Halomonas profundi sp. nov., isolated from deep-sea sediment of the Mariana Trench.

Two novel Gram-stain-negative, facultative anaerobic, non-flagellated, rod-shaped bacterial strains, designated MT13T and MT32, were isolated from sediment samples collected from the Mariana Trench at a depth of 8300 m. The two strains grew at -2-30 °C (optimum, 25 °C), at pH 5.5-10.0 (optimum, pH 7.5-8.0) and with 0-15 % (w/v) NaCl (optimum, 3-6 %). They did not reduce nitrate to nitrite nor hydrolyse Tweens 40 and 80, aesculin, casein, starch and DNA. The genomic G+C contents of draft genomes of strain MT13T and MT32 were 52.2 and 54.1 m ol%, respectively. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strains MT13T and MT32 were affiliated with the genus Halomonas, with the highest similarity to the type strain of Halomonas olivaria. The values of average nucleotide identity and in silico DNA-DNA hybridization between strain MT13T and MT32, and between strain MT13T and five closely related type strains of Halomonas species indicated that strains MT13T and MT32 belonged to the same species, but represented a novel species in the genus of Halomonas. The major cellular fatty acids of strains MT13T and MT32 were C16 : 0, summed feature 3(C16 : 1 ω7c/ω6c) and summed feature 8 (C18 : 1 ω7c/ω6c). Major polar lipids of strains MT13T and MT32 included phosphatidylglycerol, phosphatidylethanolamine and diphosphatidylglycerol. Ubiquinone-9 was the predominant respiratory quinone. Based on data from the present polyphasic study, strains MT13T and MT32 represent a novel species of the genus Halomonas, for which the name Halomonas profundi sp. nov. is proposed. The type strain is MT13T (=MCCC 1K06389T=KCTC 82923T).

Structural characteristics and immune-enhancing activity of an extracellular polysaccharide produced by marine Halomonas sp. 2E1.

Microbial polysaccharides from extreme environments, such as cold seeps and hydrothermal vents, usually exhibit novel structural features and diverse biological activities. In this study, an exopolysaccharide (EPS2E1) was isolated from cold-seep bacterium Halomonas sp. 2E1 and its immune-enhancing activity was evaluated. The total sugar content and protein content were determined as 83.1% and 7.9%, respectively. EPS2E1 contained mannose and glucose with the molar ratio of 3.76: 1. The molecular weight was determined to be 47.0 kDa. Structural analysis indicated that EPS2E1 was highly branched, the backbone mainly consisted of →2)-Man-(α-1→ and →2, 6)-Man-(α-1→ with the ratio of 2.45: 1.00. The chain also contained →4)-Glc-(α-1→, →6)-Man-(α-1→ and →3)-Glc-(ß-1→. EPS2E1 could significantly increase the production of NO, COX-2, TNF-α, IL-1ß and IL-6 by activating the MAPK and NF-κB pathways on RAW264.7 macrophages. EPS2E1 exhibits the potential to be an immunopotentiator in the near future.

Halomonas diversa sp. nov., isolated from deep-sea sediment of the Pacific Ocean.

A novel Gram-stain-negative, facultatively anaerobic, rod-shaped bacterium, designated as D167-6-1T, was isolated from deep-sea sediment collected from the Pacific Ocean. The cells were catalase- and oxidase-positive, and motile by means of peritrichous flagella. Growth occurred at NaCl concentrations ranging from 0 to 19 % (optimum, 2-8 %, w/v), from pH 6 to 11 (optimum, 7-8) and at temperatures between 4 and 45 °C (optimum, 33 °C). Phylogenetic analysis based on 16S rRNA, gyrB and rpoD gene sequences and its genome sequence revealed that strain D167-6-1T formed a monophyletic branch within the genus Halomonas and was most closely related to Halomonas saliphila, Halomonas pellis, Halomonas kenyensis, Halomonas daqingensis, Halomonas desiderata and Halomonas lactosivorans (with 98.5, 98.5, 98.4, 98.1, 97.5 and 97.8 % 16S rRNA sequence similarity, respectively). The complete genome size of strain D167-6-1T was 4.49 Mb, with a DNA G+C content of 62.8 mol%. The estimated averagenucleotide identity and DNA-DNA hybridization values between strain D167-6-1T and other closely related species were 77.59-85.35 % and 22.0-30.6 %, respectively. The principal cellular fatty acids (>5 %) were C18 : 1 ω7c, C16 : 0, C19 : 0 cyclo ω8c, summed feature 3 (C16 : 1 ω7c/C16 : 1 ω6c) and C17 : 0 cyclo. The polar lipids were identified as diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, an unidentified aminolipid, aminophospholipid and two unidentified phospholipids. The predominant respiratory quinones were Q-9 and Q-8. The combined genotypic and phenotypic data show that strain D167-6-1T represents a novel species of the genus Halomonas, for which the name Halomonas diversa sp. nov. is proposed, with the type strain D167-6-1T (=MCCC 1A13316T=KCTC 72441T).

Halomonas maris sp. nov., a moderately halophilic bacterium isolated from sediment in the southwest Indian Ocean.

A halophilic, Gram-staining-negative, rod-shaped, flagellated and motile bacterium, strain QX-1 T, was isolated from deep-sea sediment at a depth of 3332 m in the southwestern Indian Ocean. Strain QX-1 T growth was observed at 4-50 °C (optimum 37 °C), pH 5.0-11.0 (optimum pH 7.0), 3-25% NaCl (w/v; optimum 7%), and it did not grow without NaCl. A phylogenetic analysis based on the 16S rRNA gene placed strain QX-1 T in the genus Halomonas and most closely related to Halomonas sulfidaeris (97.9%), Halomonas zhaodongensis (97.8%), Halomonas songnenensis (97.6%), Halomonas hydrothermalis (97.4%), Halomonas subterranea (97.3%), Halomonas salicampi (97.1%), and Halomonas arcis (97.0%). DNA-DNA hybridization (< 26.5%) and average nucleotide identity values (< 83.5%) between strain QX-1 T and the related type strains meet the accepted criteria for a new species. The principal fatty acids (> 10%) of strain QX-1 T are C16:0 (25.5%), C17:0 cyclo (14.0%), C19:0 cyclo ω8c (18.7%), and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c, 18.1%). The polar lipids of strain QX-1 T are mainly diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, unidentified phospholipid, unidentified aminophospholipid, and five unidentified lipids. The main respiratory quinone is Q-9. The G + C content of its chromosomal DNA is 54.4 mol%. Its fatty acid profile, respiratory quinones, and G + C content also support the placement of QX-1 T in the genus Halomonas. These phylogenetic, phenotypic, and chemotaxonomic analyses indicate that QX-1 T is a novel species, for which the name Halomonas maris is proposed. The type strain is QX-1 T (= MCCC 1A17875T = KCTC 82198 T = NBRC 114670 T).

Genomic analysis of a novel species Halomonas shambharensis isolated from hypersaline lake in Northwest India.

Genome analysis of Halomonas shambharensis, a novel species, was performed to understand the osmoprotectant strategies used by the strain to overcome the salinity stress and to explore the prospective industrial uses. It will also help to better understand the ecological roles of Halomonas species in hypersaline habitats. Ultrastructure of the cell was determined by using transmission electron microscopy. Standard microbiological methods were used to find out growth parameters and heterotrophic mode of nutrition. For Genome analysis, complete bacterial genome sequencing was performed using the Oxford Nanopore MinION DNA Sequencer. Assembly, annotation and finishing of the obtained sequence were done by using a Prokaryotic Genome Annotation Pipeline (PGAP) (SPAdes v. 3.10.1). Predicted Coading sequences (CDSs) obtained through the PGAP were used for functional annotation using Clusters of Orthologous Groups and Kyoto Encyclopedia of Genes and Genomes (KEGG) platforms. The H. shambharensis was found to be a Gram-stain-negative, rod-shaped bacterium, motile with a peritrichous flagella. The H. shambharensis bacterium can grow in a wide range of temperature (from 25 to 65 °C), pH (pH 4 to pH 12.0) and salt concentration (5.0% NaCl to 30.0% NaCl). After annotation and assembly, the total genome size obtained was 1,533,947 bp, which revealed 146 subsystems, 3847 coding sequences, and 19RNAs with G+C content of 63.6%. Gene annotation identified the genes related to various metabolic pathways, including carbohydrate metabolism, fatty acid metabolism and stress tolerance. The genomic dataset of H. shambharensis will be useful for analysis of protein-coding gene families and how these coding genes are significant for the survival and metabolism among the different species of Halomonas. The complete genome sequence presented here will help to unravel the biotechnological potential of H. shambharensis for production of the high-value products such as betaine, or as a source of gene-mining for individual enzymes.

Halomonas azerbaijanica sp. nov., a halophilic bacterium isolated from Urmia Lake after the 2015 drought.

A novel, slightly halophilic bacterium, designated TBZ202T, was isolated from water of Urmia Lake, in the Azerbaijan region of north-west Iran. The strain was facultatively anaerobic, Gram-stain-negative, rod-shaped and motile. Colonies were creamy, circular, convex and shiny. It grew at NaCl concentrations of 0-12 % (w/v) (optimum 3-5 % w/v), at temperatures of 20-45 °C (optimum 30 °C) and at pH 5.0-10.0 (optimum pH 7.0). Based on the 16S rRNA gene sequence, strain TBZ202T belongs to the genus Halomonas in the Halomonadaceae and the most closely related species are Halomonas gudaonensis CGMCC 1.6133T (98.6 % similarity), Halomonas ventosae Al12T (96.8 %) and Halomonas rambilicola RS-16T (96.6%). The G+C content was 67.9 % and the digital DNA-DNA hybridization and average nucleotide identity values with H. gudaonensis were 35.8 and 83.8 %, respectively, indicating that the isolate differs from all species described. The major fatty acids were C18 : 1 ω7c, C16 : 0 and C16 : 1 ω7c. The only respiratory quinone detected was Q-9 and polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, aminophospholipid and three unknown phospholipids. On the basis of a polyphasic taxonomic analysis, the isolate is considered to represent a novel species of the genus Halomonas, for which the name Halomonas azerbaijanica sp. nov. is proposed. The type strain is TBZ202T (=KCTC 62817T=CECT 9693T).

Halomonas icarae sp. nov., a moderately halophilic bacterium isolated from beach soil in India.

A moderately halophilic, Gram-stain-negative, aerobic bacterium, strain D1-1T, belonging to the genus Halomonas, was isolated from soil sampled at Pentha beach, Odisha, India. Phylogenetic trees reconstructed based on 16S rRNA genes and multilocus sequence analysis of gyrB and rpoD genes revealed that strain D1-1T belonged to the genus Halomonas and was most closely related to Halomonas alimentaria YKJ-16T (98.1 %) followed by Halomonas ventosae Al12T (97.5 %), Halomonas sediminicola CPS11T (97.5 %), Halomonas fontilapidosi 5CRT (97.4 %) and Halomonas halodenitrificans DSM 735T (97.2 %) on the basis of 16S rRNA gene sequence similarity. Sequence identities with other species within the genus were lower than 97.0 %. The digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values of 22.4-30 % and 79.5-85.4 % with close relatives of H. halodenitrificans DSM 735T, H. alimentaria YKJ-16T, H. ventosae Al12T and H. fontilapidosi 5CRT were lower than the threshold recommended for species delineation (70 % and 95-96 % for dDDH and ANI, respectively). Further, strain D1-1T formed yellow-coloured colonies; cells were rod-shaped, motile with optimum growth at 30 °C (range, 4-45 °C) and 2-8 % NaCl (w/v; grew up to 24 % NaCl). The major fatty acids were summed feature 8 (C18 : 1 ω7c/C18 : 1 ω6c), summed feature 3 (C16 : 1 ω7c/C16 : 1 ω6c) and C16 : 0 and the main respiratory quinone was ubiquinone Q-9 in line with description of the genus. Based on its chemotaxonomic and phylogenetic characteristics and genome uniqueness, strain D1-1T represents a novel species in the genus Halomonas, for which we propose the name Halomonas icarae sp. nov., within the family Halomonadaceae. The type strain is D1-1T (=JCM 33602T=KACC 21317T=NAIMCC-B-2254T).

Halomonas lysinitropha sp. nov., a novel halophilic bacterium isolated from a hypersaline wetland.

We carried out a polyphasic taxonomic study on a new halophilic strain designated 3(2)T, isolated from Meighan wetland, Iran. Cells of the novel strain were Gram-stain-negative, non-hemolytic, catalase- and oxidase-positive, rod-shaped, non-endospore-forming and motile. Cell growth occurred at 3-15 % NaCl (w/v; optimum, 5 %), pH 7.0-9.0 (optimum, pH 7.5-8.0) and 15-35 °C (optimum, 30 °C). 16S rRNA gene sequence comparisons confirmed the affiliation of strain 3(2)T to the class Gammaproteobacteria and the genus Halomonas with highest similarity to Halomonas daqiaonensis YCSA28T (98.4 %) and Halomonas ventosae Al12T (97.9 %). Experimental and in silico DNA-DNA hybridization values were 42.7 and 35.1% with H. daqiaonensis IBRC-M 10931T and 48 and 35.2% with H. ventosae IBRC-M 10566T, respectively, and indicated that they are different members of the same genus. The genome of the type strain was characterized by a size of 3.83 Mbp with 63 scaffolds and a G+C content of 64.8 mol%. Moreover, the average nucleotide identity values against H. ventosae Al12T and H. daqiaonensis YCSA28T were 88.8 and 88.5 %, respectively. The predominant respiratory quinone was Q-9 (92 %) with Q-8 (8 %) as a minor component. Major fatty acids were C16 : 0 cyclo, C19 : 0 ω8c, C16 : 1 ω7c and/or iso-C15:0 2-OH, C12 : 0 3-OH and C18 : 1 ω7c. The polar lipid profile of the strain contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphoaminoglycolipid and four unidentified phospholipids. According to our results, strain 3(2)T could be classified as a novel species in the genus Halomonas for which the name Halomonas lysinitropha sp. nov. is proposed. The type strain is 3(2)T (=IBRC M 10929T=LMG 29450T=CIP 111708T).

Halomonas pellis sp. nov., a moderately halophilic bacterium isolated from wetsalted hides.

A Gram-stain-negative, moderately halophilic strain, designated strain L5T, was isolated from wetsalted hides collected from Chengdu, south-west PR China. The cells were motile, facultative aerobic, short rod-shaped and non-endospore-forming. Growth of strain L5T occurred at pH 6-10 (optimum, pH 8), 10-45 °C (optimum, 30 °C) and in the presence of 1-17 % (w/v) NaCl (optimum, 10 %). Results of phylogenetic analyses based on 16S rRNA, gyrB and rpoD gene sequences and its genome revealed that strain L5T belonged to the genus Halomonas. Strain L5T was found to be most closely related to the type strains of Halomonas saliphila, Halomonas lactosivorans, Halomonas kenyensis, Halomonas daqingensis and Halomonas desiderata (98.8, 98.6, 98.3, 97.9 and 97.4 % 16S rRNA gene sequence similarity, respectively). The draft genome was approximately 4.2 Mb in size with a G+C content of 63.5 mol%. The average nucleotide identity (ANI) and digital DNA-DNA hybridization values among strain L5T and the selected Halomonas species were 83.3-88.9 % (ANIm), 71.1-87.3 % (ANIb) and 20.2-34.6 %, which are below the recommended cutoff values. Major fatty acids were C16 : 0, C16 : 1 ω7c, C18 : 1 ω7c and C19 : 0 cyclo ω8c and the predominant ubiquinone was Q-9, with minor ubiquinone Q-8 also present. The phospholipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, four unidentified aminophospholipids and three unidentified phospholipids. Based on the mentioned polyphasic taxonomic evidence, strain L5T represents a novel species within the genus Halomonas, for which Halomonas pellis sp. nov. is proposed. The type strain is L5T (=CGMCC 1.17335T=KCTC 72573T).

Polyhydroxyalkanoates (PHA) from Halomonas pacifica ASL10 and Halomonas salifodiane ASL11 isolated from Mariout salt lakes.

Two novel PHA producing bacterial strains were chosen out of 12 strains collected from Mariout salt lakes. Analysis of 16srRNA gene sequence of the two new strains revealed 95.38% and 98.78% similarity to that of Halomonas pacifica and Halomonas salifodiane, respectively. A maximum polymer productivity of 6.9 g/l and 7.1 g/l was recorded by ASL10 and ASL11, respectively. Furthermore, a pH of 7 contributed to the highest polymer production for both strains. Interestingly, both ASL10 and ASL11showed a great ability to tolerate salinity up to 17 g/l NaCL. Moreover, both promising isolates were able to degrade crude oil efficiently by degradation percentages of 69.2% and 67.3% for ASL10 and ASL11, respectively. GCMS, FTIR, NMR, XRD and thermal properties were performed for poly (3 HV-co-3HB) characterization.

Halomonas rituensis sp. nov. and Halomonas zhuhanensis sp. nov., isolated from natural salt marsh sediment on the Tibetan Plateau.

Two novel Gram-stain-negative, aerobic and non-motile rods bacteria, designated TQ8ST and ZH2ST, were isolated from salt marsh sediment collected from the Tibetan Plateau. Strain TQ8ST was found to grow at 10-40 °C (optimum, 30 °C), pH 6.0-11.0 (optimum, pH 8.0-9.0) and in the presence of 2-12 % (w/v) NaCl (optimum, 6-8 %). Strain ZH2ST was found to grow at 15-40 °C (optimum, 30 °C), pH 7.0-10.0 (optimum pH 9.0) and in the presence of 2-10 % (w/v) NaCl (optimum, 4-6 %). Phylogenetic analysis based on the 16S rRNA gene sequences showed that strains TQ8ST and ZH2ST shared 99.07 % sequence similarity between each other and were affiliated with the genus Halomonas, sharing 97.48 % and 97.41 % of sequence similarity to their closest neighbour Halomonas sulfidaeris Esulfide1T, respectively. DNA-DNA hybridization analyses showed 61.0 % relatedness between strains TQ8ST and ZH2ST. The average nucleotide identity and the average amino acid identity values between the two genomes were 92.33 and 92.84 %, respectively. The values between the two strains and their close phylogenetic relatives were all below 95 %. The major respiratory quinones of strain TQ8ST were Q-9 and Q-8, while that of ZH2ST was Q-9. The main fatty acids shared by the two strains were C18 : 1 ω6c and/or C18 : 1 ω7c, C16 : 1 ω6c and/or C16 : 1 ω7c, C16 : 0 and C12 : 0 3-OH. Strain ZH2ST can be distinguished from TQ8ST by a higher proportion of C19 : 0 cyclo ω8c. The G+C content of the genomic DNA of strains TQ8ST and ZH2ST were 57.20 and 57.14 mol%, respectively. On the basis of phenotypic distinctiveness and phylogenetic divergence, the two isolates are considered to represent two novel species of the genus Halomonas, for which the names Halomonas rituensis sp. nov (type strain TQ8ST=KCTC 62530T=CICC 24572T) and Halomonas zhuhanensis sp. nov (type strain ZH2ST=KCTC 62531T=CICC 24505T) are proposed.

Halomonas lactosivorans sp. nov., isolated from salt-lake sediment.

A bacteria strain, designated CFH 90008T, was isolated from a salt lake sediment sample collected from Yuncheng city, Shanxi Province, PR China. Strain CFH 90008T was Gram-stain-negative, strictly aerobic, motile with lateral flagella and rod-shaped. Colonies were yellow, circular and smooth. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain CFH 90008T belonged to the genus Halomonas, showing highest sequence similarity to Halomonas daqingensis DQD2-30T (98.6 %), Halomonas saliphila LCB169T (98.5 %), Halomonas desiderata FB2T (98.1 %) and Halomonas kenyensis AIR-2T (98.0 %). Good growth was observed at 10-50 °C, pH 6.0-9.0 and with NaCl concentration from 1.0 to 12.0 % (w/v). The predominant quinone was Q9. The major fatty acid (>10 %) was C18 : 1 ω7c, C16 : 0 and C16 : 1 ω7c. The genome of strain CFH 90008T was 4.36 Mbp with a genomic DNA G+C content of 66.7 mol%. Based on low average nucleotide identity and DNA-DNAhybridization results, chemotaxonomic characteristics, and differential physiological properties, strain CFH 90008T could not be classified into any recognized species of the genus Halomonas. Therefore, a new species, for which the name Halomonas lactosivorans sp. nov. is proposed. The type strain is CFH 90008T (=DSM 103220T=KCTC 52281T).

Halomonas montanilacus sp. nov., isolated from hypersaline Lake Pengyanco on the Tibetan Plateau.

A Gram-stain-negative, catalase- and oxidase-positive, aerobic, rod-shaped, motile strain (PYC7WT) was isolated from Lake Pengyanco on the Tibetan Plateau. Comparisons based on 16S rRNA gene sequences showed that strain PYC7WT belongs to the genus Halomonas, with Halomonas malpeensis YU-PRIM-29T and Halomonas johnsoniae T68687T as its closest neighbours (96.8 and 96.6 % 16S rRNA gene sequence similarity, respectively), and only 93.1 % 16S rRNA gene sequence similarity to Halomonas elongata ATCC 33173T. The predominant respiratory quinone of strain PYC7WT is Q-9, with Q-8 as a minor component. The major fatty acids are C18 : 1 ω6c and / or C18 : 1 ω7c, C16 : 0, C16 : 1 ω6c and/or C16 : 1 ω7c, and C12 : 0 3OH. The polar lipids of strain PYC7WT include phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol and two unidentified phospholipids. Genome sequencing revealed a genome size of 4.79 Mbp and a G+C content of 62.9 mol%. DNA-DNA hybridization values of strain PYC7WT showed 45, 30 and 38 % relatedness with Halomonas johnsoniae DSM 21197T, Halomonas hamiltonii DSM 21196T and Halomonas stevensii DSM 21198T, respectively. Combining phenotypic, biochemical, genotypic and DNA-DNA hybridization data, we propose that strain PYC7WT represents a novel species within the genus Halomonas and to have the name Halomonas montanilacus sp. nov.; PYC7WT (=CICC 24506T= KCTC 62529T) is the type strain.

Fructose based hyper production of poly-3-hydroxybutyrate from Halomonas sp. YLGW01 and impact of carbon sources on bacteria morphologies.

Polyhydroxyalkanoates (PHA), such as poly (3-hydroxybutyrate) (PHB), have emerged as potential alternatives to petroleum-based plastics and can be produced through the appropriate selection of marine bacteria that are already adapted to high salt and low temperature conditions without the requirement of antibiotic treatment. The present study, thus, aimed to screen and characterize thirteen PHA-producing microbial strains isolated from the Gwangalli beach in Busan, South Korea. Among them, Halomonas sp. YLGW01 produced the highest amount of PHB (94.6 ± 1.8% (w/w)) using fructose. Interestingly Halomonas sp. YLGW01 showed increase in cell size (8.39 ± 3.63 µm) with fructose as carbon source as compared to glucose (2.34 ± 0.44 µm). Fructose syrup was investigated as carbon source under unsterilized conditions and 95.26 ± 1.78% of PHB was produced. Overall, this strain showed the highest PHB contents in halotolerant bacteria.

Halomonas piezotolerans sp. nov., a multiple-stress-tolerant bacterium isolated from a deep-sea sediment sample of the New Britain Trench.

A piezotolerant, H2O2-tolerant, heavy-metal-tolerant, slightly halophilic bacterium (strain NBT06E8T) was isolated from a deep-sea sediment sample collected from the New Britain Trench at depth of 8900 m. The strain was aerobic, motile, Gram-stain-negative, rod-shaped, oxidase-positive and catalase-positive. Growth of the strain was observed at 4-45 °C (optimum, 30 °C), at pH 5-11 (optimum, pH 8-9) and in 0.5-21 % (w/v) NaCl (optimum, 3-7 %). The optimum pressure for growth was 0.1-30 MPa with tolerance up to 60 MPa. Under optimum growth conditions, the strain could tolerate 15 mM H2O2. Resuls of 16S rRNA gene sequence analysis showed that strain NBT06E8T is closely related to Halomonas aquamarina DSM 30161T (99.5%), Halomonas meridiana DSM 5425T (99.43%) and Halomonas axialensis Althf1T (99.35%). The digital DNA-DNA hybridization values between strain NBT06E8T and the three related type strains, H. aquamarina, H. meridiana and H. axialensis, were 30.5±2.4 %, 30.7±2.5% and 31.5±2.5 %, respectively. The average nucleotide identity values between strain NBT06E8T and the three related type strains were 86.26, 86.26 and 83.63 %, respectively. The major fatty acids were summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and C16 : 0. The predominant respiratory quinone detected was ubiquinone-9 (Q-9). Based on its phenotypic and phylogenetic characteristics, we conclude that strain NBT06E8T represents a novel species of the genus Halomonas, for which the name Halomonas piezotolerans sp. nov. is proposed (type strain NBT06E8T= MCCC 1K04228T=KCTC 72680T).

Halomonas sambharensis sp. nov., a Moderately Halophilic Bacterium Isolated from the Saltern Crystallizer Ponds of the Sambhar Salt Lake in India.

Two moderately halophilic strains SBS 10T and SSO 06 were isolated from the saltern crystallizer ponds of the hypersaline Sambhar Salt Lake in India. Strains were aerobic, Gram-stain-negative, and rod shaped. Phylogenetic analysis based on the 16S rRNA gene sequences indicated that two strains belong to the genus Halomonas in the Gammaproteobacteria, with highest 16S rRNA gene sequence similarities with Halomonas gudaonensis LMG 23610T (98.2% similarity) and Halomonas campaniensis 5AGT (99.0% similarity). Strains grew optimally at 37 °C, pH 7.5-8.0 in the presence of 5-8% (w/v) NaCl. The major fatty acids of the strain SBS 10T were C18:1ω7c (54.37%), C16:0 (25.69%), C16:1 × 7c/C16:1 × 6c (13.28%), and C12:0 (1.21%). The G+C content was 63.6 mol % (Tm). Phenotypic features, fatty acids profile, and DNA G+C content supported placement of the strain SBS 10T in the genus Halomonas having distinct characteristics with related strains. Analysis of the housekeeping genes: gryB and rpoD and in silico DNA-DNA hybridization between the strain SBS 10T and its type strain Halomonas gudaonensis (LMG 23610T) further revealed the strain SBS 10T to be a distinct species. On the basis of the phenotypic, chemotaxonomic and phylogenetic analysis, the strain SBS 10T is considered to represent a novel species for which the name Halomonas sambharensis is proposed. The type strain is SBS 10T (= MTCC 12313T = LMG 30344T).

Halomonas urmiana sp. nov., a moderately halophilic bacterium isolated from Urmia Lake in Iran.

In the course of screening halophilic bacteria in Urmia Lake in Iran, which is being threatened by dryness, a novel Gram-negative, moderately halophilic, heterotrophic and short rod-shaped bacteria was isolated and characterized. The bacterium was isolated from a water specimen and designated as TBZ3T. Colonies were found to be creamy yellow, with catalase- and oxidase-positive activities. The growth of strain TBZ3T was observed to be at 10-45 °C (optimum, 30 °C), at pH 6.0-9.0 (optimum, pH 7.0) and in the presence of 0.5-20 % (w/v) NaCl (optimum, 7.5 %). Strain TBZ3T contained C16 : 0, cyclo-C19 : 0 ω8c, summed feature 3 (comprising C16 : 1 ω7c and/or C16 : 1 ω6c) and summed feature 8 (comprising C18 : 1 ω7c and/or C18 : 1 ω6c) as major fatty acids and ubiquinone-9 as the only respiratory isoprenoid quinone. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, glycolipid, unidentified phospholipid and unidentified polar lipids were detected as the major polar lipids. Strain TBZ3T was found to be most closely related to Halomonas saccharevitans AJ275T , Halomonas denitrificans M29T and Halomonas sediminicola CPS11T with the 16S rRNA gene sequence similarities of 98.93, 98.15 and 97.60 % respectively and in phylogenetic analysis strain TBZ3T grouped with Halomonas saccharevitans AJ275T contained within a large cluster within the genus Halomonas. Based on phenotypic, chemotaxonomic and molecular properties, strain TBZ3T represents a novel species of the Halomonas genus, for which the name Halomonas urmiana sp. nov. is proposed. The type strain is TBZ3T (=DSM 22871T=LMG 25416T).

Halomonas radicis sp. nov., isolated from Arthrocnemum macrostachyum growing in the Odiel marshes(Spain) and emended descriptions of Halomonas xinjiangensis and Halomonas zincidurans.

Strain EAR18T was isolated as an endophyte from the roots of a halophyte plant, Arthrocnemum macrostachyum, growing in the Odiel marshes (Huelva, Spain). Cells of strain EAR18T were Gram- stain-negative, motile, non-spore-forming aerobic rods. It grew optimally on tryptic soy agar supplemented with 2.5 % NaCl (w/v), at pH 7 and 30 °C for 48 h. It tolerated NaCl from 0 to 25 % (w/v). It presented Q9 as the major quinone and C19 : 0 cyclo ω8c, summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) and C16 : 0 as the predominant fatty acids. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and four unidentified phospholipids. The results of phylogenetic analysis based on 16S rRNA gene sequences revealed that strain EAR18T formed a well-supported clade with species Halomonas zincidurans B6T and Halomonas xinjiangensis TRM 0175T (similarities of 98.3 and 96.1 % respectively). Furthermore, digital DNA-DNA hybridization analysis resulted in values of 20.4 % with H. xinjiangensis TRM 0175T and 35.50 % with H. zincidurans B6T, and ANIb/ANIm results in values of 73.8 %/84.2 % with H. xinjiangensis TRM 0175T and 86.8 %/89.4 % with H. zincidurans B6T. Based on phylogeny and differential phenotypic properties in comparison with its closest related species, strain EAR18T is suggested to represent a new species in the genus Halomonas, for which the name Halomonas radicis sp. nov. is proposed. The type strain is EAR18T (=CECT 9077T=LMG 29859T). The whole genome was sequenced, and it had a total length of 4.6 Mbp and a G+C content of 64.9 mol%.

Halomonas borealis sp. nov. and Halomonas niordiana sp. nov., two new species isolated from seawater.

Two Gram-negative strains obtained from tank water in a scallop hatchery in Norway, were phenotypically and genotypically characterized in order to clarify their taxonomic position. On the basis of 16S rRNA gene sequence analysis, these isolates, ATF 5.2T and ATF 5.4T, were included in the genus Halomonas, being their closest relatives H. smyrnensis and H. taeanensis, with similarities of 98.9% and 97.7%, respectively. Sequence analysis of the housekeeping genes atpA, ftsZ, gyrA, gyrB, mreB, rpoB, rpoD, rpoE, rpoH, rpoN and rpoS clearly differentiated the isolates from the currently described Halomonas species, and the phylogenetic analysis using concatenated sequences of these genes located them in two robust and independent branches. DNA-DNA hybridization (eDDH) percentage, together with average nucleotide identity (ANI), were calculated using the complete genome sequences of the strains, and demonstrate that the isolates constitute two new species of Halomonas, for which the names of Halomonas borealis sp. nov. and Halomonas niordiana sp. nov. are proposed, with type strains ATF 5.2T (=CECT 9780T=LMG 31367T) and ATF 5.4T (=CECT 9779T=LMG 31227T), respectively.

Evaluation of the production of exopolysaccharides by newly isolated Halomonas strains from Tunisian hypersaline environments.

Ten Halomonas strains were screened from different Tunisian hypersaline environments for the production of exopolysaccharides (EPS), characterized and identified basing on 16S rRNA gene sequencing. EPS production was therefore studied using two different culture media M1 (complex medium) and M2 (semi-complex medium). Selected isolates produced different EPS amounts ranging from 86 to 170 mg L-1 and 26 to 105 mg L-1 when grown on M1 and M2, respectively. The use of M1 encouraged stronger bacterial growth associated with greater EPS production compared to M2. Nevertheless, the highest EPS yield (YEPS/X) was observed for strains grown on M2. When cultivated on M1, all isolates produced EPS exhibiting almost the same monosaccharide profile with mannose, glucose and arabinose being the main monomers. However, the produced EPS on M2 were characterized by heterogeneous monosaccharide profiles among the different species, mostly consisting of glucomannan that could be a versatile material used for many further applications.