Effect of the Aeration Strategy on NOB Suppression in Activated Sludge and Biofilm in a Hybrid Reactor with Nitrification/Denitrification

The purpose of the study was to analyse the impact of aeration strategies defined by the changes in the duration of aerated sub-phases, the ratio between non-aerated and aerated sub-phase times (R), and dissolved oxygen concentrations (DO) on the suppression of nitrite-oxidizing bacteria (NOB) in activated sludge and biofilm developing in a hybrid reactor with nitrification/denitrification. The primary factor causing NOB suppression both in biofilm and in activated sludge was an increase in the R-value (from 0 to 1/4 and from 1/4 to 1/3). After reducing the DO from 3 to 2 mg O2/L, there were no changes in the frequency of NOB occurrence, and no reduction in the nitrite oxidation rate was recorded. The abundance of Comammox bacteria was considerably affected by the change from continuous to intermittent aeration. Activated sludge showed a substantial increase in the quantity of clade A and B, whereas the quantity considerably decreased in biofilm.

Substantial oxygen consumption by aerobic nitrite oxidation in oceanic oxygen minimum zones

Oceanic oxygen minimum zones (OMZs) are globally significant sites of biogeochemical cycling where microorganisms deplete dissolved oxygen (DO) to concentrations <20 µM. Amid intense competition for DO in these metabolically challenging environments, aerobic nitrite oxidation may consume significant amounts of DO and help maintain low DO concentrations, but this remains unquantified. Using parallel measurements of oxygen consumption rates and 15N-nitrite oxidation rates applied to both water column profiles and oxygen manipulation experiments, we show that the contribution of nitrite oxidation to overall DO consumption systematically increases as DO declines below 2 µM. Nitrite oxidation can account for all DO consumption only under DO concentrations <393 nM found in and below the secondary chlorophyll maximum. These patterns are consistent across sampling stations and experiments, reflecting coupling between nitrate reduction and nitrite-oxidizing Nitrospina with high oxygen affinity (based on isotopic and omic data). Collectively our results demonstrate that nitrite oxidation plays a pivotal role in the maintenance and biogeochemical dynamics of OMZs.

Giant sulfur bacteria (Beggiatoaceae) from sediments underlying the Benguela upwelling system host diverse microbiomes

Due to their lithotrophic metabolisms, morphological complexity and conspicuous appearance, members of the Beggiatoaceae have been extensively studied for more than 100 years. These bacteria are known to be primarily sulfur-oxidizing autotrophs that commonly occur in dense mats at redox interfaces. Their large size and the presence of a mucous sheath allows these cells to serve as sites of attachment for communities of other microorganisms. But little is known about their individual niche preferences and attached microbiomes, particularly in marine environments, due to a paucity of cultivars and their prevalence in habitats that are difficult to access and study. Therefore, in this study, we compare Beggiatoaceae strain composition, community composition, and geochemical profiles collected from sulfidic sediments at four marine stations off the coast of Namibia. To elucidate community members that were directly attached and enriched in both filamentous Beggiatoaceae, namely Ca. Marithioploca spp. and Ca. Maribeggiatoa spp., as well as non-filamentous Beggiatoaceae, Ca. Thiomargarita spp., the Beggiatoaceae were pooled by morphotype for community analysis. The Beggiatoaceae samples collected from a highly sulfidic site were enriched in strains of sulfur-oxidizing Campylobacterota, that may promote a more hospitable setting for the Beggiatoaceae, which are known to have a lower tolerance for high sulfide to oxygen ratios. We found just a few host-specific associations with the motile filamentous morphotypes. Conversely, we detected 123 host specific enrichments with non-motile chain forming Beggiatoaceae. Potential metabolisms of the enriched strains include fermentation of host sheath material, syntrophic exchange of H2 and acetate, inorganic sulfur metabolism, and nitrite oxidation. Surprisingly, we did not detect any enrichments of anaerobic ammonium oxidizing bacteria as previously suggested and postulate that less well-studied anaerobic ammonium oxidation pathways may be occurring instead.

Controls on nitrite oxidation in the upper Southern Ocean: insights from winter kinetics experiments in the Indian sector

Across the Southern Ocean in winter, nitrification is the dominant mixed-layer nitrogen cycle process, with some of the nitrate produced therefrom persisting to fuel productivity during the subsequent growing season, potentially weakening the spring/summer biological CO2 sink. To better understand the controls on Southern Ocean nitrification, we conducted nitrite oxidation kinetics experiments in surface waters across the western Indian sector in winter. While all experiments (seven in total) yielded a Michaelis-Menten relationship with substrate concentration, the nitrite oxidation rates only increased substantially once the nitrite concentration exceeded 115±2.3 to 245±18 nM, suggesting that nitrite oxidizing bacteria (NOB) require a minimum (i.e., "threshold") nitrite concentration to produce nitrate. The half-saturation constant ranged from 134±8 to 403±24 nM, indicating a relatively high affinity of Southern Ocean NOB for nitrite, in contrast to results from culture experiments. Despite the high affinity of NOB for nitrite, its concentration rarely declines below 150 nM in the Southern Ocean's mixed layer, regardless of season. In the upper mixed layer, we measured ammonium oxidation rates that were two- to seven-fold higher than the coincident rates of nitrite oxidation, indicating that nitrite oxidation is the rate-limiting step for nitrification in the winter Southern Ocean. The decoupling of ammonium and nitrite oxidation, combined with a possible nitrite concentration threshold for NOB, may explain the non-zero nitrite that persists throughout the Southern Ocean's mixed layer year-round. We hypothesize that the apparent threshold nitrite requirement of NOB indicates nitrite undersaturation of the heme-rich nitrite oxidoreductase enzyme, perhaps driven by the limited availability of iron in surface waters.

Relevance of Candidatus Nitrotoga for nitrite oxidation in technical nitrogen removal systems

Many biotechnological applications deal with nitrification, one of the main steps of the global nitrogen cycle. The biological oxidation of ammonia to nitrite and further to nitrate is critical to avoid environmental damage and its functioning has to be retained even under adverse conditions. Bacteria performing the second reaction, oxidation of nitrite to nitrate, are fastidious microorganisms that are highly sensitive against disturbances. One important finding with relevance for nitrogen removal systems was the discovery of the mainly cold-adapted Cand. Nitrotoga, whose activity seems to be essential for the recovery of nitrite oxidation in wastewater treatment plants at low temperatures, e.g., during cold seasons. Several new strains of this genus have been recently described and ecophysiologically characterized including genome analyses. With increasing diversity, also mesophilic Cand. Nitrotoga representatives have been detected in activated sludge. This review summarizes the natural distribution and driving forces defining niche separation in artificial nitrification systems. Further critical aspects for the competition with Nitrospira and Nitrobacter are discussed. Knowledge about the physiological capacities and limits of Cand. Nitrotoga can help to define physico-chemical parameters for example in reactor systems that need to be run at low temperatures. Key points • Characterization of the psychrotolerant nitrite oxidizer Cand. Nitrotoga • Comparison of the physiological features of Cand. Nitrotoga with those of other NOB • Identification of beneficial environmental/operational parameters for proliferation

Physiology of the Nitrite-Oxidizing Bacterium Candidatus Nitrotoga sp. CP45 Enriched From a Colorado River

Nitrogen cycling microbes, including nitrite-oxidizing bacteria (NOB), perform critical ecosystem functions that help mitigate anthropogenic stresses and maintain ecosystem health. Activity of these beneficial nitrogen cycling microbes is dictated in part by the microorganisms’ response to physicochemical conditions, such as temperature, pH, and nutrient availability. NOB from the newly described Candidatus Nitrotoga genus have been detected in a wide range of habitats across the globe, yet only a few organisms within the genus have been physiologically characterized. For freshwater systems where NOB are critical for supporting aquatic life, Ca. Nitrotoga have been previously detected but little is known about the physiological potential of these organisms or their response to changing environmental conditions. Here, we determined functional response to environmental change for a representative freshwater species of Ca. Nitrotoga (Ca. Nitrotoga sp. CP45, enriched from a Colorado river). The physiological findings demonstrated that CP45 maintained nitrite oxidation at pH levels of 5–8, at temperatures from 4 to 28°C, and when incubated in the dark. Light exposure and elevated temperature (30°C) completely halted nitrite oxidation. Ca. Nitrotoga sp. CP45 maintained nitrite oxidation upon exposure to four different antibiotics, and potential rates of nitrite oxidation by river sediment communities were also resilient to antibiotic stress. We explored the Ca. Nitrotoga sp. CP45 genome to make predictions about adaptations to enable survival under specific conditions. Overall, these results contribute to our understanding of the versatility of a representative freshwater Ca. Nitrotoga sp. Identifying the specific environmental conditions that maximize NOB metabolic rates may ultimately direct future management decisions aimed at restoring impacted systems.

A Cu(II)-MOF Based on a Propargyl Carbamate-Functionalized Isophthalate Ligand as Nitrite Electrochemical Sensor

This paper investigates the electrochemical properties of a new Cu(II)-based metal-organic framework (MOF). Noted as Cu-YBDC, it is built upon a linker containing the propargyl carbamate functionality and immobilized on a glassy carbon electrode by drop-casting (GC/Cu-YBDC). Afterward, GC/Cu-YBDC was treated with HAuCl4 and the direct electro-deposition of Au nanoparticles was carried at 0.05 V for 600 s (GC/Au/Cu-YBDC). The performance of both electrodes towards nitrite oxidation was tested and it was found that GC/Au/Cu-YBDC exhibited a better electrocatalytic behavior toward the oxidation of nitrite than GC/Cu-YBDC with enhanced catalytic currents and a reduced nitrite overpotential from 1.20 to 0.90 V. Additionally GC/Au/Cu-YBDC showed a low limit of detection (5.0 μM), an ultrafast response time (<2 s), and a wide linear range of up to 8 mM in neutral pH.