Biofilm Kinetics Research Articles

Diamond-like carbon (DLC) surface treatment decreases biofilm burden by S. aureus on titanium alloy in vitro- a pilot study.

Periprosthetic joint infection is a complication of total joint arthroplasty with treatment costs over $1.6billion dollars per year in the US with high failure rates. Therefore, generation of coatings that can prevent infection is paramount. Diamond-like carbon (DLC) is an ideal coating for implants as they are wear-resistant, corrosion-resistant, inert, and have a low friction coefficient. The purpose of this study was to test the efficacy of DLC surface treatment in prevention of biofilm on titanium discs infected with Staphylococcus aureus in vitro. Titanium alloy discs (n = 4 non-coated and n = 4 DLC-coated) were infected with 5 × 105 colony-forming units (CFU) of S. aureus for 2 weeks then analysed via crystal violet and scanning electron microscopy (SEM). Crystal violet analysis yielded differences in the appearance of biofilm on implant surface where DLC-coated had a clumpier appearance but no difference in biofilm quantification. Interestingly, this clumpy appearance did lead to differences in SEM biofilm coverage where significantly less biofilm coverage was found on DLC-coated discs (81.78% vs. 54.17%, p < 0.003). DLC-coated titanium alloy implants may have preventative properties in S. aureus infection. Observing differences in biofilm coverage does warrant additional testing including CFU titration and biofilm kinetics with eventual use in an animal model of periprosthetic joint infection.

Paracoccidioides spp.: the structural characterization of extracellular matrix, expression of glucan synthesis and associated genes and adhesins during biofilm formation.

The genus Paracoccidioides includes Paracoccidioides lutzii and the Paracoccidioides brasiliensis complex, which comprises four phylogenetic species. A key feature distinguishing planktonic growth from biofilm is the presence of a 3D extracellular matrix (ECM). Therefore, in this study, we analyzed biofilm formation in different species of Paracoccidioides yeast phase, characterized the structural elements of the matrix of P. brasiliensis (Pb18), P. lutzii (Pl01 and 8334) and P. restrepiensis (339 and 192) and evaluated the expression of glucan genes, according to the stage of biofilm evolution for P. brasiliensis. The strains were cultivated in planktonic and biofilm form for 24-144 h. The fungi biomass and metabolic activity were determined by crystal violet and tetrazolium salt reduction (XTT) tests and colony-forming unit (CFU) by plating. The biofilm structure was designed using scanning electron microscopy and confocal laser scanning microscopy techniques. The extracellular matrix of P. brasiliensis and P. lutzii biofilms was extracted by sonication, and polysaccharides, proteins, and extracellular DNA (eDNA) were quantified. The RNA was extracted with the Trizol® reagent and quantified; then, the cDNA was synthesized to analyze the enolase expression, 14-3-3, FKS1, AGS1, GEL3, and KRE6 genes by real-time PCR. All strains of Paracoccidioides studied form a biofilm with more significant metabolic activity and biomass values in 144 h. The extracellular matrix of P. brasiliensis and P. lutzii had a higher content of polysaccharides in their composition, followed by proteins and eDNA in smaller quantities. The P. brasiliensis biofilm kinetics of formation showed greater expression of genes related to glucan's synthesis and its delivery to the external environment in addition adhesins during the biofilm's adhesion, initiation, and maturation. The GEL3 and enolase genes increased in expression within 24 h and during the biofilm maturation period, there was an increase in 14-3-3, AGS1, and FKS1. Furthermore, at 144 h, there was a decrease in KRE6 expression and an increase in GEL3. This study highlights the potential for biofilm formation for three species of Paracoccidioides and the main components of the extracellular matrix that can contribute to a better understanding of biofilm organization.

A three-phase model for biofilm formation on a porous solid surface

We investigate the growth kinetics of bacterial biofilms on porous substrates. A three-phase model is developed, which accounts explicitly for the cell phase, extracellular matrix (ECM), and nutrient-rich aqueous phase. We use the thin-film approximation as the characteristic height of the biofilm is much smaller than its characteristic radius. We use the 2D axisymmetric model to capture biofilm growth on a porous agar substrate. Our model accounts for osmotic flux and predicts the spatiotemporal variations of the volume fractions of the different phases and the nutrient concentrations in the biofilm and the substrate. An increase in surface tension helps redistribute biomass radially. Our model captures the behavior of different kinds of biofilms: films characterized by low (yeast) and high (bacterial) ECM content. The predictions of our model are quantitatively validated with the experimental data from the literature. Our model provides insights on the role of different parameters on biofilm growth, which can be used to develop strategies to prevent or accelerate biofilm formation on surfaces.

Interrogating Salmonella Typhi biofilm formation and dynamics to understand antimicrobial resistance

AimsSalmonella Typhi biofilm-mediated infections are globally rising. Due to the emergence of drug resistance antibiotics did not show effective results against S. Typhi biofilm. Therefore, there is an urgent need for an in-depth interrogation of S. Typhi biofilm to understand its formation kinetics, compositions, and surface charge value. MethodsThis study utilized the S. Typhi MTCC-733 strain from a microbial-type culture collection in India. The S. Typhi biofilm was formed on a glass slide in a biofilm development apparatus. Typhoidal biofilm analysis was done with the help of various assays such as a crystal violet assay, SEM analysis, FTIR analysis, Raman analysis, and zeta potential analysis. Key findingThis article contained a comprehensive assessment of the typhoid biofilm formation kinetics, biofilm compositions, and surface charge which revealed that cellulose was a major molecule in the typhoidal biofilm which can be used as a major biofilm drug target against typhoidal biofilm. SignificanceThis study provided interrogations about typhoidal biofilm kinetics which provided ideas about the biofilm composition. The cellulose molecule showed a major component of S. Typhi biofilm and it could potentially involved in drug resistance, and offer a promising avenue for developing a new antibiofilm therapeutic target to conquer the big obstacle of drug resistance. The obtained information can be instrumental in designing novel therapeutic molecules in the future to combat typhoidal biofilm conditions effectively for overcoming antibiotic resistance against bacterial infection Salmonella.

Efficacy of Pectobacterium carotovorum subsp. carotovorum inactivation using ultraviolet C light-emitting diodes: Investigation of inactivation efficacy toward planktonic and biofilm cells on the stainless steel surfaces of cutting machines

The presence of Pectobacterium carotovorum subsp. carotovorum (P. carotovorum) in kimchi cabbage (Brassica rapa L. subsp. pekinensis) negatively influences kimchi quality. Accordingly, inactivation processes to effectively control spoilage caused by plant bacteria, including P. carotovorum, are crucial for extending the shelf life of kimchi. In this regard, ultraviolet C light-emitting diode (UVC-LED) technologies are increasingly attracting attention as non-thermal and non-contact sterilization technologies and are also considered as alternatives to chemical disinfectants. In this study, we investigated the efficacy of the inactivation of P. carotovorum planktonic and biofilm cells that cause cabbage soft rot disease using the UVC-LED treatment technology as a potential disinfection method for a safe kimchi manufacturing process. A biofilm was formed on the surface of a stainless-steel cutter knife using the recommended CDC biofilm reactor system and thereafter, exposed to a range of UVC-LED intensities. Our results indicated that P. carotovorum planktonic and biofilm cells to UVC-LED irradiation at 9.0 mJ/cm2 resulted in >5.0 and >2.0 log10 inactivation, respectively. Additionally, the P. carotovorum planktonic and biofilm cell inactivation kinetics fitted the non-linear reduction curve. The UVC-LED treatment was also found to be highly efficient in eliminating planktonic cells, showing consistency with the Weibull and Weibull tailing models for planktonic cells.

Temporal modelling of the biofilm lifecycle (TMBL) establishes kinetic analysis of plate-based bacterial biofilm dynamics

Bacterial biofilms are critical to pathogenesis and infection. They are associated with rising rates of antimicrobial resistance. Biofilms are correlated with worse clinical outcomes, making them important to infectious diseases research. There is a gap in knowledge surrounding biofilm kinetics and dynamics which makes biofilm research difficult to translate from bench to bedside. To address this gap, this work employs a well-characterized crystal violet biomass accrual and planktonic cell density assay across a clinically relevant time course and expands statistical analysis to include kinetic information in a protocol termed the TMBL (Temporal Mapping of the Biofilm Lifecycle) assay. TMBL's statistical framework quantitatively compares biofilm communities across time, species, and media conditions in a 96-well format. Measurements from TMBL can reliably be condensed into response features that inform the time-dependent behavior of adherent biomass and planktonic cell populations. Staphylococcus aureus and Pseudomonas aeruginosa biofilms were grown in conditions of metal starvation in nutrient-variable media to demonstrate the rigor and translational potential of this strategy. Significant differences in single-species biofilm formation are seen in metal-deplete conditions as compared to their controls which is consistent with the consensus literature on nutritional immunity that metal availability drives transcriptomic and metabolomic changes in numerous pathogens. Taken together, these results suggest that kinetic analysis of biofilm by TMBL represents a statistically and biologically rigorous approach to studying the biofilm lifecycle as a time-dependent process. In addition to current methods to study the impact of microbe and environmental factors on the biofilm lifecycle, this kinetic assay can inform biological discovery in biofilm formation and maintenance.

Vertical and seasonal variations in biofilm formation on plastic substrates in coastal waters of the Black Sea

Plastic contamination of the marine environment is an increasing concern worldwide. Therefore, it is important to understand the kinetics of biofilms on plastics to study their behavior, fate, and transport pathways in the ocean. In this study, the vertical and seasonal variations in biofouling formation on transparent polyethylene terephthalate (PET) plastic fragments in the Southwest Crimea coastal waters of the Black Sea were investigated. Biofilms were identified in the transient light as ‘dark spots’ on the plastic surface, for which the numbers, size, and area were measured using specialized software. The rate of biofouling in the surface water layer was lower than those found in the middle and near-bottom water column, which could be due to a damaging effect of turbulent mixing on the biofilm. The highest rates of biofouling and diverse community were observed during the summer. The epibiotic assembly was represented by diatoms (11 taxa), dinoflagellates (3 taxa), green algae, filamentous cyanobacteria, small flagellates, and ciliates. Significant differences between the biofouling rates observed in different seasons made it difficult to estimate the period of time the plastic substrate has been in the marine environment. It was proposed to use the green alga Phycopeltis arundinacea (Montgn) De Tender et al., 2015 as a bioindicator to study the age of the biofouling community. Discoid thalli were identified at all stages of colonization of the plastic fragments in different seasons. Results obtained in this study demonstrate that biofouling organisms may be good model organisms in revealing age of biofilm formation and longevity of plastic debris in the ocean. Consequently, it is proposed that such biofouling organisms could be used as target species to monitor the biodegradation of plastic debris.

Modeling biofilm and development of rate law expressions for biofilm kinetics

Biofilm processes are gaining significance in the field of wastewater treatment. Biofilm modeling has undergone immense development in the last few decades. The modeling involves developing a mass balance equation for nutrient transport across the biofilm matrix composed of microbes distributed within. The diffusion of nutrients within the biofilm matrix and their consumption by microbes govern the biofilm kinetics. For steady-state, the model becomes independent of time. The solution of steady-state Biofilm is used to calculate the mass flux entering the biofilm surface. This flux can be referred to as the biofilm reaction rate. A dimensionless number named 'Biofilm Number'(B) is formulated, which is used to develop the rate law expressions for the flux in terms of a function Ψ(S*), which measures the quality of transport across the biofilm matrix. The Biofilm reaction rate at Monod's half-saturation concentration is calculated, which is superior to the microbial reaction rate in suspended cultures. The Biofilm's effectiveness factor has been developed based on Ψ(S*) and B. The biofilm number gives the knowledge about the nutrient-consuming capacity of the Biofilm. The model presented in this work can be a fundamental tool for designing a typical biofilm-based reactor. The Biofilm model has been validated using the experimental data from the studies by Shukla 2021.

Sulfide Generation in Force Mains and Its Control Using Nitrate Dosing

Sulfide generation rates in the force mains of four pump stations in Edmonton, Alberta, Canada, were evaluated in the field, and a continuous nitrate dosing was applied in the pump wet well of the Big Lake pump station to investigate its effectiveness on sulfide control. The sulfide production rates at these force mains were estimated to be 0.08–0.15 g/m2 h under 20°C, which could be modeled by using the 1/2-order biofilm kinetics of dissolved chemical oxygen demand (DCOD). The modeled rate constant a was obtained at around 0.006 using the field monitoring data. It was found that 20 mg/L sulfide was formed in the force main of the Big Lake pump station, which resulted in 400–500 parts per million (ppm) H2S gas in the discharge manhole. It was found that a nitrate dosing concentration of 85 mg/L could completely suppress the sulfide generation in the force main. A constant dosing rate resulted in nitrate overdosage at midnight and underdosage in peak hours. Injecting nitrate in the wet well led to a large portion of nitrate consumption where a heterotrophic denitrification process happened. The appropriate dosing location for this dosing strategy is at the beginning of force main instead of the wet well. The corresponding dosing rate is a flow-paced dosing rate instead of a constant dosing rate. A more cost-effective dosing strategy is to add the nitrate at downstream section of the force main where sulfide generated upstream could be removed in the presence of nitrate when passing downstream. This dosing strategy could significantly reduce the nitrate demand.

Visualization of initial bacterial colonization on dentin using fluorescence activating headlight for fluorescence enhanced theragnosis

ObjectiveTo evaluate accuracy of caries detection and the application-sensitivity of the new Designs for Vision's REVEAL™ utilizing a fluorescence activating headlight for excitation purpose. Materials and methodsREVEAL dental fluorescence loupes and headlight system were used. Occlusal enamel was removed, and mid-coronal dentine was exposed. Carious artificial lesion was created. Streptococcus mutans, Actinomyces naeslundii, and Streptococcus sanguis were used. The assessment was performed using two diagnostic methods: naked eye and Design for Vision Glasses with inter examiner blinding using two calibrated examiners. After 7 days, Raman measurements were made on dentin disc specimens with 785 nm wavelength. The bacterial counts in colony-forming units (CFU) were used to examine the growth kinetics of biofilms. The collagen fibril structure within the discs was performed using Transmission Electron Microscope. Scanning Electron Microscope was used to image samples at various magnifications. FISH was performed with specimens fixed in 4% paraformaldehyde in phosphate-buffered saline. Reproducibility was measured by Cohen kappa scores, values of which range from 0 for less than chance agreement to 1 for almost perfect agreement (p < 0.05). ResultsA significant kappa score of 0.706 showing significant reproducibility for the given diagnostic techniques, as all the teeth included in the study were spotted with the lesions. Most bacteria were detected using the CFU technique. The Raman bands scanned across the dentin surface at 960 cm−1 (P–O peak) are assigned to hydroxyapatite phosphate vibrations. FISH identified nearly all stained bacteria as days and time and dental hard tissue had a significant impact on the number of adherent bacteria. Scanning electron micrographs of polished cross sections of demineralized and non-demineralized specimens with perpendicular each tubule orientation (zone of demineralized dentin inset. ConclusionFluorescent enhanced theragnosis through Reveal vision glasses can ensure constant monitoring and diagnosis of caries progress . This may allow for a better clinical outcome.

Reaction kinetics of anodic biofilms under changing substrate concentrations: Uncovering shifts in Nernst-Monod curves via substrate pulses.

In the present study, it is shown that the concentration dependency of undefined mixed culture anodic biofilms does not follow a single kinetic curve, such as the Nernst‐Monod curve. The biofilms adapt to concentration changes, which inevitably have to be applied to record kinetic curves, resulting in strong shifts of the kinetic parameters. The substrate concentration in a continuously operated bioelectrochemical system was changed rapidly via acetate pulses to record Nernst‐Monod curves which are not influenced by biofilm adaptation processes. The values of the maximum current density j max and apparent half‐saturation rate constant K s increased from 0.5 to 1 mA cm−2 and from 0.5 to 1.6 mmol L−1, respectively, within approximately 5 h. Double pulse experiments with a starvation phase between the two acetate pulses showed that j max and K s decrease reversibly through an adaptation process when no acetate is available. Pseudo‐capacitive charge values estimated from non‐turnover cyclic voltammograms (CV) led to the hypothesis that biofilm adaptation and the observed shift of the Nernst‐Monod curves occurred due to changes in the concentration of active redox proteins in the biofilm. It is argued that concentration‐related parameters of kinetic models for electroactive biofilms are only valid for the operating points where they have been determined and should always be reported with those conditions.

Modeling Biofilm and Development of Rate Law Expressions for Biofilm Kinetics

Modeling Biofilm and Development of Rate Law Expressions for Biofilm Kinetics

Unaerated feeding alters the fate of dissolved methane during aerobic wastewater treatment

In municipal wastewater treatment plants, some dissolved methane can enter the aerobic bioreactors. This greenhouse gas originates from sewers and return flows from anaerobic sludge treatment. In well-mixed conventional activated sludge reactors, methane emissions are largely avoided because methane oxidizing bacteria consume a large fraction, even without optimizing for this purpose. In this work, the fate of dissolved methane is studied in aerobic granular sludge reactors, as they become increasingly popular. The influence of the characteristic design and operating conditions of these reactors are studied with a mathematical model with apparent conversion kinetics and stripping: the separation of feeding and aeration in time, a higher substrate transport resistance, a high retention time of granular biomass and a taller water column. Even for a best-case scenario combining an unrealistically low intragranule substrate transport resistance, a high retention time, a tall reactor, an extremely high influent methane concentration and no oxygen limitation, the methane conversion efficiency was only 12% when feeding and aeration were separated in time, which is lower than for continuous activated sludge reactors under typical conditions. A more rigorous model was used to confirm the limited conversion, considering the multi-species and multi-substrate biofilm kinetics, anoxic methane consumers and the high substrate concentration at the bottom during upward plug flow feeding. The observed limited methane conversion is mainly due to the high concentration that accumulates during unaerated feeding phases, which favours stripping more than conversion in the subsequent aeration phase. Based on these findings, strategies were proposed to mitigate methane emissions from wastewater treatment plants with sequentially operated reactors.

Applicaion of Moving Bed Biofilm Reactor (MBBR) for Treatment of Industrial Wastewater: A mini Review

This research aims at presenting Moving Bed Biofilm Reactor (MBBR) technology as one of the alternatives and efficient methods for treating industrial wastewaters at a variety of conditions. In the last decade, this technology became more popular and widely utilized worldwide, due to the fact that the need for clean water keeps to rapidly increase with the increase in the population of the world grows year by year, which is why a high number of the waste-water treatment facilities are necessary expand, for the purpose of providing additional capacity with the minimal costs. The present study aimed at covering the most significant MBBR processes, like the fundamental process of the treatment, biofilm kinetics, and MBBR operation principles. This review includes as well a large number of the relevant researches that have been performed at the lab and pilot scales, in addition to cover the important procedures on the basic process of the treatment, which affects the influent types as well as the carrier type. None-the-less, this review concluded yet has been compiled herein and reported to be acquiring more sufficient insights and outlook upon the theme with a view to meet the new method. For that purpose, the most feasible technology may be the advanced biological process (i.e., the bio-reactor systems), which include the MBBR system

A multi-component model for granular activated carbon filters combining biofilm and adsorption kinetics

Along with the rise of biological active granular activated carbon (bGAC) filtration as advanced treatment technology for wastewater treatment plant (WWTP) effluents, the mathematical representation of such systems is gaining increasing importance. This work introduces a model that describes the performance of bGAC-filters for Dissolved Organic Carbon (DOC) removal from a WWTP effluent. The DOC removal within bGAC-filters is accomplished by two mechanisms: adsorptive removal and biological transformation. An appropriate representation of the adsorptive removal requires the DOC to be divided into fictive fractions according to its adsorbability. Likewise, a further DOC classification according to its biodegradability is necessary. Modeling a bGAC-filter then becomes a multi-component adsorption problem, with the simultaneous occurrence of DOC degradation within a biofilm. For dealing with this modeling task, this work integrated the Ideal Adsorbed Solution (IAS) theory into a traditional biofilm model compatible with the Activated Sludge Model (ASM) Framework. For the description of the adsorption dynamics, a Freundlich isotherm for the equilibrium and a pseudo first order model for the kinetics were selected. The biofilm consisted of heterotrophic bacteria able to oxidize DOC using oxygen as electron acceptor. The correctness of the model was evaluated using experimental data from a pilot plant. The predicted DOC breakthrough curve satisfactorily fitted the experimental measurements for empty bed contact times (EBCT) of 6, 12, 24 and 33 min. Moreover, the model predicted the relationship between EBCT, DOC removal and bGAC-filter lifespan. The developed model is the first that combines multi-component adsorption and biofilm kinetics in a wastewater treatment context.

Use of a novel 9.3-μm carbon dioxide laser and silver diamine fluoride: Prevention of enamel demineralisation and inhibition of cariogenic bacteria

ObjectiveTo investigate the effects of a 9.3-μm carbon dioxide (CO2) laser and silver diamine fluoride (SDF) on the prevention of enamel demineralisation and inhibition of cariogenic bacteria. MethodsEnamel blocks were applied with Laser (Group-1), SDF (Group-2), Laser + SDF (Group-3) and no treatment (Group-4), and then subjected to an 8-day pH-cycling for cariogenic challenge. Lesion depth and cross-sectional micro-hardness were assessed. Surface morphological and chemical changes were studied using scanning electron microscope (SEM) with energy dispersive spectroscopy (EDS). For the antibacterial activity, treated enamel blocks were incubated with Streptococcus mutans. The biofilm morphology, kinetics and viability were assessed by SEM, colony-forming units (CFUs) and confocal laser scanning microscope (CLSM), respectively. ResultsLesion depths (μm) for Group-1 to Group-4 were 88 ± 21, 26 ± 11, 13 ± 9 and 115 ± 25, respectively (p < 0.001; Group-2 and Group-3 < Group-1 < Group-4). Group-3 had a significantly higher cross-sectional micro-hardness than the other three groups. EDS determined that Group-4 had the lowest calcium-to-phosphorus molar ratio among the groups (p < 0.001). SEM images showed apparent bacteria accumulation on enamel surfaces in Group-4, but not in other groups. Log CFUs for Group-1 to Group-4 were 6.2 ± 0.6, 2.9 ± 0.8, 2.2 ± 1.1 and 7.3 ± 0.3, respectively (p < 0.001; Group-2 and Group-3 < Group-1 < Group-4). CLSM images revealed that live bacteria dominated in Group-4, but not in other groups. SignificanceThe irradiation with a 9.3-μm CO2 laser alone can prevent the demineralisation of enamel and reduce the adhesion of cariogenic bacteria. Moreover, adding SDF can significantly increase the preventive effect and antibacterial ability.

Effect of antimicrobials on Stenotrophomonas maltophilia biofilm.

Aim: To evaluate the activity of five antimicrobials against young and mature Stenotrophomonas maltophilia biofilms. Materials &methods: Nineteen clinical strains from hemoculture of hemodialysis patients were tested for biofilm kinetics, MIC and minimum biofilm inhibitory concentration (MBIC) in young and mature biofilms. Results: All strains were moderate biofilm producers. MIC showed total susceptibility to levofloxacin and trimethoprim-sulfamethoxazole and partial resistance to ceftazidime (63.2%) and gentamicin (21%). Young and mature biofilms showed the lowest MBIC/MIC ratio for gentamicin, chloramphenicol and levofloxacin, respectively. The highest MBIC/MIC was for trimethoprim-sulfamethoxazole (young) and ceftazidime (mature). Conclusion: Gentamicin displayed surprising activity against S. maltophilia biofilms. Chloramphenicol was indicated as a good option against young S. maltophilia biofilms, and trimethoprim-sulfamethoxazole showed limited antibiofilm activity.

Characterizing the biofilm stoichiometry and kinetics on the media in situ based on pulse-flow respirometer coupling with a new breathing reactor

Biofilm based systems and the hybrid between activated sludge and biofilms have been popularly applied for wastewater treatment. Unlike the suspended biomass, the biofilm concentration and kinetics on the media cannot be easily measured. In this study, a novel and easy-to-use approach has been developed based on pulse-flow respirometer to characterize the biofilm stoichiometry and kinetics in situ. With the new designed breathing reactor, the mutual interference between the magnetic stirring and biofilm media that happened in the conventional breathing reactor was solved. Moreover, Microsoft Excel based programs had been developed to fit the oxygen uptake rate curves with dynamic nonlinear regression. With this new approach, the yield coefficient, maximum oxidation capacity, and half-saturation constant of substrate for the heterotrophic biofilms in a fix bed reactor were determined to be 0.46 g-VSS/g-COD, 67.0 mg-COD/(h·L-media), and 4.4 mg-COD/L, respectively. Those parameters for biofilm ammonia oxidizers from a moving bed biofilm reactor were determined to be 0.17 g-VSS/g-N, 18.6 mg-N/(h·L-media), and 1.2 mg-N/L, respectively, and they were 0.11 g-VSS/g-N, 20.9 mg-N/(h·L-media), and 0.98 mg-N/L for nitrite oxidizers in the same biofilms. This study also found that the maximum specific substrate utilization rate for detached biofilms increased by 3.2 times, indicating that maintaining biofilm integrity was very important in the kinetic tests. Using this approach, the biofilm kinetics on the media can be regularly measured for treatment optimization.

Antimicrobial activity and toxicity of glass ionomer cement containing an essential oil.

The aim of this study was to evaluate the antimicrobial activity and toxicity of glass ionomer cement (GIC) modified with 5-methyl-2-(1-methylethyl)phenol (thymol) against Streptococcus mutans in silico and in vitro. The antimicrobial activity of thymol on GIC modified with concentrations of 2% (GIC-2) and 4% (GIC-4) was evaluated in a model of planktonic cell biofilm using agar diffusion test, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), dynamic biofilm (continuous flow cell parallel), and bacterial kinetics. Conventional GIC (GIC-0) was used as a control. Thymol toxicity was evaluated in Artemia salina and in silico using Osiris® software. Differences between groups were estimated by analysis of variance, followed by Tukey post hoc test, with a 5% significance level. The results of the agar diffusion test between groups were not significantly different (P≥0.05). Thymol had potential bacteriostatic and bactericidal activity against Streptococcus mutans with respect to planktonic growth, with MIC of 100 µg/mL and MBC of 400 µg/mL. The groups GIC-0, GIC-2, and GIC-4 reduced the biofilm by approximately 10, 85, and 95%, respectively. Bacterial kinetics showed efficiency of the modified GICs for up to 96 h. GIC with thymol was effective against S. mutans, with significant inhibition of the biofilms. Analyses in silico and using Artemia salina resulted in no relevant toxicity, suggesting potential for use in humans. GIC-2 was effective against S. mutans biofilm, with decreased cell viability.

Influence of Laboratory Culture Media on in vitro Growth, Adhesion, and Biofilm Formation of Pseudomonas aeruginosa and Staphylococcus aureus

Objective: Pseudomonas aeruginosa and Staphylococcus aureus dual-species biofilm infections are notoriously difficult to manage. This study aimed at investigating the influence of four different culture media on the planktonic growth, adhesion, and biofilm formation of P. aeruginosa and S. aureus. Materials and Methods: We monitored four different culture media including Nutrient Broth, Brain Heart Infusion (BHI) broth, Luria-Bertani broth, and RPMI 1640 medium on the planktonic growth, adhesion, and biofilm formation of P. aeruginosa (ATCC 27853) and S. aureus (ATCC 25923) using MTT assay and scanning electron microscopy (SEM). Results: The most robust growth of the mono- and dual-species cultures was noted in BHI broth. On the contrary, RPMI 1640 medium promoted maximal initial adhesion of both the mono- and dual-species, but BHI broth fostered the maximal biofilm growth. SEM images showed profuse extracellular polysaccharide production in biofilms, particularly in coculture, in BHI medium. Conclusion: Our data demonstrate that BHI broth, relative to the other tested media, is the most conducive for in vitro evaluation of biofilm and planktonic growth kinetics of these two pathogens, both in mono- and coculture.