Two metagenome-assembled genomes (MAGs), obtained from laboratory-scale enhanced biological phosphorus removal bioreactors, were analyzed. The values of 16S rRNA gene sequence identity, average nucleotide identity, and average amino acid identity indicated that these genomes, designated as RT and SSD2, represented two novel species within the genus Thiothrix, ‘Candidatus Thiothrix moscowensis’ and ‘Candidatus Thiothrix singaporensis’. A complete set of genes for the tricarboxylic acid cycle and electron transport chain indicates a respiratory type of metabolism. A notable feature of RT and SSD2, as well as other Thiothrix species, is the presence of a flavin adenine dinucleotide (FAD)-dependent malate:quinone oxidoreductase instead of nicotinamide adenine dinucleotide (NAD)-dependent malate dehydrogenase. Both MAGs contained genes for CO₂ assimilation through the Calvin–Benson–Bassam cycle; sulfide oxidation (sqr, fccAB), sulfur oxidation (rDsr complex), direct (soeABC) and indirect (aprBA, sat) sulfite oxidation, and the branched Sox pathway (SoxAXBYZ) of thiosulfate oxidation to sulfur and sulfate. All these features indicate a chemoorganoheterotrophic, chemolithoautotrophic, and chemolithoheterotrophic lifestyle. Both MAGs comprise genes for nitrate reductase and NO-reductase, while SSD2 also contains genes for nitrite reductase. The presence of polyphosphate kinase and exopolyphosphatase suggests that RT and SSD2 could accumulate and degrade polyhosphates during the oxic-anoxic growth cycle in the bioreactors, such as typical phosphate-accumulating microorganisms.

For ammonium removal from wastewater, anammox technologies are among the most efficient and rapidly developing ones. Due to the low growth rate of anammox bacteria and their sensitivity to various inhibitors, technologies using attached biocenosis carriers (ABCs) provide for reliable operation. The goal of the present work was to investigate a new ABC type, ETEK biochips based on a nonwoven fibrous material. The work involved the techniques of materials science (design of a new ABC type) and physical modeling of the anammox process (in a laboratory bioreactor), as well as electron microscopy and molecular profiling of activated sludge communities. Comparison of the ETEK biochips with the ABCs of foamed polyethylene BF33 and Mutag revealed more rapid accumulation (5-fold) of the activated sludge biomass on ETEK biochips upon reactor launching, as well as comparable buoyancy and reactor productivity regarding N removal. The specific rate of nitrogen removal obtained with ETEK biochips considerably exceeded that for foamed polyethylene with a filler: 1.5–3 times higher per chip and 1.5 times higher per activated sludge biomass unit. The studied ABC shared the same issue of floating to the surface due to the active formation of gas (N₂). The algorithm for calculating the downward flows in bioreactors with rapidly surfacing ABC is proposed, and a new hydrodynamic type of a bioreactor (with hybrid hydrodynamics) is described, a moving bed–sequencing batch reactor (MB-SBR).

Train Z-0024, a psychrotolerant aerobic heterotrophic representative of the prosthecate bacteria of the genus Caulobacter, was isolated from a methanotrophic enrichment obtained from Russian polar tundra soil. The cells of the new isolate are vibrios (0.5–0.6 × 1.3–1.8 mm) with a polar stalk. The organism grows in a temperature range from 5 to 36°C, with an optimum at 20°C. The pH range for growth is from 4.5 to 7.0 with an optimum at pH 6.0. Strain Z-0024 utilizes a wide range of organic compounds: sugars, amino acids, volatile fatty acids, and primary alcohols. It tolerates a NaCl concentration in the medium of up to 15 g/l. The G+C content of DNA is 66.6 mol %. The 16S rRNA gene sequence analysis revealed that strain Z-0024 belongs to the cluster of Caulobacter species, showing a 98.8–99.2% sequence similarity to them. DNA-DNA hybridization revealed a low level of homology (24%) between strain Z-0024 and C. vibrioides ATCC 15252. The new isolate is described as Caulobacter sp. Z-0024.

Candidatus Accumulibacter belongs to phosphate-accumulating organisms (PAOs) which exhibit a cyclic metabolism and are capable of intracellular polyphosphate accumulation and their hydrolysis under feast-famine anaerobic-aerobic cycling. In consortia of activated sludge microorganisms, these bacteria are responsible for enhanced biological phosphorus removal (EBPR). The spectrum of the substrates used by Ca. Accumulibacter remains insufficiently studied. It was investigated by measuring the oxygen uptake rates (OUR) of Ca. Accumulibacter-enriched culture supplemented with 17 different organic substrates. The highest oxygen uptake rate values were observed in the presence of tryptone, volatile fatty acids (acetate, propionate, and butyrate), succinate, pyruvate, and amino acids (aspartate and glutamate). Phosphate dynamics in the medium under shifts from anaerobic to aerobic cultivation in batch experiments were studied for these compounds (except for tryptone). All tested substrates were shown to cause phosphate cycling (release in the anaerobic phase and uptake in the aerobic one), with OURs for the substrates correlating with the number of phosphates consumed during the aerobic phase. It was concluded that OUR may be used as an indicator of the monosubstrates used by Ca. Accumulibacter in the anaerobic/aerobic cycle. The possible pathways for substrate transport and metabolism by Ca. Accumulibacter are discussed using stoichiometric data and the results of metagenomic analysis.

The psychrophilic aerobic heterotrophic bacterium, strain 1639^T, was isolated from the low-temperature Lake Untersee in Antarctica. The bacterium was Gram-positive, non-motile, yellow–green-pigmented, non-spore-forming, and a pleomorphic rod. Growth was observed at temperatures of 0–25 °C with an optimum at 10 °C. The strain used urea as a nitrogen source. The major fatty acids were i-C_16:0 (49.69%), ai-C_15:0 (17.59%), and C_16:1 branched (12.03%). Identified polar lipids were phosphatidylglycerols and a glycolipid. The respiratory quinone was determined to be MK-10. The genomic DNA G+C content was 68.03 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain 1639^T was a member of the genus Cryobacterium, with the highest sequence similarity to C. arcticum SK1^T (98.4%), C. soli GCJ02^T (98.4%), C. lactosi Sr59^T (98.3%), C. zongtaii TMN-42^T (98.2%), and C. adonitolivorans RHLS22-1^T (98.1%). The ANI and the DNA–DNA hybridization estimate values between strain 1639^T and all type strains of species of the genus Cryobacterium were in the range of 84.3–87.8% and 20.5–40.3%, respectively. The combined genotypic and phenotypic data indicate that strain 1639^T represents a novel species within the genus Cryobacterium, for which the name Cryobacterium inferilacus sp. nov. is proposed with the type strain 1639^T (=KCTC 59142^T, =VKM Ac-2907^T, UQM 41460^T).

During long-term cultivation in a laboratory sequencing batch reactor in the phosphorus removal mode, granule-like aggregates of two morphotypes, differing in their physical structure and microbial composition, with predominance of different microbial physiological groups, were formed out of the activated sludge floccules. Phosphate-accumulating bacteria Candidatus “Accumulibacter” prevailed in one morphotype, and glycogen-accumulating bacteria Candidatus “Competibacter,” in the other. These are aerobic microorganisms, known to compete for the substrate during the anaerobic period of the anaerobic/aerobic cultivation cycle. Aggregate formation paralleled development of phosphate-accumulating microorganisms, increase in their relative abundance, and higher phosphorus removal efficiency.