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The biofilm architecture is significantly influenced by external environmental conditions. Biofilms grown on drinking water distribution systems (DWDS) are exposed to environmental contaminants, including parabens, and disinfection strategies, such as chlorine. Although changes in biofilm density and culturability from chemical exposure are widely reported, little is known about the effects of parabens and chlorine on biofilm morphology and architecture. This is the first study evaluating architectural changes in Stenotrophomonas maltophilia colony biofilms (representatives of bacterial communities presented in DWDS) induced by the exposure to methylparaben (MP) at environmental (15 μg/L) and in-use (15 mg/L) concentrations, and chlorine at 5 mg/L, using widefield epi-fluorescence mesoscopy with Mesolens. The GFP fluorescence of colony biofilms allowed the visualization of internal structures and Nile Red fluorescence permitted the inspection of the distribution of lipids. Our data show that exposure to MP triggers physiological and morphological adaptation in mature colony biofilms by increasing the complexity of internal structures, which may confer protection to embedded cells from external chemical molecules. These architectural modifications include changes in lipid distribution as an adaptive response to MP exposure. Although chlorine exposure affected colony biofilm diameter and architecture, the colony roundness was completely affected by the simultaneous presence of MP and chlorine. This work is pioneer in using Mesolens to highlight the risks of exposure to emerging environmental contaminants (MP), by affecting the architecture of biofilms formed by drinking water (DW) bacteria, even when combined with routine disinfection strategies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Parabens are markedly present in products of daily use, considered emerging environmental contaminants that can harm human health and aquatic life, due to their release into aquatic sources. The impact of the exposure of microbial communities to parabens remains unclear. This study investigates aspects of the mode of action of methylparaben (MP), propylparaben (PP), butylparaben (BP), and MIX at environmental (15 μg/L) and in-use (15000 μg/L) concentrations, against two bacterial strains of Acinetobacter calcoaceticus and Stenotrophomonas maltophilia previously isolated from drinking water (DW). BP showed the strongest antimicrobial activity, while MP exhibited the weakest. The mechanism of action of parabens at the selected concentrations was found to be related to perturbations on physicochemical bacterial cell surface properties and charge, by causing an increase of bacterial cell envelope hydrophilicity and zeta potential values. In addition, parabens may activate osmotic regulation mechanisms as observed by the increase in vacuole area for MP-exposed A. calcoaceticus. The bacterial metabolic activity as well as bacterial size was also affected by parabens exposure. MP exposure further enhanced the biofilm formation ability and increased bacterial tolerance to antibiotics. The results raise environmental implications, particularly concerning water quality and public health, as parabens exposure can potentiate the virulence of DW bacteria, increasing the risk of human exposure to harmful microorganisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Methylparaben (MP) is a preservative considered an environmental contaminant of emerging concern due to its persistence in water sources, including drinking water (DW). This study assesses the interaction between MP and dual-species biofilms of Acinetobacter calcoaceticus and Stenotrophomonas maltophilia. These biofilms were grown under realism-based conditions in a multiple-cylinder biofilm reactor on polypropylene (PPL) surfaces, for 7 days, and then exposed to MP at 0.5 mg/L for three consecutive days. S. maltophilia predominantly succeeds within these biofilms compared to A. calcoaceticus. Exposure to MP resulted in a 4-fold increase in the number of culturable cells and a 1.4-fold rise in polysaccharide content, suggesting that bacterial cells may utilize MP as a carbon source to enhance biofilm fitness. MP was found to adsorb to PPL with biofilms following a pseudo-second-order kinetic model. Circa 37 % of MP adsorbed to PPL after 3 days of exposure. Besides that, MP was biodegraded by biofilms following an apparent first-order kinetic model. Part (25 %) of the MP was biodegraded whereas only 0.02 % bioaccumulated on the biofilm matrix. Biodegradation was related to esterase and lipase activity. The results provide new insights into the interaction between MP with biofilms and materials used in DW industries., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Bacterial coaggregation is a highly specific type of cell-cell interaction, well-documented among oral bacteria, and involves specific characteristics of the cell surface of the coaggregating strains. However, the understanding of the mechanisms promoting coaggregation in aquatic systems remains limited. This gap is critical to address, given the broad implications of coaggregation for multispecies biofilm formation, water quality, the performance of engineered systems, and diverse biotechnological applications. Therefore, this study aims to comprehensively characterize the cell surface of the coaggregating strain Delftia acidovorans 005P, isolated from drinking water, alongside a non-coaggregating strain, D. acidovorans 009P. By analyzing two strains of the same species, we aim to identify the factors contributing to the coaggregation ability of strain 005P. To achieve this, we employed a combination of physicochemical characterization, Fourier-transform infrared spectroscopy (FTIR), and advancing imaging techniques [transmission electron microscopy and cryo-electron tomography (cryo-ET)]. The coaggregating strain (005P) exhibited higher surface hydrophobicity, negative surface charge, and cell surface and co-adhesion energies than the non-coaggregating strain (009P). The chemical characterization of bacterial surfaces through FTIR revealed subtle differences, particularly in spectral regions linked to carbohydrates and phosphodiesters/amide III of proteins (860-930 cm -1 and 1212-1240 cm -1 , respectively). Cryo-ET highlighted significant differences in pili structures between the strains, such as variations in length, frequency, and arrangement. The pili in the 005P strain, identified as pili-like adhesins, serve as key mediators of coaggregation. By integrating physicochemical analyses and high-resolution imaging techniques, this study conclusively links the coaggregation ability of D. acidovorans 005P to its unique pili characteristics, emphasizing their crucial role in microbial coaggregation in aquatic environments., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Biofilms in drinking water (DW) systems persistently challenge traditional disinfection methods due to intricate microbial interactions, with coaggregation playing a crucial role in forming multispecies biofilms. This study examined the implications of coaggregation on tolerance towards sodium hypochlorite (NaOCl) disinfection. Dual-species biofilms were formed for seven days on polyvinyl chloride coupons, comprising a strain of the emerging pathogen Stenotrophomonas maltophilia and the coaggregating strain Delftia acidovorans 005 P. For comparison, dual-species biofilms were also formed with a non-coaggregation strain (D. acidovorans 009 P). The minimum bactericidal concentration (MBC) for each planktonic strain varied (D. acidovorans: 1 mg/L, S. maltophilia: 1.5 mg/L) below the safe DW treatment limits. However, high NaOCl doses (10 ×MBC and 100 ×MBC,) showed low efficacy against dual-species biofilms, indicating significant biofilm tolerance to disinfection. Membrane damage occurred at sub-MBC without culturability loss, underscoring biofilm resilience. The biofilm analysis revealed a complex interplay between the composition of extracellular polymeric substances and the architecture, which was influenced by the presence of the coaggregating strain. Overall, coaggregation significantly influenced biofilm formation and resilience, impacting NaOCl disinfection. These findings underscore the challenges of microbial interactions in biofilms, emphasizing the need for improved disinfection strategies to control biofilms in drinking water systems., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Parabens are emerging contaminants that have been detected in drinking water. Their presence in DW distribution systems (DWDS) can alter bacterial behaviour, characteristics, and structure, which may compromise DW disinfection. This work provides insights into the impact of methylparaben (MP) on the tolerance to chlorine disinfection and antibiotics from dual-species biofilms formed by Acinetobacter calcoaceticus and Stenotrophomonas maltophilia isolated from DW and grown on high-density polyethylene (HDPE) and polypropylene (PPL). Results showed that dual-species biofilms grown on PPL were more tolerant to chlorine disinfection, expressing a decrease of over 50 % in logarithmic reduction values of culturable cells in relation to non-exposed biofilms. However, bacterial tolerance to antibiotics was not affected by MP presence. Although MP-exposed dual-species biofilms grown on HDPE and PPL were metabolically more active than non-exposed counterparts, HDPE seems to be the material with lower impact on DW risk management and disinfection, if MP is present. Overall, results suggest that MP presence in DW may compromise chlorine disinfection, and consequently affect DW quality and stability, raising potential public health issues., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The presence of biofilms in drinking water distribution systems (DWDS) is responsible for water quality deterioration and a possible source of public health risks. Different factors impact the biological stability of drinking water (DW) in the distribution networks, such as the presence and concentration of nutrients, water temperature, pipe material composition, hydrodynamic conditions, and levels of disinfectant residual. This review aimed to evaluate the current state of knowledge on strategies for DW biofilm disinfection through a qualitative and quantitative analysis of the literature published over the last decade. A systematic review method was performed on the 562 journal articles identified through database searching on Web of Science and Scopus, with 85 studies selected for detailed analysis. A variety of disinfectants were identified for DW biofilm control such as chlorine, chloramine, UV irradiation, hydrogen peroxide, chlorine dioxide, ozone, and others at a lower frequency, namely, electrolyzed water, bacteriophages, silver ions, and nanoparticles. The disinfectants can impact the microbial communities within biofilms, reduce the number of culturable cells and biofilm biomass, as well as interfere with the biofilm matrix components. The maintenance of an effective residual concentration in the water guarantees long-term prevention of biofilm formation and improves the inactivation of detached biofilm-associated opportunistic pathogens. Additionally, strategies based on multi-barrier processes by optimization of primary and secondary disinfection combined with other water treatment methods improve the control of opportunistic pathogens, reduce the chlorine-tolerance of biofilm-embedded cells, as well as decrease the corrosion rate in metal-based pipelines. Most of the studies used benchtop laboratory devices for biofilm research. Even though these devices mimic the conditions found in real DWDS, future investigations on strategies for DW biofilm control should include the validity of the promising strategies against biofilms formed in real DW networks., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The contamination of indoor areas is a global health problem that can cause the dispersion of infectious diseases. In that sense, it is urgent to find new strategies applying a lower concentration of the traditional chemicals used for cleaning and disinfection. Ultraviolet radiation (UV), in particular far-UV-C (200-225 nm), has emerged as a successful, powerful, easy-to-apply, and inexpensive approach for bacterial eradication that still requires scientific assessment. This study investigated new strategies for disinfection based on far-UV-C (222 nm) combined with chlorine and mechanical cleaning, providing an innovative solution using low doses. The bactericidal activity of far-UV-C (222 nm) was tested at an intensity of irradiation from 78.4 μW/cm 2 to 597.7 μW/cm 2 (for 1 min) against Escherichia coli and Staphylococcus epidermidis adhered on polystyrene microtiter plates. It was further tested in combination with mechanical cleaning (ultrasounds for 1 min) and free chlorine (0.1, 0.5, and 1 mg/L for 5 min). The triple combination consisting of mechanical cleaning + free chlorine (0.5 mg/L) + far-UV-C (54 mJ/cm 2 ) was tested against cells adhered to materials found in hospital settings and other public spaces: polyvinyl chloride (PVC), stainless steel (SS), and polyetheretherketone (PEEK). Disinfection with far-UV-C (54 mJ/cm 2 ) and free chlorine at 0.5 mg/L for 5 min allowed a total reduction of culturable E. coli cells and a logarithmic reduction of 2.98 ± 0.03 for S. epidermidis. The triple combination of far-UV-C, free chlorine, and mechanical cleaning resulted in a total reduction of culturable cells for both adhered bacteria. Bacterial adhesion to PVC, SS, and PEEK occurred at distinct extents and influenced the bactericidal activity of the triple combination, with logarithmic reductions of up to three. The overall results highlight that, based on culturability assessment, far-UV-C (54 mJ/cm 2 ) with chlorine (0.5 mg/L; 5 min) and mechanical cleaning (1 min) as an efficient disinfection strategy using mild conditions. The combination of culturability and viability assessment of disinfection is recommended to detect regrowth events and increase the effectiveness in microbial growth control., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests or conflicts of interest in the writing of this article., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Among different pollutants of emerging concern, parabens have gained rising interest due to their widespread detection in water sources worldwide. This occurs because parabens are used in personal care products, pharmaceuticals, and food, in which residues are generated and released into aquatic environments. The regulation of the use of parabens varies across different geographic regions, resulting in diverse concentrations observed globally. Concentrations of parabens exceeding 100 μg/L have been found in wastewater treatment plants and surface waters while drinking water (DW) sources typically exhibit concentrations below 6 μg/L. Despite their low levels, the presence of parabens in DW is a potential exposure route for humans, raising concerns for both human health and environmental microbiota. Although a few studies have reported alterations in the functions and characteristics of microbial communities following exposure to emerging contaminants, the impact of the exposure to parabens by microbial communities, particularly biofilm colonizers, remains largely understudied. This review gathers the most recent information on the occurrence of parabens in water sources, as well as their effects on human health and aquatic organisms. The interactions of parabens with microbial communities are reviewed for the first time, filling the knowledge gaps on the effects of paraben exposure on microbial ecosystems and their impact on disinfection tolerance and antimicrobial resistance, with potential implications for public health., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The potential of pentapotassium bis(peroxymonosulphate) bis(sulphate) (OXONE) to control biofilms in drinking water distribution systems (DWDS) was evaluated and compared to chlorine disinfection. Mature biofilms of drinking water (DW)-isolated Stenotrophomonas maltophilia were formed using a simulated DWDS with a rotating cylinder reactor (RCR). After 30 min of exposure, OXONE at 10 × minimum bactericidal concentration (MBC) caused a significant 4 log reduction of biofilm culturability in comparison to the unexposed biofilms and a decrease in the number of non-damaged cells below the detection limit (4.8 log cells/cm 2 ). The effects of free chlorine were restricted to approximately 1 log reduction in both biofilm culturability and non-damaged cells. OXONE in synthetic tap water (STW) at 25 ºC was more stable over 40 days than free chlorine in the same conditions. OXONE solution exhibited a disinfectant decrease of about 10% of the initial concentration during the first 9 days, and after this time the values remained stable. Whereas possible reaction of chlorine with inorganic and organic substances in STW contributed to free chlorine depletion of approximately 48% of the initial concentration. Electron paramagnetic resonance (EPR) spectroscopy studies confirmed the presence of singlet oxygen and other free radicals during S. maltophilia disinfection with OXONE. Overall, OXONE constitutes a relevant alternative to conventional DW disinfection for effective biofilm control in DWDS., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Shake flask cultivation is a routine technique in microbiology and biotechnology laboratories where cell growth can be affected by the hydrodynamic conditions, which depend on the agitation velocity, shaking diameter, and shake flask size. Liquid agitation is implemented inherently to increase aeration, substrate transfer to the cells, and prevent sedimentation, disregarding the role of hydrodynamics in microbial growth and metabolism. Here, we present a simple approach to help standardize the hydrodynamic forces in orbital shakers to increase the experimental accuracy and reproducibility and give students a better knowledge of the significance of the agitation process in microbial growth., Competing Interests: The authors declare no conflict of interest., (Copyright © 2023 Simões et al.)

The activity of two chlorinated isocyanurates (NaDCC and TCCA) and peroxymonosulphate (OXONE) was evaluated against biofilms of Stenotrophomonas maltophilia , an emerging pathogen isolated from drinking water (DW), and for the prevention of biofilm regrowth. After disinfection of pre-formed 48 h-old biofilms, the culturability was reduced up to 7 log, with OXONE, TCCA, and NaDCC showing more efficiency than free chlorine against biofilms formed on stainless steel. The regrowth of biofilms previously exposed to OXONE was reduced by 5 and 4 log CFU cm -2 in comparison to the unexposed biofilms and biofilms exposed to free chlorine, respectively. Rheometry analysis showed that biofilms presented properties of viscoelastic solid materials, regardless of the treatment. OXONE reduced the cohesiveness of the biofilm, given the significant decrease in the complex shear modulus (G*). AFM analysis revealed that biofilms had a fractured appearance and smaller bacterial aggregates dispersed throughout the surface after OXONE exposure than the control sample. In general, OXONE has been demonstrated to be a promising disinfectant to control DW biofilms, with a higher activity than chlorine. The results also show the impact of the biofilm mechanical properties on the efficacy of the disinfectants in biofilm control.

Ultraviolet irradiation C (UVC) has emerged as an effective strategy for microbial control in indoor public spaces. UVC is commonly applied for air, surface, and water disinfection. Unlike common 254 nm UVC, far-UVC at 222 nm is considered non-harmful to human health, being safe for occupied spaces, and still effective for disinfection purposes. Therefore, and allied to the urgency to mitigate the current pandemic of SARS-CoV-2, an increase in UVC-based technology devices appeared in the market with levels of pathogens reduction higher than 99.9 %. This environmentally friendly technology has the potential to overcome many of the limitations of traditional chemical-based disinfection approaches. The novel UVC-based devices were thought to be used in public indoor spaces such as hospitals, schools, and public transport to minimize the risk of pathogens contamination and propagation, saving costs by reducing manual cleaning and equipment maintenance provided by manpower. However, a lack of information about UVC-based parameters and protocols for disinfection, and controversies regarding health and environmental risks still exist. In this review, fundamentals on UVC disinfection are presented. Furthermore, a deep analysis of UVC-based technologies available in the market for the disinfection of public spaces is addressed, as well as their advantages and limitations. This comprehensive analysis provides valuable inputs and strategies for the development of effective, reliable, and safe UVC disinfection systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Coaggregation plays an important role in the development of multispecies biofilms in different environments, often serving as an active bridge between biofilm members and other organisms that, in their absence, would not integrate the sessile structure. The ability of bacteria to coaggregate has been reported for a limited number of species and strains. In this study, 38 bacterial strains isolated from drinking water (DW) were investigated for their ability to coaggregate, in a total of 115 pairs of combinations. Among these isolates, only Delftia acidovorans (strain 005P) showed coaggregating ability. Coaggregation inhibition studies have shown that the interactions mediating D. acidovorans 005P coaggregation were both polysaccharide-protein and protein-protein, depending on the interacting partner bacteria. Dual-species biofilms of D. acidovorans 005P and other DW bacteria were developed to understand the role of coaggregation on biofilm formation. Biofilm formation by Citrobacter freundii and Pseudomonas putida strains highly benefited from the presence of D. acidovorans 005P, apparently due to the production of extracellular molecules/public goods favouring microbial cooperation. This was the first time that the coaggregation capacity of D. acidovorans was demonstrated, highlighting its role in providing a metabolic opportunity for partner bacteria., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The failure of current sanitation practices requires the development of effective solutions for microbial control. Although combinations using antibiotics have been extensively studied to look for additive/synergistic effects, biocide combinations are still underexplored. This study aims to evaluate the antimicrobial effectiveness of dual biocide and triple biocide/phytochemical combinations, where phytochemicals are used as quorum sensing (QS) inhibitors. The biocides selected were benzalkonium chloride (BAC) and peracetic acid (PAA) - as commonly used biocides, and glycolic acid (GA) and glyoxal (GO) - as alternative and sustainable biocides. Curcumin (CUR) and 10-undecenoic acid (UA) were the phytochemicals selected, based on their QS inhibition properties. A checkerboard assay was used for the screening of chemical interactions based on the cell growth inhibitory effects against Bacilluscereus and Pseudomonasfluorescens. It was observed that dual biocide combinations resulted in indifference, except the PAA + GA combination, which had a potential additive effect. PAA + GA + CUR and PAA + GA + UA combinations also triggered additive effects. The antimicrobial effects of the combinations were further evaluated on the inactivation of planktonic and biofilm cells after 30 min of exposure. These experiments corroborated the checkerboard results, in which PAA + GA was the most effective combination against planktonic cells (additive/synergistic effects). The antimicrobial effects of triple combinations were species- and biocide-specific. While CUR only potentiate the antimicrobial activity of GA against B.cereus, GA + UA and PAA + GA + UA combinations promoted additional antimicrobial effects against both bacteria. Biofilms were found to be highly tolerant, with modest antimicrobial effects being observed for all the combinations tested. However, this study demonstrated that low doses of biocides can be effective in bacterial control when combining biocides with a QS inhibitor, in particular, the combination of the phytochemical UA (as a QS inhibitor) with GA and PAA., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Parabens have been detected in drinking water (DW) worldwide, however, their impact on DW microbial communities remains to be explored. Microorganisms can easily adapt to environmental changes. Therefore, their exposure to contaminants of emerging concern, particularly parabens, in DW distribution systems (DWDS) may affect the microbiological quality and safety of the DW reaching the consumers tap. This work provides a pioneer evaluation of the effects of methylparaben (MP), propylparaben (PP), butylparaben (BP), and their combination (MIX), in bacterial biofilms formed on different surfaces, representative of DWDS materials - high-density polyethylene (HDPE), polypropylene (PPL) and polyvinyl chloride (PVC). Acinetobacter calcoaceticus and Stenotrophomonas maltophilia, isolated from DW, were used to form single and dual-species biofilms on the surface materials selected. The exposure to MP for 7 days caused the most significant effects on biofilms, by increasing their cellular culturability, density, and thickness up to 233%, 150%, and 224%, respectively, in comparison to non-exposed biofilms. Overall, more pronounced alterations were detected for single biofilms than for dual-species biofilms when HDPE and PPL, demonstrating that the surface material used affected the action of parabens on biofilms. Swimming motility and the production of virulence factors (protease and gelatinase) by S. maltophilia were increased up to 141%, 41%, and 73%, respectively, when exposed to MP for 7 days. The overall results highlight the potential of parabens to interfere with DW bacteria in planktonic state and biofilms, and compromise the DW microbiological quality and safety., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The quorum sensing (QS) system is related to cell-to-cell communication as a function of population density, which regulates several physiological functions including biofilm formation and virulence gene expression. QS inhibitors have emerged as a promising strategy to tackle virulence and biofilm development. Among a wide variety of phytochemicals, many of them have been described as QS inhibitors. Driven by their promising clues, this study aimed to identify active phytochemicals against LuxS/autoinducer-2 (AI-2) (as the universal QS system) from Bacillus subtilis and LasI/LasR (as a specific QS system) of Pseudomonas aeruginosa, through in silico analysis followed by in vitro validation. The optimized virtual screening protocols were applied to screen a phytochemical database containing 3479 drug-like compounds. The most promising phytochemicals were curcumin, pioglitazone hydrochloride, and 10-undecenoic acid. In vitro analysis corroborated the QS inhibitory activity of curcumin and 10-undecenoic acid, however, pioglitazone hydrochloride showed no relevant effect. Inhibitory effects on LuxS/AI-2 QS system triggered reduction of 33-77% by curcumin (at 1.25-5 µg/mL) and 36-64% by 10-undecenoic acid (at 12.5-50 µg/mL). Inhibition of LasI/LasR QS system was 21% by curcumin (at 200 µg/mL) and 10-54% by 10-undecenoic acid (at 15.625-250 µg/mL). In conclusion, in silico analysis allowed the identification of curcumin and, for the first time, 10-undecenoic acid (showing low cost, high availability, and low toxicity) as alternatives to counteract bacterial pathogenicity and virulence, avoiding the imposition of selective pressure usually related to classic industrial disinfection and antibiotics therapy., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Pharmaceuticals are present as pollutants in several ecosystems worldwide. Despite the reduced concentrations at which they are detected, their negative impact on natural biota constitutes a global concern. The consequences of pharmaceuticals' presence in water sources and food have been evaluated with a higher detail for human health. However, although most of the pharmaceuticals detected in the environment had not been designed to act against microorganisms, it is of utmost importance to understand their impact on the environmental native microbiota. Microbial communities can suffer serious consequences from the presence of pharmaceuticals as pollutants in the environment, which may directly impact public health and ecosystem equilibrium. Among this class of pollutants, the ones that have been studied in more detail are antibiotics. This work aims to provide an overview of the impacts of different pharmaceuticals on environmental biofilms, more specifically in biofilms from aquatic ecosystems and engineered water systems. The alterations caused in the biofilm function and characteristics, as well as bacteria antimicrobial tolerance and consequently the associated risks for public health, are also reviewed. Despite the information already available on this topic, the need for additional data urges the assessment of emerging pollutants on microbial communities and the potential public health impacts.

The advent of antimicrobial resistance has added considerable impact to infectious diseases both in the number of infections and healthcare costs. Furthermore, the relentless emergence of multidrug-resistant bacteria, particularly in the biofilm state, has made mandatory the discovery of new alternative antimicrobial therapies that are capable to eradicate resistant bacteria and impair the development of new forms of resistance. Amongst the therapeutic strategies for treating biofilms, antimicrobial photodynamic therapy (aPDT) has shown great potential in inactivating several clinically relevant micro-organisms, including antibiotic-resistant 'priority bacteria' declared by the WHO as critical pathogens. Its antimicrobial effect is centred on the basis that harmless low-intensity light stimulates a non-toxic dye named photosensitizer, triggering the production of reactive oxygen species upon photostimulation. In addition, combination therapies of aPDT with other antimicrobial agents (e.g. antibiotics) have also drawn considerable attention, as it is a multi-target strategy. Therefore, the present review highlights the recent advances of aPDT against biofilms, also covering progress on combination therapy., (© 2022 Society for Applied Microbiology.)

Musk fragrances have been detected in drinking water (DW) at trace concentrations. However, their impact on the microbial quality of DW has been disregarded. This work provides a pioneer evaluation of the effects of two synthetic musks contaminants, tonalide (AHTN) and galaxolide (HHCB), in microbial biofilms formed on two different surfaces, polyvinyl chloride (PVC) and stainless steel AISI 316 (SS316). Three bacterial species isolated from DW (Acinetobacter calcoaceticus, Burkholderia cepacia and Stenotrophomonas maltophilia), were used to develop 7-day-old single and mixed species biofilms. The impact of musks was assessed directly on biofilms but also on the bacteria motility, biofilm formation ability and biofilm susceptibility to chlorination. AHTN musk caused the most remarkable effects by increasing the cellular density and viability of mixed biofilms, and the extracellular polysaccharides content of biofilms on SS316. Most of the alterations caused by the direct exposure of biofilms to musks were observed when SS316 was used as an adhesion surface. In contrast, the ability to form biofilms and their susceptibility to chlorine were more affected for bacteria from HHCB-exposed biofilms on PVC. The overall results demonstrate that the presence of musks at residual concentrations influences DW bacterial dynamics, with the potential to impact the DW quality and safety. The type of plumbing material may further impact the effects of musks., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The understanding of microbial susceptibility to disinfectants is an important step to provide drinking water (DW) of adequate microbiological quality. In drinking water distribution systems (DWDS) the application of disinfectants is the main approach to control microorganisms. Although chlorine has been commonly used for DW treatment, the increase of microbial resistance and the production of harmful disinfection by-products promote the necessity to seek new alternatives. This study evaluated the antimicrobial activity of sodium dichloroisocyanurate (NaDCC), trichloroisocyanuric acid (TCCA), and pentapotassium bis(peroxymonosulphate) bis(sulphate) (OXONE) against two emerging pathogens isolated from DW, Acinetobacter calcoaceticus and Stenotrophomonas maltophilia. Free chlorine from calcium hypochlorite was used for comparison. The dose and time-responses against planktonic bacteria were performed as well as the assessment of the effects on membrane integrity. Moreover, the effects against 48 h-old biofilms formed on polyvinyl chloride and stainless steel were evaluated in terms of biofilm culturability and removal. Minimum bactericidal concentrations of 2.1 and 3.1 mg/L for NaDCC, 2.5 and 3.8 mg/L for TCCA, 340 and 690 mg/L for OXONE, and 0.80 and 1.0 mg/L for free chlorine alone were obtained against S. maltophilia and A. calcoaceticus, respectively. The kinetic modeling revealed that NaDCC and TCCA caused similar inactivation rates and the time for first log reduction by OXONE was less than 10 min, for both bacteria. All the disinfectants triggered significant bacterial cytoplasmic membrane destabilization, even at sub-lethal concentrations. A 30 min treatment with the disinfectants allowed a reduction in the biofilm culturability up to 5 log. OXONE was the disinfectant with the best efficiency against both bacterial biofilms. However, none of the disinfectants caused significant biofilm removal (reduction < 1 log cells/cm 2 ). This study highlights NaDCC, TCCA, and OXONE as promising alternatives to free chlorine for DW disinfection, particularly for planktonic growth control and biofilm culturability reduction., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Biofilms on food-contact surfaces act as potential reservoirs of microbial pathogens and can cause operational problems. The search for effective biofilm control agents is a significant research need. In this study, glycolic acid (GA) and glyoxal (GO) were tested in the control of biofilms formed by Bacilluscereus and Pseudomonasfluorescens. Benzalkonium chloride (BAC) and peracetic acid (PAA) were used as reference biocides for industrial surface disinfection. The action of the selected biocides was assessed in bacterial motility, culturability, biofilm removal and inactivation, interference with biofilm components and limitation of biocide penetration through the biofilms (reaction-diffusion interactions). Bacterial motility was not affected by the exposure to sub-inhibitory biocide concentrations. In terms of antibiofilm activity, B.cereus biofilms were tolerant to the action of BAC, GA, and GO, with reductions of circa 2-log CFU/cm 2 . Even 10000 µg/mL of PAA had modest effects against B. cereus biofilms (5-log CFU/cm 2 ). On the other hand, P.fluorescens biofilms were more susceptible to the biocides, except BAC which was not effective. The minimum concentrations to cause 3-log CFU/cm 2 reduction in P. fluorescens biofilms were 10000 µg/mL of PAA and GA, and 20000 µg/mL of GO. GO and BAC were the most actives for biofilm removal, while high biofilm inactivation was caused by GA and PAA. In general, biofilm components affected the antimicrobial activity of all the biocides. The activity of GA and GO was not notably diffusion-reaction limited in contrast to PAA. Overall, this study demonstrated that GA and GO had potential antibiofilm activity, being limited at a low level by physicochemical interactions with biofilm components., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The present study evaluates the antimicrobial susceptibility of persister cells of Bacillus cereus and Pseudomonas fluorescens after their regrowth in suspension and as biofilms. Two conventional (benzalkonium chloride-BAC and peracetic acid-PAA) and two emerging biocides (glycolic acid-GA and glyoxal-GO) were selected for this study. Persister cells resulted from biofilms subjected to a critical treatment using the selected biocides. All biocide treatments developed B. cereus persister cells, except PAA that effectively reduced the levels of vegetative cells and endospores. P. fluorescens persister cells comprise viable and viable but non-culturable cells. Afterwards, persister cells were regrown in suspension and in biofilms and were subjected to a second biocide treatment. In general, planktonic cultures of regrown persister cells in suspension lost their antimicrobial tolerance, for both bacteria. Regrown biofilms of persister cells had antimicrobial susceptibility close to those regrown biofilms of biocide-untreated cells, except for regrown biofilms of persister P. fluorescens after BAC treatment, which demonstrated increased antimicrobial tolerance. The most active biocide against persister cells was PAA, which did not promote changes in susceptibility after their regrowth. In conclusion, persister cells are ubiquitous within biofilms and survive after critical biocide treatment. The descendant planktonic and biofilms populations showed similar properties as the original ones.

The effect that the hydrodynamic conditions under which biofilms are formed has on their persistence is still unknown. This study assessed the behaviour of Pseudomonas fluorescens biofilms, formed on stainless steel under different shear stress ( τ w ) conditions (1, 2 and 4   Pa), to chemical (benzalkonium chloride - BAC, glutaraldehyde - GLUT and sodium hypochlorite - SHC) and mechanical (20   Pa) treatments (alone and combined). The biofilms formed under different τ w   showed different structural characteristics. Those formed under a higher τ w were invariably more tolerant to chemical and mechanical stresses. SHC was the biocide which caused the highest biofilm killing and removal, followed by BAC. The sequential exposure to biocides and mechanical stress was found to be insufficient for effective biofilm control. A basal layer containing biofilm cells mostly in a viable state remained on the surface of the cylinders, particularly for the 2 and 4   Pa-generated biofilms.

Background: Until a few years ago, keratoconus was defined as a noninflammatory degenerative disease. However, recent studies have shown that the altered balance between inflammatory cytokines, proteases, and protease inhibitors, as well as free radicals and oxidants, have a crucial role in the pathogenesis of this disease. The aim of this study is to investigate whether interleukin 17 A G197A (rs2275913) and interleukin 17 F T7488C (rs763780) polymorphisms are associated with keratoconus in patients from a population of the northwestern region of the State of São Paulo, Brazil., Methods and Results: 35 patients and 61 controls were enrolled. Genotyping of interleukin 17 A G197A and interleukin 17 F T7488C polymorphisms was carried out using the polymerase chain reaction-restriction fragment length polymorphism technique. Statistical analyses were conducted using the chi-square test, and an odds ratio with a 95% confidence interval was also calculated to evaluate the association between polymorphisms and disease. Evaluating interleukin 17 F T7488C, we found that the TT genotype is associated as a risk factor for keratoconus (P = 0.04; OR = 3.01; CI 1.11-8.14). As for evaluating interleukin 17 A G197A, the allele and genotype frequencies between patients and controls were compared and no statistically significant differences were found., Conclusions: Our data showed that the interleukin 17 F T7488C polymorphisms may exert an influence in keratoconus., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Biofilm control is mainly based on chemical disinfection, without a clear understanding of the role of the biocides and process conditions on biofilm removal. This study aims to understand the effects of a biocide (benzyldimethyldodecyl ammonium chloride-BDMDAC) and mechanical treatment (an increase of shear stress -τw) on single- and dual-species biofilms formed by Bacillus cereus and Pseudomonas fluorescens on high-density polyethene (HDPE). BDMDAC effects were initially assessed on bacterial physicochemical properties and initial adhesion ability. Then, mature biofilms were formed on a rotating cylinder reactor (RCR) for 7 days to assess the effects of chemical and mechanical treatments, and the combination of both on biofilm removal. The results demonstrated that the initial adhesion does not predict the formation of mature biofilms. It was observed that the dual-species biofilms were the most susceptible to BDMDAC exposure. The exposure to increasing τw emphasised the mechanical stability of biofilms, as lower values of τw&nbsp;(1.66 Pa) caused high biofilm erosion and higher τw values (17.7 Pa) seem to compress the remaining biofilm. In general, the combination of BDMDAC and the mechanical treatment was synergic in increasing biofilm removal. However, these were insufficient to cause total biofilm removal (100%; an average standard deviation of 11% for the method accuracy should be considered) from HDPE.

The establishment of a sessile community is believed to occur in a sequence of steps where genetically distinct bacteria can become attached to partner cells via specific molecules, in a process known as coaggregation. The presence of bacteria with the ability to autoaggregate and coaggregate has been described for diverse aquatic systems, particularly freshwater, drinking water, wastewater, and marine water. In these aquatic systems, coaggregation already demonstrated a role in the development of complex multispecies sessile communities, including biofilms. While specific molecular aspects on coaggregation in aquatic systems remain to be understood, clear evidence exist on the impact of this mechanism in multispecies biofilm resilience and homeostasis. The identification of bridging bacteria among coaggregating consortia has potential to improve the performance of wastewater treatment plants and/or to contribute for the development of strategies to control undesirable biofilms. This study provides a comprehensive analysis on the occurrence and role of bacterial coaggregation in diverse aquatic systems. The potential of this mechanism in water-related biotechnology is further described, with particular emphasis on the role of bridging bacteria., Competing Interests: Declaration of Competing Interest Authors declare no conflict of interests., (Copyright © 2021. Published by Elsevier Ltd.)

Biofilms are structures comprising microorganisms associated to surfaces and enclosed by an extracellular polymeric matrix produced by the colonizer cells. These structures protect microorganisms from adverse environmental conditions. Biofilms are typically associated with several negative impacts for health and industries and no effective strategy for their complete control/eradication has been identified so far. The antimicrobial properties of copper are well recognized among the scientific community, which increased their interest for the use of these materials in different applications. In this review the use of different copper materials (copper, copper alloys, nanoparticles and copper-based coatings) in medical settings, industrial equipment and plumbing systems will be discussed considering their potential to prevent and control biofilm formation. Particular attention is given to the mode of action of copper materials. The putative impact of copper materials in the health and/or products quality is reviewed taking into account their main use and the possible effects on the spread of antimicrobial resistance.

The development of effective disinfection strategies, including the search for new active biocides, is a priority research for the food industry. Glycolic acid (GA) and glyoxal (GO) are two glycolysis by-products approved as biocides for surface disinfection, whose antimicrobial action remains to be understood. This study focused on the understanding of the antimicrobial activity of GA and GO against two foodborne pathogens, Bacillus cereus and Pseudomonas fluorescens. Benzalkonium chloride (BAC) and peracetic acid (PAA) were used as reference biocides for comparison. The influence of biocide concentration and exposure time on the antimicrobial activity of the selected biocides was evaluated based on the European Standard EN 1276. The mode of action of each biocide was characterized based on their effects on the cell envelope and cell replication. According to the Chick-Watson model, all biocides interacted chemically with cell targets of both bacteria, except GO that only stablished physicochemical interactions with P. fluorescens. The survival curves revealed that high concentrations of biocide induced readily effects on cell culturability. The susceptibility profile of P. fluorescens was constant over time. In general, B. cereus increased tolerance to BAC and became more susceptible to cumulative damages of GA. Overall, this study demonstrates that the biocidal activity was species-, dose- and time-dependent. GA, similarly to BAC, was a membrane-active and oxidant agent. GO had no effect on the bacterial surface as well as PAA. GO was mainly categorized as a cell replication inhibitor. For the first time, the antimicrobial activity of GA and GO were characterized revealing their potential for rational combination with other biocides commonly used in the food industry., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

This work aimed to evaluate the action of materials with different copper content (0, 57, 96 and 100%) on biofilm formation and control by chlorination and mechanical stress. Stenotrophomonas maltophilia isolated from drinking water was used as a model microorganism and biofilms were developed in a rotating cylinder reactor using realism-based shear stress conditions. Biofilms were characterized phenotypically and exposed to three control strategies: 10 mg l -1 of free chlorine for 10 min, an increased shear stress (a fluid velocity of 1.5 m s -1 for 30s), and a combination of both treatments. These shock treatments were not effective in biofilm control. The benefits from the use of copper surfaces was found essentially in reducing the numbers of non-damaged cells. Copper materials demonstrated better performance in biofilm prevention than chlorine. In general, copper alloys may have a positive public health impact by reducing the number of non-damaged cells in the water delivered after chlorine exposure.

The presence of pharmaceuticals in water sources, including in drinking water (DW), is increasingly being recognized as an emerging and global concern for the environment and public health. Based on the principles of the "One Health" initiative, the present work aims to understand the effects of clofibric acid (CA), a lipid regulator, on the behavior of a selected bacterium isolated from drinking water (DW). Biofilms of the opportunistic pathogen Stenotrophomonas maltophilia were exposed to CA for 12 weeks at 170 and 17000 ng/L. The effects of CA were evaluated on planktonic S. maltophilia susceptibility to chlorine and antibiotics (amoxicillin, ciprofloxacin, erythromycin, kanamycin, levofloxacin, oxacillin, spectinomycin, tetracycline and trimethoprim-sulfamethoxazole), biofilm formation, motility, siderophores production and on the adhesion and internalization of the human colon adenocarcinoma cell line (HT-29). It was found that CA did not affect planktonic S. maltophilia tolerance to chlorine exposure. Additionally, no effects were observed on biofilm formation, motility and siderophores production. However, biofilms formed after CA exposure were more tolerant to chlorine disinfection and lower CFU reductions were obtained. Of additional concern was the effect of CA exposure on S. maltophilia increased tolerance to erythromycin. CA exposure also slightly reduced S. maltophilia ability to invade HT-29 cells. In conclusion, this work reinforces the importance of studying the effects of non-antibiotic contaminants on the behavior of environmental microorganisms, particularly their role as drivers affecting resistance evolution and selection., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Copper pipes are conventionally used to supply tap water. Their role in biofilm prevention remains to be understood. This study evaluates the ability of selected surface materials with different copper contents (0, 57, 79, 87, 96, 100% of copper) to control biofilm formation and regrowth. Further experiments were performed to assess copper leaching and corrosion under conditions mimicking real plumbing systems. Acinetobacter calcoaceticus and Stenotrophomonas maltophilia isolated from a drinking water distribution system were used as model bacteria. All the copper materials showed positive results on the control of single and dual species biofilms presenting high reductions of bacterial culturability > 4 log CFU per cm 2 . The antimicrobial action of the selected materials seem not to be related to copper leaching or to the formation of reactive oxygen species. However, bacterial-copper contact demonstrated damage to bacterial membranes. The alloy containing 96% copper was the most promising surface in reducing biofilm culturability and viability, and was the only surface able to avoid the regrowth of single species biofilms when in contact with high nutrient concentrations. The alloy with 87% copper was shown to be unsuitable for use in chlorinated systems due to the high copper leaching observed when exposed to free chlorine. The presence of viable but non-culturable bacteria was remarkable, particularly in dual species biofilms. The overall results provide novel data on the role of copper alloys for use under chlorinated and unchlorinated conditions. Copper alloys demonstrated comparable or even higher biofilm control effects than elemental copper surfaces., Competing Interests: Authors declare no conflict of interest., (This journal is © The Royal Society of Chemistry.)

Concerns on the presence of emerging contaminants (ECs) in water sources have increased in recent years. The lack of efficient technologies to remove ECs from residual waters contributes for their appearance in drinking water distribution systems (DWDS). Therefore, sessile microorganisms on DWDS pipes are continuously exposed to trace concentrations of ECs. However, no data exists on the role of ECs on the resident microbiota. The present work aims to understand the effects of prolonged exposure of a bacterial strain of Stenotrophomonas maltophilia, isolated from a DWDS, in both planktonic and biofilm states, to trace concentrations of selected ECs (antipyrine-ANTP; diclofenac sodium salt-DCF; ibuprofen-IBP; galaxolide-GAL; tonalide-TON; carbamazepine-CBZ; clofibric acid-CA; tylosin-TY) on its tolerance to sodium hypochlorite (NaOCl) and resistance to antibiotics. Pre-established S. maltophilia biofilms were exposed to ECs for 26 d. Subsequently, the planktonic behaviour of the biofilm cells grown in the presence of ECS was characterized in terms of susceptibility to NaOCl and to selected antibiotics (levofloxacin and trimethoprim-sulfamethoxazole). Moreover, S.maltophilia was tested on its biofilm productivity in the presence of ECs (alone and mixed). These biofilms were challenged by NaOCl in order to assess the role of ECs on biofilm susceptibility. The results did not evidence remarkable effects of ECs on planktonic S. maltophilia susceptibility to NaOCl and antibiotics. However, S. maltophilia biofilm production and susceptibility to NaOCl was affected from ECs pre-exposure, particularly by the combination of different ECs (CA + CBZ, CA + IBP, CA + CBZ + IBP). S. maltophilia biofilms became more resistant to removal by NaOCl when developed in the presence of mixtures of CA + CBZ and CA + CBZ + IBP. Also, biofilm production was significantly affected. CA was present in all the combinations that altered biofilm behaviour. The overall results propose that exposure to ECs for 26 days had not a huge impact on S. maltophilia planktonic antimicrobial susceptibility. Nevertheless, the prolonged exposure to some ECs altered biofilm production and tolerance to NaOCl, with a potential practical outcome of hindering DWDS disinfection. The simultaneous presence of different ECs in the environment may amplify biofilm resilience., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Biofilms can cause severe problems to human health due to the high tolerance to antimicrobials; consequently, biofilm science and technology constitutes an important research field. Growing a relevant biofilm in the laboratory provides insights into the basic understanding of the biofilm life cycle including responses to antibiotic therapies. Therefore, the selection of an appropriate biofilm reactor is a critical decision, necessary to obtain reproducible and reliable in vitro results. A reactor should be chosen based upon the study goals and a balance between the pros and cons associated with its use and operational conditions that are as similar as possible to the clinical setting. However, standardization in biofilm studies is rare. This review will focus on the four reactors (Calgary biofilm device, Center for Disease Control biofilm reactor, drip flow biofilm reactor, and rotating disk reactor) approved by a standard setting organization (ASTM International) for biofilm experiments and how researchers have modified these standardized reactors and associated protocols to improve the study and understanding of medical biofilms.

The presence of biofilms in drinking water distribution systems (DWDS) is a global public health concern as they can harbor pathogenic microorganisms. Sodium hypochlorite (NaOCl) is the most commonly used disinfectant for microbial growth control in DWDS. However, its effect on biofilm removal is still unclear. This work aims to evaluate the effects of the combination of chemical (NaOCl) and mechanical stresses on the removal of single and dual species biofilms of two bacteria isolated from DWDS and considered opportunistic, Acinectobacter calcoaceticus and Stenotrophomonas maltophilia. A rotating cylinder reactor was successfully used for the first time in drinking water biofilm studies with polyvinyl chloride as substratum. The single and dual species biofilms presented different characteristics in terms of metabolic activity, mass, density, thickness and content of proteins and polysaccharides. Their complete removal was not achieved even when a high NaOCl concentrations and an increasing series of shear stresses (from 2 to 23Pa) were applied. In general, NaOCl pre-treatment did not improve the impact of mechanical stress on biofilm removal. Dual species biofilms were colonized mostly by S. maltophilia and were more susceptible to chemical and mechanical stresses than these single species. The most efficient treatment (93% biofilm removal) was the combination of NaOCl at 175mg·l -1 with mechanical stress against dual species biofilms. Of concern was the high tolerance of S. maltophilia to chemical and mechanical stresses in both single and dual species biofilms. The overall results demonstrate the inefficacy of NaOCl on biofilm removal even when combined with high shear stresses., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Introduction Percutaneous balloon valvuloplasty is the primary treatment for critical pulmonary valve stenosis in neonates. Thus far, a few studies have reported long-term results of this technique in neonatal critical pulmonary valve stenosis., Methods: We carried out a retrospective study of all consecutive newborns with critical pulmonary valve stenosis subjected to percutaneous balloon valvuloplasty at a single centre, between 1994 and 2014, to assess its immediate and long-term safety and efficacy., Results: A total of 24 neonates presented with critical pulmonary valve stenosis. The mean diameter of the pulmonary annulus was 7 mm (±1.19); 33.3% had a dysplastic pulmonary valve, and 92% were started on prostaglandin E1 treatment. Percutaneous balloon valvuloplasty was performed at a mean age of 4.0±4.3 days using, on average, a balloon-to-pulmonary annulus ratio of 1.18 mm (with a range from 0.9 to 1.43). Immediate success was achieved in 22/24 patients (92%) with a reduction in the pulmonary transvalvular peak gradient (p<0.05) and in the right ventricle/systemic pressure ratio (p<0.05). There was one death (4%) 6 days after the procedure, and 29.2% of them had transient rhythm complications. For a mean follow-up time of 8.4 years, the re-intervention rate was 42.9%. In total, 14 re-interventions were performed in nine neonates, including surgery in six. Freedom from re-intervention was 50% at 8 years and 43% at 10 and 15 years., Conclusion: This series, to the best of our knowledge, has had the longest follow-up of neonates with critical pulmonary valve stenosis. Percutaneous balloon valvuloplasty is a safe and effective treatment, and in our study 75% of the patients were exclusively treated using this technique.

New biocidal solutions are needed to combat effectively the evolution of microbes developing antibiotic resistance while having a low or no environmental toxicity impact. This work aims to assess the efficacy of commonly used biocides and natural-based compounds on the disinfection of silicone and stainless steel (SS) surfaces seeded with differentStaphylococcus aureusstrains. Minimum inhibitory concentration was determined for synthetic (benzalkonium chloride-BAC, glutaraldehyde-GTA,ortho-phthalaldehyde-OPA and peracetic acid-PAA) and natural-based (cuminaldehyde-CUM), eugenol-EUG and indole-3-carbinol-I3C) biocides by the microdilution method. The efficacy of selected biocides at MIC, 10 × MIC and 5500 mg/L (representative in-use concentration) on the disinfection of sessileS. aureuson silicone and SS was assessed by viable counting. Silicone surfaces were harder to disinfect than SS. GTA, OPA and PAA yielded complete CFU reduction of sessile cells for all test concentrations as well as BAC at 10 × MIC and 5500 mg/L. CUM was the least efficient compound. EUG was efficient for SS disinfection, regardless of strains and concentrations tested. I3C at 10 × MIC and 5500 mg/L was able to cause total CFU reduction of silicone and SS deposited bacteria. Although not so efficient as synthetic compounds, the natural-based biocides are promising to be used in disinfectant formulations, particularly I3C and EUG., (© FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Objective: The gait speed test (GST) is a physical test that can predict falls and aid in the diagnosis of sarcopenia in the elderly. However, to our knowledge, there have been no studies evaluating its reproducibility in hospitalized elderly patients. The objective of this study was to evaluate the safety and reproducibility of the six-meter GST (6GST) in hospitalized elderly patients., Methods: This repeated measures study involved hospitalized elderly patients (≥ 60 years of age) who underwent the 6GST by the fifth day of hospitalization, were able to walk without assistance, and presented no signs of dyspnea or pain that would prevent them from performing the test. The 6GST was performed three times in sequence, with a rest period between each test, in a level corridor. Gait speed was measured in meters/second. Reproducibility was assessed by comparing the means, intraclass correlation coefficients (ICCs) and Bland-Altman plots., Results: We evaluated 110 elderly patients in a total of 330 tests. All participants completed all of the tests. The comparisons between the speeds obtained during the three tests showed high ICCs and a low mean bias (Bland-Altman plots). The correlation and accuracy were greatest when the mean maximum speed was compared with that obtained in the third test (1.26 ± 0.44 m/s vs. 1.22 ± 0.44 m/s; ICC = 0.99; p = 0.001; mean bias = 0.04; and limits of agreement = -0.27 to 0.15)., Conclusions: The 6GST was proven to be safe and to have good reproducibility in this sample of hospitalized elderly patients. The third measurement seems to correspond to the maximum speed, since the first two measurements underestimated the actual performance., Objetivo: O teste de velocidade de marcha (TVM) é um teste físico que pode predizer quedas e auxiliar no diagnóstico de sarcopenia em idosos da comunidade. Entretanto, pelo que sabemos, não há estudos que avaliaram sua reprodutibilidade em idosos hospitalizados. O objetivo deste estudo foi avaliar a segurança e a reprodutibilidade do TVM de seis metros (TVM6) em idosos hospitalizados., Métodos: Estudo com medidas repetidas envolvendo idosos hospitalizados (idade ≥ 60 anos) que realizaram o TVM6 até o quinto dia de hospitalização, sendo capazes de andar sem auxílio e não apresentando dispneia ou dor que os incapacitasse para a realização dos testes. O TVM6 foi realizado sequencialmente três vezes, com período de descanso, em um corredor plano, e a velocidade de marcha foi calculada em metros/segundo. A reprodutibilidade foi avaliada pela comparação das médias, coeficientes de correlação intraclasse (CCI) e disposições gráficas de Bland-Altman., Resultados: Foram avaliados 110 idosos avaliados com um total de 330 testes. Todos os participantes completaram todos os testes. As comparações entre as velocidades obtidas nos três testes realizados indicaram CCIs elevados e viés médio baixo pela disposição gráfica de Bland-Altman. Em relação à maior velocidade aferida, a correlação e a precisão foram maiores quando comparadas à obtida no terceiro teste (1,26 ± 0,44 m/s vs. 1,22 ± 0,44 m/s; CCI = 0,99; p = 0,001; viés médio = 0,04; e limites de concordância = -0,27 a 0,15)., Conclusões: O TVM6 mostrou-se seguro e teve boa reprodutibilidade nessa amostra de idosos hospitalizados. A terceira aferição parece corresponder à velocidade máxima, já que duas primeiras subestimaram o desempenho real.

The effect of nitrification, nitratation and heterotrophic conditions on the biotransformation of several pharmaceuticals in a highly enriched nitrifying activated sludge was evaluated in this study by selective activation of ammonia oxidizing bacteria (AOB), nitrite oxidizing bacteria (NOB) and heterotrophic bacteria. Nitrifiers displayed a noticeable capacity to process ibuprofen due to hydroxylation by ammonia monooxygenase (AMO) to produce 2-hydroxy-ibuprofen. Naproxen was also biotransformed under nitrifying conditions. On the other hand, heterotrophic bacteria present in the nitrifying activated sludge (NAS) biotransformed sulfamethoxazole. In contrast, both nitrifying and heterotrophic activities were ineffective against diclofenac, diazepam, carbamazepine and trimethoprim. Similar biotransformation rates of erythromycin, roxithromycin and fluoxetine were observed under all conditions tested. Overall, results from this study give more evidence on the role of the different microbial communities present in activated sludge reactors on the biological removal of pharmaceuticals., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Aims: Diabetic nephropathy (DN) is one of the most important causes of chronic renal disease, and the incidence of DN is increasing worldwide. Considering our previous report (Gomes et al., 2014) indicating that chronic treatment with oral low-dose quercetin (10 mg/Kg) demonstrated anti-oxidative, anti-apoptotic and renoprotective effects in the C57BL/6J model of DN, we investigated whether this flavonoid could also have beneficial effects in concurrent DN and spontaneous atherosclerosis using the apolipoprotein E-deficient mouse (apoE(-/-))., Methods: Streptozotocin was used to induce diabetes (100 mg/kg/day, 3 days) in male apoE(-/-) mice (8 week-old). After 6 weeks, the mice were randomly separated into DQ: diabetic apoE(-/-) mice treated with quercetin (10 mg/kg/day, 4 weeks, n = 8), DV: diabetic ApoE(-/-) mice treated with vehicle (n = 8) and ND: non-treated non-diabetic mice (n = 8)., Results: Quercetin treatment diminished polyuria (~30%; p < 0.05), glycemia (~25%, p < 0.05), normalized the hypertriglyceridemia. Moreover, this bioflavonoid diminished creatininemia (~30%, p < 0.01) and reduced proteinuria but not to normal levels. We also observed protective effects on the renal structural changes, including normalization of the index of glomerulosclerosis and kidney weight/body weight., Conclusions: Our data revealed that quercetin treatment significantly reduced DN in hypercholesterolemic mice by inducing biochemical changes (decrease in glucose and triglycerides serum levels) and reduction of glomerulosclerosis. Thus, this study highlights the relevance of quercetin as an alternative therapeutic option for DN, including in diabetes associated with dyslipidemia.

Background: Sarcopenia is an important public health problem that affects mainly elders, and has negative consequences, such as disability and even death. Due to the lack of studies evaluating sarcopenia in elderly persons hospitalized in Brazil, the aim of the present study was to describe the frequency of sarcopenia and associated factors among elders in a hospital in the city of Salvador-Brazil., Methods: This cross-sectional study included 110 hospitalized elderly patients in a multi-specialty hospital in Salvador-BA, Brazil. Inclusion criteria: were elders aged ≥60 years between the first and fifth day of hospitalization; who were able to walk without external assistance; with medical permission to walk, and who did not take vasoactive and inotropic drugs. The diagnosis of sarcopenia was determined by combining the reduction in skeletal muscle mass with muscle weakness (women, <20 kg; men, <30 kg) and/or poor physical performance (gait speed ≤0.8 m/s). To obtain reduced skeletal muscle mass, the skeletal muscle mass index ≤6.37 kg/m(2) for women and ≤8.90 kg/m(2) for men was used. Cognitive function, Charlson index, admission profile (clinical and surgical), smoking, falls suffered in the last year and physical inactivity prior to admission were also evaluated. The frequency of sarcopenia was described in percentages with their respective confidence intervals and logistic regression was performed for multivariate analysis of factors associated with sarcopenia., Results: Among the 110 patients included, the frequency of sarcopenia was 21.8%, with 10.0% being of the severe type. There was a predominance of clinical profile (59.1%), such as heart disease (20.0%), pneumonia (13.6%) and skin infections (9.1%), with a Charlson index of 5.4 ± 1.8. The factors associated with sarcopenia were age (OR = 1.14; 95% CI = 1.06 to 1.23), clinical profile on admission (OR = 5.15; 95% CI = 1.16-22.9) and smoking (OR = 7.8; 95% CI = 1.53-39.9)., Conclusions: The frequency of sarcopenia in elderly hospitalized patients was high (1 in 5 elderly) and anthropometric equation can be a viable and inexpensive alternative to screening and programming intervention in this population.

Objectives: The ability of the Timed Up and Go test to predict sarcopenia has not been evaluated previously. The objective of this study was to evaluate the accuracy of the Timed Up and Go test for predicting sarcopenia in elderly hospitalized patients., Methods: This cross-sectional study analyzed 68 elderly patients (≥60 years of age) in a private hospital in the city of Salvador-BA, Brazil, between the 1st and 5th day of hospitalization. The predictive variable was the Timed Up and Go test score, and the outcome of interest was the presence of sarcopenia (reduced muscle mass associated with a reduction in handgrip strength and/or weak physical performance in a 6-m gait-speed test). After the descriptive data analyses, the sensitivity, specificity and accuracy of a test using the predictive variable to predict the presence of sarcopenia were calculated., Results: In total, 68 elderly individuals, with a mean age 70.4±7.7 years, were evaluated. The subjects had a Charlson Comorbidity Index score of 5.35±1.97. Most (64.7%) of the subjects had a clinical admission profile; the main reasons for hospitalization were cardiovascular disorders (22.1%), pneumonia (19.1%) and abdominal disorders (10.2%). The frequency of sarcopenia in the sample was 22.1%, and the mean length of time spent performing the Timed Up and Go test was 10.02±5.38 s. A time longer than or equal to a cutoff of 10.85 s on the Timed Up and Go test predicted sarcopenia with a sensitivity of 67% and a specificity of 88.7%. The accuracy of this cutoff for the Timed Up and Go test was good (0.80; IC=0.66-0.94; p=0.002)., Conclusion: The Timed Up and Go test was shown to be a predictor of sarcopenia in elderly hospitalized patients.

In translational medicine, the discovery of new drugs or new potential uses for currently available drugs is crucial for treating the resistant hypertension associated with renal artery stenosis. The phosphodiesterase 5 inhibitor sildenafil has been shown to reduce blood pressure and to improve the endothelium-dependent relaxation in the two kidney, one clip (2K1C) mouse model of renovascular hypertension. In the present study, we evaluated the effects of sildenafil (40 mg/kg/day for two weeks) on the endothelial structure and contractile function in mesenteric resistance arteries 28 days after clipping the renal artery. The data showed an enhanced vascular contractile response to norepinephrine in 2K1C hypertensive mice (56%) when compared with Sham mice, which was associated with increased oxidative stress and with a thinning of endothelial cells. Sildenafil treatment caused a significant amelioration in the enhanced contractile responsiveness (18%), which was associated to the recovery of the endothelial surface and abolishment of the oxidative stress. These data suggest that sildenafil could be considered a promising therapeutic option to manage endothelial dysfunction and hypertension in resistant patients.

Background: Diabetic nephropathy (DN) is one of the major causes of end-stage renal disease in diabetic patients. Increasing evidence from studies in the rodents has suggested that this disease is associated with increased oxidative stress due to hyperglycemia. In the present study, we evaluated the renoprotective, anti-oxidative and anti-apoptotic effects of the flavonoid quercetin in C57BL/6J model of DN., Methods: DN was induced by streptozotocin (STZ, 100 mg/kg/day, for 3 days) in adult C57BL/6J mice. Six weeks later, mice were divided into the following groups: diabetic mice treated with quercetin (DQ, 10 mg/kg/day, 4 weeks), diabetic mice treated with vehicle (DV) or non-treated non-diabetic (ND) mice., Results: Quercetin treatment caused a reduction in polyuria (~45%) and glycemia (~35%), abolished the hypertriglyceridemia and had significant effects on renal function including, decreased proteinuria and high plasma levels of uric acid, urea and creatinine, which were accompanied by beneficial effects on the structural changes of the kidney including glomerulosclerosis. Flow cytometry showed a decrease in oxidative stress and apoptosis in DN mice., Conclusion: Taken together, these data show that quercetin effectively attenuated STZ-induced cytotoxicity in renal tissue. This study provides convincing experimental evidence and perspectives on the renoprotective effects of quercetin in diabetic mice and outlines a novel therapeutic strategy for this flavonoid in the treatment of DN.

The development of biofilms in drinking water distribution systems (DWDS) can cause pipe degradation, changes in the water organoleptic properties but the main problem is related to the public health. Biofilms are the main responsible for the microbial presence in drinking water (DW) and can be reservoirs for pathogens. Therefore, the understanding of the mechanisms underlying biofilm formation and behavior is of utmost importance in order to create effective control strategies. As the study of biofilms in real DWDS is difficult, several devices have been developed. These devices allow biofilm formation under controlled conditions of physical (flow velocity, shear stress, temperature, type of pipe material, etc), chemical (type and amount of nutrients, type of disinfectant and residuals, organic and inorganic particles, ions, etc) and biological (composition of microbial community - type of microorganism and characteristics) parameters, ensuring that the operational conditions are similar as possible to the DWDS conditions in order to achieve results that can be applied to the real scenarios. The devices used in DW biofilm studies can be divided essentially in two groups, those usually applied in situ and the bench top laboratorial reactors. The selection of a device should be obviously in accordance with the aim of the study and its advantages and limitations should be evaluated to obtain reproducible results that can be transposed into the reality of the DWDS. The aim of this review is to provide an overview on the main reactors used in DW biofilm studies, describing their characteristics and applications, taking into account their main advantages and limitations., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Background: The clipping of an artery supplying one of the two kidneys (2K1C) activates the renin-angiotensin (Ang) system (RAS), resulting in hypertension and endothelial dysfunction. Recently, we demonstrated the intrarenal beneficial effects of sildenafil on the high levels of Ang II and reactive oxygen species (ROS) and on high blood pressure (BP) in 2K1C mice. Thus, in the present study, we tested the hypothesis that sildenafil improves endothelial function in hypertensive 2K1C mice by improving the NO/ROS balance., Methods: 2K1C hypertension was induced in C57BL/6 mice. Two weeks later, they were treated with sildenafil (40 mg/kg/day, via oral) or vehicle for 2 weeks and compared with sham mice. At the end of the treatment, the levels of plasma and intrarenal Ang peptides were measured. Endothelial function and ROS production were assessed in mesenteric arterial bed (MAB)., Results: The 2K1C mice exhibited normal plasma levels of Ang I, II and 1-7, whereas the intrarenal Ang I and II were increased (~35% and ~140%) compared with the Sham mice. Sildenafil normalized the intrarenal Ang I and II and increased the plasma (~45%) and intrarenal (+15%) Ang 1-7. The 2K1C mice exhibited endothelial dysfunction, primarily due to increased ROS and decreased NO productions by endothelial cells, which were ameliorated by treatment with sildenafil., Conclusion: These data suggest that the effects of sildenafil on endothelial dysfunction in 2K1C mice may be due to interaction with RAS and restoring NO/ROS balance in the endothelial cells from MAB. Thus, sildenafil is a promising candidate drug for the treatment of hypertension accompanied by endothelial dysfunction and kidney disease.

Background: Atherosclerosis is an inflammatory process of the arterial walls and is initiated by endothelial dysfunction accompanied by an imbalance in the production of reactive oxygen species (ROS) and nitric oxide (NO). Sildenafil, a selective phosphodiesterase-5 (PDE5) inhibitor used for erectile dysfunction, exerts its cardiovascular effects by enhancing the effects of NO. The aim of this study was to investigate the influence of sildenafil on endothelial function and atherosclerosis progression in apolipoprotein E knockout (apoE-/-) mice., Methods: ApoE-/- mice treated with sildenafil (Viagra®, 40 mg/kg/day, for 3 weeks, by oral gavage) were compared to the untreated apoE-/- and the wild-type (WT) mice.Aortic rings were used to evaluate the relaxation responses to acetylcholine (ACh) in all of the groups. In a separate set of experiments, the roles of NO and ROS in the relaxation response to ACh were evaluated by incubating the aortic rings with L-NAME (NO synthase inhibitor) or apocynin (NADPH oxidase inhibitor). In addition, the atherosclerotic lesions were quantified and superoxide production was assessed., Results: Sildenafil restored the vasodilator response to acetylcholine (ACh) in the aortic rings of the apoE-/- mice. Treatment with L-NAME abolished the vasodilator responses to ACh in all three groups of mice and revealed an augmented participation of NO in the endothelium-dependent vasodilation in the sildenafil-treated animals. The normalized endothelial function in sildenafil-treated apoE-/- mice was unaffected by apocynin highlighting the low levels of ROS production in these animals. Moreover, morphological analysis showed that sildenafil treatment caused approximately a 40% decrease in plaque deposition in the aorta., Conclusion: This is the first study demonstrating the beneficial effects of chronic treatment with sildenafil on endothelial dysfunction and atherosclerosis in a model of spontaneous hypercholesterolemia. These data indicate that the main mechanism of the beneficial effect of sildenafil on the endothelial function appears to involve an enhancement of the NO pathway along with a reduction in oxidative stress.

Purpose: Dyslipidemia is characterized by high lipid blood levels that are risk factors for cardiovascular diseases, which are leading causes of death. However, it is unclear whether dyslipidemia is a cause of the dry eye syndrome (DES). Therefore we determined in transgenic mice models of dyslipidemia, whether there is an association with DES development., Methods: Dyslipidemic models included male and female adult mice overexpressing apolipoprotein CIII (Apo CIII), LDL receptor knockout (LDLR-KO) and ApoE knockout (ApoE-KO). They were compared with age- and gender-matched C57BL/6 mice. Ocular health was evaluated based on corneal slit lamp assessment, phenol red thread test (PRT) and impression cytology. Blood lipid profiles and histology of meibomian and lacrimal glands were also evaluated. Effects of high-fat diet and aging were observed in LDLR-KO and ApoCIII strains, respectively., Results: Body weight and lacrimal gland weight were significantly higher in male mice compared to females of the same strain (P < 0.05). Body weight was significantly lower in LDLRKO mice receiving high lipid diet compared to their controls (P = 0.0043). ApoE-KO were hypercholesterolemic and ApoCIII hypertriglyceridemic while LDLR-KO showed increases in both parameters. The PRT test was lower in male LDLR-KO mice with high-fat diet than control mice with standard diet (P = 0.0273). Aging did not affect lacrimal structural or functional parameters of ApoCIII strain., Conclusions: DES development is not solely dependent on dyslipidemia in relevant mice models promoting this condition. On the other hand, lacrimal gland structure and function are differentially impacted by lipid profile changes in male and female mice. This dissociation suggests that other factors beside dyslipidemia impact on tear film dysfunction and DES development.