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Four strains of lactic acid bacteria isolated from cachaça and alcohol fermentation vats in Brazil were characterised in order to determine their taxonomic position. Phylogenetic analysis revealed that they belong to the genus Oenococcus and should be distinguished from their closest neighbours. The 16S rRNA gene sequence similarity against the type strains of the other two species of the genus was below 94.76 % (Oenococcus kitaharae) and 94.62 % (Oenococcus oeni). The phylogeny based on pheS gene sequences also confirmed the position of the new taxon. DNA-DNA hybridizations based on in silico genome-to-genome comparison, Average Amino Acid Identity, Average Nucleotide Identity and Karlin genomic signature confirmed the novelty of the taxon. Distinctive phenotypic characteristics are the ability to metabolise sucrose but not trehalose. The name Oenococcus alcoholitolerans sp. nov. is proposed for this taxon, with the type strain UFRJ-M7.2.18(T) ( = CBAS474(T) = LMG27599(T)). In addition, we have determined a draft genome sequence of the type strain.

Three novel isolates (A-354(T), A-328, and A-384) were retrieved from apparently healthy scleractinian Madracis decactis in the remote St Peter & St Paul Archipelago, Mid-Atlantic Ridge, Brazil. The novel isolates formed a distinct lineage based on the phylogenetic reconstruction using the 16S rRNA and pyrH gene sequences. They fell into the Mediterranei clade and their closest phylogenetic neighbour was V. mediterranei species, sharing upto 98.1 % 16S rRNA gene sequence similarity. Genomic analysis including in silico DDH, MLSA, AAI and genomic signature distinguished A-354(T) from V. mediterranei LMG 19703 (=AK1) with values of 33.3, 94.2, 92 %, and 11.3, respectively. Phenotypically, the novel isolates can be differentiated from V. mediterranei based on the four following features. They do not grow at 8 % NaCl; use D-gluconic acid but not L-galactonic acid lactone as carbon source; and do not have the fatty acid C18:0. Differentiation from both the other Mediterranei clade species (V. maritimus and V. variabilis) is supported by fifteen features. The novel species show lysine decarboxylase and tryptophan deaminase, but not gelatinase and arginine dihydrolase activity; produce acetoin; use α-D-lactose, N-acetyl-D-galactosamine, myo-Inositol, D-gluconic acid, and β-hydroxy-D,L-butyric acid; and present the fatty acids C14:0 iso, C15:0 anteiso, C16:0 iso, C17:0 anteiso, and C17:1x8c . Whole-cell protein profiles, based on MALDI-TOF, showed that the isolates are not clonal and also distinguished them from the closes phylogenetic neighbors. The name Vibrio madracius sp. nov. is proposed to encompass these novel isolates. The G+C content of the type strain A-354(T) (=LMG 28124(T)=CBAS 482(T)) is 44.5 mol%.

Background: Salinivibrios are moderately halophilic bacteria found in salted meats, brines and hypersaline environments. We obtained three novel conspecific Salinivibrio strains closely related to S. costicola, from Socompa Lake, a high altitude hypersaline Andean lake (approx. 3,570 meters above the sea level)., Results: The three novel Salinivibrio spp. were extremely resistant to arsenic (up to 200 mM HAsO42-), NaCl (up to 15%), and UV-B radiation (19 KJ/m2, corresponding to 240 minutes of exposure) by means of phenotypic tests. Our subsequent draft genome ionsequencing and RAST-based genome annotation revealed the presence of genes related to arsenic, NaCl, and UV radiation resistance. The three novel Salinivibrio genomes also had the xanthorhodopsin gene cluster phylogenetically related to Marinobacter and Spiribacter. The genomic taxonomy analysis, including multilocus sequence analysis, average amino acid identity, and genome-to-genome distance revealed that the three novel strains belong to a new Salinivibrio species., Conclusions: Arsenic resistance genes, genes involved in DNA repair, resistance to extreme environmental conditions and the possible light-based energy production, may represent important attributes of the novel salinivibrios, allowing these microbes to thrive in the Socompa Lake.

We report on the first characterization of the culturable heterotrophic bacteria of the scleractinian Madracis decactis. In addition, we characterized the culturable bacteria associated with the fireworm Hermodice carunculata, observed predating partially bleached coral colonies. Our study was carried out in the remote St. Peter and St. Paul Archipelago (SPSPA), Mid-Atlantic Ridge, Brazil. We constituted a 403 isolates collection and subsequently characterized it by means of pyrH and 16S rRNA partial sequences. We identified Photobacterium, Bacillus, and Vibrio species as members of the culturable microbiota of healthy M. decactis. V. campbellii, V. harveyi, V. communis, and V. maritimus were the most commonly found Vibrio species in healthy corals, representing more than 60 % of all vibrio isolates. Most of the vibrios isolated from the fireworm's tissues (n = 143; >90 %) were identified as V. shiloi. However, we did not recover V. shiloi from bleached M. decactis. Instead, we isolated V. communis, a novel Photobacterium species, Bacillus, Kocuria, and Pseudovibrio, suggesting a possible role of other facultative anaerobic bacteria and/or environmental features (such as water quality) in the onset of bleaching in SPSPA's M. decactis.

Two novel vibrio isolates (R-40492(T) and R-40493(T)) originating from the zoanthid Palythoa caribaeorum in Brazil in 2005 were taxonomically characterized by means of a polyphasic approach comprising multilocus sequence analysis (MLSA), DNA-DNA hybridization (DDH), ΔT(m) analysis and phenotypic characterization. Phylogenetic analysis based on 16S rRNA gene sequences showed that R-40492(T) and R-40493(T) fell within the genus Vibrio and were most closely related to each other with 99% similarity; similarities of these two novel isolates towards Vibrio neptunius LMG 20536(T), Vibrio coralliilyticus LMG 20984(T), Vibrio nigripulchritudo LMG 3896(T), Vibrio sinaloensis LMG 25238(T) and Vibrio brasiliensis LMG 20546(T) varied between 97.1 and 98.5%. DDH experiments showed that the two isolates had less than 15% relatedness to the phylogenetically most closely related Vibrio species. R-40492(T) and R-40493(T) had 55-57% relatedness to each other. The ΔT(m) between R-40492(T) and R-40493(T) was 6.12 °C. In addition, MLSA of concatenated sequences (16S rRNA, ftsZ, gyrB, recA, rpoA, topA, pyrH and mreB; 6035 bp in length) showed that the two novel isolates formed a separate branch with less than 92% concatenated gene sequence similarity towards known species of vibrios. Two novel species are proposed to accommodate these novel isolates, namely Vibrio variabilis sp. nov. (type strain, R-40492(T)=LMG 25438(T)=CAIM 1454(T)) and Vibrio maritimus sp. nov. (type strain, R-40493(T)=LMG 25439(T)=CAIM 1455(T)).

A Gram-negative, aerobic bacterium, designated strain R-40503(T), was isolated from mucus of the reef-builder coral Mussismilia hispida, located in the São Sebastião Channel, São Paulo, Brazil. Phylogenetic analyses revealed that strain R-40503(T) belongs to the genus Marinomonas. The 16S rRNA gene sequence similarity of R-40503(T) was above 97 % with the type strains of Marinomonas vaga, M. basaltis, M. communis and M. pontica, and below 97 % with type strains of the other Marinomonas species. Strain R-40503(T) showed less than 35 % DNA-DNA hybridization (DDH) with the type strains of the phylogenetically closest Marinomonas species, demonstrating that it should be classified into a novel species. Amplified fragment length polymorphism (AFLP), chemotaxonomic and phenotypic analyses provided further evidence for the proposal of a novel species. Concurrently, a close genomic relationship between M. basaltis and M. communis was observed. The type strains of these two species showed 78 % DDH and 63 % AFLP pattern similarity. Their phenotypic features were very similar, and their DNA G+C contents were identical (46.3 mol%). Collectively, these data demonstrate unambiguously that Marinomonas basaltis is a later heterotypic synonym of Marinomonas communis. Several phenotypic features can be used to discriminate between Marinomonas species. The novel strain R-40503(T) is clearly distinguishable from its neighbours. For instance, it shows oxidase and urease activity, utilizes l-asparagine and has the fatty acid C(12 : 1) 3-OH but lacks C(10 : 0) and C(12 : 0). The name Marinomonas brasilensis sp. nov. is proposed, with the type strain R-40503(T) ( = R-278(T)  = LMG 25434(T)  = CAIM 1459(T)). The DNA G+C content of strain R-40503(T) is 46.5 mol%.

Six Gram-negative coccobacilli were isolated from Pacific oysters (Crassostrea gigas) from Mexico and haemolymph of spider crabs (Maja brachydactyla) from Spain. All of the isolates grew as small green colonies on thiosulphate-citrate-bile salts-sucrose (TCBS) agar and were facultatively anaerobic, oxidase-positive and sensitive to the vibriostatic agent O/129. Repetitive palindromic PCR analysis revealed a high degree of genomic homogeneity among the isolates. Several phenotypic traits differentiated the isolates from the type strains of species of the genus Photobacterium. DNA-DNA relatedness between two representative isolates and their closest phylogenetic neighbours by 16S rRNA gene sequence similarity, Photobacterium aplysiae CAIM 14(T) and Photobacterium frigidiphilum CAIM 20(T), was 44.01-53.85 %. We propose a novel species of the genus Photobacterium to accommodate the six isolates, with the name Photobacterium swingsii sp. nov. The type strain is CAIM 1393(T) (=CECT 7576(T)).

Eight Vibrio isolates originating from the marine corals Mussismilia hispida and Phyllogorgia dilatata and the zoanthids Palythoa caribaeorum and Palythoa variabilis in Brazil and the Pacific white shrimp (Litopenaeus vannamei) in Ecuador were studied by means of a polyphasic approach. The novel isolates formed a tight monophyletic group in the genus Vibrio and were closely related to species of the Vibrio harveyi group, to which they showed more than 99 % 16S rRNA gene sequence similarity. Analysis based on concatenated sequences of the following seven genes, 16S rRNA, gyrB, recA, rpoA, topA, pyrH and mreB (5633 bp in length), showed clear separation between the isolates and species of the V. harveyi group. Amplified fragment length polymorphism (AFLP) analysis, performed previously, revealed that a representative isolate of this group, LMG 20370, was clearly separate from known Vibrio species (it belonged to the so-called AFLP cluster A31). DNA-DNA hybridization (DDH) experiments with representative isolates and type strains of the V. harveyi species group revealed high DDH between the novel isolates (more than 74 %) and less than 70 % DDH towards type strains of related Vibrio species, proving the novel species status of the isolates. Phenotypically, the novel species belongs to the arginine dihydrolase (A)-negative, lysine decarboxylase (L)-positive and ornithine decarboxylase (O)-positive (A-/L+/O+) cluster reported previously. Most species of the V. harveyi group (i.e. Vibrio rotiferianus, V. harveyi, V. parahaemolyticus and V. alginolyticus) also belong to this A-/L+/O+ cluster. However, several phenotypic features can be used for the identification of the novel species. In contrast to its closest phylogenetic neighbours, the novel species exhibits esterase (C4) and N-acetyl-β-glucosaminidase activities, but it does not produce acetoin, does not use citrate, α-ketoglutaric acid or propionic acid and does not ferment melibiose. The novel species can also be differentiated on the basis of the presence of the fatty acids C(17 : 0,) C(17 : 1)ω8c, iso-C(17 : 0) and iso-C(13 : 0) and the absence of the fatty acid C(18 : 0). The name Vibrio communis sp. nov. is proposed for this taxon. Strain R-40496(T) (=LMG 25430(T) =CAIM 1816(T)) is the type strain.

A Gram-negative, aerobic bacterium, designated R-40509(T), was isolated from mucus of the reef builder coral (Mussismilia hispida) located in the São Sebastião Channel, São Paulo, Brazil. The strain was oxidase-positive and catalase-negative, and required Na(+) for growth. Its phylogenetic position was in the genus Marinobacterium and the closest related species were Marinobacterium sediminicola, Marinobacterium maritimum and Marinobacterium stanieri; the isolate exhibited 16S rRNA gene sequence similarities of 97.5-98.0 % with the type strains of these species. 16S rRNA gene sequence similarities with other type strains of the genus Marinobacterium were below 96 %. DNA-DNA hybridizations between strain R-40509(T) and the type strains of the phylogenetically closest species of the genus Marinobacterium revealed less than 70 % DNA-DNA relatedness, supporting the novel species status of the strain. Phenotypic characterization revealed that the strain was able to grow at 15-42 °C and in medium containing up to 9 % NaCl. The isolate could be differentiated from phenotypically related species by several features, including its ability to utilize d-alanine, l-alanine, bromosuccinic acid, β-hydroxybutyric acid and α-ketovaleric acid, but not acetate or l-arabinose. It produced acetoin (Voges-Proskauer), but did not have esterase lipase (C8) or catalase activities. It possessed C(18 : 1)ω7c (35 %), summed feature 3 (iso-C(15 : 0) 2-OH and/or C(16 : 1)ω7c; 25 %) and C(16 : 0) (22 %) as major cellular fatty acids. The DNA G+C content was 58.5 mol%. The name Marinobacterium coralli sp. nov. is proposed to accommodate this novel isolate; the type strain is R-40509(T) (=LMG 25435(T) =CAIM 1449(T)).

Four novel isolates (R-40508(T), R-40507, R-40903 and R-21419) were obtained from different cnidarian species (Phyllogorgia dilatata, Merulina ampliata and Palythoa caribaeorum) from different places in Brazil and Australia. The novel isolates formed a tight phylogenetic group based on 16S rRNA, recA, topA, ftsZ, mreB and rpoA gene sequences. Their closest phylogenetic neighbours were the type strains of Photobacterium leiognathi, P. rosenbergii and P. halotolerans, sharing 97.1-97.5 % 16S rRNA gene sequence similarity. DNA-DNA hybridization between a representative strain (R-40508(T)) and the type strains of these Photobacterium species revealed less than 20 % relatedness, showing that the new isolates belong to a novel species. Several phenotypic features allow the differentiation of the novel species from its closest phylogenetic neighbours. It has gelatinase and lipase activity and can utilize melibiose, but it cannot grow on 6 % NaCl. In addition, the novel species has the fatty acid iso-C(16 : 0), but lacks the fatty acids C(17 : 0), C(17 : 0) cyclo, iso-C(17 : 0), C(17 : 1)ω8c and iso-C(17 : 1)ω9c. The name Photobacterium jeanii sp. nov. is proposed for this species, with the type strain R-40508(T) (=LMG 25436(T) =CAIM 1817(T)). The G+C content of the type strain is 45.5mol%.

Aims: A taxonomic survey of the vibrios associated with the Brazilian endemic coral Mussismilia hispida and the sympatric zoanthids (i.e. Palythoa caribaeorum, Palythoa variabilis and Zoanthus solanderi)., Methods and Results: Mucus of 54 cnidarian specimens collected in three different places at São Sebastião in two consecutive years (i.e. 2005 and 2006) was used for taxonomic characterization of the cnidarian microbiota. Ninety-eight of the 151 vibrio isolates fell within the vibrio core group according to partial 16S rDNA sequences. We performed the sequencing of recA and pyrH genes of all vibrio isolates. The most abundant taxa belonged to the vibrio core group (Vibrio harveyi, Vibrio rotiferianus, Vibrio campbellii and Vibrio alginolyticus), Vibrio mediterranei (=Vibrio shillonii) and Vibrio chagasii. With the exception of V. chagasii which was found only in the mucus of M. hispida, the other species appeared in different hosts with no evidence for the presence of host-specific clones or species. Using rep-PCR analysis, we observed a high genomic heterogeneity within the vibrios. Each vibrio isolate generated a different rep-PCR fingerprint pattern. There was a complete agreement between the grouping based on rep-PCR and concatenated sequences of pyrH, recA and 16S rDNA, but the pyrH gene has the highest discriminatory power for vibrio species identification., Conclusion: The vibrio core group is dominant in the mucus of these cnidarians. There is a tremendous diversity of vibrio lineages within the coral mucus. pyrH gene sequences permit a clear-cut identification of vibrios., Significance and Impact of the Study: The taxonomic resolution provided by pyrH (but not recA) appears to be enough for identifying species of vibrios and for disclosing putative new taxa. The vibrio core group appears to be dominant in the mucus of the Brazilian cnidarians. The overrepresentation of these vibrios may reflect as yet unknown ecological functions in the coral holobiont.

Taxonomic characterization was performed on the putative N(2)-fixing microbiota associated with the coral species Mussismilia hispida, and with its sympatric species Palythoa caribaeorum, P. variabilis, and Zoanthus solanderi, off the coast of São Sebastião (São Paulo State, Brazil). The 95 isolates belonged to the Gammaproteobacteria according to the 16S rDNA gene sequences. In order to identify the isolates unambiguously, pyrH gene sequencing was carried out. The majority of the isolates (n = 76) fell within the Vibrio core group, with the highest gene sequence similarity being towards Vibrio harveyi and Vibrio alginolyticus. Nineteen representative isolates belonging to V. harveyi (n = 7), V. alginolyticus (n = 8), V. campbellii (n = 3), and V. parahaemolyticus (n = 1) were capable of growing six successive times in nitrogen-free medium and some of them showed strong nitrogenase activity by means of the acetylene reduction assay (ARA). It was concluded that nitrogen fixation is a common phenotypic trait among Vibrio species of the core group. The fact that different Vibrio species can fix N(2) might explain why they are so abundant in the mucus of different coral species.