High-resolution mapping and modeling of anammox recovery from recurrent oxygen exposure.

Oxygen inhibits anammox, a bioconversion executed by anoxic ammonium oxidizing bacteria (AnAOB). Nonetheless, oxygen is mostly found in the proximity of AnAOB in nitrogen removal applications, being a substrate for nitritation. The experiments performed to date were mostly limited to batch activity tests where AnAOB activity is estimated during oxygen exposure. However, little attention has been paid to the recovery and reversibility of activity following aerobic conditions, of direct relevance for bioreactor operation. In this work, anoxic and autotrophic reactor cultivation at 20 °C yielded an enriched microbial community in AnAOB, consisting for 75% of a member of the genus Brocadia. High-resolution kinetic data were obtained with online ammonium measurements and further processed with a newly developed Python data pipeline. The experimentally obtained AnAOB response showed complete inhibition until micro-aerobic conditions were reached again (<0.02 mg O2 L-1). After oxygen inhibition, AnAOB recovered gradually, with recovery times of 5-37 h to reach a steady-state activity, dependent on the perceived inhibition. The recovery immediately after inhibition was lowest when exposed to higher oxygen concentrations (range: 0.5-8 mg O2 L-1) with long contact times (range: 9-24 h). The experimental data did not fit well with a conventional 'instant recovery' Monod-type inhibition model. Yet, the fit greatly improved by incorporating a dynamic growth rate formula accurately describing gradual activity recovery. With the upgraded model, long-term kinetic simulations for partial nitritation/anammox (PN/A) with intermittent aeration showed a decrease in growth rate compared to the instant recovery mode. These results indicate that recovery of AnAOB after oxygen exposure was previously overlooked. It is recommended to account for this effect in the intensification of partial nitritation/anammox.

Enrichment and adaptation yield high anammox conversion rates under low temperatures.

This study compared two anammox sequencing batch reactors (SBR) for one year. SBRconstantT was kept at 30 °C while temperature in SBRloweringT was decreased step-wise from 30 °C to 20 °C and 15 °C followed by over 140 days at 12.5 °C and 10 °C. High retention of anammox bacteria (AnAOB) and minimization of competition with AnAOB were key. 5-L anoxic reactors with the same inoculum were fed synthetic influent containing 25.9 mg NH4+-N/L and 34.1 mg NO2--N/L (no COD). Specific ammonium removal rates continuously increased in SBRconstantT, reaching 785 mg NH4+-N/gVSS/d, and were maintained in SBRloweringT, reaching 82.2 and 91.8 mg NH4+-N/gVSS/d at 12.5 and 10 °C respectively. AnAOB enrichment (increasing hzsA and 16S rDNA gene concentrations) and adaptation (shift from Ca. Brocadia to Ca. Kuenenia in SBRloweringT) contributed to these high rates. Rapidly settling granules developed, with average diameters of 1.2 (SBRconstantT) and 1.6 mm (SBRloweringT). Results reinforce the potential of anammox for mainstream applications.

Uncoupling the solids retention times of flocs and granules in mainstream deammonification: A screen as effective out-selection tool for nitrite oxidizing bacteria.

This study focused on a physical separator in the form of a screen to out-select nitrite oxidizing bacteria (NOB) for mainstream sewage treatment. This separation relied on the principle that the NOB prefer to grow in flocs, while anammox bacteria (AnAOB) reside in granules. Two types of screens (vacuum and vibrating) were tested for separating these fractions. The vibrating screen was preferred due to more moderate normal forces and additional tangential forces, better balancing retention efficiency of AnAOB granules (41% of the AnAOB activity) and washout of NOB (92% activity washout). This operation resulted in increased NOB out-selection (AerAOB/NOB ratio of 2.3) and a total nitrogen removal efficiency of 70% at influent COD/N ratio of 1.4. An effluent total nitrogen concentration <10mgN/L was achieved using this novel approach combining biological selection with physical separation, opening up the path towards energy positive sewage treatment.

Impact of carbon to nitrogen ratio and aeration regime on mainstream deammonification.

While deammonification of high-strength wastewater in the sludge line of sewage treatment plants has become well established, the potential cost savings spur the development of this technology for mainstream applications. This study aimed at identifying the effect of aeration and organic carbon on the deammonification process. Two 10 L sequencing bath reactors with different aeration frequencies were operated at 25°C. Real wastewater effluents from chemically enhanced primary treatment and high-rate activated sludge process were fed into the reactors with biodegradable chemical oxygen demand/nitrogen (bCOD/N) of 2.0 and 0.6, respectively. It was found that shorter aerobic solids retention time (SRT) and higher aeration frequency gave more advantages for aerobic ammonium-oxidizing bacteria (AerAOB) than nitrite oxidizing bacteria (NOB) in the system. From the kinetics study, it is shown that the affinity for oxygen is higher for NOB than for AerAOB, and higher dissolved oxygen set-point could decrease the affinity of both AerAOB and NOB communities. After 514 days of operation, it was concluded that lower organic carbon levels enhanced the activity of anoxic ammonium-oxidizing bacteria (AnAOB) over denitrifiers. As a result, the contribution of AnAOB to nitrogen removal increased from 40 to 70%. Overall, a reasonably good total removal efficiency of 66% was reached under a low bCOD/N ratio of 2.0 after adaptation.

Energy efficient treatment of A-stage effluent: pilot-scale experiences with shortcut nitrogen removal.

Energy autarky of sewage treatment plants, while reaching chemical oxygen demand (COD) and N discharge limits, can be achieved by means of shortcut N-removal. This study presents the results of a shortcut N-removal pilot, located at the biological two-'stage (high/low rate) wastewater treatment plant of Breda, The Netherlands. The pilot treated real effluent of a high-rate activated sludge (COD/N = 3), fed in a continuous mode at realistic loading rates (90-100 g N/(m(3)·d)). The operational strategy, which included increased stress on the sludge settling velocity, showed development of a semi-granular sludge, with average particle size of 280 µm (ø(4,3)), resulting in increased suppression of nitrite-oxidizing bacteria. The process was able to remove part of the nitrogen (51 ± 23%) over nitrite, with COD/N removal ratios of 3.2 ± 0.9. The latter are lower than the current operation of the full-scale B-stage in Breda (6.8-9.4), showing promising results for carbon-efficient N-removal, while producing a well settling sludge (SVI(30) < 100 mL/g).

Production of carboxylates from high rate activated sludge through fermentation.

The aim of this work was to study the key parameters affecting fermentation of high rate activated A-sludge to carboxylates, including pH, temperature, inoculum, sludge composition and iron content. The maximum volatile fatty acids production was 141mgCg(-1) VSSfed, at pH 7. Subsequently the potential for carboxylate and methane production for A-sludge from four different plants at pH 7 and 35°C were compared. Initial BOD of the sludge appeared to be key determining carboxylate yield from A-sludge. Whereas methanogenesis could be correlated linearly to the quantity of ferric used for coagulation, fermentation did not show a dependency on iron presence. This difference may enable a strategy whereby A-stage sludge is separated to achieve fermentation, and iron dosing for phosphate removal is only implemented at the B-stage.

Effective carbon and nutrient treatment solutions for mixed domestic-industrial wastewater in India.

The present study evaluates effectiveness of up-flow anaerobic sludge blanket (UASB) reactor followed by two post-anaerobic treatment options, namely free-surface, up-flow constructed wetland (FUP-CW) and oxygen-limited anaerobic nitrification/denitrification (OLAND) processes in treating sewage from the peri-urban areas in India receiving illegal industrial infiltrations. The UASB studies yielded robust results towards fluctuating strength of sewage and consistently removed 87-98% chemical oxygen demand (COD) at a hydraulic retention time of 1.5-2 d. The FUP-CW removed 68.5±13% COD, 68±3% NH4+-N, 38±5% PO4(3-)-P, 97.6±5% suspended particles and 97±13% fecal coliforms. Nutrient removal was found to be limiting in FUP-CW, especially in winter. Nitrogen removal in the OLAND process were 100 times higher than the FUP-CW process. Results show that UASB followed by FUP-CW can be an excellent, decentralized sewage treatment option, except during winter when nutrient removal is limited in FUP-CW. Hence, the study proposes bio-augmentation of FUP-CW with OLAND biomass for overall improvement in the performance of UASB followed by FUP-CW process.

When the smoke disappears: dealing with extinguishing chemicals in firefighting wastewater.

Water is not enough. Nowadays, numerous chemicals are used for fire extinction. After use, however, these may unintentionally enter sewerage systems. In order to safely treat firefighting wastewater (FFWW), knowledge of the potential effects of these chemicals on biological treatment processes is essential. This study characterized and mimicked the composition of FFWW containing two powders, three foams and one foam degrader. Nitrogen (162-370 mg NH4(+)-N L(-1)) and phosphorus (173-320 mg PO4(3-)-P L(-1)) concentrations exceeded discharge limits, whereas chemical and biological oxygen demand, suspended solids and detergent concentrations remained sufficiently low. Adequate nutrient removal could be obtained through FeCl3 addition and nitrification/denitrification with acetate as substrate. In batch tests, residual nitrifying activities of 84, 81, 89, 95 and 93% were observed in the presence of powders, foams, foam degrader, synthetic and real FFWW, respectively. All categories showed higher denitrification rates than the control. Although the powders at first seemed to inhibit anammox activity at 82%, after pH correction anammox was fully feasible, allowing nitrogen removal through oxygen-limited nitrification/denitrification (OLAND). Detailed cost calculations indicated that OLAND could save 11% of capital and 68% of operational costs compared to nitrification/denitrification, identifying OLAND as the most recommendable process for nitrogen removal from firefighting wastewaters.

Microbial resource management of one-stage partial nitritation/anammox.

About 30 full-scale partial nitritation/anammox plants are established, treating mostly sewage sludge reject water, landfill leachate or food processing digestate. Although two-stage and one-stage processes each have their advantages, the one-stage configuration is mostly applied, termed here as oxygen-limited autotrophic nitrification/denitrification (OLAND), and is the focus of this review. The OLAND application domain is gradually expanding, with technical-scale plants on source-separated domestic wastewater, pre-treated manure and sewage, and liquors from organic waste bioenergy plants. A 'microbial resource management' (MRM) OLAND framework was elaborated, showing how the OLAND engineer/operator (1: input) can design/steer the microbial community (2: biocatalyst) to obtain optimal functionality (3: output). In the physicochemical toolbox (1), design guidelines are provided, as well as advantages of different reactor technologies. Particularly the desirable aeration regime, feeding regime and shear forces are not clear yet. The development of OLAND trickling filters, membrane bioreactors and systems with immobilized biomass is awaited. The biocatalyst box (2) considers 'Who': biodiversity and its dynamic patterns, 'What': physiology, and 'Where': architecture creating substrate gradients. Particularly community dynamics and extracellular polymeric substances (EPS) still require insights. Performant OLAND (3) comprises fast start-up (storage possibility; fast growth of anammox bacteria), process stability (endured biomass retention; stress resilience), reasonable overall costs, high nitrogen removal efficiency and a low environmental footprint. Three important OLAND challenges are elaborated in detailed frameworks, demonstrating how to maximize nitrogen removal efficiency, minimize NO and N(2)O emissions and obtain through OLAND a plant-wide net energy gain from sewage treatment.

Fast start-up of a pilot-scale deammonification sequencing batch reactor from an activated sludge inoculum.

Deammonification involves the combined application of aerobic and anoxic ammonium-oxidizing bacteria (AerAOB & AnAOB) and allows to treat wastewaters with a high ammonium concentration in a sustainable and cost-efficient way. So far, it could take more than one year to start up the process, even with the addition of AnAOB enriched inocula. In contrast, we started up a deammonifying reactor for the treatment of sludge digestate in less than four months without any AnAOB enriched inoculum. In a single sequencing batch reactor (SBR) of 3 m(3), nitritation and anammox were performed without nitrite accumulation. Larger biomass aggregates (>1.0 mm) had a typical reddish colour, but FISH also showed that small aggregates (<0.25 mm) contained a considerable amount of AnAOB. The AerAOB were related to Nitrosomonas halophila, N. eutropha and N. halophila, and the AnAOB to "Candidatus Kuenenia & Brocadia", as shown by FISH. Our results show that the deammonification inoculum does not play an important role, and that the AnAOB can quickly develop under the proper aerational conditions. Nitrogen was removed stably at high nitrogen loading rates (740 mg N/L/d) and removal efficiency (90%).

Granular biomass capable of partial nitritation and anammox.

Water Science and Technology 58(5) 1113-1120. Publisher's note. We regret that an outdated version of this article was used in production; the correct final version, which incorporates several amendments and different authorship, is printed below.

Granular biomass capable of partial nitritation and anammox.

A novel and efficient way of removing nitrogen from wastewater poor in biodegradable organic carbon, is the combination of partial nitritation and anoxic ammonium oxidation (anammox), as in the one-stage oxygen-limited autotrophic nitrification/denitrification (OLAND) process. Since anoxic ammonium-oxidizing bacteria grow very slowly, maximum biomass retention in the reactor is required. In this study, a lab-scale sequencing batch reactor (SBR) was used to develop granular, rapidly settling biomass. With SBR cycles of one hour and a minimum biomass settling velocity of 0.7 m/h, OLAND granules were formed in 1.5 months and the nitrogen removal rate increased from 50 to 450 mg N L(-1) d(-1) in 2 months. The granules had a mean diameter of 1.8 mm and their aerobic and anoxic ammonium-oxidizing activities were well equilibrated to perform the OLAND reaction. Fluorescent in-situ hybridization (FISH) demonstrated the presence of both beta-proteobacterial aerobic ammonium oxidizers and planctomycetes (among which anoxic ammonium oxidizers) in the granules. The presented results show the applicability of rapidly settling granular biomass for one-stage partial nitritation and anammox.