55 degrees C, dwell time 35 seconds) and then stored in sterile phosphate buffer for 7 days at 37 degrees C. Subsequently the occlusal surface was carefully reduced exposing the dentin. Finishing on wet sand paper removed the gold flash from grinding. After sterilization the specimens were subjected to bacterial microleakage in a dual chamber apparatus for a period of 60 days. Turbidity was recorded daily. The data were analyzed using the Kaplan-Meyer survival analysis and the log-rank test (P<0.05). RESULTS: A statistical analysis demonstrated that the groups cemented with CM and RX showed no significant difference in microleakage patterns (P>0.05), while both recorded significantly lower microleakage scores (P<0.05) than the group cemented with KC.

Tuning up microbiome analysis to monitor WWTPs' biological reactors functioning.

Wastewater treatment plants (WWTPs) are necessary to protect ecosystems quality and human health. Their function relies on the degradation of organic matter and nutrients from a water influent, prior to the effluent release into the environment. In this work we studied the bacterial community dynamics of a municipal WWTP with a membrane bioreactor through 16S rRNA gene sequencing. The main phyla identified in the wastewater were Proteobacteria, Bacteroidetes, Chloroflexi, Planctomycetes and Actinobacteria. The WWTP is located in Spain and, like other studied WWTP in temperate climate zones, the temperature played a major role in community assembly. Seasonal community succession is observed along the two years sampling period, in addition to a continual annual drift in the microbial populations. The core community of the WWTP bioreactor was also studied, where a small fraction of sequence variants constituted a large fraction of the total abundance. This core microbiome stability along the sampling period and the likewise dissimilarity patterns along the temperature gradient makes this feature a good candidate for a new process control in WWTPs.

Unravelling mechanisms of bacterial quorum sensing disruption by metal-based nanoparticles.

Nanoparticles are released in the environment causing a negative impact in several ecosystems such as microbial communities. To adapt to environmental changes some bacteria use a collective behaviour ruled by a cell-to-cell communication process called quorum sensing (QS). In this study, the impact of some of the most employed metal-based nanoparticles, such as zinc oxide nanoparticles (ZnONPs), titanium dioxide nanoparticles (TiO2NPs) and silver nanoparticles (AgNPs) on bacterial QS has been assessed by using two different strains of the model organism Chromobacterium violaceum and by employing different experimental conditions. TiO2NPs were tested with and without applying a previous step of UV-irradiation while the effect of AgNPs of two diameter sizes (40 and 60 nm) and two different coating agents (PVP and citrate) was evaluated. Results evidenced that all nanoparticles produced a significant effect on violacein production and therefore, in the QS system. ZnONPs mainly disrupted the QS steps related to signal perception and response whereas TiO2NPs and AgNPs affected the autoinducer biosynthesis. AgNPs with the smallest size and citrate as capping agent produced the most deleterious effect while the impact of TiO2NPs was not affected by UV irradiation. The present study provides new insights into the mechanisms by which these commonly employed metal-based nanoparticles disturb bacterial QS-based communication and clearly evidences the potential risk of releasing nanoparticles to the environment, especially for microbial communities which play a key role in many environmental and technological processes.

Selenium and tellurium-based nanoparticles as interfering factors in quorum sensing-regulated processes: violacein production and bacterial biofilm formation.

A cell-to-cell communication system called quorum sensing (QS) promotes the transcription of certain target genes in bacterial cells leading to the activation of different cellular processes, some of them related to bacterial biofilm formation. The formation of bacterial biofilms favours antibiotic resistance, which is nowadays a significant public-health problem. In this study, the effect of selenium (SeNPs) and tellurium (TeNPs) nanoparticles was examined in two bacterial processes mediated by QS: violacein production by Chromobacterium violaceum and biofilm formation by Pseudomonas aeruginosa. For this purpose, quantification of the pigment production in the presence of these nanoparticles was monitored using the C. violaceum strain. Additionally, a combination of different microscopical imaging techniques was applied to examine the changes in the 3D biofilm structure of P. aeruginosa, which were quantified through performing architectural metric calculations (substratum area, cell area coverage and biovolume). SeNPs produce an 80% inhibition in the violacein production by C. violaceum and a significant effect on the P. aeruginosa biofilm architecture (a reduction of 80% in the biovolume of the bacterial biofilm was obtained). TeNPs similarly affect violacein production and the P. aeruginosa biofilm structure but at lower concentration levels. The results obtained suggest an important disruption of the QS signalling system by SeNPs and TeNPs, supporting nanotechnology as a promising tool to fight against the emerging problem of bacterial resistance related to bacterial biofilm formation.

Significance of moist root canal dentin with the use of methacrylate-based endodontic sealers: an in vitro coronal dye leakage study.

This in vitro study compared the effect of different levels of moisture of root canals, from none to wet, on the coronal seal after filling with resin-coated gutta-percha cones/EndoRez [RGPC/ER (groups 1-4)], Resilon/Epiphany [RE/EP (groups 5-8)], and gutta-percha/Grossman's cement [GP/G (groups 9-12)]. The length of 76 single-rooted extracted human teeth was standardized to approximately 17 mm. After instrumentation with size 10 K-Files, #2 and #3 Gates Glidden burs, and preparation to the working length with K-Type files, the smear layer was removed with 17% ethylenediaminetetraacetic acid followed by flooding with distilled water. On the basis of similarities of canal shape determined by x-rays, roots were assigned to the groups (n = 5 per group) and treated according to one of the following protocols. (I) ETOH: excess distilled water was removed with paper points followed by dehydration with 95% ethanol; (II) PAPER POINTS: the canals were blot-dried with paper points with the last one appearing dry; (III) MOIST: the canals were dried with low vacuum by using a luer adapter for 5 seconds followed by 1 paper point for 1 second; and (IV) WET: the canals remained totally flooded. The roots were then filled with one of the obturation systems outlined above. The teeth were coated with 2 layers of nail varnish and 1 layer of sticky wax, except for the coronal access. In addition, positive and negative controls were added. After immersion in 2% methylene blue dye for 7 days, the samples were embedded in clear orthodontic resin and cross-sectioned at 0.5-mm intervals along the length of the roots. Dye penetration was evaluated by an independent investigator with a stereomicroscope at 40x magnification. The results indicated that dye leakage was affected by the degree of moisture. All materials evaluated showed some evidence of dye penetration, however, root canals filled with resin-coated gutta-percha/EndoRez and Resilon/Epiphany demonstrated significantly less leakage (P < .05) when moist conditions II and III were present.

Quorum Sensing versus Quenching Bacterial Isolates Obtained from MBR Plants Treating Leachates from Municipal Solid Waste.

Quorum sensing (QS) is a mechanism dependent on bacterial density. This coordinated process is mediated by the synthesis and the secretion of signal molecules, called autoinducers (AIs). N-acyl-homoserine lactones (AHLs) are the most common AIs that are used by Gram-negative bacteria and are involved in biofilm formation. Quorum Quenching (QQ) is the interference of QS by producing hydrolyzing enzymes, among other strategies. The main objective of the present study was to identify QS and QQ strains from MBR wastewater treatment plants. A total of 99 strains were isolated from two Spanish plants that were intended to treat leachate from municipal solid waste. Five AHL producers were detected using AHL biosensor strains (Chromobacterium violaceum CV026 and Agrobacterium tumefaciens NT1). Fifteen strains of seventy-one Gram-positive were capable of eliminating or reducing at least one AHL activity. The analysis of 16S rRNA gene sequence showed the importance of the Pseudomonas genus in the production of biofilms and the relevance of the genus Bacillus in the disruption of the QS mechanism, in which the potential activity of lactonase or acylase enzymes was investigated with the aim to contribute to solve biofouling problems and to increase the useful lifespan of membranes.