Novosphingobium decolorationis sp. nov., an aniline blue-decolourizing bacterium isolated from East Pacific sediment.

Aniline blue-decolourizing bacterial strain 502str22T, isolated from sediment collected in the East Pacific, was subjected to characterization by a polyphasic taxonomic approach. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain 502str22T belongs to the genus Novosphingobium, with closely related type strains 'Novosphingobium profundi' F72T (97.6%), N. mathurense SM117T (97.1%) and N. arvoryzae Jyi-02T (97.0%). Digital DNA-DNA hybridization and average nucleotide identity values between strain 502str22T and closely related type strains were 20.3-24.8% and 74.1-81.9%, respectively. The major cellular fatty acid (>10%) was C18:1 ω7c. The polar lipid profile consisted of a mixture of phosphatidylcholine, one sphingoglycolipid, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidylmonomethylethanolamine. The DNA G+C content of strain 502str22T was 65.5 mol%. The polyphasic taxonomic results indicated that strain 502str22T represents a novel species of the genus Novosphingobium, for which the name Novosphingobium decolorationis sp. nov is proposed. The type strain is 502str22T (=KCTC 82134T= MCCC 1K04799 T).

Devosia indica sp. nov., isolated from surface seawater in the Indian Ocean.

A Gram-stain-negative bacterium, designated IO390501T, was isolated from a sea water sample from the Indian Ocean and taxonomically characterized using a polyphasic approach. The strain is phylogenetically close to 'Devosia lucknowensis' L15 and Devosia chinhatensis IPL18T, with 16S rRNA gene sequence similarity of 97.7 and 97.4 %, respectively. The genome of IO390501T has a DNA G+C content of 61.9 mol% for the 3.9 Mb chromosome. Genome-based phylogenetic trees indicated that IO390501T clusters as an independent lineage with 'D. lucknowensis' L15. Genomic relatedness of in silico DNA-DNA hybridization between IO390501T and phylogenetic neighbours ranged from 18.8 to 21.5 %, below the cutoff of 70 %, and corresponding average nucleotide identity values were between 71.4 and 79.0 %, lower than the 95.0 % threshold. The predominant cellular fatty acids of IO390501T are summed feature 8 (C18 : 1ω7c/C18  : 1ω6c) and C16  :  0. IO390501T contains ubiquinone-10 as the sole respiratory quinone, and phosphatidylglycerol and glycolipids as the major polar lipids. On the basis of the results of phenotypic, chemotaxonomic and genetic analyses, strain IO390501T represents a novel species of the genus Devosia for which the name Devosia indica sp. nov. is proposed. The type strain is IO390501T.

Jiella pacifica sp. nov., isolated from the West Pacific Ocean.

A Gram-stain-negative bacterium, designated strain 40Bstr34T, isolated from a sediment sample from the West Pacific Ocean, was taxonomically characterized by using a polyphasic approach. The strain was phylogenetically close to Jiella aquimaris LZB041T and Jiella endophytica CBS5Q-3T, with 16S rRNA gene sequence similarities of 98.5 and 97.1 %, respectively. The genome of strain 40Bstr34T featured a G+C content of 65.7 % for a 5.8 Mb chromosome. Up-to-date bacterial core gene set analysis revealed that strain 40Bstr34T represents one independent lineage with J.aquimaris LZB041T. In silico DNA-DNA hybridization values between strain 40Bstr34T and its phylogenetic neighbours ranged from 30.3-34.2 %, below the cutoff of 70 %. In addition, the corresponding average nucleotide identity values were between 81.8-83.7 %, which are lower than 95 % threshold. The predominant cellular fatty acids of strain 40Bstr34T were summed feature 8 (C18 : 1 ω6c and/or C18 : 1 ω7c), cyclo-C19 : 0 ω8c and iso-C17 : 0 3-OH, and ubiquinone-10 as the predominant respiratory quinone. The major polar lipids included phosphatidylethanolamine, phosphatidylmonomethylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, two unidentified aminolipids and two unidentified lipids. Based on the results of phenotypic, chemotaxonomic and genetic analyses, strain 40Bstr34T is identified as representing a novel species of the genus Jiella for which the name Jiella pacifica sp. nov. is proposed. The type strain is 40Bstr34T (=JCM 33903T=MCCC 1K04569T).

Alteromonas profundi sp. nov., isolated from the Indian Ocean.

A Gram-staining-negative bacterium, designated 345S023T, was isolated from a sea water sample from the Indian Ocean. The results of 16S rRNA gene sequence analysis revealed that 345S023T represents a member of the genus Alteromonas, with closely related type strains Alteromonas fortis 1T (98.7 %), Alteromonas hispanica F-32T (98.6 %) and Alteromonas genovensis LMG 24078T (98.6 %). Up-to-date bacterial core gene set analysis revealed that 345S023T formed a phyletic lineage with Alteromonas australica H 17T. The case for 345S023T representing a novel species was supported by genomic results. Pairwise in silico DNA-DNA hybridization and average nucleotide identity values were much lower than the proposed and generally accepted species boundaries. Strain 345S023T contains ubiquinone-8 (Q-8) as the sole isoprenoid quinone, summed featured 3 (C16 : 1ω7c and/or C16 : 1ω6c), C16 : 0 and C18 : 1ω7c as the dominant cellular fatty acids (>10 %), and phosphatidylglycerol and phosphatidylethanolamine as the major polar lipids. The genome of strain 345S023T consisted of a 4.4 Mb chromosome with a DNA G+C content of 44.4 %. On the basis of these genomic, chemotaxonomic and phenotypic characteristics, we propose a novel species: Alteromonas profundi sp. nov. The type strain is 345S023T(=JCM 33893T=MCCC 1K04570T).

Muricauda oceani sp. nov., isolated from the East Pacific Ocean.

A Gram-stain-negative bacterium, designated strain 501str8T, was isolated from a sediment sample collected from the East Pacific Ocean. 16S rRNA gene sequence analysis revealed that strain 501str8T belonged to the genus Muricauda, with closely related type strains Muricauda aquimarina SW-63T (98.5 %), Muricauda lutimaris SMK-108T (98.3 %) and Muricauda ruestringensis B1T (97.9 %). Up-to-date bacterial core gene set analysis revealed that strain 501str8T represented one independent lineage with M. aquimarina SW-63T. The average nucleotide identity values of strain 501str8T with M. aquimarina SW-63T and M. lutimaris SMK-108T were 80.2 and 81.3 %, respectively. In silico DNA-DNA hybridization values between strain 501str8T and M. aquimarina SW-63T and M. lutimaris SMK-108T were 22.8 and 32.9 %, respectively. The predominant isoprenoid quinone was menaquinone-6, and iso-C15 : 0, iso-C17 : 0 3-OH and iso-C15 : 1 G were the dominant cellular fatty acids. The G+C content of the genomic DNA was 42.8 mol%. Differential phylogenetic distinctiveness and chemotaxonomic differences, together with the phenotypic properties observed in this study, revealed that strain 501str8T could be differentiated from closely related species. Therefore, we propose that strain 501str8T represents a novel species of the genus Muricauda, for which the name Muricauda oceani sp. nov. is suggested. The type strain is 501str8T (=JCM 33902T=MCCC 1K04567T).

Devosia naphthalenivorans sp. nov., isolated from East Pacific Ocean sediment.

A Gram-stain-negative bacterium, designated CM5-1T, was isolated from a sediment sample collected from the East Pacific Ocean. 16S rRNA gene sequence analysis revealed that strain CM5-1T belongs to the genus Devosia, with closely related type strains Devosia submarina KMM 9415T (98.6 %), Devosia psychrophilaCr7-05T (98.6 %) and Devosia psychrophilaE84T (98.2 %). Up-to-date bacterial core gene set analysis revealed that strain CM5-1T represents one independent lineage with D. submarina KMM 9415T. The average nucleotide identity values of CM5-1T with D. submarina KMM 9415T and D. psychrophila Cr7-05T are 80.1 and 77.9 %, respectively. In silico DNA-DNA hybridization values between strain CM5-1T and D. submarina KMM 9415T and D. psychrophila Cr7-05T are 23.8 and 21.9 %, respectively. Strain CM5-1T contains diphosphatidylglycerol, phosphatidylglycerol and glycolipid as major polar lipids. The sole isoprenoid quinone is ubiquinone-10, and C18 : 1ω7c and 11-methyl C18 : 1ω7c are the dominant cellular fatty acids. The G+C content of the genomic DNA is 61.4 mol%. Differential phylogenetic distinctiveness and chemotaxonomic differences, together with the phenotypic properties observed in this study, revealed that strain CM5-1T could be differentiated from closely related species. Therefore, we propose strain CM5-1T as a novel species of the genus Devosia, for which the name Devosia naphthalenivorans sp. nov. is suggested. The type strain is CM5-1T (=JCM32509T=CGMCC 1.13553T).

Muricauda sediminis sp. nov., isolated from western Pacific Ocean sediment.

A Gram-stain-negative bacterium, designated strain 40Bstr401T, was isolated from a sediment sample collected from the western Pacific Ocean. Analysis of its 16S rRNA gene sequence revealed that strain 40Bstr401T belongs to the genus Muricauda and is closely related to type strains Muricauda antarctica Ar-22T (98.2 %), Muricauda taeanensis 105T (98.2 %) and Muricauda beolgyonensis BB-My12T (97.4 %). The average nucleotide identity values for 40Bstr401T with M. antarctica Ar-22T and M. taeanensis 105T are 79.3 % and 78.8 %, respectively. The in silico DNA-DNA hybridization values between strain 40Bstr401T and M. antarctica Ar-22T and M. taeanensis 105T are 26.7 and 26.6 %, respectively. The major isoprenoid quinone of 40Bstr401T is MK-6, and iso-C17 : 0 3-OH and iso-C15 : 0 are the dominant cellular fatty acids. The major polar lipids are phosphatidylethanolamine, four unidentified amino lipids and two unidentified lipids. The G+C content of the genomic DNA is 42.9 mol%. Its phylogenetic distinctiveness and chemotaxonomic differences, together with the phenotypic properties observed in this study, indicate that strain 40Bstr401T can be differentiated from closely related species. Therefore, we propose strain 40Bstr401T represents a novel species in the genus Muricauda, for which the name Muricauda sediminis sp. nov. is suggested. The type strain is 40Bstr401T (=MCCC 1K04568T=KCTC 82139T).

Paracoccus indicus sp. nov., isolated from surface seawater in the Indian Ocean.

Strain IO390502T, isolated from surface seawater in the Indian Ocean, was characterised using a polyphasic taxonomy approach. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain IO390502T belongs to the genus Paracoccus and is closely related to Paracoccus seriniphilus DSM 14827T (97.6%), followed by P. zeaxanthinifaciens JCM 21774T (97.5%), P. homiensis DSM 17862T (97.3%), P. marcusii DSM 11574T (97.2%), P. haeundaensis BC 74171T (97.0%) and P. carotinifaciens E-396T (97.0%). Cells are Gram-negative, aerobic, poly-ß-hydroxybutyrate-accumulating, motile, rod-shaped, and forms creamy-white colonies. Optimal growth occurred at 25-30 °C, pH 5-8, and in the presence of 3-8% NaCl. The genome of strain IO390502T has a G+C content of 64.9 mol% and a 3.5 Mb chromosome. The in silico DNA-DNA hybridisation and average nucleotide identity values between strain IO390502T and the three closely related taxa, P. seriniphilus DSM 14827T, P. zeaxanthinifaciens JCM 21774T and P. homiensis DSM 17862T, are 19.6%, 21.9% and 20.6%, and 76.0%, 79.9% and 77.8%, respectively. Phosphatidylglycerol is the major lipid present, ubiquinone-10 (Q-10) is the sole isoprenoid quinone, and the major cellular fatty acid is C18:1ω7c. Based on data from phenotypic tests and genotypic differences between strain IO390502T and its close phylogenetic relatives, strain IO390502T represents a new species belonging to the genus Paracoccus, for which the name Paracoccus indicus sp. nov. is proposed. The type strain is IO390502T (= JCM 32637T = CCTCC AB 2018071T).

Potential TSPO Ligand and Photooxidation Quencher Isorenieratene from Arctic Ocean Rhodococcus sp. B7740.

Due to its special aromatic structure, isorenieratene is thought to be an active natural antioxidant and photo/UV damage inhibitor. In this work, isorenieratene that was extracted from Rhodococcus sp. B7740 isolated from the Arctic Ocean, showed excellent scavenging ability of both singlet oxygen and hydroxyl radical in the UVB-induced auto-oxidation process using the EPR method. Within an ARPE-19 cell model damaged by UVB radiation, isorenieratene showed fine protective effects (1.13 ± 0.03 fold) compared with macular xanthophylls (MXs) through upregulating of tspo. The molecular docking was firstly performed to investigate the interaction of isorenieratene with TSPO as a special ligand. Results showed isorenieratene might form a better binding conformation (S-score -8.5438) than MXs and indicate that isorenieratene not only can function as a direct antioxidant but also activate tspo in ARPE-19 cells. Thus, isorenieratene might ease the UV-related damages including age-related macular degeneration (AMD).

Alteromonas indica sp. nov., isolated from surface seawater from the Indian Ocean.

A Gram-stain-negative bacterium, designated strain IO390401T, was isolated from a seawater sample from the sulphide region of the Indian Ocean. Phylogenetic trees based on 16S rRNA gene sequences showed that strain IO390401T is a member of the genus Alteromonas, sharing 97.0-97.4 % 16S rRNA gene sequence similarity with Alteromonas additaR10SW13T, A. stellipolaris LMG 21861T, A. naphthalenivorans JCM 17741T, A. gracilis 9a2T and A. australica H17T, and 94.8-96.8 % with the type strains of other species of the genus Alteromonas. Strain IO390401T contained ubiquinone-8 (Q-8) as the sole isoprenoid quinone, C16:0 and C16:1ω7c/C16:1ω6c as the dominant cellular fatty acids, and phosphatidylglycerol and phosphatidylethanolamine as the major polar lipids. The genome of strain IO390401T consists of a 4.4 Mb chromosome with a G+C content of 48.2 mol%. Average nucleotide identity values between strain IO390401T and the three closest phylogenetic neighbours, namely A. additaR10SW13T, A. stellipolaris LMG 21861T and A. naphthalenivorans JCM 17741T, were 70.5, 70.4 and 70.6 %, respectively, and the corresponding in silico DNA-DNA hybridization values were 20.6, 20.7 and 21.1 %. Phylogenetic distinctiveness and chemotaxonomic differences, together with phenotypic properties determined in this study, revealed that strain IO390401T could be differentiated from closely related species. It is therefore proposed as representing a novel species in the genus Alteromonas, for which the name Alteromonas indica sp. nov. is suggested. The type strain is IO390401T (=JCM 32638T=CGMCC 1.13554T=CCTCC AB 2018072T).

Carnobacterium antarcticum sp. nov., a psychrotolerant, alkaliphilic bacterium isolated from sandy soil in Antarctica.

A novel, alkaliphilic, psychrotolerant, facultative anaerobe, designated CP1T, was isolated from sandy soil near the Davis Station in Antarctica. The short-rod-shaped cells displayed Gram-positive staining and did not form spores. Strain CP1T was able to grow at temperatures between 4 and 36 °C, pH 6.0-9.5, and in the presence of up to 5.0 % (w/v) NaCl. 16S rRNA gene and multilocus (pheS, rpoA, and atpA) sequence analysis revealed Carnobacterium mobile DSM 4848T and Carnobacterium iners LMG 26642T as the closest relatives (97.4 and 97.1 % 16S rRNA gene sequence similarity, respectively). The genomic G+C content was 38.1 mol%, and DNA-DNA hybridization with DSM 4848T revealed 32.4±3.4 % similarity. The major fatty acid components were C14 : 0 and C16 : 1ω9c. The cell wall contained meso-diaminopimelic acid and was of peptidoglycan type A1γ. Based on physiological, genotypic and biochemical characteristics, strain CP1T represents a novel species of the genus Carnobacterium for which the name Carnobacterium antarcticum sp. nov. is proposed. The type strain is CP1T (=DSM 103363T=CGMCC 1.15643T).

Characterization of MK8(H2) from Rhodococcus sp. B7740 and Its Potential Antiglycation Capacity Measurements.

Menaquinone (MK) has an important role in human metabolism as an essential vitamin (VK2), which is mainly produced through the fermentation of microorganisms. MK8(H2) was identified to be the main menaquinone from Rhodococcus sp. B7740, a bacterium isolated from the arctic ocean. In this work, MK8(H2) (purity: 99.75%) was collected through a convenient and economic extraction process followed by high-speed countercurrent chromatography (HSCCC) purification. Additionally, high-resolution mass spectrometry (HRMS) was performed for further identification and the hydrogenation position of MK8(H2) (terminal unit) was determined using nuclear magnetic resonance (NMR) for the first time. MK8(H2) showed a superior antioxidant effect and antiglycation capacity compared with ubiquinone Q10 and MK4. High-performance liquid chromatography⁻mass spectrometer (HPLC-MS/MS) and molecular docking showed the fine interaction between MK8(H2) with methylglyoxal (MGO) and bull serum albumin (BSA), respectively. These properties make MK8(H2) a promising natural active ingredient with future food and medicine applications.

Identification and characterization of host cell proteins interacting with Scylla serrata reovirus non-structural protein p35.

We have previously shown that non-structural protein p35, encoded by Scylla serrata reovirus (SsRV) S10, may act as a viroporin. To characterize the role of p35 protein in the modulation of cellular function, a yeast two-hybrid system was used to screen a cDNA library derived from S. serrata to find its interacting partner. Protein interactions were confirmed in vitro by GST pull-down. Full cDNAs of p35 interactors were cloned by the rapid amplification of cDNA ends. After two-hybrid library screening, we isolated partial cDNAs encoding hemocyanin, cryptocyanin, and TAX1BP1. Interaction of p35 with each of hemocyanin, cryptocyanin, and TAX1BP1 was confirmed by GST pull-down. The full-length cDNA fragments of hemocyanin, cryptocyanin, and TAX1BP1 were 2287, 2422, and 3437 bp, respectively, and they encoded three putative proteins with molecular masses of ~76.9, ~79.2, and ~107.2 kDa, respectively. This study casts new light on the function and physiological significance of p35 during the SsRV replication cycle.

Identification and RNA segment assignment of six structural proteins of Scylla serrata reovirus.

Scylla serrata reovirus (SsRV) is one of the most prevalent viral pathogens of the mud crab (S. serrata). The virus represents an unassigned novel genus in the Reoviridae family, and contains 12 double-stranded RNA genomic segments. Previous analysis of virion proteins concluded that SsRV contains at least eight structural proteins, ranging from 25 to 160 kDa. Here, tandem time-of-flight mass spectrometry and Western blotting were used to re-identify the structural proteins. The results indicate that proteins encoded by SsRV segments S1, S3, S6, S9, S11, and S12 are structural proteins.

[Bacterial diversity in deep-sea sediments from two stations in the east Pacific polymetallic nodule province].

Objective: The objective of this study was to assess bacterial diversity in sediment samples from two stations (WBC1305 and WBC1316A) in the Pacific polymetallic nodule province. Methods: The environmental total DNAs were extracted, and 6 bacterial 16S rRNA gene libraries were generated from 6 sediment layers. The Shannon diversity index and Simpson dominance index were calculated for each bacterial community and then compared. The bacterial community structure of each sediment sample was analyzed, and the results were used to construct phylogenetic trees. Results: In total, 533 bacterial clones were obtained from 6 bacterial clone libraries. Among these 533 clones, 472 clones could be assigned to 16 phylogenetic groups (Acidobacteria, Actinobacteria, Alpha, Beta, Delta, gamma-Proteobacteria, Bacteroidetes, Chloroflexi, Firmicutes, Gemmatimonadetes, Nitrospirae, Planctomycetes, Elusimicrobia, Hydrogenedentes, Chlorobi, and Nitrospinae), whereas the remaining 61 clones could not be classified into any known groups. Conclusion: The bacterial communities in sediments from WBC1305 are dominated mainly by gamma-Proteobacteria and from WBC1316A by Firmicutes. In addition, the bacterial community structure at WBC1316A is more abundant and complex than that at WBC1305.

Scylla serrata reovirus p35 protein expressed in Escherichia coli cells alters membrane permeability.

To promote viral entry, replication, release, and spread to neighboring cells, many cytolytic animal viruses encode proteins responsible for modification of host cell membrane permeability and for formation of ion channels in host cell membranes. Scylla serrata reovirus (SsRV) is a major pathogen that can severely damage mud crab (S. serrata) aquaculture. Protein p35, which is encoded by segment 10 of SsRV, contains two transmembrane domains. In this study, we found that SsRV p35 can induce membrane permeability changes when expressed in Escherichia coli. SsRV p35 expressed in bacterial cells existed as monomers under reducing conditions but formed homodimers and homotrimers under non-reducing conditions. These findings demonstrate that p35 may act as a viroporin; further studies are needed to elucidate the detailed structure-function relationships of this protein.

Molecular characterization of eight segments of Scylla serrata reovirus (SsRV) provides the complete genome sequence.

Scylla serrata reovirus (SsRV) is one of the most prevalent viral pathogens of mud crabs (S. serrata). Of the 12 double-stranded RNA (dsRNA) genomic segments (S1-S12), the three largest (S1-S3) and S7 were sequenced previously and were shown to have no or only low sequence homology to known members within the family Reoviridae. The sequences of the remaining segments, S4-S6 and S8-S12, are reported here. With the exception of S4, all have single open reading frames (ORFs) on their positive strands, and the terminal sequences 5'-AUAAA(U)/(C) (A)/(U)…G(A)/(G) (A)/(U) (A)/(C)AAC(G)/(U)AU-3' are conserved among currently and previously sequenced segments. S4 contains two out-of-phase ORFs on the positive strand, suggesting that this segment is bicistronic. The ORFs of segments S4-S6 and S8-S12 have low or no homology to other reovirus genes, with the exception that all of the SsRV segments have high sequence similarity to those of mud crab reovirus (MCRV) and share the same 5'- and 3'-terminal nucleotide sequences, suggesting that the two viruses belong to the same species in the family Reoviridae. Analysis of virion proteins revealed that SsRV contains at least eight structural proteins, with sizes ranging from 25 to 160 kDa.

Complete Genome Sequence of Cellulomonas sp. Strain C5510, Isolated from Pacific Ocean Seawater.

We report the complete genome sequence of Cellulomonas sp. strain C5510, obtained by Oxford Nanopore and Illumina sequencing. The approximately 4.15-Mb genome sequence consists of one circular chromosome and one circular plasmid, with an overall G+C content of 74.9%.

Nucleotide sequences of four RNA segments of a reovirus isolated from the mud crab Scylla serrata provide evidence that this virus belongs to a new genus in the family Reoviridae.

This is the first sequence-based characterization of mud crab (Scylla serrata) reovirus (SsRV), which causes severe disease of cultured mud crabs in southern China. We sequenced and analyzed genome segments S1, S2, S3, and S7, which were 4,327, 2,721, 2,715, and 1,517 nucleotides long, respectively. Conserved motifs were found at the 5' (AUAAAU) and 3' (AACGAU) ends of each segment. RNA segments S1, S2, S3, and S7 each contained a single open reading frame (ORF) that encoded predicted proteins of 160, 100, 96, and 46 kDa, respectively. The ORFs of segments S1 and S2 showed distant homologies (< 25%) with cognate genes of other reoviruses, whereas the ORFs of segments S3 and S7 had no homologies with any other viral genes. Based on these observations, we propose that SsRV should be considered a member of a new genus in the family Reoviridae.

Isolation and Identification of Acholeplasma sp. from the Mud Crab, Scylla serrata.

For the first time, a mollicute-like organism (MLO) was cultured from moribund mud crabs (Scylla serrata) during an outbreak of clearwater disease in Zhejiang Province, China. The MLO displayed a fried-egg colony morphology in culture, did not possess a cell wall, and was not retained by 0.45 µm and 0.2 µm filters. It was able to ferment glucose, sucrose, lactose, and maltose, but it did not utilize arginine and urea. The MLO grew in the absence of bovine serum and was not susceptible to digitonin. Sequence analysis of the 16S rRNA gene revealed that this MLO had 99% identity with Acholeplasma laidlawii PG-8A, which indicates that the organism isolated from mud crabs is a member of the genus Acholeplasma.