Metabolic network analysis reveals microbial community interactions in anammox granules.

Microbial communities mediating anaerobic ammonium oxidation (anammox) represent one of the most energy-efficient environmental biotechnologies for nitrogen removal from wastewater. However, little is known about the functional role heterotrophic bacteria play in anammox granules. Here, we use genome-centric metagenomics to recover 17 draft genomes of anammox and heterotrophic bacteria from a laboratory-scale anammox bioreactor. We combine metabolic network reconstruction with metatranscriptomics to examine the gene expression of anammox and heterotrophic bacteria and to identify their potential interactions. We find that Chlorobi-affiliated bacteria may be highly active protein degraders, catabolizing extracellular peptides while recycling nitrate to nitrite. Other heterotrophs may also contribute to scavenging of detritus and peptides produced by anammox bacteria, and potentially use alternative electron donors, such as H2, acetate and formate. Our findings improve the understanding of metabolic activities and interactions between anammox and heterotrophic bacteria and offer the first transcriptional insights on ecosystem function in anammox granules

A unique bacteriohopanetetrol stereoisomer of marine anammox

Anaerobic ammonium oxidation (anammox) is a major process of bioavailable nitrogen removal from marine systems. Previously, a bacteriohopanetetrol (BHT) isomer, with unknown stereochemistry, eluting later than BHT using high performance liquid chromatography (HPLC), was detected in ‘Ca. Scalindua profunda’ and proposed as a biomarker for anammox in marine paleo-environments. However, the utility of this BHT isomer as an anammox biomarker is hindered by the fact that four other, non-anammox bacteria are also known to produce a late-eluting BHT stereoisomer. The stereochemistry in Acetobacter pasteurianus, Komagataeibacter xylinus and Frankia sp. was known to be 17β, 21β(H), 22R, 32R, 33R, 34R (BHT-34R). The stereochemistry of the late-eluting BHT in Methylocella palustris was unknown. To determine if marine anammox bacteria produce a unique BHT isomer, we studied the BHT distributions and stereochemistry of known BHT isomer producers and of previously unscreened marine (‘Ca. Scalindua brodeae’) and freshwater (‘Ca. Brocadia sp.’) anammox bacteria using HPLC and gas chromatographic (GC) analysis of acetylated BHTs and ultra high performance liquid chromatography (UHPLC)-high resolution mass spectrometry (HRMS) analysis of non-acetylated BHTs. The 34R stereochemistry was confirmed for the BHT isomers in Ca. Brocadia sp. and Methylocella palustris. However, ‘Ca. Scalindua sp.’ synthesise a stereochemically distinct BHT isomer, with still unconfirmed stereochemistry (BHT-x). Only GC analysis of acetylated BHT and UHPLC analysis of non-acetylated BHT distinguished between late-eluting BHT isomers. Acetylated BHT-x and BHT-34R co-elute by HPLC. As BHT-x is currently only known to be produced by ‘Ca. Scalindua spp.’, it may be a biomarker for marine anammox

Vertical segregation among pathways mediating nitrogen loss (N2 and N2O production) across the oxygen gradient in a coastal upwelling ecosystem

Indexación: ScopusThe upwelling system off central Chile (36.5 S) is seasonally subjected to oxygen (O2)-deficient waters, with a strong vertical gradient in O2 (from oxic to anoxic conditions) that spans a few metres (30-50€m interval) over the shelf. This condition inhibits and/or stimulates processes involved in nitrogen (N) removal (e.g. anammox, denitrification, and nitrification). During austral spring (September 2013) and summer (January 2014), the main pathways involved in N loss and its speciation, in the form of N2 and/or N2O, were studied using 15N-tracer incubations, inhibitor assays, and the natural abundance of nitrate isotopes along with hydrographic information. Incubations were developed using water retrieved from the oxycline (25€m depth) and bottom waters (85€m depth) over the continental shelf off Concepción, Chile. Results of 15N-labelled incubations revealed higher N removal activity during the austral summer, with denitrification as the dominant N2-producing pathway, which occurred together with anammox at all times. Interestingly, in both spring and summer maximum potential N removal rates were observed in the oxycline, where a greater availability of oxygen was observed (maximum O2 fluctuation between 270 and 40€μmol€L'1) relative to the hypoxic bottom waters ( < €20€μmol€O2€L'1). Different pathways were responsible for N2O produced in the oxycline and bottom waters, with ammonium oxidation and dissimilatory nitrite reduction, respectively, as the main source processes. Ammonium produced by dissimilatory nitrite reduction to ammonium (DNiRA) could sustain both anammox and nitrification rates, including the ammonium utilized for N2O production. The temporal and vertical variability of /15N-NO3' confirms that multiple N-cycling processes are modulating the isotopic nitrate composition over the shelf off central Chile during spring and summer. N removal processes in this coastal system appear to be related to the availability and distribution of oxygen and particles, which are a source of organic matter and the fuel for the production of other electron donors (i.e. ammonium) and acceptors (i.e. nitrate and nitrite) after its remineralization. These results highlight the links between several pathways involved in N loss. They also establish that different mechanisms supported by alternative N substrates are responsible for substantial accumulation of N2O, which are frequently observed as hotspots in the oxycline and bottom waters. Considering the extreme variation in oxygen observed in several coastal upwelling systems, these findings could help to understand the ecological and biogeochemical implications due to global warming where intensification and/or expansion of the oceanic OMZs is projected.https://www.biogeosciences.net/14/4795/2017

Nitrogen removal in marine environments: recent findings and future research challenges

Respiratory reduction of nitrate (denitrification) is recognized as the most important process converting biologically available (fixed) nitrogen to N2. In current N cycle models, a major proportion of global marine denitrification (50–70%) is assumed to take place on the sea floor, particularly in organic rich continental margin sediments. Recent observations indicate that present conceptual views of denitrification and pathways of nitrate reduction and N2 formation are incomplete. Alternative N cycle pathways, particularly in sediments, include anaerobic ammonium oxidation to nitrite, nitrate and N2 by Mn-oxides, and anaerobic ammonium oxidation coupled to nitrite reduction and subsequent N2 mobilization. The discovery of new links and feedback mechanisms between the redox cycles of, e.g., C, N, S, Mn and Fe casts doubt on the present general understanding of the global N cycle. Recent models of the oceanic N budget indicate that total inputs are significantly smaller than estimated fixed N removal. The occurrence of alternative N reaction pathways further exacerbates the apparent imbalance as they introduce additional routes of N removal. In this contribution, we give a brief historical background of the conceptual understanding of N cycling in marine ecosystems, emphasizing pathways of aerobic and anaerobic N mineralization in marine sediments, and the implications of recently recognized metabolic pathways for N removal in marine environments

Physiological function and catalytic versatility of bacterial multihaem cytochromescinvolved in nitrogen and sulfur cycling

Bacterial MCCs (multihaem cytochromes c) represent widespread respiratory electron-transfer proteins. In addition, some of them convert substrates such as nitrite, hydroxylamine, nitric oxide, hydrazine, sulfite, thiosulfate or hydrogen peroxide. In many cases, only a single function is assigned to a specific MCC in database entries despite the fact that an MCC may accept various substrates, thus making it a multifunctional catalyst that can play diverse physiological roles in bacterial respiration, detoxification and stress defence mechanisms. The present article briefly reviews the structure, function and biogenesis of selected MCCs that catalyse key reactions in the biogeochemical nitrogen and sulfur cycles

Nutrient removal from UASB effluent in agro-industries

Phosphorus and nitrogen are important elements, making a major contribution to agricultural and industrial development, but their release to natural water bodies are the main causes of eutrophication. Anaerobic digestion yields effluents rich in ammonium and phosphate and poor in biodegradable organic carbon, thereby making them less suitable for conventional biological nitrogen and phosphorus removal. In addition, the demand for fertilizers is increasing, energy prices are rising and global phosphate reserves are declining. This requires both changes in wastewater treatment technologies and implementation of new processes. In this contribution the combination of an ureolytic MAP (magnesium ammonium phosphate) precipitation and autotrophic nitrogen removal is described on the anaerobic effluent of a potato processing company to obtain a more sustainable and cheaper method than conventional wastewater treatment processes. The results obtained during this experiment (6 weeks period) show that it is possible to recover phosphate as struvite and remove nitrogen with the autotrophic nitrogen process from wastewater after anaerobic digestion coming from a potato processing company. However further research is necessary to obtain stable results during several months, especially for the nitrite:ammonium ratio produced by the partial nitritation reactor

Effect of Tidal Cycling Rate on the Distribution and Abundance of Nitrogen-Oxidizing Bacteria in a Bench-Scale Fill-and-Drain Bioreactor

Most domestic wastewater can be effectively treated for secondary uses by engineered biological systems. These systems rely on microbial activity to reduce nitrogen (N) content of the reclaimed water. Such systems often employ a tidal-flow process to minimize space requirements for the coupling of aerobic and anaerobic metabolic processes. In this study, laboratory-scale tidal-flow treatment systems were studied to determine how the frequency and duration of tidal cycling may impact reactor performance. Fluorescent in situ hybridization and epifluorescence microscopy were used to enumerate the key functional groups of bacteria responsible for nitrification and anaerobic ammonium oxidation (anammox), and N-removal efficiency was calculated via a mass-balance approach. When water was cycled (i.e., reactors were filled and drained) at high frequencies (16–24 cycles day−1), nitrate accumulated in the columns—presumably due to inadequate periods of anoxia that limited denitrification. At lower frequencies, such as 4 cycles day−1, nearly complete N removal was achieved (80–90%). These fill-and-drain systems enriched heavily for nitrifiers, with relatively few anammox-capable organisms. The microbial community produced was robust, surviving well through short (up to 3 h) anaerobic periods and frequent system-wide perturbation

Construction, start-up and operation of a continuously aerated laboratory-scale SHARON reactor in view of coupling with an Anammox reactor

In this study practical experiences during start-up and operation of a laboratory-scale SHARON reactor are discussed, along with the construction of the reactor. Special attention is given to the start-up in view of possible toxic effects of high nitrogen concentrations (up to 4 000 mgN(.)l(-1)) on the nitrifier population and because the reactor was inoculated with sludge from an SBR reactor operated under completely different conditions. Because of these considerations, the reactor was first operated as an SBR to prevent biomass washout and to allow the selection of a strong nitrifying population. A month after the inoculation the reactor was switched to normal chemostat operation. As a result the nitrite oxidisers were washed out and only the ammonium oxidisers persisted in the reactor. In this contribution also some practical considerations concerning the operation of a continuously aerated SHARON reactor, such as mixing, evaporation and wall growth are discussed. These considerations are not trivial, since the reactor will be used for kinetic characterisation and modelling studies. Finally the performance of the SHARON reactor under different conditions is discussed in view of its coupling with an Anammox unit. Full nitrification was proven to be feasible for nitrogen loads up to 1.5 gTAN-N(.)l(-1.)d(-1), indicating the possibility of the SHARON process to treat highly loaded nitrogen streams. Applying different influent concentrations led to different effluent characteristics indicating the need for proper control of the SHARON reactor