Cable bacteria are multicellular sulfide oxidizing bacteria that display a unique metabolism based on long-distance electron transport. Cells in deeper sediment layers perform the sulfide oxidizing half-reaction whereas cells in the surface layers of the sediment perform the oxygen-reducing half-reaction. These half-reactions are coupled via electron transport through a conductive fiber network that runs along the shared cell envelope. Remarkably, only the sulfide oxidizing half-reaction is coupled to biosynthesis and growth whereas the oxygen reducing half-reaction serves to rapidly remove electrons from the conductive fiber network and is not coupled to energy generation and growth. Cells residing in the oxic zone are believed to (temporarily) rely on storage compounds of which polyphosphate (poly-P) is prominently present in cable bacteria. Here we investigate the role of poly-P in the metabolism of cable bacteria within the different redox environments. To this end, we combined nanoscale secondary ion mass spectrometry with dual-stable isotope probing (¹³C-DIC and ¹⁸O-H₂O) to visualize the relationship between growth in the cytoplasm (¹³C-enrichment) and poly-P activity (¹⁸O-enrichment). We found that poly-P was synthesized in almost all cells, as indicated by ¹⁸O enrichment of poly-P granules. Hence, poly-P must have an important function in the metabolism of cable bacteria. Within the oxic zone of the sediment, where little growth is observed, ¹⁸O enrichment in poly-P granules was significantly lower than in the suboxic zone. Thus, both growth and poly-P metabolism appear to be correlated to the redox environment. However, the poly-P metabolism is not coupled to growth in cable bacteria, as many filaments from the suboxic zone showed poly-P activity but did not grow. We hypothesize that within the oxic zone, poly-P is used to protect the cells against oxidative stress and/or as a resource to support motility, while within the suboxic zone, poly-P is involved in the metabolic regulation before cells enter a non-growing stage.

Cable bacteria are multicellular, Gram-negative filamentous bacteria that display a unique division of metabolic labor between cells. Cells in deeper sediment layers are oxidizing sulfide, while cells in the surface layers of the sediment are reducing oxygen. The electrical coupling of these two redox half reactions is ensured via long-distance electron transport through a network of conductive fibers that run in the shared cell envelope of the centimeter-long filament. Here we investigate how this unique electrogenic metabolism is linked to filament growth and cell division. Combining dual-label stable isotope probing (¹³C and ¹⁵N), nanoscale secondary ion mass spectrometry, fluorescence microscopy and genome analysis, we find that the cell cycle of cable bacteria cells is highly comparable to that of other, single-celled Gram-negative bacteria. However, the timing of cell growth and division appears to be tightly and uniquely controlled by long-distance electron transport, as cell division within an individual filament shows a remarkable synchronicity that extends over a millimeter length scale. To explain this, we propose the “oxygen pacemaker” model in which a filament only grows when performing long-distance transport, and the latter is only possible when a filament has access to oxygen so it can discharge electrons from its internal electrical network.

Eutrophication of water bodies has been a problem causing severe degradation of water quality in cities. To gain mechanistic understanding of the temporal dynamics of nitrogen (N) and phosphorus (P) in a groundwater-fed lowlying urban polder, we applied high-frequency monitoring in Geuzenveld, a polder in the city of Amsterdam. The highfrequency monitoring equipment was installed at the pumping station where water leaves the polder. From March 2016 to June 2017, total phosphorus (TP), ammonium (NH4), turbidity, electrical conductivity (EC), and water temperature were measured at intervals of less than 20 min. This paper discusses the results at three timescales: Annual scale, rain event scale, and single pumping event scale. Mixing of upwelling groundwater (main source of N and P) and runoff from precipitation on pavements and roofs was the dominant hydrological process governing the temporal pattern of the EC, while N and P fluxes from the polder were also regulated by primary production and iron transformations. In our groundwater-seepage controlled catchment, NH4 appeared to be the dominant form of N with surface water concentrations in the range of 2-6 mgNL-1, which stems from production in an organic-rich subsurface. The concentrations of NH4 in the surface water were governed by the mixing process in autumn and winter and were reduced down to 0.1 mgNL-1 during the algal growing season in spring. The depletion of dissolved NH4 in spring suggests uptake by primary producers, consistent with high concentrations of chlorophyll a, O2, and suspended solids during this period. Total P and turbidity were high during winter (range 0.5-2.5 mg P L-1 and 200-1800 FNU, respectively, where FNU represents Formazin Nephelometric Unit) due to the release of P and reduced iron from anoxic sediment to the water column, where Fe2C was rapidly oxidized and precipitated as iron oxides which contributed to turbidity. In the other seasons, P is retained in the sediment by sorption to precipitated iron oxides. Nitrogen is exported from the polder to the receiving waters throughout the whole year, mostly in the form of NH4 but in the form of organic N in spring. P leaves the polder mainly during winter, primarily associated with Fe(OH)3 colloids and as dissolved P. Based on this new understanding of the dynamics of N and P in this low-lying urban catchment, we suggested management strategies that may effectively control and reduce eutrophication in urban polders and receiving downstream waters.

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 reprecipitation. However, at present, it is unknown whether and how cable bacteria play an active or direct role in the mineral reprecipitation 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 microscopy and spectroscopic techniques. Our observations reveal the formation of polyphosphate granules within the cells and two different types of biomineral formation directly associated with multicellular filaments of these cable bacteria: (i) the attachment and incorporation of clay particles in a coating surrounding the bacteria and (ii) encrustation of the cell envelope by iron minerals. 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. In particular, 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.