Yasuaki Hotta

Characterization of a filamentous biofilm community established in a cellulose-fed microbial fuel cell

BackgroundMicrobial fuel cells (MFCs) are devices that exploit microorganisms to generate electric power from organic matter. Despite the development of efficient MFC reactors, the microbiology of electricity generation remains to be sufficiently understood.ResultsA laboratory-scale two-chamber microbial fuel cell (MFC) was inoculated with rice paddy field soil and fed cellulose as the carbon and energy source. Electricity-generating microorganisms were enriched by subculturing biofilms that attached onto anode electrodes. An electric current of 0.2 mA was generated from the first enrichment culture, and ratios of the major metabolites (e.g., electric current, methane and acetate) became stable after the forth enrichment. In order to investigate the electrogenic microbial community in the anode biofilm, it was morphologically analyzed by electron microscopy, and community members were phylogenetically identified by 16S rRNA gene clone-library analyses. Electron microscopy revealed that filamentous cells and rod-shaped cells with prosthecae-like filamentous appendages were abundantly present in the biofilm. Filamentous cells and appendages were interconnected via thin filaments. The clone library analyses frequently detected phylotypes affiliated with Clostridiales, Chloroflexi, Rhizobiales and Methanobacterium. Fluorescence in-situ hybridization revealed that the Rhizobiales population represented rod-shaped cells with filamentous appendages and constituted over 30% of the total population.ConclusionBacteria affiliated with the Rhizobiales constituted the major population in the cellulose-fed MFC and exhibited unique morphology with filamentous appendages. They are considered to play important roles in the cellulose-degrading electrogenic community.

Coaggregation Facilitates Interspecies Hydrogen Transfer between Pelotomaculum thermopropionicum and Methanothermobacter thermautotrophicus

ABSTRACT A thermophilic syntrophic bacterium, Pelotomaculum thermopropionicum strain SI, was grown in a monoculture or coculture with a hydrogenotrophic methanogen, Methanothermobacter thermautotrophicus strain ΔH. Microscopic observation revealed that cells of each organism were dispersed in a monoculture independent of the growth substrate. In a coculture, however, these organisms coaggregated to different degrees depending on the substrate; namely, a large fraction of the cells coaggregated when they were grown on propionate, but relatively few cells coaggregated when they were grown on ethanol or 1-propanol. Field emission-scanning electron microscopy revealed that flagellum-like filaments of SI cells played a role in making contact with ΔH cells. Microscopic observation of aggregates also showed that extracellular polymeric substance-like structures were present in intercellular spaces. In order to evaluate the importance of coaggregation for syntrophic propionate oxidation, allowable average distances between SI and ΔH cells for accomplishing efficient interspecies hydrogen transfer were calculated by using Ficks diffusion law. The allowable distance for syntrophic propionate oxidation was estimated to be approximately 2 μm, while the allowable distances for ethanol and propanol oxidation were 16 μm and 32 μm, respectively. Considering that the mean cell-to-cell distance in the randomly dispersed culture was approximately 30 μm (at a concentration in the mid-exponential growth phase of the coculture of 5 × 107 cells ml−1), it is obvious that close physical contact of these organisms by coaggregation is indispensable for efficient syntrophic propionate oxidation.

Methanogenesis versus Electrogenesis : Morphological and Phylogenetic Comparisons of Microbial Communities

Two H-type microbial fuel cells were prepared. The anaerobic chambers were inoculated with rice paddy field soil and fed cellulose as an energy source. In one reactor, the anode and cathode were connected with a wire (closed circuit, CC), while they were not connected in the other reactor (open circuit, OC). The OC reactor actively produced methane. In the CC reactor, however, an electric current of 0.2 to 0.3 mA was constantly generated, and methane production was almost completely suppressed. Electron microscopy revealed that rod-shaped cells with long prosthecae-like filaments were specifically enriched in the CC reactor. Comparisons of 16S rRNA gene clone libraries revealed entirely different phylogenetic compositions in the CC and OC communities; phylotypes related to Rhizobiceae, Desulfovibrio, and Ethanoligenens were specifically enriched in the CC community. The results indicate that electrogenesis resulted in the enrichment of distinctive microbial populations and suppressed methanogenesis from cellulose.

Simulating the contribution of coaggregation to interspecies hydrogen fluxes in syntrophic methanogenic consortia

ABSTRACT A simple model (termed the syntrophy model) for simulating the contribution of coaggregation to interspecies hydrogen fluxes between syntrophic bacteria and methanogenic archaea is described. We applied it to analyzing partially aggregated syntrophic cocultures with various substrates, revealing that large fractions of hydrogen molecules were fluxed in aggregates.

Formation of filamentous appendages by Acinetobacter sp. Tol 5 for adhering to solid surfaces.

The toluene-degrading bacterium Acinetobacter sp. Tol 5 is highly adhesive to solid surfaces owing to two filamentous cell appendages, namely, anchors and peritrichate fibrils. When growing this bacterium in the presence of a carrier made of polyurethane foam, almost all the cells adhered to the surface of the carrier. In contrast, when Tol 5 cells were grown in the absence of the polyurethane carrier, the cells were suspended as aggregated cells or individually dispersed cells. The aggregated cells possessed the cell appendages and showed an adhesiveness similar to that of cells grown in the presence of the carrier, while the dispersed cells scarcely produced the cell appendages and showed a low level of adhesiveness. The dispersed cells started to adhere to the polyurethane carrier by producing the filamentous appendages within 30 min of the addition of the carrier as a substratum and toluene as a carbon source. Peritrichate fibrils just sprouting and growing anchors longer than 3 microm were observed when the cells started to adhere. This suggests that the presence of surface areas sufficient for adhesion might trigger cell appendage formation in Tol 5 cells for adhesion by increasing the amount of cell contact with the surfaces.

Examination of Six Orthodontic Adhesives with Electron Microscopy, Hardness Tester and Energy Dispersive X-ray Microanalyzer

OBJECTIVE To examine the ultrastructure of six light-cure orthodontic adhesives with scanning electron microscope (SEM) and transmission electron microscope (TEM), microhardness tester, and energy dispersive X-ray microanalyzer (EDX). MATERIALS AND METHODS The orthodontic adhesives evaluated were Transbond XT, Light Bond, BeautyOrtho Bond, Kurasper F, Heliosit Orthodontic, and Salivatect. Specimens of each adhesive were carefully prepared for observation under SEM and TEM. Furthermore, the Vickers hardness was tested, and the adhesives were evaluated with EDX. RESULTS SEM and TEM images illustrated great diversity of the adhesives ultrastructure. The Vickers hardness test showed significant differences among all the adhesives (except Transbond XT and Salivatect). Although some similar elements were detected with EDX, the concentration was different in each adhesive. CONCLUSION Orthodontic brackets can be bonded to the enamel surface with the adhesives available on the market. However, orthodontists might achieve better results identifying their properties and compositions.

Ultrastructural Alteration of Pulmonary Capillary Endothelial Glycocalyx During Endotoxemia

Background The most recent diagnostic criteria for sepsis include organ failure. Microvascular endothelial injury is believed to lead to the multiple organ failure seen in sepsis, although the precise mechanism is still controversial. ARDS is the primary complication during the sequential development of multiple organ dysfunction in sepsis, and endothelial injury is deeply involved. Sugar‐protein glycocalyx coats all healthy vascular endothelium, and its disruption is one factor believed to contribute to microvascular endothelial dysfunction during sepsis. The goal of this study was to observe the three‐dimensional ultrastructural alterations in the pulmonary capillary endothelium, including the glycocalyx, during sepsis‐induced pulmonary vasculitis. Methods This study investigated the three‐dimensional ultrastructure of pulmonary vascular endothelial glycocalyx in a mouse lipopolysaccharide‐induced endotoxemia model. Lungs were fixed with lanthanum‐containing alkaline fixative to preserve the glycocalyx. Results On both scanning and transmission electron microscopic imaging, the capillary endothelial glycocalyx appeared as a moss‐like structure entirely covering the endothelial cell surface in normal mice. In the septic lung following liposaccharide injection, however, this structure was severely disrupted; it appeared to be peeling away and coagulated. In addition, syndecan‐1 levels were significantly reduced in the septic lung, and numerous spherical structures containing glycocalyx were observed on the endothelial surface. Conclusions It appears that endothelial glycocalyx in the lung is markedly disrupted under experimental endotoxemia conditions. This finding supports the notion that disruption of the glycocalyx is causally related to the microvascular endothelial dysfunction that is characteristic of sepsis‐induced ARDS.

Ultrastructural aspects of vacuolar degeneration of cardiomyocytes in human endomyocardial biopsies

Vacuolar degeneration of cardiomyocytes is a histological finding commonly encountered during routine light microscopic examination of human endomyocardial biopsy specimens. The vacuoles appear as intracellular clear areas lacking myofibers. By itself, this finding has little diagnostic value, but may have important clinical implications when the vacuolar contents are of etiological significance (e.g., accumulation of abnormal metabolites), and the clinical importance is increased when the disease is treatable. Thanks to its great resolving power, electron microscopy can often reveal the contents of the vacuoles and lead to a correct diagnosis. It can be used to differentially diagnose lysosomal storage diseases such as Fabry, Danon, and Pompe disease, doxorubicin cardiomyopathy, mitochondrial cardiomyopathy, autophagic degeneration, and accumulation of subcellular organelles (mitochondria, lipofuscin, glycogen granules, endoplasmic reticulum, etc.) as a nonspecific finding in failing cardiomyocytes. Nonetheless, undiagnosed cases certainly remain. It is strongly recommended that small pieces of tissue samples be fixed for electron microscopy at every endomyocardial biopsy procedure, and electron microscopic examination should be performed when a marked vacuolar degeneration is found.