Ammonia-oxidizing archaea possess a wide range of cellular ammonia affinities

Nitrification, the oxidation of ammonia to nitrate, is an essential process in the biogeochemical nitrogen cycle. The first step of nitrification, ammonia oxidation, is performed by three, often co-occurring guilds of chemolithoautotrophs: ammonia-oxidizing bacteria (AOB), archaea (AOA), and complete ammonia oxidizers (comammox). Substrate kinetics are considered to be a major niche-differentiating factor between these guilds, but few AOA strains have been kinetically characterized. Here, the ammonia oxidation kinetic properties of 12 AOA representing all major cultivated phylogenetic lineages were determined using microrespirometry. Members of the genus Nitrosocosmicus have the lowest affinity for both ammonia and total ammonium of any characterized AOA, and these values are similar to previously determined ammonia and total ammonium affinities of AOB. This contrasts previous assumptions that all AOA possess much higher substrate affinities than their comammox or AOB counterparts. The substrate affinity of ammonia oxidizers correlated with their cell surface area to volume ratios. In addition, kinetic measurements across a range of pH values supports the hypothesis that—like for AOB—ammonia and not ammonium is the substrate for the ammonia monooxygenase enzyme of AOA and comammox. Together, these data will facilitate predictions and interpretation of ammonia oxidizer community structures and provide a robust basis for establishing testable hypotheses on competition between AOB, AOA, and comammox.

Discrepancy in exchangeable and soluble ammonium-induced effects on aerobic methane oxidation: a microcosm study of a paddy soil

Ammonium-induced stimulatory, inhibitory, and/or neutral effects on soil methane oxidation have been attributable to the ammonium concentration and mineral forms, confounded by other edaphic properties (e.g., pH, salinity), as well as the site-specific composition of the methanotrophic community. We hypothesize that this inconsistency may stem from the discrepancy in the cation adsorption capacity of the soil. We postulate that the effects of ammonium on the methanotrophic activity in soil are more accurately portrayed by relating methane uptake rates to the soluble ammonium (bioavailable), rather than the exchangeable (total) ammonium. To reduce adsorption (exchangeable) sites for ammonium in a paddy soil, two successive pre-incubation steps were introduced resulting in a 1000-fold soil dilution (soil enrichment), to be compared to a soil slurry (tenfold dilution) incubation. Ammonium was supplemented as NH4Cl at 0.5–4.75gL−1 after pre-incubation. While NH4Cl significantly stimulated the methanotrophic activity at all concentrations in the soil slurry incubation, methane uptake showed a dose-dependent effect in the soil enrichment. The trend in methane uptake could be explained by the soluble ammonium concentration, which was proportionate to the supplemented ammonium in the soil enrichment. In the soil slurry incubation, a fraction (36–63%) of the supplemented ammonium was determined to be adsorbed to the soil. Accordingly, Methylosarcina was found to predominate the methanotrophic community after the incubation, suggesting the relevance of this methanotroph at elevated ammonium levels (< 3.25gL−1 NH4Cl). Collectively, our results showed that the soluble, rather than the exchangeable ammonium concentration, is relevant when determining the effects of ammonium on methane oxidation, but this does not exclude other (a)biotic factors concurrently influencing methanotrophic activity.

Investigation on Recycling Dry Toilet Generated Blackwater by Anaerobic Digestion: From Energy Recovery to Sanitation

Anaerobic digestion (AD) has been widely adapted for blackwater treatment, however, the effect of water-conserving toilet generated blackwater on the AD process is still unknown. In this study, the anaerobic digestion process of dry toilet generated blackwater was investigated by means of a biomethane potential test. It was demonstrated that anaerobic digestion was inhibited and then adapted because of a high total ammonium nitrogen (TAN) level (3673.3 mg/L). The start-up period was 14.04 days and the biomethane potential of dry toilet blackwater was 402.36 mLCH4/gVS (55 days, 38 °C). Inhabitation and adaptation could be described as the increase of free ammonia nitrogen content and acetic acid concentration, followed by an enhancement of the relative abundance of acetic acid-type methanogens (from 33.53–61.52%). The main pathogen in dry toilet blackwater fermentation broth, Pseudomonas aeruginosa, kept multiplying in the first 8 days and then stabilized at a higher level than that of the beginning. This work showed the self-adjustment process and pathogen dynamics of dry toilet blackwater anaerobic digestion and highlights the significance of dry toilet blackwater characteristics when designing and maintaining anaerobic digestion sanitary treatment and reuse systems.

Microbiome of Seven Full-Scale Anaerobic Digestion Plants in South Korea: Effect of Feedstock and Operational Parameters

In this study, the microbiomes linked with the operational parameters in seven mesophilic full-scale AD plants mainly treating food waste (four plants) and sewage sludge (three plants) were analyzed. The results obtained indicated lower diversity and evenness of the microbial population in sludge digestion (SD) plants compared to food digestion (FD) plants. Candidatus Accumulibacter dominated (up to 42.1%) in SD plants due to microbial immigration from fed secondary sludge (up to 89%). Its potential activity in SD plants was correlated to H2 production, which was related to the dominance of hydrogenotrophic methanogens (Methanococcus). In FD plants, a balance between the hydrogenotrophic and methylotrophic pathways was found, while Flavobacterium and Levilinea played an important role during acidogenesis. Levilinea also expressed sensitivity to ammonia in FD plants. The substantial differences in hydraulic retention time (HRT), organic loading rate (OLR), and total ammonium nitrogen (TAN) among the studied FD plants did not influence the archaeal methane production pathway. In addition, the bacterial genera responsible for acetate production through syntrophy and homoacetogenesis (Smithella, Treponema) were present in all the plants studied.

Application of the Anammox Process for Treatment of Liquid Phase Digestate

The liquid phase of the digestate (LPD) contains a relatively high concentration of nitrogen, with total ammonium nitrogen being the dominant form of nitrogen, as well as other essential nutrients such as phosphorus and potassium. Consequently, it must be treated before it is released into the environment. However, there are no reports of co-purification of LPD in the anammox process in sequencing batch reactor with granular sludge, which is a novelty for the presented research. The main objective of this paper is to assess the possibility of nitrogen removal in the anammox process with LPD from biogas plants conducting the co-fermentation process along with the participation of agricultural products (cattle slurry). This publication presents the research results of the efficiency of the anammox process, accounting for the effect of dissolved organic matter. The conducted experiments revealed the potential of LPD purification, which co-ferments waste activated sludge and bovine slurry for the anammox process. In the reactor ammonium utilization rate (AUR) process with LPD addition increased from 2.3 mg N/(g VSS∙h) with 0.5% LPD addition to 8.5 mg N/(g VSS∙h) with 7.5% LPD addition. SAA in the reactor with LPD addition increased from 5.3 mg N/(g VSS∙h) with 0.5% LPD addition to 18.5 mg N/(g VSS∙h) with 4 and 5% LPD addition. With the addition of 7.5% LPD, SAA dropped to a value of 18.1 mg N/(g VSS∙h) in the LPD reactor.

An example of creating a biological filter Cyclobio with digital prototyping

It is important to note that biofilters are more cost-effective in comparison with other types of purification, which is due to the low concentration of pollutants entering fish tanks to bred objects. The best filters of this type are structures of more than four meters and with a circulating liquid, having as the main material fiberglass, little susceptible to chemical environments and the effects of microorganisms, which determines its durability and performance. The principle of operation of such a filter is based on the purification of water through a material covered with a biofilm of microbial colonies. The study was conducted to determine the algorithm for digital prototyping and designing the biofilter capacity for pseudo-liquefied sand layers. The stages of setting and solving the problem are defined. The selection of the amount of biological sand loading is described using the equation for predicting total ammonium nitrogen in the system (TAN). To process 67 kg per day of extruded feed with 56 % protein, 5 m3 of quartz sand with a D10 size of 0.1–0.3 mm is required. The technology of 3D modeling used for designing buildings, structures and equipment is described. Modeling and rendering of the biofilter capacity can be performed in the SketchUp program. The principle of selecting sand for creating pseudo-liquefied layers is described. Prospects for further application of the technology are outlined. The research results presented in the article have shown that when predicting the release of total ammonium nitrogen (TAN), they help in solving problems with the conversion of fish and aquatic aquatic life products into nitrates, allowing you to select the sand layer in the required amount. Modern methods of mathematical modeling for designing for the development and use of fish-breeding equipment, which is actively used in agriculture, are proposed. In this case, it is necessary to take into account the idealization of the model by conducting empirical research at the stage of meaningful modeling.

Effect of ammonia on fine-particle pH in agricultural regions of China: comparison between urban and rural sites

Particle acidity is a fundamental property that affects atmospheric particulate chemistry. Synchronous online monitoring was performed at two urban sites, Zhengzhou (U-ZZ) and Anyang (U-AY), and three rural sites, Anyang (R-AY), Xinxiang (R-XX), and Puyang (R-PY) in Henan Province, during a haze episode to investigate the pH value and its driving factors in the agricultural regions of China. The pH values of particles calculated by ISORROPIA-II model at rural sites were slightly higher than those at urban sites, with the median (min–max) values of 5.2 (4.8–6.9, R-PY), 5.1 (4.7–6.5, R-AY), 4.9 (4.1–6.8, R-XX), 4.8 (3.9–5.9, U-AY), and 4.5 (3.8–5.2, U-ZZ). Sensitivity tests show that TNHx (total ammonium, gas + aerosol) followed by total sulfate were the most important factors that influenced the predicted pH. Generally, particle pH increased with a cation increase and decreases in anions, temperature, and relative humidity. Similar pH values (∼3.0) at the required NHx concentrations for the five sites indicated that the presence of excess NHx was likely important for the lower acidity of PM2.5 during the severe haze episodes in this region. Moreover, the concentrations of excess NHx may drive the higher pH values at rural sites, because of the higher excess NHx concentrations at rural sites than those at urban sites. The underlying influence of regional transport on local particle pH cannot be neglected, as it differs the chemical components of PM2.5 and meteorological conditions. Air masses transported from rural and agricultural regions may enhance the particle pH value in urban aerosols given the high pH of particles and high ammonia levels. These results suggest that ammonia is urgently needed to be involved in the regional strategy for the improvement of air quality in China.

Effect of a Profound Feedstock Change on the Structure and Performance of Biogas Microbiomes

In this study the response of biogas-producing microbiomes to a profound feedstock change was investigated. The microbiomes were adapted to the digestion of either 100% sugar beet, maize silage, or of the silages with elevated amounts of total ammonium nitrogen (TAN) by adding ammonium carbonate or animal manure. The feedstock exchange resulted in a short-range decrease or increase in the biogas yields according to the level of chemical feedstock complexity. Fifteen taxa were found in all reactors and can be considered as generalists. Thirteen taxa were detected in the reactors operated with low TAN and six in the reactors with high TAN concentration. Taxa assigned to the phylum Bacteroidetes and to the order Spirochaetales increased with the exchange to sugar beet silage, indicating an affinity to easily degradable compounds. The recorded TAN-sensitive taxa (phylum Cloacimonetes) showed no specific affinity to maize or sugar beet silage. The archaeal community remained unchanged. The reported findings showed a smooth adaptation of the microbial communities, without a profound negative impact on the overall biogas production indicating that the two feedstocks, sugar beet and maize silage, potentially do not contain chemical compounds that are difficult to handle during anaerobic digestion.

Nitrogen Removal Performance and Metabolic Pathways Analysis of a Novel Aerobic Denitrifying Halotolerant Pseudomonas balearica Strain RAD-17

An aerobic denitrification strain, Pseudomonas balearica RAD-17, was identified and showed efficient inorganic nitrogen removal ability. The average NO3−-N, NO2−-N, and total ammonium nitrogen (TAN) removal rate (>95% removal efficiency) in a batch test was 6.22 mg/(L∙h), 6.30 mg/(L∙h), and 1.56 mg/(L∙h), respectively. Meanwhile, optimal incubate conditions were obtained through single factor experiments. For nitrogen removal pathways, the transcriptional results proved that respiratory nitrate reductases encoded by napA, which was primarily performed in aerobic denitrification and cell assimilation, were conducted by gluS and gluD genes for ammonium metabolism. In addition, adding the strain RAD-17 into actual wastewater showed obvious higher denitrification performance than in the no inoculum group (84.22% vs. 22.54%), and the maximum cell abundance achieved 28.5 ± 4.5% in a ratio of total cell numbers. Overall, the efficient nitrogen removal performance plus strong environmental fitness makes the strain RAD-17 a potential alternative for RAS (recirculating aquaculture system) effluent treatment.