Natronosporangium hydrolyticum gen. nov., sp. nov., a haloalkaliphilic polyhydrolytic actinobacterium from a soda solonchak soil in Central Asia.

During a cultural diversity survey on hydrolytic bacteria in saline alkaline soils, a hydrolytic actinobacterium strain ACPA39T was enriched and isolated in pure culture from a soda solonchak soil in southwestern Siberia. It forms a substrate mycelium with rod-shaped sporangia containing 1-3 exospores. The isolate is obligately alkaliphilic, growing at pH 7.5-10.3 (optimum at 8.5-9.0) and moderately halophilic, tolerating up to 3 M total Na+ in the form of sodium carbonates. It is an obligately aerobic, organoheteroterophic, saccharolytic bacterium, utilizing various sugars and alpha/beta-glucans as growth substrates. According to the 16S rRNA gene-based phylogenetic analysis, strain ACPA39T forms a distinct branch within the family Micromonosporaceae, with the sequence identities below 94.5% with type strains of other genera. This is confirmed by phylogenomic analysis based on the 120 conserved single copy protein-based markers and genomic indexes (ANI, AAI). The cell-wall of ACPA39T contained meso-DAP, glycine, glutamic acid and alanine in a equimolar ratio, characteristic of the peptidoglycan type A1γ'. The whole-cell sugars include galactose and xylose. The major menaquinone is MK-10(H4). The identified polar lipids consist of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. The polar lipid fatty acids were dominated by anteiso-C17:0, iso-C16:0, iso-C17:0, 10 Me-C18:0 and C18:1ω9. Based on the distinct phylogeny, the chemotaxonomy features and unique phenotypic properties, strain ACPA39T (DSM 106523T = VKM 2772T) is classified into a new genus and species in the family Micromonosporaceae for which the name Natronosporangium hydrolitycum gen. nov., sp. nov. is proposed.

The Pathways to Create Containers for Bacteriophage Delivery.

Antimicrobial resistance is a global public health threat. One of the possible ways to solve this problem is phage therapy, but the instability of bacteriophages hinders the development of this approach. A bacteriophage delivery system that stabilizes the phage is one of the possible solutions to this problem. This study is dedicated to exploring methods to create encapsulated forms of bacteriophages for delivery. We studied the effect of proteolytic enzymes on the destruction of the polyelectrolyte microcapsule shell and revealed that protease from Streptomyces griseus was able to destroy the membrane of the microcapsule (dextran sulfate/polyarginine)3 ((DS/PArg)3). In addition, the protease decreased the activity of the bacteriophage in the second hour of incubation, and the phage lost activity after 16 h. It was found that a medium with pH 9.02 did not affect the survival of the bacteriophage or E. coli. The bacteriophages were encapsulated into polyelectrolyte microcapsules (DS/PArg)3. It was established that it is impossible to use microcapsules as a means of delivering bacteriophages since the bacteriophages are inactivated. When bacteriophages were included inside a CaCO3 core, it was demonstrated that the phage retained activity before and after the dissolution of the CaCO3 particle. From the results of this study, we recommend using CaCO3 microparticles as a container for bacteriophage delivery through the acidic stomach barrier.

'Candidatus Xiphinematincola pachtaicus' gen. nov., sp. nov., an endosymbiotic bacterium associated with nematode species of the genus Xiphinema (Nematoda, Longidoridae).

An intracellular bacterium, strain IAST, was observed to infect several species of the plant-parasitic nematode genus Xiphinema (Xiphinema astaregiense, Xiphinema incertum, Xiphinema madeirense, Xiphinema pachtaicum, Xiphinema parapachydermum and Xiphinema vallense). The bacterium could not be recovered on axenic medium. The 16S rRNA gene sequence of IAST was found to be new, being related to the family Burkholderiaceae, class Betaproteobacteria. Fungal endosymbionts Mycoavidus cysteinexigens B1-EBT (92.9 % sequence identity) and 'Candidatus Glomeribacter gigasporarum' BEG34 (89.8 % identity) are the closest taxa and form a separate phylogenetic clade inside Burkholderiaceae. Other genes (atpD, lepA and recA) also separated this species from its closest relatives using a multilocus sequence analysis approach. These genes were obtained using a partial genome of this bacterium. The localization of the bacterium (via light and fluorescence in situ hybridization microscopy) is in the X. pachtaicum females clustered around the developing oocytes, primarily found embedded inside the epithelial wall cells of the ovaries, from where they are dispersed in the intestine. Transmission electron microscopy (TEM) observations supported the presence of bacteria inside the nematode body, where they occupy ovaries and occur inside the intestinal epithelium. Ultrastructural analysis of the bacterium showed cells that appear as mostly irregular, slightly curved rods with rounded ends, 0.8-1.2 µm wide and 2.5-6.0 µm long, possessing a typical Gram-negative cell wall. The peptidoglycan layer is, however, evident only occasionally and not detectable by TEM in most cells. Another irregularly occurring shell surrounding the endosymbiont cells or the cell clusters was also revealed, probably originating from the host cell membrane. Flagella or spore-like cells do not occur and the nucleoid is diffusely distributed throughout the cell. This endosymbiont is transmitted vertically through nematode generations. These results support the proposal of IAST as a new species, although its obligate intracellular and obligate endosymbiont nature prevented isolation of a definitive type strain. Strain IAST is therefore proposed as representing 'Candidatus Xiphinematincola pachtaicus' gen. nov., sp. nov.

Draft Genome Sequence of Coralloluteibacterium stylophorae LMG 29479T.

Here, we report a draft genome sequence of the strain Coralloluteibacterium stylophorae LMG 29479T, acquired from the Belgian Coordinated Collections of Microorganisms. The genus Coralloluteibacterium currently includes only one species with a validly published name. These genome sequencing data are important for the phylogeny of the Lysobacteraceae family.

Natronoglycomyces albus gen. nov., sp. nov, a haloalkaliphilic actinobacterium from a soda solonchak soil.

A haloalkaliphilic hydrolytic actinobacterium, strain ACPA22T, was enriched and isolated in pure culture from saline alkaline soil (soda solonchak) in northeastern Mongolia. The isolate was facultatively alkaliphilic, growing at pH 6.5-10.5 (optimum at 7.3-9.0) and highly salt-tolerant, tolerating up to 3 M total Na+ as carbonates. The hydrolytic nature of ACPA22T was confirmed by two different growth-dependent methods and by the presence of multiple glycosidase-encoding genes in the genome. The 16S rRNA gene-based phylogenetic analysis demonstrated that strain ACPA22T formed a deep-branching lineage within the family Glycomycetaceae, with the highest sequence similarity value to Glycomyces buryatensis 18T (92.1 %) and Salininema proteolyticum Miq-4T (91.8 %). The average amino acid identity values (56.1-61.5 %) between ACPA22T and other Glycomycetaceae members with available genomes did not exceed the threshold reported for different genera. The cell wall of ACPA22T contained meso-diaminopimelic acid, glycine, glutamic acid and alanine in a molar ratio, characteristic of the peptidoglycan type A1γ'. The whole-cell sugars included mannose, galactose, arabinose, ribose and xylose. The major menaquinones were MK-10(Н4) and MK-11(Н4). The identified polar lipids were represented by phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides. In addition, the strain had a few unidentified characteristic polar lipids, including an amine-containing phospholipid with chromatographic mobility similar to that of phosphatidylinositol. The polar lipid fatty acids were dominated by anteiso-C17 : 0 and iso-C16 : 0. The genome included a chromosome of 3.94 Mbp (G+C content 61.5 mol%) encoding 3285 proteins and two plasmids of 59.8 and 14.8 kBp. Based on the data obtained in this study, a new genus and species, Natronoglycomyces albus gen. nov., sp. nov, is proposed with the type strain ACPA22T (=DSM 106290T=VKM Ac-2771T).

The Influence of Polyanions and Polycations on Bacteriophage Activity.

Phage therapy is a great alternative to antibiotic drugs, but it can't effectively overcome the over-acidic medium of the stomach. We offer the use of polyelectrolyte microcapsules as a protective means of bacteriophage. It is necessary to understand the influence of polyelectrolytes on bacteriophage survival. The work studied the effect of polyanions and polycations on the coliprotetic bacteriophage's viability. We have shown that polyallylamine decreased bacteriophage's viability during increasing polyelectrolyte concentration and polyarginine had a lower inhibitory effect (then PAH) on the activity of the bacteriophage due to polyelectrolyte concentration from 0.05 to 5 mg/mL. It was shown that the inhibition of the bacteriophage by polyallylamine had an electrostatic nature and the use of high ionic strength prevented the formation of the PAH-protein capsid complex. Polystyrene sulfonate does not affect bacteriophage viability during increasing polyelectrolyte concentration from 0.05 mg/mL to 1 mg/mL. Polystyrene sulfonate decreases the viability of bacteriophage from 5 mg/mL of polyelectrolyte concentration. Dextran sulfate inhibits bacteriophage activity at 20-30%. Dextran inhibits bacteriophage activity by 80% at diapason concentration from 0.05 to 5 mg/mL and loses the inhibition effect from a concentration of 5 mg/mL.

Whole-Genome Sequencing of Xanthomonadaceae Strain Alg18-2.2, Isolated from the Saline Lake Gudzhirganskoe in the Republic of Buryatia, Russia.

A draft genome sequence of the bacterial isolate Alg18-2.2, recovered from the highly saline and alkaline lake Gudzhirganskoe (Buryatia, Russia), was determined. The results of bacterial identification using 16S rRNA gene sequence and whole-genome analyses suggest that the bacterium belongs to a novel genus. Some genomic features are discussed here.

Cohnella kolymensis sp. nov., a novel bacillus isolated from Siberian permafrost.

A facultative anaerobic, rod-shaped, endospore-forming and non-motile bacterium was isolated from permafrost sediment cores in the Kolyma lowland, Siberia, Russia. The permafrost isolate clustered with members of the genus Cohnella on the basis of 16S rRNA gene sequence analysis and showed the highest sequence similarity to Cohnella saccharovorans CJ22T (96.3 %), followed by Cohnella cellulosilytica FCN3-3T (96.0 %) and Cohnella panacarvi KCTC 13060T (96.0 %). The chemotaxonomic characteristics (quinone system, cellular fatty acids and polar lipid profile) of strain 20.16T were consistent with members of the genus Cohnella. The peptidoglycan diaminoacids included meso-diaminopimelic acid and a small amount of ll-diaminopimelic acid. The molar ratio and composition of major amino acids (meso-diaminopimelic acid, alanine, and glutamic acid) correspond to the peptydoglycan type A1γ. The estimated genome size of strain 20.16T is 4.34 Mb (lower than those in other Cohnella species). The genome has a G+C content of 50.5 mol% and encodes 4843 predicted genes, of these 4740 are protein-coding ones. The results of chemotaxonomic, physiological and biochemical characterization allowed clear differentiation of strain 20.16T from the closest Cohnella species. Based on data provided, a new species Cohnella kolymensis sp. nov. is proposed, with 20.16T (=VKM B-2846T=DSM 104983T) as the type strain.

Anoxynatronum buryatiense sp. nov., an anaerobic alkaliphilic bacterium from a low mineralization soda lake in Buryatia, Russia.

An anaerobic alkaliphilic, proteolytic bacterium, strain Su22T, was isolated from the bottom sediment of the alkaline low mineralization lake Sulphatnoe (Selenginsky district, Buryatia, Russia). A comparative analysis of the 16S rRNA gene sequence revealed that this bacterium was closely related to Anoxynatronum sibiricum Z-7981T with a similarity of 98.1 %. Strain Su22T differed from A. sibiricum Z-7981T in its inability to use carbohydrates, peptone and amino acids as carbon sources. Strain Su22T grew over a temperature range of 20-40 °C with an optimum at 30 °C and within the pH range 7.4-11.0 with an optimum at pH 9.6. Sodium cations stimulated the growth of the strain considerably with an optimal concentration at 0.76-1.09 M. The whole-cell fatty acid profile included C16 : 1ω7c, C16 : 0 and C16 : 0 ALDE. The G+C content was 46.1 mol%. Based on the DNA-DNA hybridization level (53.2 %) and phenotypical differences between strains Su22T and Z-7981T, the new isolate is thus considered to represent a novel species, for which the name Anoxynatronumburyatiense sp. nov. is proposed. The type strain is Su22Т (=VKM B-2510T=CECT 8731T).

Draft Genome Sequence of Rathayibacter sp. Strain VKM Ac-2630 Isolated from Leaf Gall Induced by the Knapweed Nematode Mesoanguina picridis on Acroptilon repens.

A draft genome sequence of Rathayibacter sp. strain VKM Ac-2630 was derived using Ion Torrent sequencing technology. The genome size of this strain is 3.88 Mb, with an average G+C content of 72.0%. Genomic evidence of an aerobic mode of respiration and a heterotrophic lifestyle of this bacterium was obtained.

Alkaliphilus namsaraevii sp. nov., an alkaliphilic iron- and sulfur-reducing bacterium isolated from a steppe soda lake.

A novel alkaliphilic spore-forming bacterium was isolated from the benthic sediments of the highly mineralized steppe Lake Khilganta (Transbaikal Region, Russia). Cells of the strain, designated Ð¥-07-2T, were straight to slightly curved rods, Gram-stain-positive and motile. Strain Ð¥-07-2T grew in the pH range from 7.0 to 10.7 (optimum pH 9.6-10.3). Growth was observed at 25-47 °C (optimum 30 °C) and at an NaCl concentration from 5 to 150 g l-1 with an optimum at 40 g l-1. Strain Ð¥-07-2T was a chemo-organoheterotroph able to reduce amorphous ferric hydroxide, Fe(III) citrate and elemental sulfur in the presence of yeast extract as the electron donor. It used tryptone, peptone and trypticase with Fe(III) citrate as the electron acceptor. The predominant fatty acids in cell walls were C16:1ω8, iso-C15:0, C14 : 0 3-OH and C16 : 0. The DNA G+C content was 32.6 mol%. 16S rRNA gene sequence analysis revealed that strain Ð¥-07-2T was related most closely to members of the genus Alkaliphilus within the family Clostridiaceae. The closest relative was Alkaliphilus peptidifermentans Z-7036T (96.4 % similarity). On the basis of the genotypic, chemotaxonomic and phenotypic data, strain Ð¥-07-2T represents a novel species in the genus Alkaliphilus, for which the name Alkaliphilus namsaraevii sp. nov. is proposed. The type strain is Ð¥-07-2T (=VKM В-2746Т=DSM 26418Т).

Draft Genome Sequence of "Cohnella kolymensis" B-2846.

A draft genome sequence of "Cohnella kolymensis" strain B-2846 was derived using IonTorrent sequencing technology. The size of the assembly and G+C content were in agreement with those of other species of this genus. Characterization of the genome of a novel species of Cohnella will assist in bacterial systematics.

Belliella buryatensis sp. nov., isolated from alkaline lake water.

Two bacterial isolates from water of the alkaline brackish Lake Solenoe (Buryatia, Russia), 2C and 5CT, were characterized by using a polyphasic taxonomic approach. The strains were small, non-motile, Gram-stain-negative rods that formed small orange-red colonies on the surface of marine agar. Studies based on 16S rRNA gene sequences showed that the strains were related closely to Belliella pelovolcani CC-SAL-25T (98.7 % sequence similarity). The G+C content of the DNA was 38-40 mol%. DNA-DNA hybridization values between strains 2C and 5CT and B. pelovolcani CC-SAL-25T were 56-58 mol%. A menaquinone with seven isoprene units (MK-7) was the major respiratory quinone. The fatty acid profiles were slightly different from that of B. pelovolcani CC-SAL-25T. The novel strains could be distinguished from the phylogenetically closest species B. pelovolcani CC-SAL-25T based on matrix-assisted laser desorption ionization time-of-flight mass spectra of whole cells and a range of physiological and biochemical characteristics. The data obtained suggest that strains 2C and 5CT represent a novel species of the genus Belliella, for which the name Belliella buryatensis sp. nov. is proposed. The type strain is 5CT ( = VKM B-2724T = KCTC 32194T).

Sphaerochaeta associata sp. nov., a spherical spirochaete isolated from cultures of Methanosarcina mazei JL01.

An anaerobic, saccharolytic bacterial strain designated GLS2T was isolated from aggregates of the psychrotolerant archaeon Methanosarcina mazei strain JL01 isolated from arctic permafrost. Bacterial cells were non-motile, spherical, ovoid and annular with diameter 0.2-4 µm. They were chemoorganoheterotrophs using a wide range of mono-, di- and trisaccharides as carbon and energy sources. The novel isolate required yeast extract and vitamins for growth. The bacteria exhibited resistance to a number of ß-lactam antibiotics, rifampicin, streptomycin and vancomycin. Optimum growth was observed between 30 and 34 °C, at pH 6.8-7.5 and with 1-2 g NaCl l- 1. Isolate GLS2T was a strict anaerobe but it tolerated oxygen exposure. On the basis of 16S rRNA gene sequence similarity, strain GLS2T was shown to belong to the genus Sphaerochaeta within the family Spirochaetaceae. Its closest relatives were Sphaerochaeta globosa BuddyT (99.3 % 16S rRNA gene sequence similarity) and Sphaerochaeta pleomorpha GrapesT (95.4 % similarity). The G+C content of DNA was 47.2 mol%. The level of DNA-DNA hybridization between strains GLS2T and BuddyT was 34.7 ± 8.8 %. Major polar lipids were phosphoglycolipids, phospholipids and glycolipids; major fatty acids were C14 : 0, C16 : 0, C16 : 0 3-OH, C16 : 0 dimethyl acetal (DMA), C16 : 1n8 and C16 : 1 DMA; respiratory quinones were not detected. The results of DNA-DNA hybridization, physiological and biochemical tests demonstrated genotypic and phenotypic differentiation of strain GLS2T from the four species of the genus Sphaerochaeta with validly published names that allowed its separation into a new lineage at the species level. Strain GLS2T therefore represents a novel species, for which the name Sphaerochaeta associata sp. nov. is proposed, with the type strain GLS2T ( = DSM 26261T = VKM B-2742T).

New sulfate-reducing bacteria isolated from Buryatian alkaline brackish lakes: description of Desulfonatronum buryatense sp. nov.

New strains of sulfate-reducing bacteria were isolated from two alkaline brackish lakes located in the Siberian region of Russia, namely in the Southern Transbaikalia, Buriatia. The article presents data describing morphology, physiology, and biochemical characteristics of the isolated strains. These strains Ki4, Ki5, and Su2 were mesophilic and alkaliphilic with optimal growth at pH 8.9, 9.4, and 10.0, respectively. All isolated strains utilized lactate, formate, and ethanol in the presence of sulfate for growth and sulfidogenesis accompanied with formation of acetate and CO2. Strains Ki5 and Su2 were able to reduce Fe(III). The DNA G + C content in strains Ki4, Ki5 and Su2 was 56.3, 48.8 and 59.6 mol%, respectively. According to phylogenetic analysis of 16S rRNA sequences, the new strains were clustered within the genus Desulfonatronum, and the closest relative D. lacustre Z-7951(T) (=DSM 10312(T)) showed 99.3-99.6 % similarity. DNA-DNA relatedness values of the strains Ki4, Ki5, and Su2 with D. lacustre Z-7951(T) were 89, 53, and 79 %, respectively. Polyphasic taxonomy data suggest that strain Ki5(T) is representative of the proposed novel species Desulfonatronum buryatense sp. nov.