Supplementation with Amino Acid Sources Facilitates Fermentative Growth of Shewanella oneidensis MR-1 in Defined Media.

Shewanella oneidensis MR-1 is a facultative anaerobe that grows by respiration using a variety of electron acceptors. This organism serves as a model to study how bacteria thrive in redox-stratified environments. A glucose-utilizing engineered derivative of MR-1 has been reported to be unable to grow in glucose minimal medium (GMM) in the absence of electron acceptors, despite this strain having a complete set of genes for reconstructing glucose to lactate fermentative pathways. To gain insights into why MR-1 is incapable of fermentative growth, this study examined a hypothesis that this strain is programmed to repress the expression of some carbon metabolic genes in the absence of electron acceptors. Comparative transcriptomic analyses of the MR-1 derivative were conducted in the presence and absence of fumarate as an electron acceptor, and these found that the expression of many genes involved in carbon metabolism required for cell growth, including several tricarboxylic acid (TCA) cycle genes, was significantly downregulated in the absence of fumarate. This finding suggests a possibility that MR-1 is unable to grow fermentatively on glucose in minimal media owing to the shortage of nutrients essential for cell growth, such as amino acids. This idea was demonstrated in subsequent experiments that showed that the MR-1 derivative fermentatively grows in GMM containing tryptone or a defined mixture of amino acids. We suggest that gene regulatory circuits in MR-1 are tuned to minimize energy consumption under electron acceptor-depleted conditions, and that this results in defective fermentative growth in minimal media. IMPORTANCE It is an enigma why S. oneidensis MR-1 is incapable of fermentative growth despite having complete sets of genes for reconstructing fermentative pathways. Understanding the molecular mechanisms behind this defect will facilitate the development of novel fermentation technologies for the production of value-added chemicals from biomass feedstocks, such as electro-fermentation. The information provided in this study will also improve our understanding of the ecological strategies of bacteria living in redox-stratified environments.

Improved electrochemical properties of graphite electrodes incubated with iron powders in rice-paddy fields boost power outputs from microbial fuel cells.

Studies have shown that the supplementation of anode-surrounding soil with zero-valent iron (ZVI) boosts power outputs from rice paddy-field microbial fuel cells (RP-MFCs). In order to understand mechanisms by which ZVI boosts outputs from RP-MFCs, the present study operated RP-MFCs with and without ZVI, and compositions of anode-associated bacteria and electrochemical properties of graphite anodes were analyzed after 3-month operation. Metabarcoding using 16S rRNA gene fragments showed that bacterial compositions did not largely differ among these RP-MFCs. Cyclic voltammetry showed improved electrochemical properties of anodes recovered from ZVI-supplemented RP-MFCs, and this was attributed to the adhesion of iron-oxide films onto graphite surfaces. Bioelectrochemical devices equipped with graphite anodes recovered from ZVI-supplemented RP-MFCs generated higher currents than those with fresh graphite anodes. These results suggest that ZVI is oxidized to iron oxides in paddy-field soil and adheres onto graphite anodes, resulting in the boost of power outputs from RP-MFCs.

A survey of physical and occupational therapists' views on lumbar loading movements.

[Purpose] To identify the lumbar loading movements necessary in clinical practice. [Participants and Methods] A questionnaire survey was conducted among physical and occupational therapists in Japan. There were no exclusion criteria regarding the number of years of experience, age, or field of employment. The participants were randomly selected and administered the questionnaire. They were asked to list and rank the lumbar loadings they considered necessary. [Results] A total of 739 respondents participated in the survey. The results of this nationwide survey indicated that the lifting movement of heavy objects in the trunk flexion position was the most common movement (for 354 participants). [Conclusion] The main loading movements of the lumbar spine were reported to be heavy lifting movements (in the trunk flexion position) and trunk rotation movements. As perspectives, we aim to conduct an analytical study of some of lumbar spine loading movements outlined in this study, using a musculoskeletal simulator and electromyography.

Identification of a Diguanylate Cyclase That Facilitates Biofilm Formation on Electrodes by Shewanella oneidensis MR-1.

In many bacteria, cyclic diguanosine monophosphate (c-di-GMP), synthesized by diguanylate cyclase (DGC), serves as a second messenger involved in the regulation of biofilm formation. Although studies have suggested that c-di-GMP also regulates the formation of electrochemically active biofilms (EABFs) by Shewanella oneidensis MR-1, DGCs involved in this process remained to be identified. Here, we report that the SO_1646 gene, hereafter named dgcS, is upregulated under medium flow conditions in electrochemical flow cells (EFCs), and its product (DgcS) functions as a major DGC in MR-1. In vitro assays demonstrated that purified DgcS catalyzed the synthesis of c-di-GMP from GTP. Comparisons of intracellular c-di-GMP levels in the wild-type strain and a dgcS deletion mutant (ΔdgcS mutant) showed that production of c-di-GMP was markedly reduced in the ΔdgcS mutant when cells were grown in batch cultures and on electrodes in EFCs. Cultivation of the ΔdgcS mutant in EFCs also revealed that the loss of DgcS resulted in impaired biofilm formation and decreased current generation. These findings demonstrate that MR-1 uses DgcS to synthesize c-di-GMP under medium flow conditions, thereby activating biofilm formation on electrodes.IMPORTANCE Bioelectrochemical systems (BESs) have attracted wide attention owing to their utility in sustainable biotechnology processes, such as microbial fuel cells and electrofermentation systems. In BESs, electrochemically active bacteria (EAB) form biofilms on electrode surfaces, thereby serving as effective catalysts for the interconversion between chemical and electric energy. It is therefore important to understand mechanisms for the formation of biofilm by EAB grown on electrodes. Here, we show that a model EAB, S. oneidensis MR-1, expresses DgcS as a major DGC, thereby activating the formation of biofilms on electrodes via c-di-GMP-dependent signal transduction cascades. The findings presented herein provide the molecular basis for improving electrochemical interactions between EAB and electrodes in BESs. The results also offer molecular insights into how Shewanella regulates biofilm formation on solid surfaces in the natural environment.

Identification of an extracytoplasmic function sigma factor that facilitates c-type cytochrome maturation and current generation under electrolyte-flow conditions in Shewanella oneidensis MR-1.

Shewanella oneidensis MR-1 was cultured on electrodes in electrochemical flow cells (EFCs), and transcriptome profiles of electrode-attached cells grown under electrolyte-flow conditions were compared with those under static (nonflow) conditions. Results revealed that, along with genes related to c-type cytochrome maturation (e.g., dsbD), the SO_3096 gene encoding a putative extracytoplasmic function (ECF) sigma factor was significantly upregulated under electrolyte-flow conditions. Compared to wild-type MR-1 (WT), an SO_3096-deletion mutant (∆SO_3096) showed impaired biofilm formation and decreased current generation in EFCs, suggesting that SO_3096 plays critical roles in the interaction of MR-1 cells with electrodes under electrolyte-flow conditions. We also compared transcriptome profiles of WT and ∆SO_3096 grown in EFCs, confirming that many genes upregulated under the electrolyte-flow conditions, including dsbD, are regulated by SO_3096. LacZ reporter assays showed that transcription from a promoter upstream of dsbD is activated by SO_3096. Measurement of current generated by a dsbD-deletion mutant revealed that this gene is essential for the transfer of electrons to electrodes. These results indicate that the SO_3096 gene product facilitates c-type cytochrome maturation and current generation under electrolyte-flow conditions. The results also offer ecophysiological insights into how Shewanella regulates extracellular electron transfer to solid surfaces in the natural environment.

Linewidth Narrowing with Ultimate Confinement of an Alkali Multipole Plasmon by Modifying Surface Electronic Wave Functions with Two-Dimensional Materials.

This work demonstrates significant line narrowing of a surface multipole plasmon (MP) by modifying the surface electronic wave function with two-dimensional materials (2DMs): graphene and hexagonal boron nitride. This is found in an optical reflectivity of alkali atoms (Cs or K) on an Ir(111) surface covered with the 2DMs. The reduction in reflectivity induced by deposition of the alkali atoms becomes as large as 20% at ∼2 eV, which is ascribed to a MP of a composite of alkali/2DM/alkali/Ir multilayer structure. The linewidth of the MP band becomes as narrow as 0.2 eV by the presence of the 2DM between the two alkali layers. A numerical simulation by time-dependent density functional theory with a jellium model reveals that the density of states of the surface localized state is sharpened remarkably by the 2DMs that decouple the outermost alkali layer from the Ir bulk. Consequently, a local field enhancement of an order of 10^{5} is achieved by ultimate confinement of the MP within the outermost alkali layer. This work leads to a novel strategy for reducing plasmon dissipation in an atomically thin layer via atomic scale modification of surface structure.

Thickness dependent homogeneous crystallization of ultrathin amorphous solid water films.

The crystallization mechanism and kinetics of amorphous materials are of paramount importance not only in basic science but also in the application field because they are closely related to their thermal stability. In the case of amorphous nanomaterials, thermal stability distinctively different from that of bulk materials often emerges. Despite intensive studies in the past, a thorough understanding of the stability at the molecular level has not been reached particularly on how crystallization processes depend on size and are influenced by their surface and interface. In this article, we report the film-size-dependent crystallization of thermally relaxed nonporous ASW ultrathin films on a Pt(111) surface as a benchmark system of amorphous molecular films. The crystallization processes at the surface and interior of the ASW ultrathin films are monitored simultaneously with thermal desorption and infrared reflection absorption, respectively, as a function of the film thickness. Here, we demonstrate that the crystallization is initiated solely by "homogeneous nucleation" irrespective of the film thickness while the crystallization rate remarkably depends on the thickness; the rate of 5-layer (∼1.5 nm) ASW films is one order of magnitude higher than that of 20-layer (∼6 nm) films. Moreover, we found a clear correlation between the film-thickness-dependent crystallization kinetics and microscopic structural disorder associated with the broad distribution of hydrogen-bond lengths between water molecules.

Differences in the Clinical Characteristics of Persistent Idiopathic Facial Pain (Atypical Odontalgia) Patients with or Without Neurovascular Compression of the Trigeminal Nerve.

BACKGROUND: Persistent idiopathic facial pain (PIFP) is the unexplained pain along the territory of the trigeminal nerve, including nonorganic tooth pain called atypical odontalgia (AO). Though PIFP is debilitating to patients' livelihood and well-being, its pathophysiology remains poorly understood. Although neurovascular compression (NVC) of the trigeminal nerve is known to be associated with trigeminal neuralgia (TN), the relationship between NVC and other orofacial pains has not been fully elucidated. METHODS: In this study, we investigated the differences in the characteristics of PIFP (primarily AO) patients in the presence or absence of NVC. A retrospective analysis was performed on data from 121 consecutive patients who had been diagnosed with unilateral PIFP according to the criteria of the International Classification of Headache Disorders (ICHD)-3 and underwent magnetic resonance imaging scans of the head. RESULTS: In the group without NVC, characteristic findings were significant for psychiatric morbidity, somatization, and pain disability, when compared with the group with NVC. Furthermore, the group without NVC exhibited significant headache, noncardiac chest pain, shortness of breath, and pain catastrophizing. CONCLUSIONS: These results suggest that PIFP patients can be divided into two groups: one consistent with a neuropathic pain phenotype when NVC is present and a functional somatic symptom phenotype when presenting without NVC. Our findings may enable a more precise understanding of pathophysiology of PIFP and lead to better treatment strategies.

Metatranscriptomic Evidence for Magnetite Nanoparticle-Stimulated Acetoclastic Methanogenesis under Continuous Agitation.

Conductive nanomaterials have been reported to accelerate methanogenesis by promoting direct interspecies electron transfer (DIET), while their effects seem to vary depending on operational conditions. The present study examined the effects of magnetite nanoparticles (MNPs) on methanogenesis from acetate by soil-derived anaerobic cultures under continuous agitation. We found that MNPs accelerated methanogenesis in agitated cultures, as has been observed previously for static cultures. Metabarcoding of 16S rRNA gene amplicons showed that Methanosarcina substantially increased in the presence of MNPs, while DIET-related Geobacter did not occur. Metagenomic and metatranscriptomic analyses confirmed the predominance of Methanosarcina in MNP-supplemented agitated cultures. In addition, genes coding for acetoclastic methanogenesis, but not those for hydrogenotrophic methanogenesis, were abundantly expressed in the dominant Methanosarcina in the presence of MNPs. These results suggest that MNPs stimulate acetoclastic methanogenesis under continuous agitation.IMPORTANCE Previous studies have shown that conductive nanoparticles, such as MNPs, accelerate methanogenesis and suggested that MNPs facilitate DIET between exoelectrogenic bacteria and methanogenic archaea. In these methanogens, electrons thus obtained are considered to be used for hydrogenotrophic methanogenesis. However, the present work provides evidence that shows that MNPs accelerate DIET-independent acetoclastic methanogenesis under continuous agitation. Since most of previous studies have examined effects of MNPs in static or weakly agitated methanogenic cultures, results obtained in the present work suggest that hydraulic conditions definitively determine how MNPs accelerate methanogenesis. In addition, the knowledge obtained in this study is useful for engineers operating stirred-tank anaerobic digesters, since we show that MNPs accelerate methanogenesis under continuous agitation.

Roles of d-Lactate Dehydrogenases in the Anaerobic Growth of Shewanella oneidensis MR-1 on Sugars.

Shewanella oneidensis MR-1 is a facultative anaerobe that respires using a variety of electron acceptors. Although this organism is incapable of fermentative growth in the absence of electron acceptors, its genome encodes LdhA (a putative fermentative NADH-dependent d-lactate dehydrogenase [d-LDH]) and Dld (a respiratory quinone-dependent d-LDH). However, the physiological roles of LdhA in MR-1 are unclear. Here, we examined the activity, transcriptional regulation, and traits of deletion mutants to gain insight into the roles of LdhA in the anaerobic growth of MR-1. Analyses of d-LDH activity in MR-1 and the ldhA deletion mutant confirmed that LdhA functions as an NADH-dependent d-LDH that catalyzes the reduction of pyruvate to d-lactate. In vivo and in vitro assays revealed that ldhA expression was positively regulated by the cyclic-AMP receptor protein, a global transcription factor that regulates anaerobic respiratory pathways in MR-1, suggesting that LdhA functions in coordination with anaerobic respiration. Notably, we found that a deletion mutant of all four NADH dehydrogenases (NDHs) in MR-1 (ΔNDH mutant) retained the ability to grow on N-acetylglucosamine under fumarate-respiring conditions, while an additional deletion of ldhA or dld deprived the ΔNDH mutant of this growth ability. These results indicate that LdhA-Dld serves as a bypass of NDH in electron transfer from NADH to quinones. Our findings suggest that the LdhA-Dld system manages intracellular redox balance by utilizing d-lactate as a temporal electron sink under electron acceptor-limited conditions.IMPORTANCE NADH-dependent LDHs are conserved among diverse organisms and contribute to NAD+ regeneration in lactic acid fermentation. However, this type of LDH is also present in nonfermentative bacteria, including members of the genus Shewanella, while their physiological roles in these bacteria remain unknown. Here, we show that LdhA (an NADH-dependent d-LDH) works in concert with Dld (a quinone-dependent d-LDH) to transfer electrons from NADH to quinones during sugar catabolism in S. oneidensis MR-1. Our results indicate that d-lactate acts as an intracellular electron mediator to transfer electrons from NADH to membrane quinones. In addition, d-lactate serves as a temporal electron sink when respiratory electron acceptors are not available. Our study suggests novel physiological roles for d-LDHs in providing nonfermentative bacteria with catabolic flexibility under electron acceptor-limited conditions.

Enhanced structural disorder at a nanocrystalline ice surface.

Detailed knowledge of the structure and dynamics of the surface of ice particles is of considerable importance for understanding catalytic reactions in the upper atmosphere. Here we report the enhanced structural disorder specific at a nanoscale ice island studied by using heterodyne-detected vibrational sum-frequency generation spectroscopy under ultrahigh vacuum. Ultrathin films of isotopically diluted HOD crystalline ice are grown on Rh(111), whose average height (≥1.4 nm) is controlled by varying the nominal film thickness. The Im χ(2) spectrum of the hydrogen-bonded O-H stretching band shows a bipolar line shape reflecting the orientation-dependent hydrogen bond length alternation in the subsurface of the ice island. The peak splitting and the bandwidth of the bipolar spectrum increase with a decrease in the nominal film thickness. This is ascribed to the significant enhancement of structural disorder at the surface of the ice island as the terrace size is decreased. Temperature dependence of the Im χ(2) spectra of the hydrogen bonded O-H stretching band indicates that the thermal expansivity of the top layer increases upon decreasing the island size. In addition, the stretching frequency of the dangling OD band at the island surface with the average height less than 18 nm shows a systematic blue shift with increasing temperature from 100 to 145 K: this is in stark contrast to thick ice films and bulk ice showing a negligible peak shift at a temperature lower than 180 K. These findings indicate that the anharmonicity of the intermolecular potential at the top layer of the ice island is strongly enhanced upon decreasing its terrace size, providing valuable insights for understanding the properties of ice particles in the outer atmosphere including polar mesospheric clouds.

Conductive carbon nanoparticles inhibit methanogens and stabilize hydrogen production in microbial electrolysis cells.

Nanosized conductive carbon materials have been reported to stimulate methanogenesis by anaerobic microbiomes, while other studies have shown their antimicrobial activities. The present study examined effects of conductive carbon nanoparticles (carbon black Vulcan, CB) on methanogenesis from glucose by anaerobic sludge. We found that a relatively high concentration (e.g., 2% w/v) of CB entirely inhibited the methanogenesis, where a substantial amount of acetate was accumulated after degradation of glucose. Quantitative real-time PCR assays and metabarcoding of 16S rRNA amplicons revealed that, while bacteria were stably present irrespective of the presence and absence of CB, archaea, in particular methanogens, were largely decreased in the presence of CB. Pure-culture experiments showed that methanogenic archaea were more seriously damaged by CB than fermentative bacteria. These results demonstrate that CB specifically inhibits methanogens in anaerobic sludge. We attempted to supplement cathode chambers of microbial electrolysis cells with CB for inhibiting methanogenesis from hydrogen, demonstrating that hydrogen is stably produced in the presence of CB.

Singlet fission of amorphous rubrene modulated by polariton formation.

The excited-state dynamics of molecular aggregates are governed by their potential energy landscape that can hardly be controlled artificially. However, it is possible to alter the excited state dynamics by a strong coupling between light and molecules (polariton formation) because it can decouple the electronic and vibrational degrees of freedom. Here, we demonstrate this polaron decoupling effect on the photochemical dynamics in singlet fission (SF) of amorphous rubrene thin films embedded in optical microcavities. The vibronic feature of polariton states in this system is characterized through the analysis of steady state absorption spectra by using the Holstein-Tavis-Cummings model. On the basis of this analysis, we show with time-resolved spectroscopy that the SF rate following a resonant excitation of the lowest energy polariton state is indeed modulated when the cavity photon energy is changed. A numerical simulation by using Fermi's golden rule formula with the vibronic polariton feature successfully accounts for the observed modulation of the SF rate, indicating that the polaron decoupling plays a decisive role in the nonadiabatic dynamics.

Water-Assisted Hole Trapping at the Highly Curved Surface of Nano-TiO2 Photocatalyst.

Heterogeneous photocatalysis is vital in solving energy and environmental issues that this society is confronted with. Although photocatalysts are often operated in the presence of water, it has not been yet clarified how the interaction with water itself affects charge dynamics in photocatalysts. Using water-coverage-controlled steady and transient infrared absorption spectroscopy and large-model (∼800 atoms) ab initio calculations, we clarify that water enhances hole trapping at the surface of TiO2 nanospheres but not of well-faceted nanoparticles. This water-assisted effect unique to the nanospheres originates from water adsorption as a ligand at a low-coordinated Ti-OH site or through robust hydrogen bonding directly to the terminal OH at the highly curved nanosphere surface. Thus, the interaction with water at the surface of nanospheres can promote photocatalytic reactions of both oxidation and reduction by elongating photogenerated carrier lifetimes. This morphology-dependent water-assisted effect provides a novel and rational basis for designing and engineering nanophotocatalyst morphology to improve photocatalytic performances.

Methylomusa anaerophila gen. nov., sp. nov., an anaerobic methanol-utilizing bacterium isolated from a microbial fuel cell.

Abacterial strain, designated MMFC1T, was isolated from a methanol-fed microbial fuel cell that had been inoculated with sludge obtained from a wastewater-treatmentfacility in a chemical plant. The strain grows by fermenting methanol to produce acetate under anaerobic conditions, while homoacetogenic growth is not observed. MMFC1T also grows on pyruvate and lactate but not on sugars and other organic acids. Cells are curved rods and motile, have peritrichous flagella, and form endospores. The genome sequence of strain MMFC1T supports the physiological data. Phylogenetic analysis based on the 16S rRNA gene sequence shows that strain MMFC1T is affiliated with the family Sporomusaceae, while the closest relative is Sporomusa ovata with nucleotide-sequencesimilarity of 93.5 %. Major fatty acids are iso-C13 : 0 3-OH, C16 : 1ω9 and iso-C17 : 0. On the basis of its physiological, genomic and phylogenetic features, a novel genus and species are proposed to accommodate strain MMFC1T, with the name Methylomusa anaerophila gen. nov., sp. nov. The type strain of Methylomusa anaerophila is MMFC1T (=JCM 31821T = KCTC 15592T).

CpdA is involved in amino acid metabolism in Shewanella oneidensis MR-1.

Cyclic 3',5'-adenosine monophosphate (cAMP) phosphodiesterase (CPD) is an enzyme that catalyzes the hydrolysis of cAMP, a signaling molecule affecting diverse cellular and metabolic processes in bacteria. Some CPDs are also known to function in cAMP-independent manners, while their physiological roles remain largely unknown. Here, we investigated physiological roles of CPD in Shewanella oneidensis MR-1, a model environmental bacterium, and report that CPD is involved in amino-acid metabolism. We found that a CPD-deficient mutant of MR-1 (ΔcpdA) showed decreased expression of genes for the synthesis of methionine, S-adenosylmethionine, and histidine and required these three compounds to grow in minimal media. Interestingly, deletion of adenylate cyclases in ΔcpdA did not restore the ability to grow in minimal media, indicating that the amino acid requirements were not due to the accumulation of cAMP. These results suggest that CPD is involved in the regulation of amino acid metabolism in MR-1 in a cAMP-independent manner.

Dissolved organic matter characteristics along sabo dammed streams based on ultraviolet visible and fluorescence spectral properties.

Changes in dissolved organic matter (DOM) characteristics were investigated in two mountainous streams with closed-type sabo dams. Surface water was collected from four stations along the two mountainous streams and analyzed using ultraviolet-visible spectrophotometry and excitation-emission fluorescence matrix (EEM) methods. Optical properties of DOM indicated an increase in molecular weight and aromaticity at stations near the sabo dams. Average spectral ratio values were low before and after the dam (i.e., < 0.72) compared to other sections of the stream. Specific ultraviolet absorbance (SUVA254) increased in the vicinities of the dams. While chromophoric DOM characteristics from two sites were influenced by the dam, fluorescence components, however, did not show notable changes around dams. Instead, the three chromophoric components distinguished by EEM-parallel factor analysis, that is, humic-like (C1 and C2) and protein-like (C3) increase along the stream. Fulvic-like component (C1) had a high fluorescence intensity at all stations; all the three components were more abundant in the downstream section. Chromophoric DOM properties varied along the stream based on alterations in molecular size and aromaticity. Using multivariate analysis, the studied sites were grouped into three clusters related to sabo dams and other activities. We conclude that sabo dams modify DOM characteristics which influence the behavior of DOM transported along the stream.

Conformational Planarization versus Singlet Fission: Distinct Excited-State Dynamics of Cyclooctatetraene-Fused Acene Dimers.

A set of flapping acene dimers fused with an 8π cyclooctatetraene (COT) ring showed distinct excited-state dynamics in solution. While the anthracene dimer showed a fast V-shaped-to-planar conformational change within 10 ps in the lowest excited singlet state, reminding us of extended Baird aromaticity, the tetracene dimer and the pentacene dimer underwent intramolecular singlet fission (SF) in different manners: A fast and reversible SF with a characteristic delayed fluorescence (FL), and a fast and quantitative SF, respectively. Conformational flexibility of the fused COT linkage plays an important role in these ultrafast dynamics, demonstrating the utility of the flapping molecular series as a versatile platform for designing photofunctional systems.

Structures, Compositions, and Activities of Live Shewanella Biofilms Formed on Graphite Electrodes in Electrochemical Flow Cells.

An electrochemical flow cell equipped with a graphite working electrode (WE) at the bottom was inoculated with Shewanella oneidensis MR-1 expressing an anaerobic fluorescent protein, and biofilm formation on the WE was observed over time during current generation at WE potentials of +0.4 and 0 V (versus standard hydrogen electrodes), under electrolyte-flow conditions. Electrochemical analyses suggested the presence of unique electron-transfer mechanisms in the +0.4-V biofilm. Microscopic analyses revealed that, in contrast to aerobic biofilms, current-generating biofilm (at +0.4 V) was thin and flat (∼10 µm in thickness), and cells were evenly and densely distributed in the biofilm. In contrast, cells were unevenly distributed in biofilm formed at 0 V. In situ fluorescence staining and biofilm recovery experiments showed that the amounts of extracellular polysaccharides (EPSs) in the +0.4-V biofilm were much smaller than those in the aerobic and 0-V biofilms, suggesting that Shewanella cells suppress the production of EPSs at +0.4 V under flow conditions. We suggest that Shewanella cells perceive electrode potentials and modulate the structure and composition of biofilms to efficiently transfer electrons to electrodes.IMPORTANCE A promising application of microbial fuel cells (MFCs) is to save energy in wastewater treatment. Since current is generated in these MFCs by biofilm microbes under horizontal flows of wastewater, it is important to understand the mechanisms for biofilm formation and current generation under water-flow conditions. Although massive work has been done to analyze the molecular mechanisms for current generation by model exoelectrogenic bacteria, such as Shewanella oneidensis, limited information is available regarding the formation of current-generating biofilms over time under water-flow conditions. The present study developed electrochemical flow cells and used them to examine the electrochemical and structural features of current-generating biofilms under water-flow conditions. We show unique features of mature biofilms actively generating current, creating opportunities to search for as-yet-undiscovered current-generating mechanisms in Shewanella biofilms. Furthermore, information provided in the present study is useful for researchers attempting to develop anode architectures suitable for wastewater treatment MFCs.

Effect of Water Adsorption on Carrier Trapping Dynamics at the Surface of Anatase TiO2 Nanoparticles.

Charge carrier trapping plays a vital role in heterogeneous photocatalytic water splitting because it strongly affects the dynamics of photogenerated charges and hence the photoconversion efficiency. Although hole trapping by water at water/photocatalyst interface is the first step of oxygen evolution in water splitting, little has been known on how water adsorbate itself is involved in hole trapping dynamics. To clarify this point, we have performed infrared transient and steady-state absorption spectroscopy of anatase TiO2 nanoparticles as a function of the number of water adsorbate layers. Here, we demonstrate that water molecules reversibly adsorbed in the first layer on TiO2 nanoparticles are capable to trap photogenerated holes, while water in the second layer hydrogen bonding to the first-layer water makes hole trapping less effective.