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The class Limnochordia harbors a single cultivated member, the mesophilic Limnochorda pilosa, which was isolated from a meromictic lake. Despite numerous molecular signatures reported in various ecosystems, the ecophysiological versatility of this deeply branched lineage of Firmicutes (Bacillota) remains poorly understood. The objective of this study was to use targeted cultivation, based on metagenome-assembled genomes from a deep terrestrial aquifer in Western Siberia, to isolate two new thermophilic members of the class. These isolates, described as Geochorda subterranea gen. nov. sp. nov. and Carboxydochorda subterranea gen. nov. sp. nov. within the Geochordaceae fam. nov., were capable of both anaerobic and aerobic respiration using fumarate and O2, respectively, with simple sugars as electron donors. The cultivated Geochordaceae have demonstrated fermentative growth and degradation of various polymers, including starch, maltose, maltodextrin, xylan, and chitin. The carboxydotrophic C. subterranea sp. nov. exhibited autotrophic growth via the Calvin–Benson–Bassham cycle, using CO, H2, and formate as electron donors and O2 as an electron acceptor, adding metabolic flexibility to the bacterium in the nutrient-depleted “deep biosphere” and supporting the possibility of aerobic metabolism in the deep subsurface. The broad physiological potential deciphered from physiological experiments and comparative genomic data explains the widespread distribution of uncultivated members of the class Limnochordia in various ecosystems, where they can oxidize complex organic substrates through both aerobic and anaerobic respiration, as well as pursue a chemolithotrophic lifestyle through the oxidation of H2 or CO.

Irumamycin (Iru) is a complex polyketide with pronounced antifungal activity produced by a type I polyketide (PKS) synthase. Iru features a unique hemiketal ring and an epoxide group, making its biosynthesis and the structural diversity of related compounds particularly intriguing. In this study, we performed a detailed analysis of the iru biosynthetic gene cluster (BGC) to uncover the mechanisms underlying Iru formation. We examined the iru PKS, including the domain architecture of individual modules and the overall spatial structure of the PKS, and uncovered discrepancies in substrate specificity and iterative chain elongation. Two potential pathways for the formation of the hemiketal ring, involving either an olefin shift or electrocyclization, were proposed and assessed using 18O-labeling experiments and reaction activation energy calculations. Based on our findings, the hemiketal ring is likely formed by PKS-assisted double bond migration and TE domain-mediated cyclization. Furthermore, putative tailoring enzymes mediating epoxide formation specific to Iru were identified. The revealed Iru biosynthetic machinery provides insight into the complex enzymatic processes involved in Iru production, including macrocycle sculpting and decoration. These mechanistic details open new avenues for a targeted architecture of novel macrolide analogs through synthetic biology and biosynthetic engineering.

Sulfidogenic bacteria cause numerous issues in the oil industry since they produce sulfide, corroding steel equipment, reducing oil quality, and worsening the environmental conditions in oil fields. The purpose of this work was to isolate and taxonomically identify the sulfidogenic bacteria responsible for the corrosion of steel equipment at the Karazhanbas oil field (Kazakhstan). In this study, we characterized five sulfidogenic strains of the genera Pseudodesulfovibrio, Oleidesulfovibrio, and Acetobacterium isolated from the formation water of the Karazhanbas oil field (Kazakhstan). Sulfate-reducing strain 9FUST revealed 98.9% similarity of the 16S rRNA gene sequence with the closely related strain ‘Pseudodesulfovibrio methanolicus’ 5S69T and was studied in detail to enhance the taxonomic resolution. Strain 9FUST grew optimally at 23–28 °C, pH 6.5, and 0–2% (w/v) NaCl. The strain used lactate, pyruvate, methanol, ethanol, fructose, ribose, and H2/CO2 (in the presence of acetate) as carbon and energy sources for sulfate reduction. Iso-C17:1 ω11, C15:0, iso-C15:0, and C16:0 were the predominant fatty acids. The genome is 4.20 Mbp with a G + C content of 64.0%. The average nucleotide identity and digital DNA–DNA hybridization values with Pseudodesulfovibrio spp. genomes were 72.5–91.6% (T (=VKM B-3654T = KCTC 25498T) belonged to a novel Pseudodesulfovibrio species, for which the name Pseudodesulfovibrio karagichevae sp. nov. is proposed. Pangenome analysis of sixteen Pseudodesulfovibrio species and functional annotation analysis of identified genes revealed complete modules of enzymes of the main metabolic pathways, characteristic of bacteria of this genus, and unique genes highlighting the adaptations of strain 9FUST in carbohydrate metabolism, nutrient uptake, and environmental stress response. Isolation of these strains expands our understanding of the diversity of sulfidogens in oil reservoirs and can be used to test the effectiveness of biocides used in an oil field.

In winemaking, malolactic fermentation (MLF), which converts L-malic acid to L-lactic acid, is often applied after the alcoholic fermentation stage to improve the sensory properties of the wine and its microbiological stability. MLF is usually performed by lactic acid bacteria, which, however, are sensitive to the conditions of alcoholic fermentation. Therefore, the development of wine yeast strains capable of both alcoholic fermentation and MLF is an important task. Using genome editing, we engineered a modified variant of the triploid wine yeast strain Saccharomyces cerevisiae I-328, in which the CAR1 arginase gene was replaced by the malate permease gene from Schizosaccharomyces pombe and the malolactic enzyme gene from Oenococcus oeni. Genome-wide transcriptional profiling confirmed the expression of the introduced genes and revealed a limited effect of the modification on global gene expression. Winemaking experiments show that genome editing did not affect fermentation activity and ethanol production, while use of the modified strain resulted in a tenfold reduction in malate content with simultaneous formation of lactate. The resulting wines had a softer and more harmonious taste compared to wine obtained using the parental strain. Inactivation of arginase, which forms urea and L-ornithine through the breakdown of arginine, also resulted in a twofold decrease in the content of urea and the carcinogenic ethyl carbamate in wine. Thus, the new strain with the replacement of the arginase gene with the MLF gene cassette is promising for use in winemaking.

This article presents a mathematical model that describes the movement of suspended aerosol particles in a mixed-type filter. For the carrier medium, a model of incompressible fluid flow in a porous periodic element consisting of many cylinders with diameters in the nano- and micrometer range was adopted. The Stokes boundary value problem for the flow function was solved using the boundary element method. The efficiency of the deposition of suspended particles was determined by calculating their trajectories in the identified velocity field of the carrier medium. For various ratios of mass fractions of nano- and microfibers, the dependences of the deposition efficiency due to the inertial impact and interception mechanism on the Stokes number were found.

Gold recovery from refractory pyrite-arsenopyrite concentrates using stirred tank reactor biooxidation is widely applied worldwide. Therefore, studies to address the characteristic problem of this technology are urgent. The goal of the present work was to research the possibility of counteracting the negative effects of unfavorable conditions (increasing pulp density and temperature) on the biooxidation of pyrite-arsenopyrite concentrate in laboratory-scale stirred tank reactors using additional carbon supply in the form of CO2. A refractory concentrate containing pyrite (48%) and arsenopyrite (13%) was used in biooxidation experiments. In the control experiment, biooxidation was performed under “normal conditions”: temperature 40 °C, pulp density (solid to liquid ratio, S:L) 1:10, residence time 5 days. It was shown that under “normal conditions”, additional carbon dioxide supply insignificantly affected the biooxidation rate and composition of the microbial population of biooxidation reactors. In addition, the effect of “stressful conditions” was studied. In this case, either temperature or pulp density were increased (up to 50 °C and S:L 1:5, respectively), which provided unfavorable conditions for biooxidation and led to the decrease in biooxidation rate. Under “stressful conditions”, additional carbon dioxide supply affected biooxidation to a greater extent and made it possible to increase both pyrite and arsenopyrite biooxidation rates. The analysis of microbial populations showed that additional carbon dioxide supply also changed their composition.

The phylum Verrucomicrobiota is one of the main groups of soil prokaryotes, which remains poorly represented by cultivated organisms. The major recognized role of Verrucomicrobiota in soils is the degradation of plant-derived organic matter. These bacteria are particularly abundant in peatlands, where xylan-type hemicelluloses represent one of the most actively decomposed peat constituents. The aim of this work was to characterize the microorganisms capable of hydrolyzing xylan under the anoxic conditions typical of peatland soils. The laboratory incubation of peat samples with xylan resulted in the pronounced enrichment of several phylotypes affiliated with the Verrucomicrobiota, Firmicutes, and Alphaproteobacteria. Sequencing of the metagenome of the enrichment culture allowed us to recover high-quality metagenome-assembled genomes (MAGs) assigned to the genera Caproiciproducens, Clostridium, Bacillus (Firmicutes), and Rhizomicrobium (Alphaproteobacteria), Cellulomonas (Actinobacteriota) and the uncultured genus-level lineage of the family Chthoniobacteraceae (Verrucomicrobiota). The latter bacterium, designated “Candidatus Chthoniomicrobium xylanophilum” SH-KS-3, dominated in the metagenome and its MAG was assembled as a complete closed chromosome. An analysis of the SH-KS-3 genome revealed potential endo-1,4-beta-xylanases, as well as xylan beta-1,4-xylosidases and other enzymes involved in xylan utilization. A genome analysis revealed the absence of aerobic respiration and predicted chemoheterotrophic metabolism with the capacity to utilize various carbohydrates, including cellulose, and to perform fermentation or nitrate reduction. An analysis of other MAGs suggested that Clostridium and Rhizomicrobium could play the role of primary xylan degraders while other community members probably took advantage of the availability of xylo-oligosaccharides and xylose or utilized low molecular weight organics.

The search for the microorganisms responsible for sulfide formation and corrosion of steel equipment in the oil fields of Tatarstan (Russia) resulted in the isolation of a new halotolerant strictly anaerobic sulfate-reducing bacterium, strain 5S69T. The cells were motile curved Gram-negative rods. Optimal growth was observed in the presence of 2.0–4.0% (w/v) NaCl, at pH 6.5, and at 23–28 °C under sulfate-reducing conditions. The isolate was capable of chemoorganotrophic growth with sulfate and other sulfoxides as electron acceptors, resulting in sulfide formation; and of pyruvate fermentation resulting in formation of H2 and acetate. The strain utilized lactate, pyruvate, ethanol, methanol, fumarate, and fructose, as well as H2/CO2/acetate for sulfate reduction. The genome size of the type strain 5S69T was 4.16 Mb with a G + C content of 63.0 mol%. On the basis of unique physiological properties and results of the 16S rRNA gene-based phylogenetic analysis, phylogenomic analysis of the 120 conserved single copy proteins and genomic indexes (ANI, AAI, and dDDH), assigning the type strain 5S69T ((VKM B-3653T = KCTC 25499T) to a new species within the genus Pseudodesulfovibrio, is suggested, with the proposed name Pseudodesulfovibrio methanolicus sp. nov. Genome analysis of the new isolate showed several genes involved in sulfate reduction and its sulfide-producing potential in oil fields with high saline formation water.

Создание руководства поддержано Советом по терапевтическим наукам отделения клинической медицины Российской академии наук.

Aim. To study the features of treatment of patients with various types of coronary artery disease (CAD) and type 2 diabetes (T2D) at various healthcare stages.Material and methods. The comparative clinical study included 412 patients of both sexes with acute and chronic coronary artery disease. Depending on the glycemic status and CAD type, patients were divided into four subgroups: subgroup 1a (n=100, 56,6±0,96 years, male/female 67/33) — with acute CAD and T2D; subgroup 1b (n=106, 58,7±1,01 years, male/female 75/31) — with acute CAD without T2D; subgroup 2a (n=102, 57,9±1,04 years, male/female 72/30) — with chronic CAD and T2D; subgroup 2b (n=104, 60,2±0,9 years, 69/35) — with chronic CAD without T2D. The selection and mean doses of standard and hypoglycemic therapy were analyzed.Results. Initially, 86% and 81% of patients with diabetes in subgroups 1a and 2a adhered to glucose-lowering therapy. Among patients with chronic CAD, more than 1/4 (26,4%) received insulin therapy versus 2% in the group of patients with acute coronary syndrome (ACS) and diabetes. After discharge from hospital, oral hypoglycemic therapy was prescribed to 74% of patients with ACS and 48% of patients with chronic CAD. The prescription of insulin therapy for people with ACS increased 5 times (up to 10% of the total number of patients with ACS and T2D). By the time of hospital admission, 70% and 71,5% of patients with ACS and chronic CAD with diabetes received acetylsalicylic acid as an antiplatelet agent, respectively. During hospitalization, all patients with acute CAD were prescribed dual antiplatelet therapy, as well as triple antiplatelet therapy in the case of atrial fibrillation/flutter. It is noteworthy that in the group of patients with chronic CAD, less than 70% received statins. In patients with chronic CAD and diabetes, this parameter was even lower, amounting to 59,8%. At the outpatient stage, all patients, with the exception of the group with chronic CAD without diabetes, took beta-blockers (up to 65%). In the hospital, the use of beta blockers increased to 95%. During inpatient treatment, angiotensin-converting enzyme inhibitors and sartans were prescribed 1,5-2 times more often than in the prehospital stage.Conclusion. The study indicates insufficient prescription of standard CAD and glucose-lowering therapy. Careful inpatient selection of therapy requires continuity at the outpatient stage.

Planctomycetes of the genus Singulisphaera are common inhabitants of soils and peatlands. Although described members of this genus are characterized as possessing hydrolytic capabilities, the ability to degrade chitin has not yet been reported for these bacteria. In this study, a novel Singulisphaera representative, strain Ch08, was isolated from a chitinolytic enrichment culture obtained from a boreal fen in Northern European Russia. The 16S rRNA gene sequence of this isolate displayed 98.2% similarity to that of Singulisphaera acidiphila MOB10T. Substrate utilization tests confirmed that strain Ch08 is capable of growth on amorphous chitin. The complete genome of strain Ch08 determined in this study was 10.85 Mb in size and encoded two predicted chitinases, which were only distantly related to each other and affiliated with the glycoside hydrolase family GH18. One of these chitinases had a close homologue in the genome of S. acidiphila MOB10T. The experimental verification of S. acidiphila MOB10T growth on amorphous chitin was also positive. Transcriptome analysis performed with glucose- and chitin-growth cells of strain Ch08 showed upregulation of the predicted chitinase shared by strain Ch08 and S. acidiphila MOB10T. The gene encoding this protein was expressed in Escherichia coli, and the endochitinase activity of the recombinant enzyme was confirmed. The ability to utilize chitin, a major constituent of fungal cell walls and arthropod exoskeletons, appears to be one of the previously unrecognized ecological functions of Singulisphaera-like planctomycetes.

While granulated activated sludge exhibits high productivity, the processes of granule formation are incompletely studied. The processes of granule formation and succession of communities were investigated in a laboratory sequencing batch reactor (SBR) under conditions for enhanced biological phosphorus removal (EBPR) using microbiological and molecular techniques. Active consumption of acetate, primarily by the phosphate-accumulating organisms (PAO), commenced at day 150 of cultivation. This was indicated by the high ratio of molar P-released/acetate uptake (0.73–0.77 P-mol/C-mol), characteristic of PAO. During this period, two types of granule-like aggregates formed spontaneously out of the activated sludge flocs. The aggregates differed in morphology and microbial taxonomic composition. While both aggregate types contained phosphorus-enriched bacterial cells, PAO prevailed in those of morphotype I, and glycogen-accumulating organisms (GAOs) were predominant in the aggregates of morphotype II. After 250 days, the elimination of the morphotype II aggregates from the reactor was observed. The subsequent selection of the community was associated with the development of the morphotype I aggregates, in which the relative abundance of PAO increased significantly, resulting in higher efficiency of phosphorus removal. Metagenomic analysis revealed a predominance of the organisms closely related to Candidatus Accumulibacter IС and IIС and of Ca. Accumulibacter IIB among the PAO. Based on the content of the genes of the key metabolic pathways, the genomes of potential PAO belonging to the genera Amaricoccus, Azonexus, Thauera, Zoogloea, Pinisolibacter, and Siculibacillus were selected. The patterns of physicochemical processes and the microbiome structure associated with granule formation and succession of the microbial communities were revealed.

THE PURPOSE of the article is to discuss an updated survey of the technical condition of porcelain high-voltage insulators of the type IOS 110/400, which had defects with partial discharge intensities above 4 nC, at Tatenergo substations. The experiments were carried out with simultaneous use of electromagnetic and acoustic research methods.During the analysis of the RESULTS obtained, the influence of the fields of previous powerful partial discharges on the technical condition of high-voltage insulators was established. The physical features of detecting powerful partial discharges by electromagnetic and acoustic methods have been studied. The conducted studies allow us to draw the following.CONCLUSIONS: the previous partial discharges can create additional long-lived electric fields on the dielectric surfaces of defects that exceed the field of the applied operating voltage in intensity; the necessity of using both physical methods simultaneously in the examination of high-voltage insulators in operation is shown.

In this paper, we consider an optimal control problem with a «pure», integral boundary condition. The Green’s function is constructed. Using contracting Banach mappings, a sufficient condition for the existence and uniqueness of a solution to one class of integral boundary value problems for fixed admissible controls is established. Using the functional increment method, the Pontryagin‘s maximum principle is proved. The first and second variations of the functional are calculated. Further, various necessary conditions for optimality of the second order are obtained by using variations of controls.

Burning coal seams, characterized by massive carbon monoxide (CO) emissions, the presence of secondary sulfates, and high temperatures, represent suitable environments for thermophilic sulfate reduction. The diversity and activity of dissimilatory sulfate reducers in these environments remain unexplored. In this study, using metagenomic approaches, in situ activity measurements with a radioactive tracer, and cultivation we have shown that members of the genus Desulfofundulus are responsible for the extremely high sulfate reduction rate (SRR) in burning lignite seams in the Altai Mountains. The maximum SRR reached 564 ± 21.9 nmol S cm−3 day−1 at 60°C and was of the same order of magnitude for both thermophilic (60°C) and mesophilic (23°C) incubations. The 16S rRNA profiles and the search for dsr gene sequences in the metagenome revealed members of the genus Desulfofundulus as the main sulfate reducers. The thermophilic Desulfofundulus sp. strain Al36 isolated in pure culture, did not grow at temperatures below 50°C, but produced spores that germinated into metabolically active cells at 20 and 15°C. Vegetative cells germinating from spores produced up to 0.738 ± 0.026 mM H2S at 20°C and up to 0.629 ± 0.007 mM H2S at 15°C when CO was used as the sole electron donor. The Al36 strain maintains significant production of H2S from sulfate over a wide temperature range from 15°C to 65°C, which is important in variable temperature biotopes such as lignite burning seams. Burning coal seams producing CO are ubiquitous throughout the world, and biogenic H2S may represent an overlooked significant flux to the atmosphere. The thermophilic spore outgrowth and their metabolic activity at temperatures below the growth minimum may be important for other spore-forming bacteria of environmental, industrial and clinical importance.

In this study, aerosol flow in circular and rectangular periodic cells of a regular row of porous cylinders is simulated. The hydrodynamic field of the carrier medium flow outside and in the domain of the porous cylinder is described in the approximation of the Stokes–Brinkman model for an incompressible gas. For the model of a circular periodic Kuwabara cell, an analytical solution is obtained; in the case of a rectangular cell, the boundary value problem for the flow stream function is solved numerically using the boundary element method. In the determined velocity fields of the carrier medium, the Lagrangian equations of motion of suspended particles are integrated, and the dependence of the particle capture efficiency as a result of the inertial impact and the interception mechanism on the Stokes number at different cell density and the Darcy number of a porous cylinder is calculated. The results of the calculations are found to be in a good agreement with those from other works carried out using the CFD package. Parametric studies of the efficiency of capture of aerosol particles as a function of the Stokes number for various values of the Darcy number of a porous cylinder, the periodic cell density, and the interception parameter are performed. It is shown that when a porous cylinder is flown around, the particle capture efficiency tends to a non-zero value that increases depending on how high the Darcy number and the density of the periodic cell are. In the region of small Stokes numbers, the capture of particles is provided by both the interception mechanism and the flow through the porous cylinder. A formula is derived for estimating the efficiency of particle trapping under the effect of particle interception on a porous cylinder.