Nitrincola tapanii sp. nov., a novel alkaliphilic bacterium from An Indian Soda Lake.

A novel Gram-stain-negative bacterial strain designated as MEB193T was isolated from a sediment sample collected from Lonar Lake, India. The cells were motile, non-spore-forming and rod-shaped. The strain was oxidase- and catalase-positive. It grew optimally at pH 9.0 and at 1 % (w/v) NaCl concentration at 30 °C. Based on 16S rRNA gene sequence similarity, MEB193T belongs to genus Nitrincola, with Nitrincola alkalilacustris ZV-19T (95.89 %) and Nitrincola lacisaponensis 4CAT (95.87 %) as its closest neighbours. The major fatty acid was summed feature 8 comprising C18:1ω7c/C18:1ω6c (52 %) followed by C16 : 0 (25 %). Phosphatidylglycerol (PG) and diphosphatidylglycerol (DPG) were present as the major polar lipids. The draft genome obtained in this study was 2 793 747 bp and the G+C content was 50.79 mol%. Average nucleotide identity (71.76 %) and DNA-DNA hybridization (<20 %) values between strain MEB193T and Nitrincola lacisaponensis 4CAT confirmed the novelty of this new species. Based on phenotypic including chemotaxonomic and genotypic characterization data, strain MEB193T represents a new species of the genus Nitrincola for which the name Nitrincola tapanii sp. nov. is proposed. The type strain is MEB193T (=MCC 2863T=JCM 31570 T=KCTC 52390 T).

Taxonomic description and draft genome of Pseudomonas sediminis sp. nov., isolated from the rhizospheric sediment of Phragmites karka.

The taxonomic position of a Gram-stain-negative, rod-shaped bacterial strain, designated PI11T, isolated from the rhizospheric sediment of Phragmites karka was characterized using a polyphasic approach. Strain PI11T could grow optimally at 1.0% NaCl concentration with pH 7.0 at 30°C and was positive for oxidase and catalase but negative for hydrolysis of starch, casein, and esculin ferric citrate. Phylogenetic analysis of 16S rRNA gene sequences indicated that the strain PI11T belonged to the genus Pseudomonas sharing the highest sequence similarities with Pseudomonas indoloxydans JCM 14246T (99.72%), followed by, Pseudomonas oleovorans subsp. oleovorans DSM 1045T (99.29%), Pseudomonas toyotomiensis JCM 15604T (99.15%), Pseudomonas chengduensis DSM 26382T (99.08%), Pseudomonas oleovorans subsp. lubricantis DSM 21016T (99.08%), and Pseudomonas alcaliphila JCM 10630T (99.01%). Experimental DNA-DNA relatedness between strain PI11T and P. indoloxydans JCM 14246T was 49.4%. The draft genome of strain PI11T consisted of 4,884,839 bp. Average nucleotide identity between the genome of strain PI11T and other closely related type strains ranged between 77.25-90.74%. The polar lipid pattern comprised of phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylcholine. The major (> 10%) cellular fatty acids were C18:1ω6c/ω7c, C16:1ω6c/ω7c, and C16:0. The DNA G + C content of strain PI11T was 62.4 mol%. Based on the results of polyphasic analysis, strain PI11T was delineated from other closely related type strains. It is proposed that strain PI11T represents represents a novel species of the genus Pseudomonas, for which the name Pseudomonas sediminis sp. nov. is proposed. The type strain is PI11T (= KCTC 42576T = DSMZ 100245T).

Rufibacter immobilis sp. nov., isolated from a high-altitude saline lake.

Two pinkish-red, Gram-stain-negative, non-motile aerobic bacterial strains (MCC P1(T) and MCC P2), capable of growing at low temperatures (15 °C), were isolated from water of a saline lake located in the western Himalayas of India. The strains were capable of growth in the presence of 0-2.0% NaCl and at pH 6.5-9.0. Phylogenetic analysis based on 16S rRNA gene sequences revealed the closest similarity of 96.3% to the type strain of the only species of the genus Rufibacter , Rufibacter tibetensis CCTCC AB 208084(T). Strains MCC P1(T) and MCC P2 shared 99.0% 16S rRNA gene sequence similarity and 88.6% DNA-DNA relatedness. The major cellular fatty acids were iso-C15 : 0, C17 : 1ω6c, summed feature 3 (C16 : 1ω6c/C16 : 1ω7c) and summed feature 4 (anteiso-C17 : 1 B/iso-C17 : 1 I). Predominant polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The respiratory quinone was MK-7. The DNA G+C content of the strains was 52.6-52.8 mol%. Based on morphological, physiological, chemotaxonomical and molecular characteristics, strains MCC P1(T) and MCC P2 represent a novel species of the genus Rufibacter , for which the name Rufibacter immobilis sp. nov. is proposed. The type strain is MCC P1(T) ( =MCC 2268(T) =CCTCC AB 2013351(T)).

Rhodococcus enclensis sp. nov., a novel member of the genus Rhodococcus.

A novel actinobacterial strain, designated, NIO-1009(T), was isolated from a marine sediment sample collected from Chorao Island, Goa, India. Phylogenetic analysis comparisons based on 16S rRNA gene sequences between strain NIO-1009(T) and other members of the genus Rhodococcus revealed that strain NIO-1009(T) had the closest sequence similarity to Rhodococcus kroppenstedtii DSM 44908(T) and Rhodococcus corynebacterioides DSM 20151(T) with 99.2 and 99.1%, respectively. Furthermore, DNA-DNA hybridization results showed that R. kroppenstedtii DSM 44908(T) and R. corynebacterioides DSM 20151(T) were 39.5 (3.0%) and 41.7 (2.0%) with strain NIO-1009(T), respectively, which were well below the 70% limit for any novel species proposal. Phylogenetically strain NIO-1009(T) forms a stable clade with and R. kroppenstedtii DSM 44908(T) and R. corynebacterioides DSM 20151(T) with 100% bootstrap values. Strain NIO-1009(T) contained meso-diaminopimelic acid as the diagnostic diamino acid and galactose and arabinose as the cell wall sugars. The major fatty acids were C(16 : 0), C(18 : 1)ω9c, C(16 : 1)(ω6c and/or ω7c) and 10-methyl C(18 : 0). The only menaquinone detected was MK-8(H2), while the major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside and one unknown phospholipid. The G+C content of the genomic DNA was 66.9 mol%. The phenotypic and genotypic data showed that strain NIO-1009(T) warrants recognition as a novel species of the genus Rhodococcus for which the name Rhodococcus enclensis sp. nov., is proposed; the type strain is NIO-1009(T) ( = NCIM 5452(T) = DSM 45688(T)).

Megasphaera indica sp. nov., an obligate anaerobic bacteria isolated from human faeces.

Two coccoid, non-motile, obligately anaerobic, Gram-stain-negative bacteria, occurring singly or in pairs, or as short chains, with a mean size of 1.4-2.5 µm were isolated from the faeces of two healthy human volunteers, aged 26 and 56 years, and were designated NMBHI-10(T) and BLPYG-7, respectively. Both the strains were affiliated to the sub-branch Sporomusa of the class Clostridia as revealed by 16S rRNA gene sequence analysis. The isolates NMBHI-10(T) and BLPYG-7 showed 99.1 and 99.2% 16S rRNA gene sequence similarity, respectively, with Megasphaera elsdenii JCM 1772(T). DNA-DNA hybridization and phenotypic analysis showed that both the strains were distinct from their closest relative, M. elsdenii JCM 1772(T) (42 and 53% DNA-DNA relatedness with NMBHI-10(T) and BLPYG-7, respectively), but belong to the same species (DNA-DNA relatedness of 80.9 % between the isolates). According to DNA-DNA hybridization results, the coccoid strains belong to the same genospecies, and neither is related to any of the recognized species of the genus Megasphaera. Strains NMBHI-10(T) and BLPYG-7 grew in PYG broth at temperatures of between 15 and 40 °C (optimum 37 °C), but not at 45 °C. The strains utilized a range of carbohydrates as sources of carbon and energy including glucose, lactose, cellobiose, rhamnose, galactose and sucrose. Glucose fermentation resulted in the formation of volatile fatty acids, mainly caproic acid and organic acids such as succinic acid. Phylogenetic analysis, specific phenotypic characteristics and/or DNA G+C content also differentiated the strains from each other and from their closest relatives. The DNA G+C contents of strains NMBHI-10(T) and BLPYG-7 are 57.7 and 54.9 mol%, respectively. The major fatty acids were 12 : 0 FAME and 17 : 0 CYC FAME. On the basis of these data, we conclude that strains NMBHI-10(T) and BLPYG-7 should be classified as representing a novel species of the genus Megasphaera, for which the name Megsphaera indica sp. nov. is proposed. The type strain is NMBHI-10(T) ( = DSM 25563(T) = MCC 2481(T)).

Fictibacillus enclensis sp. nov., isolated from marine sediment.

A novel Gram-positive strain, designated NIO-1003(T), was isolated from a marine sediment sample collected from the Chorao Island, Goa Provence, India. Strain NIO-1003(T) was found to be strictly aerobic, motile, endospore-forming rods. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain NIO-1003(T) belongs to the genus Fictibacillus and to be most closely related to Fictibacillus rigui KCTC 13278(T), Fictibacillus solisalsi KCTC 13181(T) and Fictibacillus barbaricus DSM 14730(T) with 98.2, 98.0 and 97.2 % similarity and 25, 28, 39 nucleotide differences respectively. Strain NIO-1003(T) was characterized by having cell-wall peptidoglycan based on meso-diaminopimelic acid and MK-7 as the predominant menaquinone. The polar lipid profile exhibited the major compounds diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. In addition, minor amounts of an aminophospholipid were detected. The major fatty acids were identified as ai-C15:0, iso-C15:0, ai-C17:0 and C16:0, supporting the grouping of strain NIO-1003(T) into the family Bacillaceae. The DNA G+C content of strain NIO-1003(T) was determined to be 42.6 mol%. On the basis of phenotypic properties, phylogeny and DNA-DNA hybridisation analysis, strain NIO-1003(T) is considered to represent a novel species of the genus Fictibacillus for which the name Fictibacillus enclensis sp. nov. is proposed. The type strain is NIO-1003(T) (= NCIM 5458(T) = DSM 25142(T)).

Bacillus enclensis sp. nov., isolated from sediment sample.

A novel bacterial strain, designated SGD-1123(T) was isolated from Chorao Island, in Goa Province, India. The strain was found to be able to grow at 15-42 °C, pH 5-12 and 0-12 % (w/v) NaCl. The whole cell hydrolysates were found to contain meso-diaminopimelic acid, galactose and arabinose. The major fatty acids were identified as iso-C15:0 and anteiso-C15:0, MK-7 was identified as the predominant menaquinone and the predominant polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified aminolipid. The genomic DNA G+C content was determined to be 44.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences placed the isolate within the genus Bacillus and further revealed that strain SGD-1123(T) had highest sequence similarity with Bacillus aquimaris, and forms a separate clade with its closest relatives i.e. B. aquimaris, Bacillus vietnamensis and Bacillus marisflavi, with which it shares 94.5, 94.1 and 94.1 % similarity respectively. The phylogenetic, chemotaxonomic and phenotypic analyses indicated that strain SGD-1123(T) represents a novel species within the genus Bacillus, for which the name Bacillus enclensis is proposed. The type strain is SGD-1123(T) (NCIM 5450(T)=CCTCC AB 2011125(T)).

Rhodomicrobium udaipurense sp. nov., a psychrotolerant, phototrophic alphaproteobacterium isolated from a freshwater stream.

Two strains (JA643(T) and JA755) of Gram-stain-negative, facultatively anaerobic phototrophic, bacteria capable of growth at low temperatures (10-15 °C) were isolated from freshwater streams from different geographical regions of India. Both strains contain bacteriochlorophyll a and carotenoids of the spirilloxanthin series. Phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified phospholipid (PL), unidentified amino lipids (AL1-AL6, AL9) and an unidentified lipid (L1) were the polar lipids present in both strains. The major cellular fatty acid was C18 : 1ω7c (76-79 % of the total). Bacteriohopane derivatives (BHD1,2), unidentified hopanoids (UH1-5), diplopterol (DPL) and diploptene (DPE) were the major hopanoids of both strains. The DNA G+C content was 64.2-64.5 mol%. 16S rRNA gene sequence-based phylogenetic analysis showed that both strains are closely related to the genus Rhodomicrobium and clustered with Rhodomicrobium vannielii DSM 162(T) (99 % sequence similarity). However, both strains exhibited only 46.1 % DNA-DNA hybridization with R. vannielii DSM 162(T). Strains JA643(T) and JA755 shared >99 % 16S rRNA gene sequence similarity and were >85 % related on the basis of DNA-DNA hybridization; they are therefore considered to represent a novel species in the genus Rhodomicrobium, for which the name Rhodomicrobium udaipurense sp. nov. is proposed. The type strain is JA643(T) ( = KCTC 15219(T) = NBRC 109057(T)).

Rhizobium subbaraonis sp. nov., an endolithic bacterium isolated from beach sand.

Two strains (JC85(T) and JC108) of Gram-stain-negative, motile bacteria were isolated from endolithic beach sand samples on an oligotrophic medium. Based on the 16S rRNA gene sequence analysis, both strains were identified as belonging to the genus Rhizobium. Strain JC108 had 16S rRNA gene sequence similarity of 100 % with Rhizobium pusense NRCPB10(T) and formed a cluster with this strain. Strain JC85(T) had 96.9 % 16S rRNA gene sequence similarity and was 18 % related (based on DNA-DNA hybridization) to Rhizobium borbori DN316(T). With other strains of the genus Rhizobium, the 16S rRNA gene sequence similarity was less than 96.3 %. Strain JC85(T) could tolerate up to 3 % salinity, fix N(2), was resistant to ampicillin (10 µg) and was positive for catalase and oxidase. The major fatty acid was C(18 : 1)ω7c (69 %) with minor amounts of C(19 : 0) cyclo ω8c (8.9 %), C(16 : 0) (6.9 %), C(12 : 0) (5.7 %) and C(19 : 1)ω7c/C(19 : 1)ω6c (2.2 %). Polar lipids of strain JC85(T) include two unidentified aminophospholipids (APL1,2), two unidentified phospholipids (PL1,2), phosphatidylcholine and four unidentified lipids (L1-4). Q-10 is the major quinone of strain JC85(T). Based on polyphasic taxonomic analysis, strain JC85(T) represents a novel species for which, the name Rhizobium subbaraonis JC85(T) is proposed. The type strain is JC85(T) ( = DSM 24765(T) = KCTC 23614(T)).

Descriptions of Rhodopseudomonas parapalustris sp. nov., Rhodopseudomonas harwoodiae sp. nov. and Rhodopseudomonas pseudopalustris sp. nov., and emended description of Rhodopseudomonas palustris.

Four strains (JA310(T), JA531(T), JA447 and JA490) of red to reddish brown pigmented, rod-shaped, motile and budding phototrophic bacteria were isolated from soil and freshwater sediment samples from different geographical regions of India. All strains contained bacteriochlorophyll a and carotenoids of the spirilloxanthin series. The major cellular fatty acid of strains JA310(T) and JA531(T) was C(18:1)ω7c, the quinone was Q-10 and polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, an aminohopanoid and an unidentified aminolipid. Phylogenetic analysis based on 16S rRNA gene sequences showed that all strains clustered with species of the genus Rhodopseudomonas in the class Alphaproteobacteria. Strains JA531(T), JA447 and JA490 were genotypically (>80% related based on DNA-DNA hybridization) and phenotypically closely related to each other and the three strains were distinct from strain JA310(T) (33% related). Furthermore, all four strains had less than 48% relatedness (DNA-DNA hybridization) with type strains of members of the genus Rhodopseudomonas, i.e. Rhodopseudomonas palustris ATCC 17001(T), Rhodopseudomonas faecalis JCM 11668(T) and Rhodopseudomonas rhenobacensis DSM 12706(T). The genomic DNA G+C contents of strains JA310(T) and JA531(T) were 63.8 and 62.4 mol%, respectively. On the basis of phenotypic, chemotaxonomic and molecular genetic evidence, it is proposed that strains JA310(T) ( = NBRC 106083(T) = KCTC 5839(T)) and JA531(T) ( = NBRC 107575(T) = KCTC 5841(T)) be classified as the type strains of two novel species of the genus Rhodopseudomonas, Rhodopseudomonas parapalustris sp. nov. and Rhodopseudomonas harwoodiae sp. nov., respectively. In addition, we propose that strain DSM 123(T) ( = NBRC 100419(T)) represents a novel species, Rhodopseudomonas pseudopalustris sp. nov., since this strain differs genotypically and phenotypically from R. palustris ATCC 17001(T) and other members of the genus Rhodopseudomonas. An emended description of R. palustris is also provided.

Marichromatium litoris sp. nov. and Marichromatium chrysaorae sp. nov. isolated from beach sand and from a jelly fish (Chrysaora colorata).

Three strains (JA349(T), JA553(T), JA439) of phototrophic sulphur bacteria were isolated from marine habitats of India. 16S rRNA gene sequence of the three strains clustered phylogenetically with members of the genus Marichromatium of the family Chromatiaceae belonging to the class Gammaproteobacteria. All the strains shared highest sequence similarity with the type strains of Marichromatium spp. (96-99% sequence similarity) and the new strains were characterized based on polyphasic taxonomy. Strains JA349(T) and JA553(T) can be distinguished from closest relative species of the genus Marichromatium with respect to distinct differences in cellular polar lipids, fatty acids and carbon/nitrogen sources utilization. Both strains were distinctly related (<50% based on DNA-DNA hybridization) with the type strains of the genus Marichromatium. Multilocus Sequence Analysis (MLSA) of the concatenated five protein coding genes (fusA, pufM, dnaK, recA, soxB) along with internal transcribed spacer (ITS; 16S-23S rRNA) had sequence similarity of less than 92% with the type strains of Marichromatium spp. Distinct phenotypic, chemotaxonomic and molecular differences allow the separation of strains JA349(T) and JA553(T) into new species of the genus Marichromatium for which, we propose the names Marichromatium litoris sp. nov. and Marichromatium chrysaorae sp. nov., respectively.