Alicyclobacillus curvatus sp. nov. and Alicyclobacillus mengziensis sp. nov., two acidophilic bacteria isolated from acid mine drainage.

Two acidophilic strains, designated as ALEF1T and S30H14T, were isolated from acid mine drainage sediment. Cells of both strains were Gram-stain-positive, aerobic, endospore-forming rods. Strains ALEF1T and S30H14T were acidophilic and mesophilic, the former grew at 20-40 °C (optimum, 30 °C) and pH 2.5-4.5 (optimum, pH 3.5), while the latter grew at 20-45 °C (optimum, 30 °C) and pH 2.0-5.5 (optimum, pH 4.5). The 16S rRNA gene-based sequence analysis revealed that strains ALEF1T and S30H14T belonged to the genus Alicyclobacillus, and were phylogenetically close to Alicyclobacillus ferrooxydans TC-34T with 97.1 and 97.4% similarity, respectively. The similarity between the two novel strains was 98.6 %. The average nucleotide identity value between the genome sequences of ALEF1T and S30H14T was 79.5 %, and that between each of the two isolates and A. ferrooxydans TC-34T were 72.0 and 74.3 %. In addition, the digital DNA-DNA hybridization value between ALEF1T and S30H14T was 24.9 %, between strain ALEF1T and A. ferrooxydans TC-34T was 21.7 %, and between S30H14T and A. ferrooxydans TC-34T was 26.3 %, far below the interspecies threshold. Both strains could utilize diverse carbon sources for heterotrophic growth; strain ALEF1T could utilize ferrous iron as the energy source for autotrophic growth. Menaquinone 7 was the only quinone detected in either strain. Both strains contained anteiso-C15 : 0 and anteiso-C17 : 0, while ω-alicyclic fatty acids were not detected. Based on their phylogenetic positions, as well as phenotypic and genomic data, it is considered that strains ALEF1T and S30H14T represent two novel species within the genus Alicyclobacillus, for which the names Alicyclobacillus curvatus sp. nov. (type strain ALEF1T=CGMCC 1.17055T=KCTC 43124T) and Alicyclobacillus mengziensis sp. nov. (S30H14T=CGMCC 1.17050T=KCTC 43125T) are proposed.

Marinobacter caseinilyticus sp. nov., Isolated from Saline Soil.

A Gram-stain-negative, motile, aerobic, rod-shaped bacterium with flagella, designated M3-13T, was isolated from a saline soil in Zhoushan, China. According to phylogenetic analysis based on 16S rRNA gene sequences, strain M3-13T was assigned to the genus Marinobacter with highest 16S rRNA gene sequence similarity of 97.7% to Marinobacter maroccanus LMG 30466T, followed by Marinobacter sediminum R65 T (97.5%) and M. salsuginis SD-14BT (97.2%). Digital DNA-DNA hybridization (DDH) and average nucleotide identity (ANI) were determined to evaluate the genomic relationship between strain M3-13T and M. maroccanus LMG 30466T. Digital DDH estimation (19.8%) as well as ANI (72.98%) proved the dissimilarity of strain M3-13T. Optimal growth of the strain M3-13T was at 28-30 °C and at pH 8.0-8.5, in the presence of 3-6% (w/v) NaCl. The major fatty acids detected in strain M3-13T were C16:1 ω7c/C16:1 ω6c, C16:0, C18:1ω7c/C18:1 ω6c and C12:03-OH, and the predominant respiratory quinone was ubiquinone-9. The major polar lipids included diphosphatidyglycerol, phosphatidylglycerol, phosphatidylethanolamine, one unidentified aminophosphoglycolipid and one unidentified phosphoglycolipid. The DNA G+C content was 56.6%. A phylogenetic analysis based on 16S rRNA gene sequences showed that strain M3-13T belongs to the genus Marinobacter. Based on the polyphasic taxonomic characterization, strain M3-13T is considered to represent a novel species of the genus Marinobacter, for which the name Marinobacter caseinlyticus sp. nov. is proposed (type strain M3-13T = MCCC 1K04560T = KCTC 72043T).

Muricauda maritima sp. nov., Muricauda aequoris sp. nov. and Muricauda oceanensis sp. nov., three marine bacteria isolated from seawater.

Three Gram-stain-negative, non-motile, rod-shaped strains, designated 72T, NH166T and 40DY170T, were isolated from seawater samples of the West Pacific Ocean, South China Sea and West Pacific Ocean, respectively. The 16S rRNA gene sequence similarity results revealed that strains 72Tand NH166T were most closely related to Muricauda antarctica Ar-22T, Muricauda taeanensis JCM 17757T, Muricauda beolgyonensis KCTC 23501T, Muricauda lutimaris KCTC 22173T and Muricauda hadalis MT-229T with 97.2-98.0% sequence similarity. 16S rRNA gene sequence analysis also indicated that strain 40DY170T was most closely related to Muricauda ruestringensis DSM 13258T, Muricauda aquimarina JCM 11811T, Muricauda lutimaris KCTC 22173T and Muricauda oceani 501str8T with 97.6-98.1% sequence similarity. The 16S rRNA gene sequence similarity values among strains 72T, NH166T and 40DY170T were 96.5-99.2%. Phylogenetic analyses indicated that three new isolates represented three novel species by forming two distinctive lineages within the genus Muricauda. The DNA G+C contents of strain 72T, NH166T and 40DY170T were 43.4, 43.4 and 42.4 mol%, respectively. The average nucleotide identity and in silico DNA-DNA hybridization values between strains 72T, NH166T, 40DY170T and the reference strains were 76.5-93.5% and 19.2-53.5%, respectively. The sole respiratory quinone in all strains was menaquinone-6. Their major fatty acids were iso-C17:0 3-OH, iso-C15:0 and iso-C15 : 1 G. The major polar lipids of strains 72T and NH166T were phosphatidylethanolamine, one unidentified aminolipid and two unidentified lipids. The major polar lipids of strain 40DY170T were phosphatidylglycerol, one unidentified phospholipid, one unidentified aminolipid and two unidentified lipids. On the basis of their distinct taxonomic characteristics, the three isolates represent three novel species of the genus Muricauda, for which the names Muricauda maritima sp. nov. (type strain 72T=KCTC 62229T=MCCC 1K03350T), Muricauda aequoris sp. nov. (NH166T=KCTC 62228T=MCCC 1K03449T) and Muricauda oceanensis sp. nov. (40DY170T=KCTC 72200T=MCCC 1K03569T) are proposed.

Mesorhizobium oceanicum sp. nov., isolated from deep seawater.

A novel Gram-staining-negative, oval-shaped (0.4-0.6×0.8-1.0 µm), non-motile strain without flagella, designated B7T, was isolated from deep seawater in the South China Sea. Strain B7T was able to grow at 25-40 °C (optimum 35 °C), at pH 5.5-9.0 (optimum pH 7.0) and with 0-8 % (w/v) NaCl (optimum 3 %). Chemotaxonomic analysis showed that the predominant isoprenoid quinone was Q-10 and the dominant fatty acids were C19 : 0 cyclo 8c and summed feature 8 (C18 : 1ω7c/C18 : 1ω6c). The polar lipids of strain B7T were diphosphatidyglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, one unknown aminophospholipid, one unknown glycolipid and three unknown lipids. The DNA G+C content of the genomic DNA was 65.1 mol%. Phylogenetic analysis of 16S rRNA gene sequences showed that strain B7T belongs to the genus Mesorhizobium with similarities ranging from 96.2 to 97.5 %. Phylogenetic analyses of housekeeping genes recA, atpD and glnII indicated that strain B7T represented a distinct evolutionary lineage with the genus Mesorhizobium. OrthoANI values between strain B7T and related strains of the genus Mesorhizobium (<80 %) were lower than the threshold value of 95 % ANI relatedness for species demarcation. Therefore, strain B7T is concluded to represent a novel species of the genus Mesorhizobium, for which the name Mesorhizobium oceanicumsp. nov. is proposed. The type strain is B7T (=KCTC 42783T=MCCC 1K02305T).

Thalassobaculum fulvum sp. nov., isolated from deep seawater.

A novel Gram-stain-negative, rod-shaped (1.0-1.2×2.0-8.0 µm), non-motile without flagella strain, designated HSF7T, was isolated from deep seawater. Strain HSF7T was able to grow at 20-40 °C (optimum 35 °C), pH 5.5-9.0 (optimum pH 6.5) and 0-10 % (w/v) NaCl (optimum 2 %). The G+C content of the genomic DNA was 69 mol%. Bacteriochlorophyll a and poly-ß-hydroxybutyrate (PHB) granules were not found. The major fatty acids were C18 : 1ω7c (69.3 %), C16 : 0 (9.1 %) and C19 : 0 cyclo ω8c (6.6 %). The polar lipids were phosphatidylglycerol, three unknown aminophospholipids, an unknown phospholipid, an unknown aminolipid and two unknown lipids. The only isoprenoid quinone was Q-10. 16S rRNA gene sequence analysis revealed that strain HSF7T was most closely related toThalassobaculum salexigens DSM 19539T, Thalassobaculum litoreum DSM 18839T, Nisaeadenitrificans DSM 18348T and Oceanibaculum indicum MCCC 1A02083Twith pairwise sequence similarities of 95.56 %, 95.21 %, 93.64 % and 92.65 %, respectively. On the basis of genotypic, phenotypic, phylogenetic and chemotaxonomic characteristics, strain HSF7T represents a novel species of the genus Thalassobaculum, or which the name Thalassobaculum fulvum sp. nov. is proposed. The type strain is HSF7T(=KCTC 42651T=MCCC 1K01158T).

Seohaeicola zhoushanensis sp. nov., isolated from seawater.

A novel, Gram-stain-negative, motile, facultatively anaerobic bacterium, designated strain NF48T, was isolated from surface seawater around Zhoushan Islands. Cells were rod-shaped (1.1-3.9×0.5-0.9 µm). Strain NF48T was able to grow at 10-40 °C (optimum 30 °C), at pH 5.5-9.0 (optimum 6.5-8.0) and with 0.5-7.0 % (w/v) NaCl (optimum 2.0 %). The genomic DNA G+C content was 65.5 mol%. Chemotaxonomic analysis showed that the main isoprenoid quinone was Q-10 and the major fatty acids were C18 : 1ω7c and C16 : 0. The major polar lipids of strain NF48T were phosphatidylcholine, phosphatidylethanolamine and phosphatidylglycerol. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain NF48T belonged to the genus Seohaeicola with similarity values of 95.5-97.2 % to members of this genus, and was most closely related to Seohaeicola nanhaiensis SS011A0-7#2-2T (97.2 %). On the basis of the phenotypic, chemotaxonomic and phylogenetic characteristics, strain NF48T is suggested to represent a novel species of the genus Seohaeicola, for which the name Seohaeicola zhoushanensis sp. nov. is proposed. The type strain is NF48T (=MCCC 1K01157T=KCTC 42650T).

Haloimpatiens lingqiaonensis gen. nov., sp. nov., an anaerobic bacterium isolated from paper-mill wastewater.

An anaerobic bacterium, strain ZC-CMC3T, was isolated from a wastewater sample in Zhejiang, China. Cells were Gram-stain-positive, peritrichous, non-spore-forming, rod-shaped (0.6-1.2 × 2.9-5.1 µm) and catalase- and oxidase-negative. Strain ZC-CMC3T was able to grow at 25-48 °C (optimum 43 °C) and pH 5.5-8.0 (optimum pH 7.0). The NaCl concentration range for growth was 0-3 % (w/v) (optimum 0 %). The major polar lipids of the isolate were diphosphatidylglycerol, phosphatidylglycerol, several phospholipids and glycolipids. Main fermentation products from PYG medium were formate, acetate, lactate and ethanol. Substrates which could be utilized were peptone, tryptone, yeast extract and beef extract. No respiratory quinone was detected. The main fatty acids were C14 : 0, C16 : 0, C16 : 1cis 7 and C16 : 1cis 9. The DNA G+C content was 30.0 mol%. 16S rRNA gene sequence analysis revealed that the isolate belonged to the family Clostridiaceae. Phylogenetically, the most closely related species were Oceanirhabdus sediminicola NH-JN4T (92.8 % 16S rRNA gene sequence similarity) and Clostridium tepidiprofundi SG 508T (92.6 %). On the basis of phylogenetic, chemotaxonomic and phenotypic characteristics, strain ZC-CMC3T represents a novel species of a new genus in the family Clostridiaceae, for which the name Haloimpatiens lingqiaonensis gen. nov., sp. nov. is proposed. The type strain of the type species is ZC-CMC3T ( = KCTC 15321T = JCM 19210T = CCTCC AB 2013104T).

Terasakiella brassicae sp. nov., isolated from the wastewater of a pickle-processing factory, and emended descriptions of Terasakiella pusilla and the genus Terasakiella.

A Gram-stain-negative, motile, polyhydroxybutyrate-accumulating, aerobic, S-shaped bacterium, designated B3T, was isolated from the wastewater of a pickle-processing factory. 16S rRNA gene sequence similarity analysis showed that it was most closely related to the type strain, Terasakiella pusilla (96.6% similarity). Strain B3T was able to grow at 4-40 °C (optimum 32-37 °C), pH 5.5-9.0 (optimum 6.5-7.5) and with 0.5-8% (w/v) NaCl present (optimum 1-2%, w/v). Chemotaxonomic analysis showed that the respiratory quinone was ubiquinone Q-10, the major fatty acids included C16:0, C18:1ω7c and C16:1ω7c and/or iso-C15:2-OH. The major polar lipids included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine, aminophospholipid and three uncharacterized phospholipids. The genomic DNA G+C content of strain B3T was 42.3 mol%. The DNA-DNA relatedness value between B3T and T. pusilla DSM 9263T was 23.9%. On the basis of the phenotypic, chemotaxonomic and genotypic characteristics of strain B3T, it represents a novel species of the genus Terasakiella, for which the name Terasakiella brassicae sp. nov. is proposed. The type strain is B3T (=KCTC 42652T=CGMCC 1.15254T). Emended descriptions of T. pusilla and the genus Terasakiella are also presented.