In silico analysis of sporulene biosynthesis pathway genes in the members of the class Bacilli.

Sporulene, a pentacyclic triterpenoid, was discovered in Bacillus subtilis and is associated with bacterial endospores. However, the study was not further extended, leaving a trail of questions. One such question is what diversity of sporulenes exists among spore-forming members? Considering the sporulene biosynthesis pathway as a fundamental tool to survey the distribution of this molecule, a genome mining study was conducted. Mining for genes encoding putative proteins of sporulene biosynthesis pathway among the class Bacilli members revealed the presence of hepS, hepT, ytpB, and sqhC genes in the members of the family Bacillaceae, Caryophanaceae, Paenibacillaceae, and Sporolactobacillaceae. However, these genes were completely absent in the members of Staphylococcaceae, Lactobacillaceae, Aerococcaceae, Carnobacteriaceae, and Leuconostocaceae. Unlike other probable pathway related proteins, a conserved amino acid domain of putative terpenoid cyclase (YtpB) appeared deep-rooted among the genus Bacillus members. In-depth analysis showed the constant gene arrangement of hepS, hepT, ytpB, and sqhC genes in these members, there by demonstrating the conserved nature of sporulene biosynthesis pathway in the members of the genus Bacillus. Our study suggests confinement of the sporulene biosynthesis pathway to spore-forming members of the class Bacilli, majorly to the genus Bacillus.

New insights into peroxide toxicology: sporulenes help Bacillus subtilis endospores from hydrogen peroxide.

AIM: The purpose of the present study was to understand the possible events involved in the toxicity of hydrogen peroxide (H2O2) to wild and sporulene-deficient spores of Bacillus subtilis, as H2O2 was previously shown to have deleterious effects. METHODS AND RESULTS: The investigation utilized two strains of B. subtilis, namely the wild-type PY79 (WT) and the sporulene-deficient TB10 (ΔsqhC mutant). Following treatment with 0.05% H2O2 (v/v), spore viability was assessed using a plate count assay, which revealed a significant decrease in cultivability of 80% for the ΔsqhC mutant spores. Possible reasons for the loss of spore viability were investigated with microscopic analysis, dipicholinic acid (DPA) quantification and propidium iodide (PI) staining. Microscopic examinations revealed the presence of withered and deflated morphologies in spores of ΔsqhC mutants treated with H2O2, indicating a compromised membrane permeability. This was further substantiated by the absence of DPA and a high frequency (50%-75%) of PI infiltration. The results of fatty acid methyl ester analysis and protein profiling indicated that the potentiation of H2O2-induced cellular responses was manifested in the form of altered spore composition in ΔsqhC B. subtilis. The slowed growth rates of the ΔsqhC mutant and the heightened sporulene biosynthesis pathways in the WT strain, both upon exposure to H2O2, suggested a protective function for sporulenes in vegetative cells. CONCLUSIONS: Sporulenes serve as a protective layer for the inner membrane of spores, thus assuming a significant role in mitigating the adverse effects of H2O2 in WT B. subtilis. The toxic effects of H2O2 were even more pronounced in the spores of the ΔsqhC mutant, which lacks this protective barrier of sporulenes.

Mesobacillus aurantius sp. nov., isolated from an orange-colored pond near a solar saltern.

An endospore producing, strict aerobic, Gram-stain-positive, orange-colored colony forming bacterium designated as strain JC1013T was isolated from an orange pond near a solar saltern of Tamil Nadu, India. Phylogenetic analysis of the 16S rRNA gene sequences indicated that strain was affiliated to the family Bacillaceae of the phylum Firmicutes. Strain showed highest 16S rRNA gene sequence identity of 98.7% with Mesobacillus selenatarsenatis SF-1 T and below 98.3% with other members of the genus Mesobacillus. Strain JC1013T produced carotenoid pigments and indole compounds. Major cellular fatty acids of strain JC1013T were iso-C15:0, anteiso-C15:0, C16:0 3-OH, iso-C17:0ω10c and summed feature 4 (iso-C17:1 I/ anteisoB). Polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unidentified aminolipids and four unidentified phospholipids. Strain JC1013T constituted m-diaminopimelic acid as diagnostic cell wall amino acids. MK-7 is the predominant menaquinone of strain JC1013T. The genome size of strain JC1013T was 4.6 Mbp and its G + C content was 42.7 mol%. For the affirmation of strain's taxonomic status, a detailed phylogenomic study was done. Based on the phylogenetic analyses, low ANI (84.6%), AAI (88.5%) values, in-silico DDH (< 29%) value, morphological, physiological and chemo-taxonomical characteristics, strain JC1013T was clearly distinguished from the nearest phylogenetic neighbor, Mesobacillus selenatarsenatis SF-1T to conclude that it is a new species of the genus Mesobacillus. We propose the name as Mesobacillus aurantius with type strain JC1013T (= NBRC 114146T = KACC 21451 T).

Crateriforma spongiae sp. nov., isolated from a marine sponge and emended description of the genus "Crateriforma".

A Gram-stain-negative, aerobic, pear to oval shaped, rosette forming bacterium with crateriform structures well distributed on the cell surface designated as strain JC647T was isolated from a sponge specimen belonging to the genus Spongia. Strain JC647T reproduces through budding. Strain JC647T shared highest 16S rRNA gene sequence identity of 99.9% with "Crateriforma conspicua" Mal65T (not a valid species name). The genome size of strain JC647T is 6.9 Mb with a G + C content of 57.8 mol %. For the resolution of the phylogenetic congruence of the novel strain, the phylogeny was also constructed with the sequences of ninety-two housekeeping genes. Based on the phylogenetic analyses, low dDDH value (51.0%), low gANI (93.2%), low AAI (94.9%) results, chemotaxonomic characteristics and differential physiological properties, strain JC647T is recognized as a new species of the genus "Crateriforma", for which we propose the name Crateriforma spongiae sp. nov. The type species is JC647T (= KCTC 72176T = NBRC 114068T).

Isoptericola sediminis sp. nov., Isolated from Chilika Lagoon.

A yellow-colored, Gram-stain-positive, rod shaped, non-motile bacterium, designated as strain JC619T, was isolated from the sediment of Chilika lagoon, India. Strain JC619T shows highest 16S rRNA gene sequence similarity (99.08%) with Isoptericola chiayiensis KCTC 19740T followed by Isoptericola halotolerans KCTC 19046T (98.6%) and other members of the genus Isoptericola (< 98%). NaCl is required for growth of strain JC619T and tolerates up to 18% (w/v) and pH up to 10. Strain JC619T grows optimally at temperature 30 °C, NaCl concentration of 3% (w/v), and at pH 7.5. The genome size of strain JC619T is 3.2 Mb with G+C content of 73.0 mol%. ANI scores of strain JC619T are 81.9% and 80.1% and dDDH values are 24.4% and 22.7% with I. chiayiensis KCTC 19740T and I. halotolerans KCTC 19046T, respectively. Respiratory quinones are MK-9(H4) and MK-9(H2). Predominant fatty acids (> 10%) are anteiso-C15:0, iso-C16:0, and iso-C15:0. Major polar lipids of strain JC619T are phosphatidylglycerol, phosphatidylinositolmannoside, diphosphatidylglycerol, and phosphatidylinositol. Strain JC619T is catalase positive but cytochrome oxidase negative and reduces nitrate. The genomic distinction of strain JC619T with its nearest related species of the genus Isoptericola is well supported with chemotaxonomic characteristics and differential physiological properties. Therefore, strain JC619T represents a new species under the genus Isoptericola for which Isoptericola sediminis sp. nov. is proposed. Type strain is JC619T (=KCTC 49244T =NBRC 114063T).

Paludisphaera soli sp. nov., a new member of the family Isosphaeraceae isolated from high altitude soil in the Western Himalaya.

A novel strain of Planctomycetes, designated JC670T, was isolated from a high altitude (~ 2900 m above sea level) soil sample collected from Garhwal region in the Western Himalaya. Colonies of this strain were observed to be light pink coloured with spherical to oval shaped cells having crateriform structures distributed all over the cell surface. The cells divide by budding. Strain JC670T was found to grow well at pH 7.0 and pH 8.0 and to tolerate up to 2% NaCl (w/v). MK6 was the only respiratory quinone identified. The major fatty acids of strain JC670T were identified as C18:1ω9c, C18:0 and C16:0, and phosphatidylcholine, two unidentified phospholipids and six unidentified lipids are present as the polar lipids. The polyamines putrescine and sym-homospermidine were detected. Strain JC670T shows high 16S rRNA gene sequence identity (95.4%) with Paludisphaera borealis PX4T. The draft genome size of strain JC670T is 7.97 Mb, with G + C content of 70.4 mol%. Based on phylogenetic analyses with the sequences of ninety-two core genes, low dDDH value (20.6%), low gANI (76.8%) and low AAI (69.1%) results, differential chemotaxonomic and physiological properties, strain JC670T (= KCTC 72850T = NBRC 114339T) is recognised as the type strain of a new species of the genus Paludisphaera, for which we propose the name Paludisphaera soli sp. nov.

Biological hydrogen production: molecular and electrolytic perspectives.

Exploration of renewable energy sources is an imperative task in order to replace fossil fuels and to diminish atmospheric pollution. Hydrogen is considered one of the most promising fuels for the future and implores further investigation to find eco-friendly ways toward viable production. Expansive processes like electrolysis and fossil fuels are currently being used to produce hydrogen. Biological hydrogen production (BHP) displays recyclable and economical traits, and is thus imperative for hydrogen economy. Three basic modes of BHP were investigated, including bio photolysis, photo fermentation and dark fermentation. Photosynthetic microorganisms could readily serve as powerhouses to successively produce this type of energy. Cyanobacteria, blue green algae (bio photolysis) and some purple non-sulfur bacteria (Photo fermentation) utilize solar energy and produce hydrogen during their metabolic processes. Ionic species, including hydrogen (H+) and electrons (e-) are combined into hydrogen gas (H2), with the use of special enzymes called hydrogenases in the case of bio photolysis, and nitrogenases catalyze the formation of hydrogen in the case of photo fermentation. Nevertheless, oxygen sensitivity of these enzymes is a drawback for bio photolysis and photo fermentation, whereas, the amount of hydrogen per unit substrate produced appears insufficient for dark fermentation. This review focuses on innovative advances in the bioprocess research, genetic engineering and bioprocess technologies such as microbial fuel cell technology, in developing bio hydrogen production.

Rhodococcus electrodiphilus sp. nov., a marine electro active actinobacterium isolated from coral reef.

An electrogenic bacterium was isolated from a marine coral, designated as strain JC435T and its taxonomic status examined by using a polyphasic approach. Results from the 16S rRNA gene sequence study showed that the isolate belonged to the genus Rhodococcus and formed a cluster with Rhodococcus ruber KCTC 9806T (99.5 % 16S rRNA gene sequence similarity) and Rhodococcus aetherivorans JCM 14343T (99.3 %), respectively. Genome relatedness based on DNA-DNA hybridization to the type strains of closest-related species was less than 30 % and the ΔTm of >7 °C, suggesting that strain represents a new species of the genus Rhodococcus. The major fatty acids were C16 : 0, C18 : 1ω9c, C18 : 010-methyl and C16 : 1ω6c and/or C16 : 1ω7c. The polar lipids of strain JC435T were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannoside, phosphatidylinositol, three unknown phospholipids and an unknown amino lipid. The major isoprenoid quinone was MK-8(H2), with 8 % of MK-7(H2) and 2 % of MK-9(H2) as minor components. Whole-cell hydrolysates contained meso-diaminopimelic acid, arabinose and galactose as the diagnostic diamino acid and sugars. Mycolic acids were detected. The genomic DNA G+C content of strain JC435T was 69.8 mol%. On the basis of phylogenetic genotypic, physiological and chemotaxonomic analysis, strain JC435T is considered to represent a novel species of the genus Rhodococcus for which the name Rhodococcuselectrodiphilus sp. nov. is proposed. The type strain is JC435T (=KCTC 39856T=LMG 29881T=MCC 3659T).

Chryseobacterium salipaludis sp. nov., isolated at a wild ass sanctuary.

A Gram-stain-negative, rod-shaped, non-motile, aerobic bacterium was isolated from a sediment sample obtained from a wild ass sanctuary in Gujarat, India. The strain designated JC490T was oxidase- and catalase-positive. The 16S rRNA gene sequence analysis and sequence comparison data indicated that strain JC490T was a member of the genus Chryseobacterium and was closely related to Chryseobacterium jeonii AT1047T (96.4 %) and with other members of the genus Chryseobacterium (<96.3 %). The DNA G+C content of strain JC490T was 34 mol%. Strain JC490T had phosphatidylethanolamine, two unidentified aminolipids, two unidentified phospholipids and five unidentified polar lipids. Menaquinone-6 was the only respiratory quinone found. Iso-C15 : 0, anteiso-C15 : 0 and iso-C17 : 0 3-OH were the major fatty acids of strain JC490T. On the basis of physiological, genotypic, phylogenetic and chemotaxonomic analyses, it is concluded that strain JC490T constitutes a novel species of the genus Chryseobacterium, for which the name Chryseobacterium salipaludis sp. nov. is proposed. The type strain is JC490T (=KCTC 52835T=LMG 30048T).

Halodesulfovibrio spirochaetisodalis gen. nov. sp. nov. and reclassification of four Desulfovibrio spp.

An antibiotic-producing, obligate anaerobic, Gram-stain-negative, catalase- and oxidase-negative strain (JC271T) was isolated from a marine habitat and identified, based on 16S rRNA gene sequence analysis, as a novel member of the family Desulfovibrionaceae. The closest phylogenetic relatives of strain JC271T were found to be Desulfovibrio marinisediminis C/L2T (99.2 %), Desulfovibrio acrylicus W218T (98.7 %), Desulfovibrio desulfuricans subsp. aestuarii (98.6 %), Desulfovibrio oceani subsp. oceani (98.0 %), Desulfovibrio oceani subsp. galatae (98.0 %) and other members of the genus Desulfovibrio (≤91.9 %). To resolve its full taxonomic position, the genomic sequence of strain JC271T was compared to available genomes of the most closely related phylogenetic members. Average Nucleotide Identity scores and DNA-DNA hybridization values confirmed that strain JC271T represents a novel genomic species. Iso-C17 : 0, iso-C17 : 1ω9c, and iso-C15 : 0 were found to be the major (comprising >10 % of the total present) fatty acids of strain JC271T. Phosphatidylglycerol, phosphatidylethanolamine and unidentified lipids (L1-8) were the polar lipids identified. The G+C content of strain JC271T was 46.2 mol%. Integrated genomic and phenotypic data supported the classification of strain JC271T as a representative of a novel genus, for which the name Halodesulfovibrio spirochaetisodalis gen. nov., sp. nov. is proposed. The type strain is JC271T (=KCTC 15474T=DSM 100016T). It is also proposed that Desulfovibrio acrylicus W218T is the latter heterotypic synonym of Desulfovibrio desulfuricans subsp. aestuarii Sylt 3T. Desulfovibrio desulfuricans subsp. aestuarii Sylt 3T should also be elevated as Halodesulfovibrio aestuarii comb. nov. and Desulfovibrio marinisediminisreclassified as Halodesulfovibrio marinisediminis comb. nov. Desulfovibrio oceani subsp. oceanishould be reclassified as Halodesulfovibrio oceani subsp. oceani comb. nov. and Desulfovibrio oceani subsp. galateae as Halodesulfovibrio oceani subsp. galateae comb. nov.

Characterisation of a newly isolated member of a candidatus lineage, Marispirochaeta aestuarii gen. nov., sp. nov.

Metagenome analysis of coastal marine habitats of Gujarat, India indicated the presence of twelve novel putative lineages of spirochaetes. Out of which a strain designated JC444T representing a novel putative lineage seven was isolated and characterized based on a polyphasic taxonomic approach. Strain JC444T was helical, Gram-stain-negative, obligate anaerobe, catalase and oxidase negative. Strain JC444T was able to grow at 15-45 °C (optimum at 30-35 °C), pH 6.5-8.6 (optimum at 7.5-8.0) and 0.6-5 % (optimum at 1.5-2.0 %) of NaCl concentration. The major end products of glucose fermentation were acetate, formate, hydrogen and carbon dioxide. C14 : 0, iso-C15 : 0, C16 : 0, C18 : 0, iso-C15 : 1H/C13 : 03OH (summed feature 1), iso-C13 : 0, anteiso-C15 : 0 and iso-C17 : 0 were present as fatty acids. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and unidentified lipids (L1-4) were the polar lipids. G+C mol% of strain JC444T was 53.6 %. 16S rRNA gene sequence comparisons indicated that strain JC444T represents a member of the family Spirochaetaceae in the order Spirochaetales. Strain JC444T has a sequence similarity of 97.1 % with 'Candidatus Marispirochaeta associata' JC231 and <90.1 % with other members of the family Spirochaetaceae. Distinct morphological, physiological and genotypic differences from the previously described taxa support the classification of strain JC444T as a representative of a new genus and species in the family Spirochaetaceae, for which the name Marispirochaeta aestuarii gen. nov., sp. nov. is proposed. Type strain is JC444T (=KCTC 15554T=DSM 103365T).

Description of Rhodobacter azollae sp. nov. and Rhodobacter lacus sp. nov.

Three strains (JA826T, JA912T and JA913), which were yellowish brown colour, rod to oval shaped, Gram-stain-negative, motile, phototrophic bacteria with a vesicular architecture of intracytoplasmic membranes, were isolated from different pond samples. The DNA G+C content of the three strains was between 64.6 and 65.5 mol%. The highest 16S rRNA gene sequence similarity of all three strains was with the type strains of the genus Rhodobacter sensu stricto in the family Rhodobacteraceae. Strain JA826T had highest sequence similarity with Rhodobacter maris JA276T (98.5 %), Rhodobacter viridis JA737T (97.5 %) and other members of the genus Rhodobacter (<97 %). Strain JA912T had highest sequence similarity with Rhodobacter viridis JA737T (99.6 %), Rhodobacter sediminis N1T (99.3 %), Rhodobacter capsulatus ATCC 11166T (98.8 %) and less than 97 % similarity with other members of the genus Rhodobacter. The 16S rRNA gene sequence similarity between strains JA826T and JA912T was 96.9 %. DNA-DNA hybridization showed that strains JA826T and JA912T (values among themselves and between the type strains of nearest members <44 %) did not belong to any of the nearest species of the genus Rhodobacter. However, strains JA912T and JA913 were closely related (DNA-DNA hybridization value >90 %). The genomic distinction was also supported by differences in phenotypic and chemotaxonomic characteristics in order to propose strains JA826T (=KCTC 15478T=LMG 28758T) and JA912T (=KCTC 15475T=LMG 28748T) as new species in the genus Rhodobacter sensu stricto with the names Rhodobacter lacus and Rhodobacter azollae, respectively.

Description of a phototrophic bacterium, Thiorhodococcus alkaliphilus sp. nov.

Strain JA878T was purified from a photoheterotrophic enrichment obtained from a sediment sample of a brown pond near Nari Salt Pan, Bhavnagar, Gujarat, India. Cells of the isolate were coccoid, motile by means of single polar flagellum and Gram-stain-negative. The internal photosynthetic membrane architecture was vesicular. Strain JA878T contained bacteriochlorophyll a and spirilloxanthin series of carotenoids with rhodopin (>85 %) as the major component. Strain JA878T grew optimally at pH 10-11, and had no requirement for NaCl (tolerated up to 6 %, w/v) or vitamins for growth. C16 : 1ω7c/C16 : 1ω6c, C18 : 1ω7c/C18 : 1ω6c and C16 : 0 were identified as the major fatty acids (>10 %). Phosphatidylglycerol, phosphatidylethanolamine, aminophospholipid and an unknown polar lipid were identified. Q8 was the predominant quinone system in strain JA878T. The DNA G+C content was 62.4 mol%. Highest 16S rRNA gene sequence similarity through EzTaxon-based blast analysis of strain JA878T was found with the type strains of Thiorhodococcus fuscus (99 %), Thiorhodococcus kakinadensis (98.6 %), Thiohalobacter thiocyanaticus (98.4 %), Thiophaeococcus fuscus (97.3 %) and other members of the class Gammaproteobacteria (<97.3 %), revealing a close affiliation to the genera Thiorhodococcus, Thiohalobacter and Thiophaeococcus. However, the phylogenetic treeing firmly placed the strain in the genus Thiorhodococcus. Phenotypic and chemotaxonomic evidence supported the affiliation of strain JA878T to the genus Thiorhodococcus and not to Thiohalobacter, Thiophaeococcus or other known genera of Chromatiaceae. Distinct physiological, genotypic and chemotaxonomic differences indicate that strain JA878T represents a novel species of the genus Thiorhodococcus, for which the name Thiorhodococcus alkaliphilus sp. nov. is proposed. The type strain is JA878T (=KCTC 15531T=JCM 31245T).

Description of Lunatimonas salinarum sp. nov.

A Gram-stain-negative, crescent-shaped, non-motile, aerobic bacterium was isolated from a saltern at Nari along the Bhavnagar coast, Gujarat, India. The strain designated JC344T was oxidase- and catalase-positive. The 16S rRNA gene sequence analysis and sequence comparison data indicated that JC344T represented a member of the genus Lunatimonas and was closely related to the only species of the genus, Lunatimonas lonarensis AK24T (95.5 %). The DNA G+C content of JC344T was 43 mol%. JC344T has phosphatidylethanolamine (PE) and four unidentified polar lipids. MK-7 is the only respiratory quinone. The major (>10 %) fatty acids of strain JC344T are iso-C15 : 0, anteiso-C15 : 0 and summed feature 9 (iso-C17 : 1ω9c and/or C16 : 0 10-methyl). On the basis of physiological, genotypic, phylogenetic and chemotaxonomic analyses, we conclude that JC344T represents a novel species of the genus Lunatimonas, for which the name Lunatimonas salinarum sp. nov. is proposed. The type strain is JC344T (=KCTC 42988T=LMG 29259T).

Description of 'Candidatus Marispirochaeta associata' and reclassification of Spirochaeta bajacaliforniensis, Spirochaeta smaragdinae and Spirochaeta sinaica to a new genus Sediminispirochaeta gen. nov. as Sediminispirochaeta bajacaliforniensis comb. nov., Sediminispirochaeta smaragdinae comb. nov. and Sediminispirochaeta sinaica comb. nov.

Strain JC231 was isolated from a coastal saline habitat of Gujarat and was identified based on 16S rRNA gene sequence analysis as a member belonging to the genus Spirochaeta and showed highest sequence similarity (<91 %) with Spirochaeta bajacaliforniensis DSM 16054T and other members of the family Spirochaetaceae. Intensive attempts to culture strain JC231 in pure culture have failed and were associated with only one species of a Desulfovibrio. However, presence of fosmidomycin inhibited the growth of Desulfovibrio sp. and strain JC231 was characterized in its presence. Strain JC231 was an obligate anaerobe, helical shaped and Gram-stain-negative with catalase and oxidase negative. Draft genome sequence analysis of strain JC231 indicated the full complement of genes for both 2-C-methyl-d-erythritol 4-phosphate and 3-hydroxy-3-methylglutaryl-CoA pathways of terpenogenesis. C14 : 0, iso-C15 : 0, C16 : 0, iso-C15 : 1H/C13 : 0 3OH and iso-C14 : 0 are the major (>5 %) fatty acids. Strain JC231 contains diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and six unidentified lipids (L1-L6). G+C content of strain JC231 was 55.7 mol%. Distinct morphological, physiological and genotypic differences from the previously described taxa support the classification of strain JC231 as a representative of a new genus and species in the family Spirochaetaceae, for which the name 'CandidatusMarispirochaeta associata' is proposed. Strain JC231 is deposited as a defined co-culture with Desulfovibrio sp. JC271 to DSMZ (DSM 29857) and KCTC (KCTC 15472). Based on phenotypic, genotypic and phylogenetic analyses, we also propose the reclassification of Spirochaeta bajacaliforniensis as Sediminispirochaeta bajacaliforniensis gen. nov., comb. nov., Spirochaeta smaragdinae as Sediminispirochaeta smaragdinae comb. nov. and Spirochaeta sinaica as Sediminispirochaeta sinaica comb. nov.

Alkalispirochaeta cellulosivorans gen. nov., sp. nov., a cellulose-hydrolysing, alkaliphilic, halotolerant bacterium isolated from the gut of a wood-eating cockroach (Cryptocercus punctulatus), and reclassification of four species of Spirochaeta as new combinations within Alkalispirochaeta gen. nov.

An obligately anaerobic spirochaete designated strain JC227T was isolated from the gut of a wood-eating cockroach, Cryptocercus punctulatus (Scudder), from the Rann of Kutch, Gujarat, India. Strain JC227T was Gram-stain-negative, mesophilic, halotolerant and alkaliphilic. Based on 16S rRNA gene sequence analysis, strain JC227T belongs to the genus Spirochaeta, with Spirochaeta sphaeroplastigenens JC133T (99.51%), S. odontotermitis JC202T (99.30%), S. alkalica Z-7491T (99.10%), S. americana (98.54%) and other members of the genus Spirochaeta (<92.7%) as its closest phylogenetic neighbours. However, DNA-DNA hybridization between strain JC227T and S. sphaeroplastigenens JC133T, S. odontotermitis JC202T, S. alkalica DSM 8900T and S. americana DSM 14872T was 62±2, 63, 58±2 and 48±4 %, respectively. Strain JC227T contained phosphatidylglycerol, diphosphatidylglycerol, an unidentified glycolipid and six unidentified lipids. Summed feature C18:1ω7c/C18:1ω6c was the predominant cellular fatty acid, with significant proportions of C16:0, C14:0, C12:0, C15:1ω6c, C16:1ω5c, C16:1ω6c/C16:1ω7c and C17:0 2-OH. The DNA G+C content of strain JC227T was 55.5 mol%. On the basis of physiological, biochemical, chemotaxonomic (including metabolomic) and genomic differences from previously described taxa, strain JC227T can be differentiated from members of the genus Spirochaeta and represents a novel species of a new genus, for which the name Alkalispirochaeta cellulosivorans gen. nov., sp. nov. is proposed. The type strain of Alkalispirochaeta cellulosivorans is JC227T (=KCTC 15343T=NBRC 110105T). We also propose the reclassification of Spirochaeta sphaeroplastigenens, Spirochaeta odontotermitis, Spirochaeta alkalica and Spirochaeta americana as Alkalispirochaeta sphaeroplastigenens comb. nov. (type strain JC133T=KCTC 15220T=NBRC 109056T), Alkalispirochaeta odontotermitis comb. nov. (type strain JC202T=KCTC 15324T=NBRC 110104T), Alkalispirochaeta alkalica comb. nov. (type strain Z-7491T=DSM 8900T=ATCC 700262T) and Alkalispirochaeta americana comb. nov. (type strain ASpG1T=ATCC BAA-392T=DSM 14872T). The type species of Alkalispirochaeta gen. nov. is Alkalispirochaeta alkalica comb. nov.

Paraclostridium benzoelyticum gen. nov., sp. nov., isolated from marine sediment and reclassification of Clostridium bifermentans as Paraclostridium bifermentans comb. nov. Proposal of a new genus Paeniclostridium gen. nov. to accommodate Clostridium sordellii and Clostridium ghonii.

Twenty-three rod-shaped, endospore-forming, Gram-stain-positive, obligately anaerobic bacteria were isolated from different marine sediment samples of Gujarat. All 23 strains shared 16S rRNA gene sequence similarity of ∼100 %. Strain JC272T was designated the type strain and shared highest sequence similarity with Clostridium bifermentans ATCC 638T (99.8 %), Clostridium ghonii JCM 1400T (98.0 %), Clostridium sordellii ATCC 9714T (97.9 %) and other members of the genus Clostridium ( < 96.4 %). C16 : 0, C18 : 0, C17 : 0, C16 : 1ω9c and iso-C16 : 0 were the major (>5 %) fatty acids. Strain JC272T contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine and two unidentified amino lipids. Genome-based analysis of average nucleotide identity (ANI) and in silico DNA-DNA hybridization (DDH) of strain JC272T with C. bifermentans ATCC 638T yielded values of 94.35 and 58.5 ± 2.8 %, respectively. The DNA G+C content of strain JC272T was 28.3 mol%. Strain JC272T together with C. bifermentans were found to fall outside Clostridium rRNA cluster I considered as Clostridium senso stricto. Based on ANI value, in-silico DDH, and distinct morphological and physiological differences from the previously described taxa, we suggest that strain JC272T represents a novel species of a new genus in the family Clostridiaceae, for which the name Paraclostridium benzoelyticum gen. nov., sp. nov. is proposed. The type strain is JC272T ( = KCTC 15476T = LMG 28745T). It is also proposed to transfer C. bifermentans to this new genus, as Paraclostridium bifermentans comb. nov. (type strain ATCC 638T = DSM 14991T = JCM 1386T). The genus Paeniclostridium gen. nov. is proposed to accommodate C. sordellii and C. ghonii as Paeniclostridium sordellii comb. nov. (type strain ATCC 9714T = LMG 15708T = JCM 3814T) and Paeniclostridium ghonii comb. nov. (type strain ATCC 25757T = DSM 15049T = JCM 1400T).

Mycobacterium oryzae sp. nov., a scotochromogenic, rapidly growing species is able to infect human macrophage cell line.

Gram-stain-positive, acid-fast-positive, rapidly growing, rod-shaped bacteria (designated as strains JC290T, JC430 and JC431) were isolated from paddy cultivated soils on the Western Ghats of India. Phylogenetic analysis placed the three strains among the rapidly growing mycobacteria, being most closely related to Mycobacterium tokaiense 47503T (98.8 % 16S rRNA gene sequence similarity), Mycobacterium murale MA112/96T (98.8 %) and a few other Mycobacterium species. The level of DNA-DNA reassociation of the three strains with M. tokaiense DSM 44635T was 23.4±4 % (26.1±3 %, reciprocal analysis) and 21.4±2 % (22.1±4 %, reciprocal analysis). The three novel strains shared >99.9 % 16S rRNA gene sequence similarity and DNA-DNA reassociation values >85 %. Furthermore, phylogenetic analysis based on concatenated sequences (3071 bp) of four housekeeping genes (16S rRNA, hsp65, rpoB and sodA) revealed that strain JC290T is clearly distinct from all other Mycobacteriumspecies. The three strains had diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositolmannosides, unidentified phospholipids, unidentified glycolipids and an unidentified lipid as polar lipids. The predominant isoprenoid quinone for all three strains was MK-9(H2). Fatty acids were C17 : 1ω7c, C16 : 0, C18 : 1ω9c, C16 : 1ω7c/C16 : 1ω6c and C19 : 1ω7c/C19 : 1ω6c for all the three strains. On the basis of phenotypic, chemotaxonomic and phylogenetic data, it was concluded that strains JC290T, JC430 and JC431 are members of a novel species within the genus Mycobacterium and for which the name Mycobacterium oryzae sp. nov. is proposed. The type strain is JC290T (=KCTC 39560T=LMG 28809T).

Description of Jeotgalibacillus alkaliphilus sp. nov., isolated from a solar salt pan, and Jeotgalibacillus terrae sp. nov., a name to replace 'Jeotgalibacillus soli' Chen et al. 2010.

A Gram-stain-positive, non-motile, rod-shaped bacterium (strain JC303T) isolated from a salt pan was identified based on 16S rRNA gene sequence analysis as a member of the genus Jeotgalibacillus. It was related most closely to Jeotgalibacillus salarius ASL-1T (99.1 % similarity), Jeotgalibacillusalimentarius YKJ-13T (97.9 %), Jeotgalibacillussoli JSM 081008 (97.9 %), Jeotgalibacillusmalaysiensis D5T (97.8 %), Jeotgalibacillusmarinus DSM 1297T (96.3 %), Jeotgalibacilluscampisalis SF-57T (96.1 %) and J. soli P9T (94.9 %). Genomic relatedness based on DNA-DNA hybridization of strain JC303T with the type strains of the closest related species was less than 40 %. Diphosphatidylglycerol, three aminophospholipids, an unidentified aminoglycolipid, two unidentified phospholipids and an unidentified lipid were the polar lipids of strain JC303T. Major (>10 %) fatty acids were anteiso-C15 : 0, iso-C15 : 0 and iso-C14 : 0. Cell-wall amino acids contained peptidoglycan with l-lysine as the diagnostic diamino acid. Strain JC303T contained MK-7 as the predominant (96 %) menaquinone with the presence of a significant amount (4 %) of MK-8. The DNA G+C content was 43 mol%. On the basis of morphological, physiological, genotypic, phylogenetic and chemotaxonomic analyses, strain JC303T is considered to represent a novel species of the genus Jeotgalibacillus, for which the name Jeotgalibacillus alkaliphilus sp. nov. is proposed. The type strain is JC303T (=KCTC 33662T=LMG 28756T). In addition, we propose to rename J. soli (Chen et al., 2010), an illegitimate homonym of the validly published name Jeotgalibacillus soli(Cunha et al., 2012) as Jeotgalibacillus terrae sp. nov. with type strain JSM 081008T (=DSM 22174T=KCTC 13528T).

Salinicoccus amylolyticus sp. nov., isolated from a saltern.

A Gram-stain-positive coccus, strain JC304T, was isolated from a saltern of Nari along the Bhavnagar Coast, Gujarat, India. The 16S rRNA gene sequence analysis and sequence comparison data indicated that JC304T represented a member of the genus Salinicoccus and was most closely related to Salinicoccus roseus 9T (99.6 %), Salinicoccus luteus YIM 70202T (97.0 %), Salinicoccus hispanicus J-82T (97.0 %) and the remaining species of the genus Salinicoccus (<97 %). Genome relatedness based on DNA-DNA hybridization of JC304T with the type strains of the most closely related species was less than 46 % and the ΔTmwas >5 °C indicating that the strain represents a novel species of the genus Salinicoccus. Independent and concatenated phylogenetic analysis of recA/fusA gene translated product showed a clear distinction of JC304T from its phylogenetic neighbors. Diphosphotidylglycerol, phosphatidylglycerol, an unidentified glycolipid and three unidentified lipids (L1, L2 and L3) were the polar lipids of JC304T. Iso-C15 : 0 and anteiso-C15 : 0 were the major (>10 %) fatty acids in strain JC304T. The cell-wall amino acids were l-lysine and d-glycine. Hopanoids were not detected. The major isoprenoid quinone was menaquinone (MK-6). The DNA G+C content of JC304T was 48 mol%. On the basis of physiological, genotypic, phylogenetic and chemotaxonomic analyses, strain JC304T is considered to represent a novel species of the genus Salinicoccus, for which the name Salinicoccusamylolyticus sp. nov. is proposed. The type strain is JC304T (=KCTC 33661T=LMG 28757T).