Vibrio ulleungensis sp. nov., isolated from Mytilus coruscus.

A Gram-stain-negative, rod-shaped, motile via polar flagellum, facultatively aerobic, light-yellow, bacterium (designated 188UL20-2T) was isolated from a mussel sample of Mytilus coruscus collected on Ulleung Island, Ulleung-gun, Gyeongsangbuk-do, Republic of Korea. On the basis of 16S rRNA gene sequencing results, strain 188UL20-2T clustered with species of the genus Vibrio and appeared closely related to Vibrio marisflavi DSM 23086T (96.59%), Vibrio variabilis DSM 26147T (96.57%), Vibrio penaeicida DSM 14398T (96.37%) and Vibrio litoralis DSM 17657T (95.97%). The average nucleotide identity and digital DNA-DNA hybridization values between strain 188UL20-2T and its closest related strain were 71.3 and 16.4%, indicating that 188UL20-2T represents a novel species of the genus Vibrio. Growth occurred at 18-37 °C on MA medium in the presence of 1-4% NaCl (w/v) and at pH 5.0-10.0. The DNA G+C content of the genomic DNA was 45.4 mol%, and ubiquinone-8 (Q-8) was the major respiratory quinone. The major cellular fatty acids (>5%) were C16:1 ω6c and/or C16:1 ω7c (summed feature 3), C18:1 ω7c and/or C18:1 ω6c (summed feature 8), C16:0, C16:0 iso, C14:0, C14:0 iso and C12:0. The polar lipids consisted of phosphatidylglycerol, phosphatidylethanolamine, two unidentified phospholipids, unidentified aminophospholipid, unidentified glycolipid and seven unidentified lipids. Physiological and biochemical characteristics indicated that strain 188UL20-2T represents a novel species of the genus Vibrio, for which the name Vibrio ulleungensis sp. nov. is proposed. The type strain is 188UL20-2T (=KACC 22258T=LMG 32202T).

PCR, in-situ hybridization, and phylogenetic analysis suggest that 'big belly' disease in barramundi, Lates calcarifer (Bloch), is caused by a novel Vibrio species.

'Big belly' disease is a chronic, granulomatous bacterial enteritis and peritonitis, first reported in 3- to 4-week-old Asian seabass or barramundi, Lates calcarifer Bloch fry. Affected fry are emaciated and have a swollen abdomen, and the condition is referred to as 'skinny pot-belly' or 'big belly' disease. In this study, histopathological examinations of diseased fish from a batch of 2-month-old, 6- to 8-cm L. calcarifer fingerlings, kept in seawater recirculating aquaculture systems, showed pathology resembling 'big belly' disease. Ethanol-fixed tissues were tested positive using specific PCR primers based on 16S rRNA genes. In situ hybridization using dioxygenin-labelled positive PCR products on formalin-fixed paraffin-embedded tissues showed positive reactions with intralesional, clusters of the large, 'big belly' coccobacilli. A phylogenetic tree constructed based on analyses of these 16S rRNA gene PCR products from five positive fish suggests that the 'big belly' bacterium is most likely a novel Vibrio species.

Vibrio ziniensis sp. nov., isolated from mangrove sediments.

A novel Gram-staining-negative, catalase- and oxidase-positive, facultatively anaerobic and rod-shaped motile bacterial strain, designated as ZWAL4003T, was isolated from mangrove sediments of the Zini Mangrove Forest, Zhangzhou City, PR China. Phylogenetic analysis based on its 16S rRNA gene sequence indicated that ZWAL4003T was grouped into a separated branch with Vibrio plantisponsor MSSRF60T (97.38% nucleotide sequence identity) and Vibrio diazotrophicus NBRC 103148T (97.27%). The major cellular fatty acids were C14 : 0 (12.6%), C16 : 0 (17.6%), and summed feature 3 (C16 : 1ω6c /C16 : 1 ω7c, 45.6%). Its genome had a length of 4650556 bp with 42.8% DNA G+C content, and contained genes involved in the biosynthesis of bacteriocin, ß-lactone, resorcinol, N-acyl amino acid, and arylpolyene. The in silico DNA-DNA hybridization and average nucleotide identity values for whole-genome sequence comparisons between ZWAL4003T and V. plantisponsor LMG 24470T were clearly below the thresholds used for the delineation of a novel species. The morphological and chemotaxonomic characteristics and the genotypic data of ZWAL4003T indicated that it represented a novel species of the genus Vibrio. Its proposed name is Vibrio ziniensis sp. nov., and the type strain is ZWAL4003T (=KCTC 72971T=MCCC 1A17474T).

Vibrio agarilyticus sp. nov., an agar-digesting marine bacterium isolated from coastal seawater in Daya Bay (Guangdong, China).

A Gram-strain-negative, facultatively anaerobic, motile, rod-shaped and flagellated marine bacterium, designated SM6T, was isolated from surface seawater collected in Daya Bay (Guangdong, China). Phylogenetic analysis based on 16S rRNA gene sequences, multilocus sequence analysis, phylogenomic analysis of single-copy gene families and whole genome data showed that strain SM6T belonged to the genus Vibrio. The closest phylogenetic relatives of SM6T were Vibrio plantisponsor MSSRF60T (97.38 % 16S rRNA gene sequence pairwise similarity), Vibrio variabilis R-40492T (97.27 %), Vibrio aestuarianus ATCC 35048T (97.21 %) and Vibrio sagamiensis LC2-047T (97.3 %). Growth of strain SM6T occurred at 10-45 °C (optimum 30 °C), at pH 6.0-9.0 (optimum 6.0) and in the presence of 0-10 % (w/v) NaCl (optimum 3-8 %). The predominant fatty acids (>10 %) were summed feature 3 (C16 : 1 ω7c or/and C16 : 1 ω6c), C16 : 0 and summed feature 8 (C18 : 1 ω7c or/and C18 : 1 ω6c). The DNA G+C content of the assembled genomic sequences was 47.37 % for strain SM6T. Average nucleotide identity values between SM6T and its reference species were lower than the threshold for species delineation (95-96 %); in silico DNA-DNA hybridization further showed that the strains shared less than 70 % similarity. On the basis of evidence from the present polyphasic study, strain SM6T is considered to represent a novel species of the genus Vibrio, for which the name Vibrio agarilyticus sp. nov. is proposed. The type strain is SM6T (=KCTC 82076T=MCCC 1K04327 T).

Vibrio sp. ArtGut-C1, a polyhydroxybutyrate producer isolated from the gut of the aquaculture live diet Artemia (Crustacea)

BACKGROUND: Vibrio species display variable and plastic fitness strategies to survive and interact with multiple hosts, including marine aquaculture species that are severely affected by pathogenic Vibrios. The culturable Vibrio sp. strain ArtGut-C1, the focus of this study, provides new evidence of such phenotypic plasticity as it accumulates polyhydroxybutyrate (PHB), a biodegradable polymer with anti-pathogen activity, particularly in the marine larviculture phase. The strain was isolated from the gut of laboratory-reared Artemia individuals, the live diet and PHB carrier used in larviculture. Its main phenotypic properties, taxonomic status and genomic properties are reported based on the whole-genome sequencing. RESULTS: Vibrio sp. ArtGut-C1 yielded 72.6% PHB of cells' dry weight at 25 C. The genomic average nucleotide identity (ANI) shows it is closely related to V. diabolicus (ANI: 88.6%). Its genome contains 5,236,997- bp with 44.8% GC content, 3,710 protein-coding sequences, 96 RNA, 9 PHB genes functionally related to PHB metabolic pathways, and several genes linked to competing and colonizing abilities. CONCLUSIONS: This culturable PHB-accumulating Vibrio strain shows high genomic and phenotypic variability. It may be used as a natural pathogen biocontrol in the marine hatchery and as a potential cell factory for PHB production.

Antitumor activity of vibrio sp. isolated biopolymers in induced breast cancer in rats. / Actividad antitumoral de biopolímeros aislados de Vibrio sp. en cáncer de mama inducido en ratas.

OBJECTIVE: To evaluate the antitumor activity of the raw extract from biopolymers isolated from the Vibrio sp. marine bacteria in breast cancer induced by N-Methyl-N-nitrosourea (MNU) in rats. MATERIALS AND METHODS: The Vibrio sp. marine bacteria was cultured for seven days, then the raw supernatant was filtered, precipitated and concentrated. MNU was administered in a single dose of 50 mg/kg to 39 Holtzman rats and were daily treated for 9 weeks orally: G1 (n = 13): 0.1 mL/100 g of saline solution; G2 (n = 13): 20 mg/kg of raw extract from Vibrio sp. biopolymers; G3 (n = 13): 100 mg/kg of tamoxifen; G4 (n = 11) received no MNU and only 0.1 mL/100 g of saline solution. Body weight and the appearance of breast tumors identified by palpation were assessed weekly, as well as histopathological examination at the end of treatment. RESULTS: Seventy-seven percent of the rats in the G1 group developed tumors from week 7 onwards in an average of 2.2 tumors per animal; in contrast to the group treated with the raw biopolymer extract and tamoxifen; where only one rat (8%) in each group developed tumors after week nine of induction (p = 0.001). The histopathological results support that all the removed tumors correspond to breast ductal adenocarcinoma with different patterns: solid, papillary and cystic. Likewise, necrotic foci were evidenced in 30% of the tumors of the G1 group. CONCLUSION: The raw extract of biopolymers isolated from Vibrio sp. present antitumor effect in breast cancer induced in rats.

OBJETIVO: Evaluar la actividad antitumoral del extracto crudo de biopolímeros aislados de la bacteria marina Vibrio sp. en cáncer de mama inducido por N-Methyl-N-nitrosourea (MNU) en ratas. MATERIALES Y MÉTODOS: Se cultivó la bacteria marina Vibrio sp. durante siete días, luego se filtró, precipitó y concentró el sobrenadante crudo. Se administró una dosis única de MNU 50 mg/kg a 39 ratas Holtzman y fueron tratadas diariamente durante nueve semanas por vía oral: G1 (n = 13): suero fisiológico 0,1 mL/100 g; G2 (n = 13): extracto crudo de biopolímeros de Vibrio sp. 20 mg/kg; G3 (n = 13): tamoxifeno 100 mg/kg. El G4 (n = 11) solo recibió suero fisiológico 0,1 mL/100 g. Se valoró semanalmente el peso corporal y la aparición de tumores mamarios identificados mediante palpación; así como el examen histopatológico al final del tratamiento. RESULTADOS: El 77% de las ratas del grupo G1 desarrollaron tumores a partir de la séptima semana en un promedio de 2,2 tumores por cada animal; en contraste al grupo tratado con el extracto crudo de biopolímeros y tamoxifeno; donde solo una rata (8%) en cada grupo desarrolló tumores y posterior a la semana nueve de la inducción (p = 0,001). Los resultados histopatológicos sostienen que todos los tumores extirpados corresponden a adenocarcinoma ductal de mama con distintos patrones: sólido, papilar y quístico. Asimismo, se evidenciaron focos necróticos en el 30% de los tumores del grupo G1. CONCLUSIÓN: El extracto crudo de biopolímeros aislados de Vibrio sp. presentan efecto antitumoral en cáncer de mama inducido en ratas.

High diversity of Vibrio spp. associated with different ecological niches in a marine aquaria system and description of Vibrio aquimaris sp. nov.

The aim of the study was to characterise the diversity and niche-specific colonization of Vibrio spp. in a marine aquaria system by a cultivation-dependent approach. A total of 53 Vibrio spp. isolates were cultured from different ecological niches in a marine aquarium including microplastic (MP) and sandy sediment particles (12 weeks after added sterile to the system), detritus, and the surrounding aquarium water. Based on the 16S rRNA gene sequence phylogeny and multilocus sequence analysis (MLSA) the isolates were assigned to seven different phylotypes. Six phylotypes were identified by high probability to the species level. The highest phylotype diversity was cultured from detritus and water (six out of seven phylotypes), while only two phylotypes were cultured from MP and sediment particles. Genomic fingerprinting indicated an even higher genetic diversity of Vibrio spp. at the strain (genotype) level. Again, the highest diversity of genotypes was recovered from detritus and water while only few partially particle-type specific genotypes were cultured from MP and sediment particles. Phylotype V-2 formed an independent branch in the MLSA tree and could not be assigned to a described Vibrio species. Isolates of this phylotype showed highest 16S rRNA gene sequence similarity to type strains of Vibrio japonicus (98.5%) and Vibrio caribbeanicus (98.4%). A representative isolate, strain THAF100T, was characterised by a polyphasic taxonomic approach and Vibrio aquimaris sp. nov., with strain THAF100T (=DSM 109633T=LMG 31434T=CIP 111709T) as type strain, is proposed as novel species.

Vibrio algicola sp. nov., isolated from the surface of coralline algae.

A Gram-stain-negative, oxidase- and catalase-positive, facultative anaerobic and rod-shaped bacterium, designated strain SM1977T, was isolated from the surface of coralline algae collected from the intertidal zone at Qingdao, PR China. The strain grew at 10-35 °C, pH 4.5-8.5 and with 1-8.5% (w/v) NaCl. It reduced nitrate to nitrite and hydrolysed Tween 20 and DNA. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain SM1977T was affiliated with the genus Vibrio, having the highest sequence similarity (97.6 %) to the type strain of Vibrio casei, followed by those of another five species (95.6-97.6 %) in the Rumoiensis clade of the genus Vibrio. However, the in silico DNA-DNA hybridization (75.3-75.9 %) and average nucleotide identity (21.6-22.8 %) values of SM1977T against these close relatives were all below the corresponding thresholds to discriminate bacterial species. The major fatty acids were summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), C16:0 and summed feature 8 (C18:1 ω6c and /or C18:1 ω7c). The predominant polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol. The sole respiratory quinone was Q-8. The genomic DNA G+C content of strain SM1977T, determined from the obtained whole genomic sequence, was 42.3 mol%. On the basis of the polyphasic results obtained in this study, strain SM1977T is considered to represent a novel species within the genus Vibrio, for which the name Vibrio algicola sp. nov. is proposed. The type strain is SM1977T (=MCCC 1K04351T=KCTC 72847T).

Actividad antitumoral de biopolímeros aislados de Vibrio sp. en cáncer de mama inducido en ratas / Antitumor activity of vibrio sp. isolated biopolymers in induced breast cancer in rats

RESUMEN Objetivo: Evaluar la actividad antitumoral del extracto crudo de biopolímeros aislados de la bacteria marina Vibrio sp. en cáncer de mama inducido por N-Methyl-N-nitrosourea (MNU) en ratas. Materiales y métodos: Se cultivó la bacteria marina Vibrio sp. durante siete días, luego se filtró, precipitó y concentró el sobrenadante crudo. Se administró una dosis única de MNU 50 mg/kg a 39 ratas Holtzman y fueron tratadas diariamente durante nueve semanas por vía oral: G1 (n=13): suero fisiológico 0,1 mL/100g; G2 (n=13): extracto crudo de biopolímeros de Vibrio sp. 20 mg/kg; G3 (n=13): tamoxifeno 100 mg/kg. El G4 (n=11) solo recibió suero fisiológico 0,1 mL/100g. Se valoró semanalmente el peso corporal y la aparición de tumores mamarios identificados mediante palpación; así como el examen histopatológico al final del tratamiento. Resultados: El 77% de las ratas del grupo G1 desarrollaron tumores a partir de la séptima semana en un promedio de 2,2 tumores por cada animal; en contraste al grupo tratado con el extracto crudo de biopolímeros y tamoxifeno; donde solo una rata (8%) en cada grupo desarrolló tumores y posterior a la semana nueve de la inducción (p=0,001). Los resultados histopatológicos sostienen que todos los tumores extirpados corresponden a adenocarcinoma ductal de mama con distintos patrones: sólido, papilar y quístico. Asimismo, se evidenciaron focos necróticos en el 30% de los tumores del grupo G1. Conclusión: El extracto crudo de biopolímeros aislados de Vibrio sp. presentan efecto antitumoral en cáncer de mama inducido en ratas.

ABSTRACT Objective: To evaluate the antitumor activity of the raw extract from biopolymers isolated from the Vibrio sp. marine bacteria in breast cancer induced by N-Methyl-N-nitrosourea (MNU) in rats. Materials and methods: The Vibrio sp. marine bacteria was cultured for seven days, then the raw supernatant was filtered, precipitated and concentrated. MNU was administered in a single dose of 50 mg/kg to 39 Holtzman rats and were daily treated for 9 weeks orally: G1 (n = 13): 0.1 mL/100 g of saline solution; G2 (n = 13): 20 mg/kg of raw extract from Vibrio sp. biopolymers; G3 (n = 13): 100 mg/kg of tamoxifen; G4 (n = 11) received no MNU and only 0.1 mL/100 g of saline solution. Body weight and the appearance of breast tumors identified by palpation were assessed weekly, as well as histopathological examination at the end of treatment. Results: Seventy-seven percent of the rats in the G1 group developed tumors from week 7 onwards in an average of 2.2 tumors per animal; in contrast to the group treated with the raw biopolymer extract and tamoxifen; where only one rat (8%) in each group developed tumors after week nine of induction (p = 0.001). The histopathological results support that all the removed tumors correspond to breast ductal adenocarcinoma with different patterns: solid, papillary and cystic. Likewise, necrotic foci were evidenced in 30% of the tumors of the G1 group. Conclusion: The raw extract of biopolymers isolated from Vibrio sp. present antitumor effect in breast cancer induced in rats.

The strains of bioluminescent bacteria isolated from the White Sea finfishes: genera Photobacterium, Aliivibrio, Vibrio, Shewanella, and first luminous Kosakonia.

Bioluminescence is a spectacular feature of some prokaryotes. In the present work, we address the distribution of bioluminescence among bacteria isolated from the White Sea finfishes. Luminous bacteria are widely distributed throughout the World Ocean. Many strains have been isolated and described for tropical latitudes, while Nordic seas still remain quite a white spot in studying bioluminescence of bacteria. We describe the strains related to the two main genera of luminous bacteria, Photobacterium and Aliivibrio, as well as Shewanella and Vibrio. They are related to families Vibrionaceae and Shewanellaceae of the Gammaproteobacteria class. Here, we at the first time, report the bioluminescence of the Enterobacteriaceae Kosakonia cowanii. Moreover, we applied the polyphasic approach to identify and describe the isolated microorganisms. The data on sequencing, diversity of cell fine structure, and light emission spectra at room temperature on the solid medium are discussed. The bacteria are characterized by features in their light emission spectra. It may reflect possible molecular mechanisms of bioluminescence as well as features of bacterial composition. The obtained data expands the existing body of knowledge about the bioluminescence spread among the bacteria of Nordic latitudes and provides complex information that is crucial for their precise identification.

Genomic taxonomy of the Mediterranei clade of the genus Vibrio (Gammaproteobacteria).

The first genomic study of Mediterranei clade using five type strains (V. mediterranei, V. maritimus, V. variabilis, V. thalassae, and V. barjaei) and fourteen reference strains isolated from marine organisms, seawater, water and sediments of the sea was performed. These bacterial strains were characterised by means of a polyphasic approach comprising 16S rRNA gene, multilocus sequence analysis (MLSA) of 139 single-copy genes, the DNA G + C content, ANI, and in silico phenotypic characterisation. We found that the species of the Mediterranei clade formed two separate clusters based in 16S rRNA gene sequence similarity, MLSA, OrthoANI, and Codon and Amino Acid usage. The Mediterranei clade species showed values between 76 and 95% for ANIb, 84 and 95% for ANIm. The core genome consisted of 2057 gene families and the pan-genome of 13,094 gene families. Based on the genomic analyses performed, the Mediterranei clade can be divided in two clusters, one with the strains of V. maritimus, V. variabilis and two potential new species, and the other cluster with the strains of V. mediterranei, V. thalassae, and V. barjaei.

Vibrio sinensis sp. nov. and Vibrio viridaestus sp. nov., two marine bacteria isolated from the East China Sea.

Two Gram-staining-negative, facultatively anaerobic, motile, short clavate and flagellated marine bacteria, designated strains BEI233T and LJC006T, were isolated from the East China Sea. On the basis of the results of 16S rRNA gene sequencing and multilocus sequence analysis, BEI233T and LJC006T should be assigned to the genus Vibrio. The closest phylogenetic relatives of BEI233T are Vibrio scophthalmi LMG 19158T (98.7 % 16S rRNA gene sequence pairwise similarity), Vibrio ichthyoenteri DSM 14397T (98.5 %), Vibrio renipiscarius KCTC 42287T (97.7 %), Vibrio aestuarianus ATCC 35048T (97.3 %) and Vibrio thalassae MD16T (96.5 %), whereas for LJC006T they were Vibrio furnissii CAIM 518T (97.1 %), Vibrio fluvialis LMG 7894T (97.0%), Vibrio tritonius JCM 16456T (96.9 %) and Vibrio xuii LMG 21346T (96.1 %). The growth of BEI233T occurred at 10-37 °C, pH 5.0-8.0 and with 1-7 % (w/v) NaCl, while the growth of LJC006T occurred at 10-37 °C, pH 6.0-9.0, and 0-8 % (w/v) NaCl. The predominant fatty acids (>10 %) were summed feature 3 (C16 : 1ω7c or/and C16 : 1ω6c), C16 : 0 and summed feature 8 (C18 : 1ω7c or/and C18 : 1ω6c), with different proportions. The DNA G+C contents of BEI233T and LJC006T are 42.41 mol% and 41.88 mol%, respectively. On the basis of the results of polyphasic analysis, BEI233T and LJC006T are considered to represent novel species of the genus Vibrio, for which the names Vibrio sinensis sp. nov. and Vibrio viridaestus sp. nov. are proposed. The type strains are BEI233T (=JCM 32692T=KCTC 62618T) and LJC006T (=JCM 32693T=KCTC 62620T), respectively.

Vibrio chemaguriensis sp. nov., from Sundarbans, Bay of Bengal.

A new species of Vibrio (annotated as SBOTS_Iso1) was isolated in August 2014 from the Stn1 located in Chemaguri creek of Sundarbans mangrove ecoregion and taxonomically characterized using a polyphasic approach. Phenotypic analysis including biochemical tests and growth across a wide range of salinities indicated the typical estuarine characteristics of this new species. The bacterium was Gram negative, rod-shaped, oxidase and catalase negative and grows in the presence of NaCl. FAME analysis indicated 31.7% of the cellular fatty acids to be made up of 16:1 ω7c/16:1 ω6c. Amplification and sequencing of 16S rRNA and multilocus sequence analysis of four loci (2040 bp; rpoA, topA, mreB, pyrH) and additional sequence data of ftsZ, atpD, ompW and rpoB genes showed this isolate to be a member of Harveyi clade of the genus Vibrio. The closest phylogenetic neighbour was Vibrio alginolyticus ATCC 17749T with 96.8% similarity. Whole-genome sequence data indicates the presence of ~ 5 Mbp genome. GGDC, orthoANIu and AAI indicated 45%, 92% and 0.962 identity respectively with genome of Vibrio alginolyticus ATCC 17749T. The isolate SBOTS_Iso1 has been named Vibrio chemaguriensis sp. nov. on the name of the site from where it was first isolated.

Vibrio echinoideorum sp. nov., isolated from an epidermal lesion on the test of a green sea urchin (Strongylocentrotus droebachiensis).

A Gram-stain-negative, facultatively anaerobic Vibrio strain, designated NFH.MB010T, was isolated from an epidermal lesion on the test (hard shell skeleton) of a green sea urchin (Strongylocentrotus droebachiensis) collected from northern Norway. Cells of strain NFH.MB010T were rod shaped and motile by means of a single, long polar flagellum. Growth was observed at 1-5% NaCl (w/v) and at 4 °C, but not above 28 °C. Phylogenetic analyses based on eight-gene multilocus sequence analysis (16S rRNA, atpA, gyrB, mreB, pyrH, recA, rpoA and rpoD) suggested novelty at the species level. In silico DNA-DNA hybridization and orthologous average nucleotide identity estimates showed percentage genomic resemblances to its closest relative, Vibrio splendidus, that were well below the established same species threshold values. Phenotypically, utilization of glycogen and gentiobiose, inability of acetoin production, and undetectable valine arylamidase and trypsin activity discriminated strain NFH.MB010T from the closely related reference strains. Protein spectra generated by maldi-tof mass spectrometry further consolidated the species level uniqueness of strain NFH.MB010T. Based on the described polyphasic approach, strain NFH.MB010T therefore appears as a novel species within the Splendidus clade of the genus Vibrio, and the name Vibrio echinoideorum sp. nov. is proposed, with NFH.MB010T (=DSM 107264T=LMG 30656T) as the type strain.

Vibrio albus sp. nov., isolated from marine sediment.

A Gram-stain-negative, facultative anaerobic, motile, slightly bent rod-shaped bacterium, designated strain E4404T, was isolated from coastal sediment sampled Weihai, China. According to phylogenetic analysis based on its 16S rRNA gene sequence, strain E4404T shared the highest similarity with the type strain of Vibriovariabilis (97.0 %), followed by the type strains of Vibriomaritimus (96.1 %) and Vibrioeuropaeus (96.0 %). Multilocus sequence analysis based on the 16S rRNA gene and eight housekeeping genes (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA and topA) showed that strain E4404T formed a unique clade in the genus Vibrio. The average nucleotide identity and digital DNA-DNA hybridization values between strain E4404T and some other species in the genus Vibrio were 71.0-72.7 % and 20.4-22.4 %, respectively. Cells grew at 15-37 °C and pH 6.0-8.5 in the presence of 1.0-6.0 % (w/v) NaCl, with optimal growth at 33 °C and pH 7.0-7.5 in the presence of 3.0-3.5 % (w/v) NaCl. No growth was observed on thiosulfate citrate bile salts sucrose medium. The major fatty acids of strain E4404T were summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH), C16 : 0 and summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c). The genomic DNA G+C content of strain E4404T was 46.1 mol%. Based on the phylogenetic, genotypic, phenotypic, biochemical and chemotaxonomic characteristics, strain E4404T represented a novel species of the genus Vibrio, for which the name Vibrioalbus sp. nov. is proposed. The type strain is E4404T (=MCCC 1H00197T=KCTC 52890T).

Litopenaeus vannamei stylicins are constitutively produced by hemocytes and intestinal cells and are differentially modulated upon infections.

Stylicins are anionic antimicrobial host defense peptides (AAMPs) composed of a proline-rich N-terminal region and a C-terminal portion containing 13 conserved cysteine residues. Here, we have increased our knowledge about these unexplored crustacean AAMPs by the characterization of novel stylicin members in the most cultivated penaeid shrimp, Litopenaeus vannamei. We showed that the L. vannamei stylicin family is composed of two members (Lvan-Stylicin1 and Lvan-Stylicin2) encoded by different loci which vary in gene copy number. Unlike the other three gene-encoded antimicrobial peptide families from penaeid shrimp, the expression of Lvan-Stylicins is not restricted to hemocytes. Indeed, they are also produced by the columnar epithelial cells lining the midgut and its anterior caecum. Interestingly, Lvan-Stylicins are simultaneously transcribed at different transcriptional levels in a single shrimp and are differentially modulated in hemocytes after infections. While the expression of both genes showed to be responsive to damage-associated molecular patterns, only Lvan-Stylicin2 was induced after a Vibrio infection. Besides, Lvan-Stylicins also showed a distinct pattern of gene expression in the three portions of the midgut (anterior, middle and posterior) and during shrimp development. We provide here the first evidence of the diversity of the stylicin antimicrobial peptide family in terms of sequence and gene expression distribution and regulation.

Vibrio gangliei sp. nov., a novel member of Vibrionaceae isolated from sawdust in a pigpen.

A Gram-stain-negative, rod-shaped, catalase- and oxidase-positive, facultatively anaerobic, and motile bacterium, designated strain SZDIS-1T, was isolated from pigpen sawdust bedding in Xiamen, Fujian Province, China. Cells grew at 10-50 °C, pH 6.0-9.0, up to 12 % (w/v) NaCl, resisted vibriostatic agent O/129 and were negative for gelatin and alginate hydrolysis. No growth on thiosulfate citrate bile salts sucrose agar medium. Based on 16S rRNA gene sequences and multilocus sequence analysis, this strain should be assigned to the genus Vibrio, with the closest relatives being Vibrio aphrogenes CA-1004T (97.7 % 16S rRNA gene sequence pairwise similarity), Vibrio algivorus SA2T (96.6 %), Vibrio casei WS 4539T (96.3 %), Vibrio rumoiensis S-1T (96.1 %) and Vibrio litoralis MANO22DT (95.5 %), but separate from them by large distances in different phylogenetic trees. Based on whole genome analysis, the orthologous average nucleotide identity and in silico DNA-DNA hybridization values against the five relatives were 76.1-78.7 and 20.1-28.7 %. The major fatty acids were summed feature 3 (C16 : 1ω7c/C16 : 1ω6c), C16 : 0, summed feature 2 (one or more of C12 : 0 aldehyde, C14 : 0 3OH and/or iso-C16 : 1) and summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c). The DNA G+C content was 43.0 mol% from whole genomic sequence analysis. Therefore, phylogenetic, genotypic, phenotypic and chemotaxonomic characteristics showed that the isolate represented a novel species of the genus Vibrio, for which the name Vibrio gangliei sp. nov. is proposed. The type strain is SZDIS-1T (=DSM 104291T=CGMCC 1.15236T).

Vibrio coralliirubri sp. nov., a new species isolated from mucus of red coral (Corallium rubrum) collected at Procida island, Italy.

Strain Corallo1T was isolated from mucus of red coral (Corallium rubrum) at Punta Pizzaco (Procida island, Naples, Italy). It was characterised as a Gram-stain negative, motile, rod-shaped bacterium. Strain Corallo1T was found to show positive responses for cytochrome-c oxidase, catalase, reduction of nitrate and nitrite, ß-galactosidase activity and hydrolysis of starch, xylan, peptone, Tween 40, Tween 80 and casein. Strain Corallo1T was found to be mesophilic, neutrophilic to alkalophilic and slightly halophilic. According to analysis of the almost-complete 16S rRNA gene, strain Corallo1T is closely related to Vibrio celticus (100% sequence similarity), Vibrio gigantis (100%), Vibrio crassostreae (99.7%), Vibrio artabrorum (99.7%) and Vibrio pomeroyi (99.6%). MLSA of five housekeeping genes (atpA, pyrH, recA, rpoA and rpoD) was performed to refine the phylogenetic relationships of strain Corallo1T. A draft genome sequence of strain Corallo1T was obtained. The DNA G+C content of this strain was determined to be 44.5 mol %. The major cellular fatty acids of strain Corallo1T are C16:1, n-C16:0 and C18:1, and the major isoprenoid ubiquinone is Q8. ANI indexes, in silico estimations of DDH values and wet lab DDH values demonstrated that strain Corallo1T represents an independent genomospecies. Based on a polyphasic taxonomic characterisation, strain Corallo1T is concluded to represent a novel species of the genus Vibrio, for which the name Vibrio coralliirubri sp. nov. is proposed. The type strain is Corallo1T (= DSM 27495T = CIP 110630T).

Vibrio hannami sp. nov., Isolated from Seawater.

A Gram-reaction negative, aerobic, motile, non-pigmented and rod-shaped bacterium, designated as 168GH5-2-16T, was isolated from seawater Jeju island. Phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that the strain formed a distinct lineage within the genus Vibrio and was most closely related to Vibrio variabilis R-40492T (96.0%). The DNA G+C content was 49.3 mol%. The major polar lipids were phosphatidylethanolamine (PE) and phosphatidylglycerol (PG). The predominant quinone was ubiquinone-8 (Q-8). The major fatty acids were C16:0, summed feature 3 (comprising C16:1 ω7c/C16:1 ω6c) and summed feature 8 (C18:1 ω7c/C18:1 ω6c) supported the affiliation of 168GH5-2-16T to the genus Vibrio. Moreover, the physiological, biochemical, and taxonomic analysis allowed the phenotypic and genotypic differentiation of strain 168GH5-2-16T from the recognized species of the genus Vibrio. Therefore, strain 168GH5-2-16T represents a novel species of the genus Vibrio, for which the name Vibrio hannami sp. nov. is proposed, with the type strain 168GH5-2-16T (=KACC 19277T = DSM105032T).

Vibrio aphrogenes sp. nov., in the Rumoiensis clade isolated from a seaweed.

A novel strain Vibrio aphrogenes sp. nov. strain CA-1004T isolated from the surface of seaweed collected on the coast of Mie Prefecture in 1994 [1] was characterized using polyphasic taxonomy including multilocus sequence analysis (MLSA) and a genome based comparison. Both phylogenetic analyses on the basis of 16S rRNA gene sequences and MLSA based on eight protein-coding genes (gapA, gyrB, ftsZ, mreB, pyrH, recA, rpoA, and topA) showed the strain could be placed in the Rumoiensis clade in the genus Vibrio. Sequence similarities of the 16S rRNA gene and the multilocus genes against the Rumoiensis clade members, V. rumoiensis, V. algivorus, V. casei, and V. litoralis, were low enough to propose V. aphrogenes sp. nov. strain CA-1004T as a separate species. The experimental DNA-DNA hybridization data also revealed that the strain CA-1004T was separate from four known Rumoiensis clade species. The G+C content of the V. aphrogenes strain was determined as 42.1% based on the genome sequence. Major traits of the strain were non-motile, halophilic, fermentative, alginolytic, and gas production. A total of 27 traits (motility, growth temperature range, amylase, alginase and lipase productions, and assimilation of 19 carbon compounds) distinguished the strain from the other species in the Rumoiensis clade. The name V. aphrogenes sp. nov. is proposed for this species in the Rumoiensis clade, with CA-1004T as the type strain (JCM 31643T = DSM 103759T).