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Biofilms formed by antibiotic-resistant bacteria in wound beds present unique challenges in terms of treating wound infections. In this work, the in vivo activity of a novel electrochemical bandage (e-bandage) composed of carbon fabric and controlled by a wearable potentiostat, designed to continuously deliver low amounts of hydrogen peroxide (H 2 O 2 ) was evaluated against methicillin-resistant Staphylococcus aureus (MRSA), multidrug-resistant Pseudomonas aeruginosa (MDR-PA) and mixed-species (MRSA and MDR-PA) wound infections. Wounds created on Swiss Webster mice were infected with the above-named bacteria and biofilms allowed to establish on wound beds for 3 days. e-Bandages, which electrochemically reduce dissolved oxygen to H 2 O 2 when polarized at -0.6 V Ag/AgCl , were placed atop the infected wound bed and polarized continuously for 48 hours. Polarized e-bandage treatment resulted in significant reductions (p <0.001) of both mono-species and mixed-species wound infections. After e-bandage treatment, electron microscopy showed degradation of bacterial cells, and histopathology showed no obvious alteration to the inflammatory host response. Blood biochemistries showed no abnormalities. Taken all together, results of this work suggest that the described H 2 O 2 -producing e-bandage can effectively reduce in vivo MRSA, MDR-PA and mixed-species wound biofilms, and should be further developed as a potential antibiotic-free strategy for treatment of wound infections.

The antibiofilm activity of a hypochlorous acid (HOCl)-producing electrochemical bandage (e-bandage) was assessed against 14 yeast isolates in vitro . The evaluated e-bandage was polarized at +1.5 V Ag/AgCl to allow continuous production of HOCl. Time-dependent decreases in the biofilm CFU counts were observed for all isolates with e-bandage treatment. The results suggest that the described HOCl-producing e-bandage could serve as a potential alternative to traditional antifungal wound biofilm treatments.

The activity of a hypochlorous acid-producing electrochemical bandage (e-bandage) in preventing methicillin-resistant Staphylococcus aureus infection (MRSA) infection and removing biofilms formed by MRSA was assessed using a porcine explant biofilm model. e-Bandages inhibited S. aureus infection (p = 0.029) after 12 h (h) of exposure and reduced 3-day biofilm viable cell counts after 6, 12, and 24 h exposures (p = 0.029). Needle-type microelectrodes were used to assess HOCl concentrations in explant tissue as a result of e-bandage treatment; toxicity associated with e-bandage treatment was evaluated. HOCl concentrations in infected and uninfected explant tissue varied between 30 and 80 µM, decreasing with increasing distance from the e-bandage. Eukaryotic cell viability was reduced by an average of 71% and 65% in fresh and day 3-old explants, respectively, when compared to explants exposed to nonpolarized e-bandages. HOCl e-bandages are a promising technology that can be further developed as an antibiotic-free treatment for wound biofilm infections., (© 2022 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC.)

Chronic wound biofilm infections represent a major clinical challenge which results in a substantial burden to patients and healthcare systems. Treatment with topical antibiotics is oftentimes ineffective as a result of antibiotic-resistant microorganisms and biofilm-specific antibiotic tolerance. Use of biocides such as hypochlorous acid (HOCl) has gained increasing attention due to the lack of known resistance mechanisms. We designed an HOCl-generating electrochemical bandage (e-bandage) that delivers HOCl continuously at low concentrations targeting infected wound beds in a similar manner to adhesive antimicrobial wound dressings. We developed a battery-operated wearable potentiostat that controls the e-bandage electrodes at potentials suitable for HOCl generation. We demonstrated that e-bandage treatment was tunable by changing the applied potential. HOCl generation on electrode surfaces was verified using microelectrodes. The developed e-bandage showed time-dependent responses against in vitro Acinetobacter baumannii and Staphylococcus aureus biofilms, reducing viable cells to non-detectable levels within 6 and 12 hours of treatment, respectively. The developed e-bandage should be further evaluated as an alternative to topical antibiotics to treat wound biofilm infections., Competing Interests: Conflict of Interests Dr. Patel reports grants from ContraFect, TenNor Therapeutics Limited, and BioFire. Dr. Patel is a consultant to PhAST, Torus Biosystems, Day Zero Diagnostics, Mammoth Biosciences, and HealthTrackRx; monies are paid to Mayo Clinic. Mayo Clinic and Dr. Patel have a relationship with Pathogenomix. Dr. Patel has research supported by Adaptive Phage Therapeutics. Mayo Clinic has a royalty-bearing know-how agreement and equity in Adaptive Phage Therapeutics. Dr. Patel is also a consultant to Netflix, Abbott Laboratories, and CARB-X. In addition, Dr. Patel has a patent on Bordetella pertussis/parapertussis PCR issued, a patent on a device/method for sonication with royalties paid by Samsung to Mayo Clinic, and a patent on an anti-biofilm substance issued. Dr. Patel receives honoraria from the NBME, Up-to-Date and the Infectious Diseases Board Review Course.

Previously, an electrochemical bandage (e-bandage) that uses a three-electrode system to produce hydrogen peroxide (H 2 O 2 ) electrochemically on its working electrode was developed as a potential strategy for treating biofilms; it showed activity in reducing biofilms in an agar biofilm model. Xanthan gum-based hydrogel, including NaCl, was used as the electrolyte. While H 2 O 2 generated at the working electrode in the vicinity of a biofilm is a main mechanism of activity, the role of the counter electrode was not explored. The goal of this research was to characterize electrochemical reactions occurring on the counter electrode of the e-bandage. Counter electrode potential varied between 1.2 and 1.5 V Ag/AgCl ; ∼125 µM hypochlorous acid (HOCl) was generated within 24 h in the e-bandage system. When HOCl was not produced on the counter electrode (achieved by removing NaCl from the hydrogel), reduction of Acinetobacter baumannii BAA-1605 biofilm was 1.08 ± 0.38 log 10 CFU/cm 2 after 24 h treatment, whereas when HOCl was produced, reduction was 3.87 ± 1.44 log 10 CFU/cm 2 . HOCl inhibited catalase activity, abrogating H 2 O 2 decomposition. In addition to H 2 O 2 generation, the previously described H 2 O 2 -generating e-bandage generates HOCl on the counter electrode, enhancing its biocidal activity., 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 © 2022 Elsevier B.V. All rights reserved.)

Aims: Effects of H 2 O 2 producing electrochemical-bandages (e-bandages) on methicillin-resistant Staphylococcus aureus colonization and biofilm removal were assessed using a porcine explant biofilm model. Transport of H 2 O 2 produced from the e-bandage into explant tissue and associated potential toxicity were evaluated., Methods and Results: Viable prokaryotic cells from infected explants were quantified after 48 h treatment with e-bandages in three ex vivo S. aureus infection models: (1) reducing colonization, (2) removing young biofilms and (3) removing mature biofilms. H 2 O 2 concentration-depth profiles in explants/biofilms were measured using microelectrodes. Reductions in eukaryotic cell viability of polarized and nonpolarized noninfected explants were compared. e-Bandages effectively reduced S. aureus colonization (p = 0.029) and reduced the viable prokaryotic cell concentrations of young biofilms (p = 0.029) with limited effects on mature biofilms (p > 0.1). H 2 O 2 penetrated biofilms and explants and reduced eukaryotic cell viability by 32-44% compared to nonpolarized explants., Conclusions: H 2 O 2 producing e-bandages were most active when used to reduce colonization and remove young biofilms rather than to remove mature biofilms., Significance and Impact of Study: The described e-bandages reduced S. aureus colonization and young S. aureus biofilms in a porcine explant wound model, supporting their further development as an antibiotic-free alternative for managing biofilm infections., (© 2022 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.)

Controlling the orientation of redox enzymes on electrode surfaces is essential in the development of direct electron transfer (DET)-based bioelectrocatalytic systems. The electron transfer (ET) distance varies according to the enzyme orientation when immobilized on an electrode surface, which influences the interfacial ET rate. We report control of the orientation of carbon monoxide dehydrogenase (CODH) as a model enzyme through the fusion of gold-binding peptide (gbp) at either the N- or the C-terminus, and at both termini to strengthen the binding interactions between the fusion enzyme and the gold surface. Key factors influenced by the gbp fusion site are described. Collectively, our data show that control of the CODH orientation on an electrode surface is achieved through the presence of dual tethering sites, which maintains the enzyme cofactor within a DET-available distance (<14 Å), thereby promoting DET at the enzyme-electrode interface., (© 2022. The Author(s).)

Wound infections are caused by bacteria and/or fungi. The presence of fungal biofilms in wound beds presents a unique challenge, as fungal biofilms may be difficult to eradicate. The goal of this work was to assess the in vitro antibiofilm activity of an H 2 O 2 -producing electrochemical bandage (e-bandage) against 15 yeast isolates representing commonly encountered species. Time-dependent decreases in viable biofilm CFU counts of all isolates tested were observed, resulting in no visible colonies with 48 h of exposure by plate culture. Fluorescence microscopic analysis showed extensive cell membrane damage of biofilm cells after e-bandage treatment. Reductions in intracellular ATP levels of yeast biofilm cells were recorded post e-bandage treatment. These results suggest that exposure to H 2 O 2 -producing e-bandages reduces in vitro viable cell counts of yeast biofilms, making this a potential new topical treatment approach for fungal wound infections.

Conventional methods for testing antibiotic susceptibility rely on bacterial growth on agar plates (diffusion assays) or in liquid culture (microdilution assays). These time-consuming assays use population growth as a proxy for cellular respiration. Herein we propose to use mediated extracellular electron transfer as a rapid and direct method to classify antibiotic-susceptible and -resistant bacteria. We tested antibiotics with diverse mechanisms of action (ciprofloxacin, imipenem, oxacillin, or tobramycin) with four important nosocomial pathogens (Acinetobacter baumannii, Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae) by adding the bacterial culture to a custom-designed electrochemical cell with a glassy-carbon electrode and growth media supplemented with a soluble electron transfer mediator, phenazine methosulfate (PMS). During cell respiration, liberated electrons reduce PMS, which is then oxidized on the electrode surface, and current is recorded. Using this novel approach, we were able to consistently classify strains as antibiotic-resistant or -susceptible in <90 min for methodology development and <150 min for blinded tests., (Copyright © 2021. Published by Elsevier B.V.)

Exosomes, powerful extracellular nanovesicles released from almost all types of living cells, are considered the communication engines (messengers) that control and reprogram physiological pathways inside target cells within a community or between different communities. The cell-like structure of these extracellular vesicles provides a protective environment for their proteins and DNA/RNA cargos, which serve as biomarkers for many malicious diseases, including infectious diseases and cancers. Cancer-derived exosomes control cancer metastasis, prognosis, and development. In addition to the unique structure of exosomes, their nanometer size and tendency of interacting with cells makes them a viable novel drug delivery solution. In recent years, numerous research efforts have been made to quantify and characterize disease-derived exosomes for diagnosis, monitoring, and therapeutic purposes. This review aims to (1) relate exosome biomarkers to their origins, (2) focus on current isolation and detection methods, (3) discuss and evaluate the proposed technologies deriving from exosome research for cancer treatment, and (4) form a conclusion about the prospects of the current exosome research.

Central line-associated bloodstream infection (CLABSI) contributes to mortality and cost. While aseptic dressings and antibiotic-impregnated catheters prevent some extraluminal infections, intraluminal infections remain a source of CLABSIs. In this proof-of-concept study, an electrochemical intravascular catheter (e-catheter) prototype capable of electrochemically generating hypochlorous acid intraluminally using platinum electrodes polarized at a constant potential of 1.5 electrode potential relative to saturated silver/silver chloride reference electrode measured in volts (V Ag/AgCl ) was developed. After 24 h of prepolarization at 1.5 V Ag/AgCl , their activity was tested against clinical isolates of Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecium, and Escherichia coli derived from catheter-related infections. e-catheters generated a mean HOCl concentration of 15.86 ± 4.03 μM and had a mean pH of 6.14 ± 0.79. E-catheters prevented infections of all four species, with an average reduction of 8.41 ± 0.61 log 10 CFU/ml at 48 h compared to controls. Polarized e-catheters which generate low amounts of HOCl continuously should be further developed to prevent intraluminal infection. IMPORTANCE Catheter-related infections constitute an economic and mortality burden in health care. Several options are available to reduce the risk of infection, but only a few focus on preventing intraluminal infection, which occurs in long-term catheters, most often used for dialysis, prolonged treatment, or chemotherapy. A prototype of a catheter called an "e-catheter" composed of three electrodes, capable of producing hypochlorous acid (HOCl) electrochemically in its lumen, was developed. When polarized at 1.5 V, chloride ions in the solution are oxidized to continuously produce low amounts of HOCl, which exhibits antibacterial activity in the lumen of the catheter. Here, this prototype was shown to be able to generate HOCl as well as prevent infection in a preliminary in vitro catheter model. This approach is a potential strategy for catheter infection prevention.

Biofilms formed by antibiotic-resistant bacteria in wound beds present unique challenges in terms of treating chronic wound infections; biofilms formed by one or more than one bacterial species are often involved. In this work, the in vitro anti-biofilm activity of a novel electrochemical bandage (e-bandage) composed of carbon fabric and controlled by a wearable potentiostat, designed to continuously deliver low amounts of hydrogen peroxide (H 2 O 2 ) was evaluated against 34 mono-species and 12 dual-species membrane bacterial biofilms formed by Staphylococcus aureus , S. epidermidis , Enterococcus faecium , E. faecalis , Streptococcus mutans , Escherichia coli , Pseudomonas aeruginosa , Acinetobacter baumannii , Klebsiella pneumoniae , Cutibacterium acnes , and Bacteroides fragilis . Biofilms were grown on polycarbonate membranes placed atop agar plates. An e-bandage, which electrochemically reduces dissolved oxygen to H 2 O 2 when polarized at -0.6 V Ag/AgCl , was then placed atop each membrane biofilm and polarized continuously for 12, 24, and 48 h using a wearable potentiostat. Time-dependent decreases in viable CFU counts of all mono- and dual-species biofilms were observed after e-bandage treatment. 48 h of e-bandage treatment resulted in an average reduction of 8.17 ± 0.40 and 7.99 ± 0.32 log 10  CFU/cm 2 for mono- and dual-species biofilms, respectively. Results suggest that the described H 2 O 2 producing e-bandage can reduce in vitro viable cell counts of biofilms grown either in mono- or dual-species forms, and should be further developed as a potential antibiotic-free treatment strategy for treating chronic wound infections., Competing Interests: This research was supported by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under the grant number R01 AI091594. The authors would like to thank Henry Chambers III (University of California, San Francisco) for providing S. aureus USA100, USA200, and USA300; Caliper Life Sciences for providing S. aureus Xen 30, S. epidermidis Xen 43, and P. aeruginosa Xen 5; Daniel Hassett (University of Cincinnati) for providing P. aeruginosa PAO1, PA14, and PA14 Δkat AB; and the Antibacterial Resistance Leadership Group (supported by a grant from the National Institutes of Health through Duke University) for providing A. baumannii ARLG-1268. R.P. reports grants from CD Diagnostics, Merck, Hutchison Biofilm Medical Solutions, Accelerate Diagnostics, ContraFect, TenNor Therapeutics Limited, and Shionogi. R.P. is a consultant for Curetis, Specific Technologies, Next Gen Diagnostics, PathoQuest, Selux Diagnostics, 1928 Diagnostics, and Qvella; monies are paid to Mayo Clinic. In addition, R.P. has patents on Bordetella pertussis/parapertussis PCR, a device/method for sonication with royalties paid by Samsung to Mayo Clinic, and an antibiofilm substance. R.P. receives travel reimbursement from ASM and IDSA, an editor's stipend from IDSA, and honoraria from the NBME, Up-to-Date, and the Infectious Diseases Board Review Course. H.B. holds a patent (US20180207301A1), “Electrochemical reduction or prevention of infections,” which refers to the electrochemical scaffold described herein., (© 2021 The Authors.)

Soil health is a complex phenomenon that reflects the ability of soil to support both plant growth and other ecosystem functions. To our knowledge, research on extracellular electron transfer processes in soil environments is limited and could provide novel knowledge and new ways of monitoring soil health. Electrochemical activities in the soil can be studied by inserting inert electrodes. Once the electrode is polarized to a favorable potential, nearby microorganisms attach to the electrodes and grow as biofilms. Biofilms are a major part of the soil and play critical roles in microbial activity and community dynamics. Our work aims to investigate the electrochemical behavior of healthy and unhealthy soils using chronoamperometry and cyclic voltammetry. We developed a bioelectrochemical soil reactor for electrochemical measurements using healthy and unhealthy soils taken from the Cook Agronomy Farm Long-Term Agroecological Research site; the soils showed similar physical and chemical characteristics, but there was higher plant growth where the healthy soil was taken. Using carbon cloth electrodes installed in these soil reactors, we explored the electrochemical signals in these two soils. First, we measured redox variations by depth and found that reducing conditions were prevalent in healthy soils. Current measurements showed distinct differences between healthy and unhealthy soils. Scanning electron microscopy images showed the presence of microbes attached to the electrode for healthy soil but not for unhealthy soil. Glucose addition stimulated current in both soil types and caused differences in cyclic voltammograms between the two soil types to converge. Our work demonstrates that we can use current as a proxy for microbial metabolic activity to distinguish healthy and unhealthy soil.

Chronic wound infections caused by biofilm-forming microorganisms represent a major burden to healthcare systems. Treatment of chronic wound infections using conventional antibiotics is often ineffective due to the presence of bacteria with acquired antibiotic resistance and biofilm-associated antibiotic tolerance. We previously developed an electrochemical scaffold that generates hydrogen peroxide (H 2 O 2 ) at low concentrations in the vicinity of biofilms. The goal of this study was to transition our electrochemical scaffold into an H 2 O 2 -generating electrochemical bandage (e-bandage) that can be used in vivo. The developed e-bandage uses a xanthan gum-based hydrogel to maintain electrolytic conductivity between e-bandage electrodes and biofilms. The e-bandage is controlled using a lightweight, battery-powered wearable potentiostat suitable for use in animal experiments. We show that e-bandage treatment reduced colony-forming units of Acinetobacter buamannii biofilms (treatment vs. control) in 12 h (7.32 ± 1.70 vs. 9.73 ± 0.09 log 10 [CFU/cm 2 ]) and 24 h (4.10 ± 12.64 vs. 9.78 ± 0.08 log 10 [CFU/cm 2 ]) treatments, with 48 h treatment reducing viable cells below the limit of detection of quantitative and broth cultures. The developed H 2 O 2 -generating e-bandage was effective against in vitro A. baumannii biofilms and should be further evaluated and developed as a potential alternative to topical antibiotic treatment of wound infections., (© 2021 Wiley Periodicals LLC.)

The goals of this work were to study the kinetics and investigate the factors controlling the scale up of oxygen reducing mixed culture cathodic biofilms. Cathodic biofilms were enriched on different electrode sizes (14.5 cm 2 , 40.3 cm 2 , 131 cm 2 and 466 cm 2 ). Biofilm enrichment shifted the oxygen reduction onset potential from -0.1 V Ag/AgCl to 0.3 V Ag/AgCl , indicating the biofilm catalyzed oxygen reduction. The kinetics of oxygen reduction were studied by varying the bulk dissolved oxygen concentration. Oxygen reduction followed a Michaelis-Menten kinetics on all electrode sizes. The maximum current density decreased with increasing electrode surface area (-97.0 ± 10.6 μA/cm 2 , -76.0 ± 8.2 μA/cm 2 , -66.3 ± 3.0 μA/cm 2 and -43.5 ± 10.5 μA/cm 2 , respectively). Cyclic voltammograms suggest that scale up was limited by ohmic resistance, likely due to the low ionic conductivity in the wastewater medium. Mathematical modeling using combined Michaelis-Menten and Butler-Volmer model supports that the decrease in current density with increasing electrode surface area is caused by ohmic losses. Analysis of the microbial community structure in different size electrodes and in multiple regions on the same electrode showed low variability, suggesting that the microbial community does not control the scale up of cathodic biofilms., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2021 Published by Elsevier B.V.)

Bio-mediated ground improvement is an attractive alternative to traditional admixtures for strength improvement of shallow surfaces because it is environmentally friendly. Since biofilms contain extracellular polymeric substances (EPS), they can be considered as an alternative to current technologies to improve soil strength. EPS containing biofilms are porous materials with charged surfaces, and therefore adsorption and capillary condensation can result in water retention. Currently, most of the literature work only tested soil improvement under the dry condition. Therefore, the influence of water retention by biofilms on the strength of improved soil remains unclear. Our goal is to evaluate the strength of a biofilm-enhanced sand over a wide range of saturations and explain the trends through the suction stress characteristic curve (SSCC), which quantifies interparticle adsorptive, capillary, and cementation forces as a function of saturation. We used homogenized anaerobic granule biofilms from an existing upflow anaerobic sludge blanket reactor and mixed it with sand to test the strength of a poorly-graded sand in a wide range of saturations, S (between 0.02  S and 0.74  S ). We found that biofilm treatment of sand increases soil strength through cementation over a wide range of saturations and through adsorptive forces among sand, biofilm surfaces, and water molecules at low saturations ( S  < ~0.3). Our results suggested that homogenized biofilms mixed with sand can be used to improve the strength of sand over a wide range of saturations., Competing Interests: The authors do not have any competing interest., (© 2021 The Authors.)

Hydrogen peroxide (H 2 O 2 ) and hypochlorous acid (HOCl) are biocides used for cleaning and debriding chronic wound infections, which often harbor drug resistant bacteria. Here, we evaluated the in vitro activity of H 2 O 2 and HOCl against 27 isolates of eight bacterial species involved in wound infections. Minimum inhibitory concentrations (MICs) and minimum biofilm bactericidal concentrations (MBBCs) were measured. When compared to their respective MICs, MBBCs of isolates exposed to H 2 O 2 were 16- to 1,024-fold higher and those exposed to HOCl were 2- to 4-fold higher. We evaluated selection of resistance after exposure of Staphylococcus aureus and Pseudomonas aeruginosa biofilms to 10 iterations of electrochemically generated HOCl or H 2 O 2 delivered using electrochemical scaffolds (e-scaffolds), observing no decrease in anti-biofilm effects with serial exposure to e-scaffold-generated H 2 O 2 or HOCl. 24-hour exposure to H 2 O 2 -generating e-scaffolds consistently decreased colony forming units (CFUs) of S. aureus and P. aeruginosa biofilms by ∼5.0-log 10 and ∼4.78-log 10 through 10 iterations of exposure, respectively. 4-hour exposure to HOCl-generating e-scaffolds consistently decreased CFUs of S. aureus biofilms by ∼4.9-log 10 , and 1-hour exposure to HOCl-generating e-scaffolds consistently decreased CFUs of P. aeruginosa biofilms by ∼1.57-log 10 These results suggest that HOCl has similar activity against planktonic and biofilm bacteria, whereas the activity of H 2 O 2 is less against biofilm than planktonic bacteria, and that repeat exposure to either biocide, generated electrochemically under the experimental conditions studied, does not lessen antibiofilm effects., (Copyright © 2021 American Society for Microbiology.)

The treatment of municipal wastewater is an energy-intensive process, with the delivery of oxygen as an electron acceptor accounting for a significant share of the total energy consumption. Microbial communities growing on polarized electrodes can facilitate wastewater treatment processes by exchanging electrons with the electrodes. As a new approach, we combined the use of polarized electrodes with microbial fuel cells (MFCs) to develop a switchable dual-mode bioelectrochemical wastewater treatment system. In this system, we first enriched microbial communities on polarized anodes and cathodes. After enrichment, the system was switched to either a self-powered MFC (SP-MFC) mode or a potentiostatically controlled (PC) mode. The system was evaluated at the laboratory scale (260 L, 4 anode and cathode pairs) and the pilot scale (1200 L, 16 anode and cathode pairs). PC and SP-MFC systems showed improved COD removal relative to control (41.6 ± 3.5 and 38.4 ± 3.1 vs 28.8 ± 2.1 mg L -1 d -1 , respectively). The laboratory-scale system was operated without biological or electrical interruption for one year. Finally, specific enrichment of active microbial communities was observed on PC anodes in comparison to mixed-operation and non-polarized control anodes. The combined PC and SP-MFC systems allowed us to develop a sustainable and failure-free bioelectrochemical wastewater treatment system., 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 B.V. All rights reserved.)

Oxidizing agents like hypochlorous acid (HOCl) have antimicrobial activity. We developed an integrated electrochemical scaffold, or e-scaffold, that delivers a continuous low dose of HOCl aimed at targeting microbial biofilms without exceeding concentrations toxic to humans as a prototype of a device being developed to treat wound infections in humans. In this work, we tested the device against 33 isolates of bacteria (including isolates with acquired antibiotic resistance) grown as in vitro biofilms alongside 12 combinations of dual-species in vitro biofilms. Biofilms were grown on the bottoms of 12-well plates for 24 h. An integrated e-scaffold was placed atop each biofilm and polarized at 1.5 V for 1, 2, or 4 h. HOCl was produced electrochemically by oxidizing chloride ions (Cl - ) in solution to chlorine (Cl 2 ); dissolved Cl 2 spontaneously dissociates in water to produce HOCl. The cumulative concentration of HOCl produced at the working electrode in each well was estimated to be 7.89, 13.46, and 29.50 mM after 1, 2, and 4 h of polarization, respectively. Four hours of polarization caused an average reduction of 6.13 log 10 CFU/cm 2 (±1.99 log 10 CFU/cm 2 ) of viable cell counts of monospecies biofilms and 5.53 log 10 CFU/cm 2 (±2.31 log 10 CFU/cm 2 ) for the 12 dual-species biofilms studied. The described integrated e-scaffold reduces viable bacterial cell counts in biofilms formed by an array of antibiotic-susceptible and -resistant bacteria alone and in combination., (Copyright © 2021 American Society for Microbiology.)

In bioelectrochemical system (BES), phosphate buffer saline (PBS) is usually used to achieve a suitable pH condition, which also increases electrolyte salinity. A series of factors that change with salinity will affect BES performance. To simplify the scenario, a three-electrode BES is used to investigate how additional salinity affects the performance of a 50 mM PBS-buffered BES. Results demonstrated that current production decreased with increasing salinity and the dominant exoelectrogens were not inhibited with the addition of 200 mM NaCl. The distribution of system resistance was analyzed by electrochemical impedance spectroscopy. Compared to the decreased solution and biofilm resistance, the increased interfacial resistance that accounted for up to 97.8% of total resistance was the dominant reason for the decreased current production with the increasing additional salinity. The effects of additional salinity on acetate degradation and columbic efficiency were also analyzed., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

'Candidatus Liberibacter asiaticus' ('Ca. L. asiaticus'), the suspected causative agent of citrus greening disease, is one of many phloem-restricted plant pathogens that have not been isolated and grown in an axenic culture. In this study, infected Asian citrus psyllids were used to prepare a host-free source of 'Ca. L. asiaticus'. Host-free mixed microbial cultures of 'Ca. L. asiaticus' were grown in the presence of various antibiotic treatments to alter the composition of the microbial communities. Our hypothesis was that the presence of selected antibiotics would enhance or reduce the presence of 'Ca. L. asiaticus' in a host-free culture composed of a mixed bacterial population through changes in the microbial community structure. We determined how 'Ca. L. asiaticus' growth changed with the various treatments. Treatment with vancomycin (50 μg/mL), streptomycin (0.02 μg/mL), or polymyxin B (4 μg/mL) was associated with an increased abundance of 'Ca. L. asiaticus' of 7.35 ± 0.27, 5.56 ± 0.15, or 4.54 ± 0.83 fold, respectively, compared to untreated mixed microbial cultures, while treatment with 100 μg/mL vancomycin; 0.5, 1, or 2 μg/mL streptomycin; or 0.5 μg/mL of polymyxin B was associated with reduced growth. In addition, the growth of 'Ca. L. asiaticus' was associated with the microbial community composition of the mixed microbial cultures. A positive relationship between the presence of the Pseudomonadaceae family and 'Ca. L. asiaticus' growth was observed, while the presence of 'Ca. L. asiaticus' was below the detection limit in cultures that displayed high abundances of Bacillus cereus. Our findings offer strategies for developing effective axenic culture conditions and suggest that enrichment of the Bacillaceae family could serve as a paratransgenic approach to controlling citrus greening disease., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Infections with bacterial biofilm communities are highly tolerant of antibiotics. This protection is attributed, in part, to a hydrated extracellular polymeric substance (EPS) that surrounds the bacterial community and that limits antibiotic diffusion. In this study, we evaluated whether it is possible to dehydrate and then re-hydrate a biofilm as a means to increase antibiotic penetration and efficacy. Acinetobacter baumannii biofilms (24 h) were exposed to hypertonic concentrations of maltodextrin, sucrose or polyethylene glycol (PEG) as the dehydration step. These biofilms were then washed with deionized water containing 10 times the concentration of antibiotics needed to kill these bacteria in broth culture (50 µg/mL tobramycin, 300 µg/mL chloramphenicol, 20 µg/mL ciprofloxacin or 100 µg/mL erythromycin) as the rehydration step. Biofilms were then harvested, and the number of viable cells was determined. Sequential treatment with PEG and tobramycin reduced cell counts 4 to 7 log ( p < 0.05) relative to combining PEG and tobramycin in a single treatment, and 3 to 7 log relative to tobramycin treatment alone ( p < 0.05). Results were variable for other osmotic compounds and antibiotics depending on the concentrations used, likely related to mass and hydrophobicity. Our findings support future clinical evaluation of sequential regimens of hypertonic and hypotonic solutions to enhance antibiotic efficacy against chronic biofilm infections.

Biomass-derived nanocarbons (BNCs) have attracted significant research interests due to their promising economic and environmental benefits. Following their extensive uses in physical and chemical research domains, BNCs are now growing in biological applications. However, their practical biological applications are still in their infancy, requiring critical evaluations and strategic directions, which are provided in this review. The carbonization of biomass sources and major types of BNCs are introduced, encompassing carbon nanodots, nanofibres, nanotubes, and graphenes. Next, essential biological uses of BNCs, antibacterial/antibiofilm materials (nanofibres and nanodots) and bioimaging agents (predominantly nanodots), are summarized. Furthermore, the future potential of BNCs, for designing wound dressing/healing materials, water and air disinfection platforms, and microbial electrochemical systems, is discussed. We reach the conclusion that a crucial challenge is the structural control of BNCs. Furthermore, a key knowledge gap for realizing practical biological applications is the lack of systematic comparisons of BNCs with nanocarbons of synthetic origin in the current literature. Although we did not attempt to perform an exhaustive literature survey, the evaluation of the existing results indicates that BNCs are promising as easily accessible materials for various biomedically and environmentally relevant applications.

Fatty acids (FAs) have emerged as a potential alternative to conventional antibiotics. Since many microbes respond differently to a variety of natural and synthetic FAs, substantial efforts have been made to understand the unique features of FAs that function as antimicrobials at high doses and biofilm inhibitors at low doses. Here, we provide an overview of the emerging antibiofilm properties of FAs and their relations to virulence and quorum sensing, such as diffusible signal factors (DSFs), acyl-homoserine lactones, and autoinducer-2 systems. Several FAs mimic DSFs and control motility, fimbriae, hyphae, and biofilm development as well as virulence characteristics of diverse microbes. The findings of FAs as antibiofilm and antivirulence agents will provide a new paradigm to cope with recalcitrant pathogens., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Parameters representing three-dimensional (3D) biofilm structure are quantified from confocal laser-scanning microscope (CLSM) images. These 3D parameters describe the distribution of biomass pixels within the space occupied by a biofilm; however, they lack a direct connection to biofilm activity. As a result, researchers choose a handful of parameters without there being a consensus on a standard set of parameters. We hypothesized that a select 3D parameter set could be used to reconstruct a biofilm image and that the reconstructed and original biofilm images would have similar activities. To test this hypothesis, an algorithm was developed to reconstruct a biofilm image with parameters identical to those of the original CLSM image. We introduced an objective method to assess the reconstruction algorithm by comparing the activities of the original and reconstructed biofilm images. We found that biofilm images with identical structural parameters showed nearly identical activities and substrate concentration profiles. This implies that the set containing all common structural parameters can successfully describe biofilm structure. This finding is significant, as it opens the door to the next step, of finding a smaller standard set of biofilm structural parameters that can be used to compare biofilm structure., (© 2020 Wiley Periodicals LLC.)

Microliter volumes are used in electrochemical detection and preconcentration of radionuclides to reduce the dose received by researchers and equipment. Unfortunately, there is a lack of analysis of radionuclides with coupled electrochemical techniques and microliter volume reactors. The goals of this work are 1) to develop a miniaturized micro-electrochemical quartz crystal microbalance (µeQCM) reactor for use in small volume (50-200 µL) electrogravimetric experiments and 2) to use this reactor to characterize the preconcentration of neptunium on carbon electrodes via electroprecipitation. We successfully deposited neptunium in the new µeQCM reactor and verified its operation. We found that preconcentration of neptunium on carbon coated electrodes was possible by chronoamperometry at -1.6 V Ag/AgCl . The mass shift of the resulting precipitate was indicative of the amount of neptunium on the electrode, although the correlation between the mass increase and activity of the preconcentrated material was not linear. Neptunium precipitate reduced electron transfer to the solution as evidenced by the increase in charge transfer resistance compared to bare electrodes.

'Candidatus Liberibacter asiaticus' is a fastidious bacterium and a putative agent of citrus greening disease (a.k.a., huanglongbing, HLB), a significant agricultural disease that affects citrus fruit quality and tree health. In citrus, 'Ca. L. asiaticus' is phloem limited. Lack of culture tools to study 'Ca. L. asiaticus' complicates analysis of this important organism. To improve understanding of 'Ca. L. asiaticus'-host interactions including parameters that affect 'Ca. L. asiaticus' replication, methods suitable for screening pathogen responses to physicochemical and nutritional variables are needed. We describe a leaf disc-based culture assay that allows highly selective measurement of changes in 'Ca. L. asiaticus' DNA within plant tissue incubated under specific physicochemical and nutritional conditions. qPCR analysis targeting the hypothetical gene CD16-00155 (strain A4) allowed selective quantification of 'Ca. L. asiaticus' DNA content within infected tissue. 'Ca. L. asiaticus' DNA replication was observed in response to glucose exclusively under microaerobic conditions, and the antibiotic amikacin further enhanced 'Ca. L. asiaticus' DNA replication. Metabolite profiling revealed a moderate impact of 'Ca. L. asiaticus' on the ability of leaf tissue to metabolize and respond to glucose., (© 2020 The Authors. Microbial Biotechnology published by Society for Applied Microbiology and John Wiley & Sons Ltd.)

The scale up of bioelectrochemical systems (BESs) is a challenging problem that limits the advancement and practical implementation of the technology. The goal of this work is to acquire an understanding of the limitations on scaling up anodic biofilms in BESs. We hypothesized that scaling up is dependent on the availability of electron donors. We tested this hypothesis by enriching anodic biofilms on electrodes of multiple sizes (15 cm 2 to 466 cm 2 ) and quantified the anodic current densities while varying the electron donor concentrations. The anodic biofilms were enriched on electrodes under two conditions: (1) in raw wastewater and (2) in wastewater supplemented with 20 mM acetate. Following anodic biofilm enrichment, the current density for each electrode was quantified in artificial wastewater medium with variable COD loadings using acetate as an electron donor. Current generated using anodic biofilms scaled up at a high COD loading (1500 mg/L), while current density decreased with increasing electrode size at lower COD loadings. Further, microbial community analysis revealed that the microbial community was independent of the electrode size but dependent on the medium composition during the enrichment phase. These results provide a practical framework for the design of large-scale BESs based on laboratory-scale measurements., 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 © 2019 Elsevier B.V. All rights reserved.)

The antibiofilm activity of a hydrogen peroxide-generating electrochemical scaffold (e-scaffold) was determined against mono- and trispecies biofilms of methicillin-resistant Staphylococcus aureus , multidrug-resistant Pseudomonas aeruginosa , and Candida albicans Significant time-dependent decreases were found in the overall CFU of biofilms of all three monospecies and the trispecies forms. Confocal laser scanning microscopy showed dramatic reductions in fluorescence intensities of biofilm matrix protein and polysaccharide components of e-scaffold-treated biofilms. The described e-scaffold has potential as a novel antibiotic-free strategy for treating wound biofilms., (Copyright © 2020 American Society for Microbiology.)

To understand electron transport in electrochemically active biofilms, it is necessary to elucidate the heterogeneous electron transport across the biofilm/electrode interface and in the interior of G. sulfurreducens biofilms bridging gaps of varying widths. The conductivity of Geobacter sulfurreducens biofilm bridging nonconductive gaps with widths of 5µm, 10µm, 20µm and 50µm is investigated. Results of electrochemical gating measurement show that biofilm conductivity peaks at the potential of -0.35V vs. Ag/AgCl. The biofilm conductivity increases with gap width (10.4±0.2µScm -1 in 5µm gap, 13.3±0.2µScm -1 in 10µm gap, 16.7±1.4µScm -1 in 20µm gap and 41.8±2.02µScm -1 in 50µm gap). These results revealed that electron transfer in G. sulfurreducens biofilm is a redox-driven. In addition, higher biofilm conductivities and lower charge transfer resistances are observed in all gaps under a turnover condition than in those under a non-turnover condition. Our results offer insights into the spatial heterogeneity of biofilm structure and extracellular electron transfer in electrochemically active biofilms., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Electroprecipitation can be used to preconcentrate lanthanum on carbon electrode surfaces. The use of complexing ligands is expected to improve the electroprecipitation of lanthanum by protecting La ions in solution from the alkaline region near the electrode surface. However, the electroprecipitation mechanism of La in the presence of a complexing ligand is not known. The goal of this work is to 1) determine the effect of the complexing ligand, α-hydroxy isobutyric acid (HIBA), on the electroprecipitation of La onto the gold electrodes, and 2) identify the changes in the mechanism of accumulation when preconcentrating in the presence of HIBA. We used an electrochemical quartz crystal microbalance (eQCM) and needle type pH microelectrodes to determine pH near the electrode surface in combination with cyclic voltammetry to understand the electroprecipitation mechanism. We used the bi-dentate ligand HIBA as a ligand and found that lanthanum electroprecipitation is hindered in the presence of HIBA. The presence of HIBA also delayed the onset of film formation during a cyclic voltammetric experiment by ~100 mV compared to experiments performed without HIBA. The shift in onset potential is attributed to the buffering action of HIBA (pK a = 3.7) since the shift is not present in subsequent scans. The precipitated film was characterized by scanning electron microscopy, X-ray photoelectron spectrometry, and Auger nanoprobe spectrometry. While we found that La(OH) 3 was the predominant chemical state of the film on electrodes in the absence of HIBA, La 2 O 3 was found for films created in the presence of HIBA. Our finding demonstrates that La(OH) 3 can be electrodeposited at room temperature., Competing Interests: Declaration of competing interest The authors have no competing interests to declare.

Plant pathogenic bacteria interact with their insect host(s)/vector(s) at the cellular and molecular levels. This interaction may alter the physiology of their insect vector, which may also promote the growth and transmission of the bacterium. Here we studied the effect of "Candidatus Liberibacter asiaticus" ("Ca. L. asiaticus") on physiochemical conditions within its insect vector, the Asian citrus psyllid (ACP), and whether these changes were beneficial for the pathogen. The local microenvironments inside ACPs were quantified using microelectrodes. The average hemolymph pH was significantly higher in infected ACPs (8.13 ± 0.21) than in "Ca. L. asiaticus"-free ACPs (7.29 ± 0.15). The average hemolymph oxygen tension was higher in "Ca. L. asiaticus"-free ACPs than in infected ACPs (67.13% ± 2.11% vs. 35.61% ± 1.26%). Oxygen tension reduction and pH increase were accompanied by "Ca. L. asiaticus" infection. Thus, oxygen tension of the hemolymph is an indicator of infection status, with pH affected by the severity of the infection.

Many bacterial and viral plant pathogens are transmitted by insect vectors, and pathogen-mediated alterations of plant physiology often influence insect vector behavior and fitness. It remains largely unknown for most plant pathogens whether, and how, they might directly alter the physiology of their insect vectors in ways that promote pathogen transmission. Here we examined whether the presence of "Candidatus Liberibacter solanacearum" ("Ca. L. solanacearum"), an obligate bacterial pathogen of plants and of its psyllid vector alters the physiochemical environment within its insect vector, the potato psyllid (Bactericera cockerelli). Microelectrodes were used to measure the local pH and oxygen tension within the abdomen of "Ca. L. solanacearum"-free psyllids and those infected with "Ca. L. solanacearum". The hemolymph of infected psyllids had higher pH at 9.09 ± 0.12, compared to "Ca. L. solanacearum"-free psyllids (8.32 ± 0.11) and a lower oxygen tension of 33.99% vs. 67.83%, respectively. The physicochemical conditions inside "Ca. L. solanacearum"-free and -infected psyllids body differed significantly with the infected psyllids having a higher hemolymph pH and lower oxygen tension than "Ca. L. solanacearum"-free psyllids. Notably, the bacterial titer increased under conditions of higher pH and lower oxygen tension values. This suggests that the vector's physiology is altered by the presence of the pathogen, potentially, resulting in a more conducive environment for "Ca. L. solanacearum" survival and subsequent transmission., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Inability to culture the phloem-restricted alpha-proteobacterium " Candidatus Liberibacter asiaticus" (" Ca. L. asiaticus") or the closely related species (" Candidatus Liberibacter americanus" and " Candidatus Liberibacter africanus") that are associated with Huanglongbing (HLB) hampers the development of effective long-term control strategies for this devastating disease. Here we report successful establishment and long-term maintenance of host-free " Ca. L. asiaticus" cultures, with the bacterium growing within cultured biofilms derived from infected citrus tissue. The biofilms were grown in a newly designed growth medium under specific conditions. The initial biofilm-based culture has been successfully maintained for over two years and has undergone over a dozen subcultures. Multiple independent cultures have been established and maintained in a biofilm reactor system, opening the door to the development of pure culture of "Ca. L. asiaticus " and the use of genetics-based methods to understand and mitigate the spread of HLB ., (© 2019 The Authors.)

In this work, contributions of extracellular polymeric substances (EPS) to the nanoscale mechanisms through which the multidrug-resistant Acinetobacter baumannii responds to antimicrobial and hyperosmotic treatments were investigated by atomic force microscopy. Specifically, the adhesion strengths to a control surface of silicon nitride (Si 3 N 4 ) and the lengths of bacterial surface biopolymers of bound and loose EPS extracted from A. baumannii biofilms were quantified after individual or synergistic treatments with hyperosmotic agents (NaCl and maltodextrin) and an antibiotic (tobramycin). In the absence of any treatment, the loose EPS were significantly longer in length and higher in adhesion to Si 3 N 4 than the bound EPS. When used individually, the hyperosmotic agents and tobramycin collapsed the A. baumannii bound and loose EPS. The combined treatment of maltodextrin with tobramycin collapsed only the loose EPS and did not alter the adhesion of both bound and loose EPS to Si 3 N 4 . In addition, the combined treatment was not as effective in collapsing the EPS molecules as when tobramycin was applied alone. Finally, the effects of treatments were dose-dependent. Altogether, our findings suggest that a sequential treatment could be effective in treating A. baumannii biofilms, in which a hyperosmotic agent is used first to collapse the EPS and limit the diffusion of nutrients into the biofilm, followed by the use of an antibiotic to kill the bacterial cells that escape from the biofilm because of starvation.

Increasing rates of chronic wound infections caused by antibiotic-resistant bacteria are a crisis in healthcare settings. Biofilms formed by bacterial communities in these wounds create a complex environment, enabling bacteria to persist, even with antibiotic treatment. Wound infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are major causes of morbidity in clinical practice. There is a need for new therapeutic interventions not based on antibiotics. Hydrogen peroxide (H 2 O 2 ) is a known antibacterial/antibiofilm agent, continuous delivery of which has been challenging. A conductive electrochemical scaffold (e-scaffold) is developed, which is composed of carbon fabric that electrochemically reduces dissolved oxygen into H 2 O 2 when polarized at -0.6 V Ag/AgCl , as a novel antibiofilm wound dressing material. In this study, the in vitro antibiofilm activity of the e-scaffold against MRSA is investigated. The developed e-scaffold efficiently eradicates MRSA biofilms, based on bacterial quantitation and ATP measurements. Moreover, imaging hinted at the possibility of cell-membrane damage as a mechanism of action. These results suggest that an H 2 O 2 -generating e-scaffold may be a novel platform for eliminating MRSA biofilms without using antibiotics and may be useful to treat chronic MRSA wound infections., Competing Interests: RP reports grants from CD Diagnostics, BioFire, Curetis, Merck, Hutchison Biofilm Medical Solutions, Accelerate Diagnostics, Allergan, and The Medicines Company. consultant to Curetis, Qvella, St. Jude, Beckman Coulter, Morgan Stanley, Heraeus Medical GmbH, CORMATRIX, Specific Technologies, Diaxonit, Selux Dx, GenMark Diagnostics, LBT Innovations Ltd, PathoQuest and Genentech; monies are paid to Mayo Clinic.

Biofilm formation causes prolonged wound infections due to the dense biofilm structure, differential gene regulation to combat stress, and production of extracellular polymeric substances. Acinetobacter baumannii, Staphylococcus aureus, and Pseudomonas aeruginosa are three difficult-to-treat biofilm-forming bacteria frequently found in wound infections. This work describes a novel wound dressing in the form of an electrochemical scaffold (e-scaffold) that generates controlled, low concentrations of hypochlorous acid (HOCl) suitable for killing biofilm communities without substantially damaging host tissue. Production of HOCl near the e-scaffold surface was verified by measuring its concentration using needle-type microelectrodes. E-scaffolds producing 17, 10 and 7 mM HOCl completely eradicated S. aureus, A. baumannii, and P. aeruginosa biofilms after 3 hours, 2 hours, and 1 hour, respectively. Cytotoxicity and histopathological assessment showed no discernible harm to host tissues when e-scaffolds were applied to explant biofilms. The described strategy may provide a novel antibiotic-free strategy for treating persistent biofilm-associated infections, such as wound infections.

A microfluidic platform for hydrodynamic electrochemical analysis was developed, consisting of a poly(methyl methacrylate) chip and three removable electrodes, each housed in 1/16" OD polyether ether ketone tube which can be removed independently for polishing or replacement. The working electrode was a 100-μm diameter Pt microdisk, located flush with the upper face of a 150 μm × 20 μm × 3 cm microchannel, smaller than previously reported for these types of removable electrodes. A commercial leak-less reference electrode was utilized, and a coiled platinum wire was the counter electrode. The platform was evaluated electrochemically by oxidizing a potassium ferrocyanide solution at the working electrode, and a typical limiting current behavior was observed after running linear sweep voltammetry and chronoamperometry, with flow rates 1-6 μL/min. While microdisk channel electrodes have been simulated before using a finite difference method in an ideal 3D geometry, here we predict the limiting current using finite elements in COMSOL Multiphysics 5.3a, which allowed us to easily explore variations in the microchannel geometry that have not previously been considered in the literature. Experimental and simulated currents showed the same trend but differed by 41% in simulations of the ideal geometry, which improved when channel and electrode imperfections were included.

The present study is focused on enhancing the rheological properties of the electrolyte and eliminating sedimentation of microorganisms/flocs without affecting the electron transfer kinetics for improved bioelectricity generation. Agar derived from polysaccharide agarose (0.05-0.2%, w/v) was chosen as a rheology modifying agent. Electroanalytical investigations showed that electrolytes modified with 0.15% agar display a nine-fold increase in current density (1.2 mA/cm 2 ) by a thermophilic strain (Geobacillus sp. 44C, 60 °C) when compared with the control. Sodium phosphate buffer (0.1 M, pH 7) electrolyte with riboflavin (0.1 mM) was used as the control. Electrolytes modified with 0.15% agar significantly improved chemical oxygen demand removal rates. This developed electrolyte will aid in improving bioelectricity generation in Bioelectrochemical Systems (BES). The developed strategy avoids the use of peristaltic pumps and magnetic stirrers, thereby improving the energy efficiency of the process., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Biofilms alter their metabolism in response to environmental stress. This study explores the effect of a hyperosmotic agent-antibiotic treatment on the metabolism of Staphylococcus aureus biofilms through the use of nuclear magnetic resonance (NMR) techniques. To determine the metabolic activity of S. aureus, we quantified the concentrations of metabolites in spent medium using high-resolution NMR spectroscopy. Biofilm porosity, thickness, biovolume, and relative diffusion coefficient depth profiles were obtained using NMR microimaging. Dissolved oxygen concentration was measured to determine the availability of oxygen within the biofilm. Under vancomycin-only treatment, the biofilm communities switched to fermentation under anaerobic condition, as evidenced by high concentrations of formate (7.4 ± 2.7 mM), acetate (13.1 ± 0.9 mM), and lactate (3.0 ± 0.8 mM), and there was no detectable dissolved oxygen in the biofilm. In addition, we observed the highest consumption of pyruvate (0.19 mM remaining from an initial 40 mM concentration), the sole carbon source, under the vancomycin-only treatment. On the other hand, relative effective diffusion coefficients increased from 0.73 ± 0.08 to 0.88 ± 0.08 under vancomycin-only treatment but decreased from 0.71 ± 0.04 to 0.60 ± 0.07 under maltodextrin-only and from 0.73 ± 0.06 to 0.56 ± 0.08 under combined treatments. There was an increase in biovolume, from 2.5 ± 1 mm 3 to 7 ± 1 mm 3 , under the vancomycin-only treatment, while the maltodextrin-only and combined treatments showed no significant change in biovolume over time. This indicated that physical biofilm growth was halted during maltodextrin-only and combined treatments., (© 2018 Wiley Periodicals, Inc.)

Biofilm-associated infections are a clinical challenge, in part because a hydrated matrix protects the bacterial community from antibiotics. Herein, we evaluated how different osmotic compounds (maltodextrin, sucrose, and polyethylene glycol [PEG]) enhance antibiotic efficacy against Acinetobacter baumannii biofilm communities. Established (24-h) test tube biofilms (strain ATCC 17978) were treated with osmotic compounds in the presence or absence of 10× the MIC of different antibiotics (50 μg/ml tobramycin, 20 μg/ml ciprofloxacin, 300 μg/ml chloramphenicol, 30 μg/ml nalidixic acid, or 100 μg/ml erythromycin). Combining antibiotics with hypertonic concentrations of the osmotic compounds for 24 h reduced the number of biofilm bacteria by 5 to 7 log ( P < 0.05). Increasing concentrations of osmotic compounds improved the effect, but there was a trade-off with increasing solution viscosity, whereby low-molecular-mass compounds (sucrose, 400-Da PEG) worked better than higher-mass compounds (maltodextrin, 3,350-Da PEG). Ten other A. baumannii strains were similarly treated with 400-Da PEG and tobramycin, resulting in a mean 2.7-log reduction in recoverable bacteria compared with tobramycin treatment alone. Multivariate regression models with data from different osmotic compounds and nine antibiotics demonstrated that the benefit from combining hypertonic treatments with antibiotics is a function of antibiotic mass and lipophilicity ( r 2 > 0.82; P < 0.002), and the relationship was generalizable for biofilms formed by A. baumannii and Escherichia coli K-12. Augmenting topical antibiotic therapies with a low-mass hypertonic treatment may enhance the efficacy of antibiotics against wound biofilms, particularly when using low-mass hydrophilic antibiotics. IMPORTANCE Biofilms form a barrier that protects bacteria from environmental insults, including exposure to antibiotics. We demonstrated that multiple osmotic compounds can enhance antibiotic efficacy against Acinetobacter baumannii biofilm communities, but viscosity is a limiting factor, and the most effective compounds have lower molecular mass. The synergism between osmotic compounds and antibiotics is also dependent on the hydrophobicity and mass of the antibiotics. The statistical models presented herein provide a basis for predicting the optimal combination of osmotic compounds and antibiotics against surface biofilms communities., (Copyright © 2017 American Society for Microbiology.)

Development of high-performance nanomaterials with not only strong ability to trap the pollutants but also good structure stability under varying environmental conditions is a critical need for air-filtration applications. However, it has been very challenging for a filtering material to simultaneously realize multifunctional air filtration and good environmental stability. Here, based on our previous studies on protein-based nanofilters, we report a cross-linked protein nanofabric to address this challenging issue. It is found that cross-linked protein nanofabrics can significantly improve the structure stability against different moisture levels and temperatures, while maintaining the multifunctional filtration performance. Moreover, it is demonstrated that the cross-linked protein nanomaterials also possess antibacterial properties, such as Shewanella oneidensis bacteria, further improving the environmental stability. The effects of cross-linking with different loadings of cross-linking agent on the structure stability and filtration performance are further investigated at different humidity levels and temperatures. This study provides a cost-effective solution for advanced "green" nanomaterials with excellent performance in both filtration functions and structure stability under varying environment.

Phototrophic mat communities are model ecosystems for studying energy cycling and elemental transformations because complete biogeochemical cycles occur over millimeter-to-centimeter scales. Characterization of energy and nutrient capture within hypersaline phototrophic mats has focused on specific processes and organisms; however, little is known about community-wide distribution of and linkages between these processes. To investigate energy and macronutrient capture and flow through a structured community, the spatial and organismal distribution of metabolic functions within a compact hypersaline mat community from Hot Lake have been broadly elucidated through species-resolved metagenomics and geochemical, microbial diversity and metabolic gradient measurements. Draft reconstructed genomes of 34 abundant organisms revealed three dominant cyanobacterial populations differentially distributed across the top layers of the mat suggesting niche separation along light and oxygen gradients. Many organisms contained diverse functional profiles, allowing for metabolic response to changing conditions within the mat. Organisms with partial nitrogen and sulfur metabolisms were widespread indicating dependence on metabolite exchange. In addition, changes in community spatial structure were observed over the diel. These results indicate that organisms within the mat community have adapted to the temporally dynamic environmental gradients in this hypersaline mat through metabolic flexibility and fluid syntrophic interactions, including shifts in spatial arrangements., (© FEMS 2017.)

Biofilms on wound surfaces are treated topically with hyperosmotic agents, such as medical-grade honey and cadexomer iodine; in some cases, these treatments are combined with antibiotics. Tissue repair requires oxygen, and a low pH is conducive to oxygen release from red blood cells and epithelialization. We investigated the variation of dissolved oxygen concentration and pH with biofilm depth and the variation in oxygen consumption rates when biofilms are challenged with medical-grade honey or cadexomer iodine combined with vancomycin or ciprofloxacin. Dissolved oxygen and pH depth profiles in Staphylococcus aureus biofilms were measured using microelectrodes. The presence of cadexomer iodine with vancomycin or ciprofloxacin on the surface of the biofilm permitted a measurable concentration of oxygen at greater biofilm depths (101.6 ± 27.3 μm, P = 0.02; and 155.5 ± 27.9 μm, P = 0.016, respectively) than in untreated controls (30.1 μm). Decreases in pH of ∼0.6 and ∼0.4 units were observed in biofilms challenged with medical-grade honey alone and combined with ciprofloxacin, respectively ( P < 0.001 and 0.01, respectively); the number of bacteria recovered from biofilms was significantly reduced (1.26 log) by treatment with cadexomer iodine and ciprofloxacin ( P = 0.002) compared to the untreated control. Combining cadexomer iodine and ciprofloxacin improved dissolved oxygen concentration and penetration depth into the biofilm, while medical-grade honey was associated with a lower pH; not all treatments established a bactericidal effect in the time frame used in the experiments. IMPORTANCE Reports about using hyperosmotic agents and antibiotics against wound biofilms focus mostly on killing bacteria, but the results of these treatments should additionally be considered in the context of how they affect physiologically important parameters, such as oxygen concentration and pH. We confirmed that the combination of a hyperosmotic agent and an antibiotic results in greater dissolved oxygen and reduced pH within an S. aureus biofilm., (Copyright © 2017 American Society for Microbiology.)

Productivity is a major determinant of ecosystem diversity. Microbial ecosystems are the most diverse on the planet yet very few relationships between diversity and productivity have been reported as compared with macro-ecological studies. Here we evaluated the spatial relationships of productivity and microbiome diversity in a laboratory-cultivated photosynthetic mat. The goal was to determine how spatial diversification of microorganisms drives localized carbon and energy acquisition rates. We measured sub-millimeter depth profiles of net primary productivity and gross oxygenic photosynthesis in the context of the localized microenvironment and community structure, and observed negative correlations between species richness and productivity within the energy-replete, photic zone. Variations between localized community structures were associated with distinct taxa as well as environmental profiles describing a continuum of biological niches. Spatial regions in the photic zone corresponding to high primary productivity and photosynthesis rates had relatively low-species richness and high evenness. Hence, this system exhibited negative species-productivity and species-energy relationships. These negative relationships may be indicative of stratified, light-driven microbial ecosystems that are able to be the most productive with a relatively smaller, even distributions of species that specialize within photic zones.

Microbial phototrophs, key primary producers on Earth, use H 2 O, H 2 , H 2 S and other reduced inorganic compounds as electron donors. Here we describe a form of metabolism linking anoxygenic photosynthesis to anaerobic respiration that we call 'syntrophic anaerobic photosynthesis'. We show that photoautotrophy in the green sulfur bacterium Prosthecochloris aestaurii can be driven by either electrons from a solid electrode or acetate oxidation via direct interspecies electron transfer from a heterotrophic partner bacterium, Geobacter sulfurreducens. Photosynthetic growth of P. aestuarii using reductant provided by either an electrode or syntrophy is robust and light-dependent. In contrast, P. aestuarii does not grow in co-culture with a G. sulfurreducens mutant lacking a trans-outer membrane porin-cytochrome protein complex required for direct intercellular electron transfer. Syntrophic anaerobic photosynthesis is therefore a carbon cycling process that could take place in anoxic environments. This process could be exploited for biotechnological applications, such as waste treatment and bioenergy production, using engineered phototrophic microbial communities.

The goal of this work was to synthesize gold nanoparticles (AuNPs) using electrode-respiring Geobacter sulfurreducens biofilms. We found that AuNPs are generated in the extracellular matrix of Geobacter biofilms and have an average particle size of 20nm. The formation of AuNPs was verified using TEM, FTIR and EDX. We also found that the extracellular substances extracted from electrode-respiring G. sulfurreducens biofilms reduce Au 3+ to AuNPs. From FTIR spectra, it appears that reduced sugars were involved in the bioreduction and synthesis of AuNPs and that amine groups acted as the major biomolecules involved in binding., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The biological synthesis of nanoparticles (NPs) by bacteria and biofilms via extracellular redox reactions has received attention because of the minimization of harmful chemicals, low cost, and ease of culturing and downstream processing. Bioreduction mechanisms vary across bacteria and growth conditions, which leads to various sizes and shapes of biosynthesized NPs. NP synthesis in biofilms offers additional advantages, such as higher biomass concentrations and larger surface areas, which can lead to more efficient and scalable biosynthesis. Although biofilms have been used to produce NPs, the mechanistic details of NP formation are not well understood. In this review, we identify three critical areas of research and development needed to advance our understanding of NP production by biofilms: 1) synthesis, 2) mechanism and 3) stabilization. Advancement in these areas could result in the biosynthesis of NPs that are suitable for practical applications, especially in drug delivery and biocatalysis. Specifically, the current status of methods and mechanisms of nanoparticle synthesis and surface stabilization using planktonic bacteria and biofilms is discussed. We conclude that the use of biofilms to synthesize and stabilize NPs is underappreciated and could provide a new direction in biofilm-based NP production., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Biofilms in chronic wounds are known to contain a persister subpopulation that exhibits enhanced multidrug tolerance and can quickly rebound after therapeutic treatment. The presence of these "persister cells" is partly responsible for the failure of antibiotic therapies and incomplete elimination of biofilms. Electrochemical methods combined with antibiotics have been suggested as an effective alternative for biofilm and persister cell elimination, yet the mechanism of action for improved antibiotic efficacy remains unclear. In this work, an electrochemical scaffold (e-scaffold) that electrochemically generates a constant concentration of H 2 O 2 was investigated as a means of enhancing tobramycin susceptibility in pre-grown Pseudomonas aeruginosa PAO1 biofilms and attacking persister cells. Results showed that the e-scaffold enhanced tobramycin susceptibility in P. aeruginosa PAO1 biofilms, which reached a maximum susceptibility at 40 µg/ml tobramycin, with complete elimination (7.8-log reduction vs control biofilm cells, P  ≤ 0.001). Moreover, the e-scaffold eradicated persister cells in biofilms, leaving no viable cells (5-log reduction vs control persister cells, P  ≤ 0.001). It was observed that the e-scaffold induced the intracellular formation of hydroxyl free radicals and improved membrane permeability in e-scaffold treated biofilm cells, which possibly enhanced antibiotic susceptibility and eradicated persister cells. These results demonstrate a promising advantage of the e-scaffold in the treatment of persistent biofilm infections.