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Cable bacteria are an emerging class of electroactive organisms that sustain unprecedented long-range electron transport across centimeter-scale distances. The local pathways of the electrical currents in these filamentous microorganisms remain unresolved. Here, the electrical circuitry in a single cable bacterium is visualized with nanoscopic resolution using conductive atomic force microscopy. Combined with perturbation experiments, it is demonstrated that electrical currents are conveyed through a parallel network of conductive fibers embedded in the cell envelope, which are electrically interconnected between adjacent cells. This structural organization provides a fail-safe electrical network for long-distance electron transport in these filamentous microorganisms. The observed electrical circuit architecture is unique in biology and can inspire future technological applications in bioelectronics. R.T.E. and R.B. contributed equally to this work. The authors thank the colleagues from X-LAB from Hasselt University and the Microbial Electricity team from University of Antwerp for discussions and feedback. Special thanks to H. T. S. Boschker, I. Cardinaletti, J. Drijkoningen, J. L. Hou, and S. Thijs for their insights and discussions. Thanks to K. Ceyssens and T. Custers for the graphics. This research was financially supported by the Research Foundation Flanders (FWO project grant G031416N to FJRM and JM, FWO aspirant grant 1180517N to RB) and Dutch Research Council (NWO Vici grant 016.VICI.170.072 to FJRM). All measurements and data analysis were performed by R.T.E. and R.B. in equal contribution. J.M. and B.C. coordinated the study. Conceptualization and discussion were done by J.M., R.V., B.C., R.C., R.T.E., and R.B. Cable bacteria enrichment and fiber sheath extraction was performed by S.H.-M., R.B., and F.J.R.M. Funding was acquired by J.M., J.V., and F.J.R.M. Writing was done by R.B., R.T.E., J.M., B.C., and F.J.R.M., with contributions from all authors.

Biological electron transport is classically thought to occur over nanometre distances, yet recent studies suggest that electrical currents can run along centimetre-long cable bacteria. The phenomenon remains elusive, however, as currents have not been directly measured, nor have the conductive structures been identified. Here we demonstrate that cable bacteria conduct electrons over centimetre distances via highly conductive fibres embedded in the cell envelope. Direct electrode measurements reveal nanoampere currents in intact filaments up to 10.1 mm long (>2000 adjacent cells). A network of parallel periplasmic fibres displays a high conductivity (up to 79 S cm−1), explaining currents measured through intact filaments. Conductance rapidly declines upon exposure to air, but remains stable under vacuum, demonstrating that charge transfer is electronic rather than ionic. Our finding of a biological structure that efficiently guides electrical currents over long distances greatly expands the paradigm of biological charge transport and could enable new bio-electronic applications., Cable bacteria’ form long multicellular filaments that can transfer electrical currents over centimetre-long distances. Here, Meysman et al. show that the electrical currents run along highly conductive fibres embedded in the cell envelope, and charge transfer is electronic rather than ionic.

Cable bacteria are multicellular, filamentous microorganisms that are capable of transporting electrons over centimeter-scale distances. Although recently discovered, these bacteria appear to be widely present in the seafloor, and when active, they exert a strong imprint on the local geochemistry. In particular, their electrogenic metabolism induces unusually strong pH excursions in aquatic sediments, which induces considerable mineral dissolution, and subsequent mineral re-precipitation. However at present, it is unknown whether and how cable bacteria play an active or direct role in the mineral re-precipitation process. To this end we present an explorative study of the formation of sedimentary minerals in and near filamentous cable bacteria using a combined approach of electron microscopic and spectroscopic techniques. Our observations reveal three different types of biomineral formation directly associated with cable bacteria: (1) the formation of intracellular polyphosphate granules, which are associated with a localized accumulation of calcium and magnesium, (2) the attachment and incorporation of clay particles in a sheath surrounding the surface of the cable bacterium filaments, and (3) the encrustation of cable bacteria filaments by newly formed solid phases containing high amounts of iron. These findings suggest a complex interaction between cable bacteria and the surrounding sediment matrix, and a substantial imprint of the electrogenic metabolism on mineral diagenesis and sedimentary biogeochemical cycling. Particularly the encrustation process leaves many open questions for further research. For example, we hypothesize that the complete encrustation of filaments might create a diffusion barrier and negatively impact the metabolism of the cable bacteria.

Cable bacteria are electroactive bacteria that form a long, linear chain of ridged cylindrical cells. These filamentous bacteria conduct centimeter-scale long-range electron transport through parallel, interconnected conductive pathways of which the detailed chemical and electrical properties are still unclear. Here, we combine time-of-flight secondary-ion mass spectrometry (ToF-SIMS) and atomic force microscopy (AFM) to investigate the structure and composition of this naturally occurring electrical network. The enhanced lateral resolution achieved allows differentiation between the cell body and the cell−cell junctions that contain a conspicuous cartwheel structure. Three ToF-SIMS modes were compared in the study of so-called fiber sheaths (i.e., the cell material that remains after the removal of cytoplasm and membranes, and which embeds the electrical network). Among these, fast imaging delayed extraction (FI-DE) was found to balance lateral and mass resolution, thus yielding the following multiple benefits in the study of structure−composition relations in cable bacteria: (i) it enables the separate study of the cell body and cell-cell junctions; (ii) by combining FI-DE with in situ AFM, the depth of Ni-containing protein - key in the electrical transport - is determined with greater precision; and (iii) this combination prevents contamination, which is possible when using an ex situ AFM. Our results imply that the interconnects in extracted fiber sheaths are either damaged during extraction, or that their composition is different from fibers, or both. From a more general analytical perspective, the proposed methodology of ToF-SIMS in the FI-DE mode combined with in situ AFM holds great promise for studying the chemical structure of other biological systems. C able bacteria are multicellular microorganisms that form long unbranched filaments and belong to the Desulfobulbaceae family. 1 They are the focus of interdisciplinary research due to their unique capability of conducting electrical currents over centimeter distances, 2,3 a process also known as long-distance electron transport (LDET). Cable bacteria have been found to thrive in different environments such as fresh water 4,5 and marine sediments, 6,7 and have also been found in different parts of the world. 7 Cable bacteria display a distinct morphology with parallel ridges running along the length of the filament. 1,8,9 Scanning electron microscopy of the cross section of a cable bacteria revealed the presence of fibers of about 50 nm in diameter under the ridges and a cartwheel structure at the junctions. 8 These fibers are embedded in the periplasm (i.e., in space between the cytoplasmic membrane and the bacterial outer membrane) and were suspected to be the conductive structures 8 (Figure 1A−C). Recently, Meysman et al. experimentally investigated the conductivity of these fibers. 9 A sequential extraction procedure was developed 8 (see the Experimental Section) from which the fiber structures can be isolated from the cable bacterium filaments. 8 After chemical removal of cytoplasm and membranes, a so-called fiber sheath remains, which embeds the periplasmic fibers. 9−11 The fiber sheath flattens when air-dried (Figure 1D), and the top part of this fiber sheath mirrors the bottom part due to its cylindrical symmetry. 8 Meysman et al. demonstrated that fiber sheaths were indeed highly conductive. 9 Fiber sheaths were placed on top of two gold pads with a nonconductive oxide spacing, and when applying a potential difference between the two pads, a flow of current indicated that the periplasmic fibers are the conductive conduits. These results were subsequently confirmed by The authors acknowledge Bart Cleuren and Robin Bonné from UHasselt for the discussion on the C-AFM experiment. The authors acknowledge the grants given by the Research Foundation − Flanders (FWO) (G031416N to R.T.E., J.V.M., and F.J.R.M.; G038819N to F.J.R.M.) and theNetherlands Organization for Scientific Research (NWO) (VICI grant 016.VICI.170.072 awarded to F.J.R.M.). ToFSIMS analysis was rendered possible, thanks to the grant awarded to imec vzw and Hasselt University by the Hercules foundation (now FWO; grant no. ZW/13/07 awarded to J.V.M. and A.F.).

Cable bacteria (Deltaproteobacteria, Desulfobulbaceae) are long filamentous sulfur-oxidizing bacteria that generate long-distance electric currents running through the bacterial filaments. This way, they couple the oxidation of sulfide in deeper sediment layers to the reduction of oxygen or nitrate near the sediment-water interface. Cable bacteria are found in a wide range of aquatic sediments, but an accurate procedure to assess their abundance is lacking. We developed a qPCR approach that quantifies cable bacteria in relation to other bacteria within the family Desulfobulbaceae. Primer sets targeting cable bacteria, Desulfobulbaceae and the total bacterial community were applied in qPCR with DNA extracted from marine sediment incubations. Amplicon sequencing of the 16S rRNA gene V4 region confirmed that cable bacteria were accurately enumerated by qPCR, and suggested novel diversity of cable bacteria. The conjoint quantification of current densities and cell densities revealed that individual filaments carry a mean current of ∼110 pA and have a cell specific oxygen consumption rate of 69 fmol O2 cell–1 day–1. Overall, the qPCR method enables a better quantitative assessment of cable bacteria abundance, providing new metabolic insights at filament and cell level, and improving our understanding of the microbial ecology of electrogenic sediments.

Significance Cable bacteria form centimeter-long, multicellular filaments whose energy metabolism involves cooperation among cells that separately perform oxidation of the electron donor and reduction of the electron acceptor. This cooperative division of labor is facilitated via long-range electrical currents that run from cell to cell along a network of conductive fibers. Here we show that biomass synthesis shows a surprising asymmetry along the filament: only the cells oxidizing the electron donor conserve energy for growth, while the other cells reduce electron acceptors without biosynthesis. Our study hence provides insights into the physiology of an unconventional chemolithotroph, which forms a multicellular electrically connected system with unique functional differentiation, integration, and coordination., Multicellularity is a key evolutionary innovation, leading to coordinated activity and resource sharing among cells, which generally occurs via the physical exchange of chemical compounds. However, filamentous cable bacteria display a unique metabolism in which redox transformations in distant cells are coupled via long-distance electron transport rather than an exchange of chemicals. This challenges our understanding of organismal functioning, as the link among electron transfer, metabolism, energy conservation, and filament growth in cable bacteria remains enigmatic. Here, we show that cells within individual filaments of cable bacteria display a remarkable dichotomy in biosynthesis that coincides with redox zonation. Nanoscale secondary ion mass spectrometry combined with 13C (bicarbonate and propionate) and 15N-ammonia isotope labeling reveals that cells performing sulfide oxidation in deeper anoxic horizons have a high assimilation rate, whereas cells performing oxygen reduction in the oxic zone show very little or no label uptake. Accordingly, oxygen reduction appears to merely function as a mechanism to quickly dispense of electrons with little to no energy conservation, while biosynthesis and growth are restricted to sulfide-respiring cells. Still, cells can immediately switch roles when redox conditions change, and show no differentiation, which suggests that the “community service” performed by the cells in the oxic zone is only temporary. Overall, our data reveal a division of labor and electrical cooperation among cells that has not been seen previously in multicellular organisms.

Recently, a new group of multicellular microorganisms was discovered, called 'cable bacteria', which are capable of generating and mediating electrical currents across centimetre-scale distances. By transporting electrons from cell to cell, cable bacteria can harvest electron donors and electron acceptors that are widely separated in space, thus providing them with a competitive advantage for survival in aquatic sediments. The underlying process of long-distance electron transport challenges some long-held ideas about the energy metabolism of multicellular organisms and entails a whole new type of electrical cooperation between cells. This review summarizes the current knowledge about these intriguing multicellular bacteria.

Electro-active sediments distinguish themselves from other sedimentary environments by the presence of microbially induced electrical currents in the surface layer of the sediment. The electron transport is generated by metabolic activity of long filamentous cable bacteria, in a process referred to as electrogenic sulfur oxidation (e-SOx). Laboratory experiments have shown that e-SOx exerts a large impact on the sediment geochemistry, but its influence on the in situ geochemistry of marine sediments has not been previously investigated. Here, we document the biogeochemical cycling associated with e-SOx in a cohesive coastal sediment in the North Sea (Station 130, Belgian Coastal Zone) during three campaigns (January, March and May 2014). Fluorescence in situ hybridization showed that cable bacteria were present in high densities throughout the sampling period, and that filaments penetrated up to 7 cm deep in the sediment, which is substantially deeper than previously recorded. High resolution microsensor profiling (pH, H2S and O2) revealed the typical geochemical fingerprint of e-SOx, with a wide separation (up to 4.8 cm) between the depth of oxygen penetration and the depth of sulfide appearance. The metabolic activity of cable bacteria induced a current density of 25–32 mA m−2 and created an electrical field of 12–17 mV m−1 in the upper centimeters of the sediment. This electrical field created an ionic drift, which strongly affected the depth profiles and fluxes of major cations (Ca2+, Fe2+) and anions (SO42−) in the pore water. The strong acidification of the pore water at depth resulted in the dissolution of calcium carbonates and iron sulfides, thus leading to a strong accumulation of iron, calcium and manganese in the pore water. While sulfate accumulated in the upper centimeters, no significant effect of e-SOx was found on ammonium, phosphate and silicate depth profiles. Overall, our results demonstrate that cable bacteria can strongly modulate the sedimentary biogeochemical cycling under in situ conditions.

Petroleum hydrocarbons reach the deep-sea following natural and anthropogenic factors. The process by which they enter deep-sea microbial food webs and impact the biogeochemical cycling of carbon and other elements is unclear. Hydrostatic pressure (HP) is a distinctive parameter of the deep sea, although rarely investigated. Whether HP alone affects the assembly and activity of oil-degrading communities remains to be resolved. Here we have demonstrated that hydrocarbon degradation in deep-sea microbial communities is lower at native HP (10 MPa, about 1000 m below sea surface level) than at ambient pressure. In long-term enrichments, increased HP selectively inhibited obligate hydrocarbon-degraders and downregulated the expression of beta-oxidation-related proteins (i.e., the main hydrocarbon-degradation pathway) resulting in low cell growth and CO2 production. Short-term experiments with HP-adapted synthetic communities confirmed this data, revealing a HP-dependent accumulation of citrate and dihydroxyacetone. Citrate accumulation suggests rates of aerobic oxidation of fatty acids in the TCA cycle were reduced. Dihydroxyacetone is connected to citrate through glycerol metabolism and glycolysis, both upregulated with increased HP. High degradation rates by obligate hydrocarbon-degraders may thus be unfavourable at increased HP, explaining their selective suppression. Through lab-scale cultivation, the present study is the first to highlight a link between impaired cell metabolism and microbial community assembly in hydrocarbon degradation at high HP. Overall, this data indicate that hydrocarbons fate differs substantially in surface waters as compared to deep-sea environments, with in situ low temperature and limited nutrients availability expected to further prolong hydrocarbons persistence at deep sea.

Cable bacteria of the family Desulfobulbaceae form centimeter-long filaments comprising thousands of cells. They occur worldwide in the surface of aquatic sediments, where they connect sulfide oxidation with oxygen or nitrate reduction via long-distance electron transport. In the absence of pure cultures, we used single-filament genomics and metagenomics to retrieve draft genomes of 3 marine Candidatus Electrothrix and 1 freshwater Ca. Electronema species. These genomes contain >50% unknown genes but still share their core genomic makeup with sulfate-reducing and sulfur-disproportionating Desulfobulbaceae, with few core genes lost and 212 unique genes (from 197 gene families) conserved among cable bacteria. Last common ancestor analysis indicates gene divergence and lateral gene transfer as equally important origins of these unique genes. With support from metaproteomics of a Ca. Electronema enrichment, the genomes suggest that cable bacteria oxidize sulfide by reversing the canonical sulfate reduction pathway and fix CO 2 using the Wood–Ljungdahl pathway. Cable bacteria show limited organotrophic potential, may assimilate smaller organic acids and alcohols, fix N 2 , and synthesize polyphosphates and polyglucose as storage compounds; several of these traits were confirmed by cell-level experimental analyses. We propose a model for electron flow from sulfide to oxygen that involves periplasmic cytochromes, yet-unidentified conductive periplasmic fibers, and periplasmic oxygen reduction. This model proposes that an active cable bacterium gains energy in the anodic, sulfide-oxidizing cells, whereas cells in the oxic zone flare off electrons through intense cathodic oxygen respiration without energy conservation; this peculiar form of multicellularity seems unparalleled in the microbial world.

Undigestible, insoluble food particles, such as wheat bran, are important dietary constituents that serve as a fermentation substrate for the human gut microbiota. The first step in wheat bran fermentation involves the poorly studied solubilization of fibers from the complex insoluble wheat bran structure. Attachment of bacteria has been suggested to promote the efficient hydrolysis of insoluble substrates, but the mechanisms and drivers of this microbial attachment and colonization, as well as subsequent fermentation remain to be elucidated. We have previously shown that an individually dependent subset of gut bacteria is able to colonize the wheat bran residue. Here, we isolated these bran-attached microorganisms, which can then be used to gain mechanistic insights in future pure culture experiments. Four healthy fecal donors were screened to account for inter-individual differences in gut microbiota composition. A combination of a direct plating and enrichment method resulted in the isolation of a phylogenetically diverse set of species, belonging to theBacteroidetes,Firmicutes,ProteobacteriaandActinobacteriaphyla. A comparison with 16S rRNA gene sequences that were found enriched on wheat bran particles in previous studies, however, showed that the isolates do not yet cover the entire diversity of wheat-bran colonizing species, comprising among others a broad range ofPrevotella,BacteroidesandClostridiumcluster XIVa species. We, therefore, suggest several modifications to the experiment set-up to further expand the array of isolated species.

Dietary modulation can alter the gut microbiota composition and activity, in turn affecting health. Particularly, dietary fibre rich foods, such as wheat bran, are an important nutrient source for the gut microbiota. Several processing methods have been developed to modify the functional, textural and breadmaking properties of wheat bran, which can affect the gut microbiota. We therefore studied the effect of enzyme treatment, particle size reduction and wheat kernel pearling on the faecal microbiota of ten healthy individuals. The most commonly studied health marker, associated to the gut microbiota activity is Short Chain Fatty Acid (SCFA) production. This study shows that modifying wheat bran physicochemical properties allows control over the extent and the rate of SCFA production by the faecal microbiota. Wheat bran pericarp fractions, depleted in starch and enriched in cellulose and highly branched arabinoxylans, were poorly fermentable compared to unmodified wheat bran, thus resulting in a reduced SCFA production with up to 20 mM. The nature of the SCFA, however, largely depends on the donor and can be linked to the individual's gut microbiota composition. The latter changed in an individually dependent manner in response to wheat bran modification. Some product dependent significant differences could still be identified across the ten donors. This product effect is more pronounced in the microbial community attached to the wheat bran residue as compared to the luminal microbial community. Generally, we find lower levels of Firmicutes, Bacteroidetes and Bifidobacterium and a higher abundance of Proteobacteria in the pericarp enriched wheat bran fractions, compared to unmodified wheat bran. ispartof: FOOD & FUNCTION vol:10 issue:1 pages:379-396 ispartof: location:England status: published

Cable bacteria represent a newly discovered group of filamentous microorganisms, which are capable of spatially separating the oxidative and reductive half-reactions of their sulfide-oxidizing metabolisms over centimeter distances. We investigated three ways that cable bacteria might interact with the nitrogen (N) cycle: (1) by reducing nitrate through denitrification or dissimilatory nitrate reduction to ammonium (DNRA) within their cathodic cells; (2) by nitrifying ammonium within their anodic cells; and (3) by indirectly affecting denitrification and/or DNRA by changing the Fe 2+ concentration in the surrounding sediment. We performed 15 N labeling laboratory experiments to measure these three processes using cable bacteria containing sediments from the Yarra River, Australia, and from Vilhelmsborg Sø, Denmark. Our results revealed that in the targeted systems, cable bacteria themselves did not perform significant rates of denitrification, DNRA, or nitrification. However, cable bacteria exhibited an important indirect effect, whereby they increased the Fe 2+ pool through iron sulfide dissolution. This elevated availability of Fe 2+ significantly increased DNRA and in some cases decreased denitrification. Thus, cable bacteria presence may affect the relative importance of DNRA in sediments and thus the extent by which bioavailable nitrogen is lost from the system.

Cable bacteria are long, multicellular, filamentous bacteria that can conduct electrons over centimeter distances in marine and freshwater sediments. Recent studies indicate that cable bacteria are widely present in many coastal environments, where they exert a major influence on the biogeochemistry of the sediment. Their energy metabolism can be based on the aerobic oxidation of sulfide, and hence to better understand their natural occurrence and distribution, we examined the growth and activity of cable bacteria in relation to bottom water oxygenation. To this end, we conducted laboratory sediment incubations at four different O2 levels in the overlying water (10%, 20%, 40%, and 100% air saturation). The abundance of cable bacteria was determined by fluorescence in situ hybridization, while their activity was assessed via microsensor profiling and geochemical pore‐water analysis. Cable bacteria did not develop in the 10% air saturation O2 incubation but were present and active at all higher O2 levels. These data show that microbial long‐distance electron transport can occur under a wide range of bottom water O2 concentrations. However, the growth rate was notably slower at lower oxygen concentrations, suggesting a reduced metabolic activity of the population when the O2 supply becomes restricted. Finally, in response to lower O2 levels, cable bacteria filaments appear to partially emerge out of the sediment and extend into the overlying water, thus likely enhancing their oxygen supply.

Cable bacteria have been reported in sediments from marine and freshwater locations, but the environmental factors that regulate their growth in natural settings are not well understood. Most prominently, the physiological limit of cable bacteria in terms of oxygen availability remains poorly constrained. In this study, we investigated the presence, activity and diversity of cable bacteria in relation to a natural gradient in bottom water oxygenation in a depth transect of the Eastern Gotland Basin (Baltic Sea). Cable bacteria were identified by FISH at the oxic and transiently oxic sites, but not at the permanently anoxic site. Three species of the candidate genus Electrothrix, i.e. marina, aarhusiensis and communis were found coexisting within one site. The highest filament density (33 m cm(-2)) was associated with a 6.3 mm wide zone depleted in both oxygen and free sulphide, and the presence of an electric field resulting from the electrogenic sulphur oxidizing metabolism of cable bacteria. However, the measured filament densities and metabolic activities remained low overall, suggesting a limited impact of cable bacteria at the basin level. The observed bottom water oxygen levels (< 5 M) are the lowest so far reported for cable bacteria, thus expanding their known environmental distribution.

Electron transport within living cells is essential for energy conservation in all respiring and photosynthetic organisms. While a few bacteria transport electrons over micrometer distances to their surroundings, filaments of cable bacteria are hypothesized to conduct electric currents over centimeter distances. We used resonance Raman microscopy to analyze cytochrome redox states in living cable bacteria. Cable-bacteria filaments were placed in microscope chambers with sulfide as electron source and oxygen as electron sink at opposite ends. Along individual filaments a gradient in cytochrome redox potential was detected, which immediately broke down upon removal of oxygen or laser cutting of the filaments. Without access to oxygen, a rapid shift toward more reduced cytochromes was observed, as electrons were no longer drained from the filament but accumulated in the cellular cytochromes. These results provide direct evidence for long-distance electron transport in living multicellular bacteria.

Seasonal hypoxia in coastal systems drastically changes the availability of electron acceptors in bottom water, which alters the sedimentary reoxidation of reduced compounds. However, the effect of seasonal hypoxia on the chemolithoautotrophic community that catalyzes these reoxidation reactions is rarely studied. Here, we examine the changes in activity and structure of the sedimentary chemolithoautotrophic bacterial community of a seasonally hypoxic saline basin under oxic (spring) and hypoxic (summer) conditions. Combined 16S rRNA gene amplicon sequencing and analysis of phospholipid-derived fatty acids indicated a major temporal shift in community structure. Aerobic sulfur-oxidizing Gammaproteobacteria ( Thiotrichales ) and Epsilonproteobacteria ( Campylobacterales ) were prevalent during spring, whereas Deltaproteobacteria ( Desulfobacterales ) related to sulfate-reducing bacteria prevailed during summer hypoxia. Chemolithoautotrophy rates in the surface sediment were three times higher in spring than in summer. The depth distribution of chemolithoautotrophy was linked to the distinct sulfur oxidation mechanisms identified through microsensor profiling, i.e., canonical sulfur oxidation, electrogenic sulfur oxidation by cable bacteria, and sulfide oxidation coupled to nitrate reduction by Beggiatoaceae . The metabolic diversity of the sulfur-oxidizing bacterial community suggests a complex niche partitioning within the sediment, probably driven by the availability of reduced sulfur compounds (H 2 S, S 0 , and S 2 O 3 2− ) and electron acceptors (O 2 and NO 3 − ) regulated by seasonal hypoxia. IMPORTANCE Chemolithoautotrophic microbes in the seafloor are dependent on electron acceptors, like oxygen and nitrate, that diffuse from the overlying water. Seasonal hypoxia, however, drastically changes the availability of these electron acceptors in the bottom water; hence, one expects a strong impact of seasonal hypoxia on sedimentary chemolithoautotrophy. A multidisciplinary investigation of the sediments in a seasonally hypoxic coastal basin confirms this hypothesis. Our data show that bacterial community structure and chemolithoautotrophic activity varied with the seasonal depletion of oxygen. Unexpectedly, the dark carbon fixation was also dependent on the dominant microbial pathway of sulfur oxidation occurring in the sediment (i.e., canonical sulfur oxidation, electrogenic sulfur oxidation by cable bacteria, and sulfide oxidation coupled to nitrate reduction by Beggiatoaceae ). These results suggest that a complex niche partitioning within the sulfur-oxidizing bacterial community additionally affects the chemolithoautotrophic community of seasonally hypoxic sediments.

Recently, a novel electrogenic type of sulphur oxidation was documented in marine sediments, whereby filamentous cable bacteria (Desulfobulbaceae) are mediating electron transport over cm-scale distances. These cable bacteria are capable of developing an extensive network within days, implying a highly efficient carbon acquisition strategy. Presently, the carbon metabolism of cable bacteria is unknown, and hence we adopted a multidisciplinary approach to study the carbon substrate utilization of both cable bacteria and associated microbial community in sediment incubations. Fluorescence in situ hybridization showed rapid downward growth of cable bacteria, concomitant with high rates of electrogenic sulphur oxidation, as quantified by microelectrode profiling. We studied heterotrophy and autotrophy by following (13)C-propionate and -bicarbonate incorporation into bacterial fatty acids. This biomarker analysis showed that propionate uptake was limited to fatty acid signatures typical for the genus Desulfobulbus. The nanoscale secondary ion mass spectrometry analysis confirmed heterotrophic rather than autotrophic growth of cable bacteria. Still, high bicarbonate uptake was observed in concert with the development of cable bacteria. Clone libraries of 16S complementary DNA showed numerous sequences associated to chemoautotrophic sulphur-oxidizing Epsilon- and Gammaproteobacteria, whereas (13)C-bicarbonate biomarker labelling suggested that these sulphur-oxidizing bacteria were active far below the oxygen penetration. A targeted manipulation experiment demonstrated that chemoautotrophic carbon fixation was tightly linked to the heterotrophic activity of the cable bacteria down to cm depth. Overall, the results suggest that electrogenic sulphur oxidation is performed by a microbial consortium, consisting of chemoorganotrophic cable bacteria and chemolithoautotrophic Epsilon- and Gammaproteobacteria. The metabolic linkage between these two groups is presently unknown and needs further study.

Recently, long filamentous bacteria, belonging to the family Desulfobulbaceae , were shown to induce electrical currents over long distances in the surface layer of marine sediments. These “cable bacteria” are capable of harvesting electrons from free sulfide in deeper sediment horizons and transferring these electrons along their longitudinal axes to oxygen present near the sediment-water interface. In the present work, we investigated the relationship between cable bacteria and a photosynthetic algal biofilm. In a first experiment, we investigated sediment that hosted both cable bacteria and a photosynthetic biofilm and tested the effect of an imposed diel light-dark cycle by continuously monitoring sulfide at depth. Changes in photosynthesis at the sediment surface had an immediate and repeatable effect on sulfide concentrations at depth, indicating that cable bacteria can rapidly transmit a geochemical effect to centimeters of depth in response to changing conditions at the sediment surface. We also observed a secondary response of the free sulfide at depth manifest on the time scale of hours, suggesting that cable bacteria adjust to a moving oxygen front with a regulatory or a behavioral response, such as motility. Finally, we show that on the time scale of days, the presence of an oxygenic biofilm results in a deeper and more acidic suboxic zone, indicating that a greater oxygen supply can enable cable bacteria to harvest a greater quantity of electrons from marine sediments. Rapid acclimation strategies and highly efficient electron harvesting are likely key advantages of cable bacteria, enabling their success in high sulfide generating coastal sediments.