Direct Observation of Trace Elements in Barium Titanate of Multilayer Ceramic Capacitors Using Atom Probe Tomography.

Accurately controlling trace additives in dielectric barium titanate (BaTiO3) layers is important for optimizing the performance of multilayer ceramic capacitors (MLCCs). However, characterizing the spatial distribution and local concentration of the additives, which strongly influence the MLCC performance, poses a significant challenge. Atom probe tomography (APT) is an ideal technique for obtaining this information, but the extremely low electrical conductivity and piezoelectricity of BaTiO3 render its analysis with existing sample preparation approaches difficult. In this study, we developed a new APT sample preparation method involving W coating and heat treatment to investigate the trace additives in the BaTiO3 layer of MLCCs. This method enables determination of the local concentration and distribution of all trace elements in the BaTiO3 layer, including additives and undesired impurities. The developed method is expected to pave the way for the further optimization and advancement of MLCC technology.

Assessment of oral hypofunction and its association with age among Korean community-dwelling older adults.

BACKGROUND: Due to the increasing proportion of older adults in Korea and growing interest in aging, the concepts of oral aging and oral hypofunction have recently been introduced. Thus, it is necessary to investigate the age-specific oral function levels of Korean older adults and develop expert intervention methods for healthy aging. METHODS: Dysphagia, independence of daily living, and oral hypofunction were assessed in 206 older adults living in Wonju, Gangwon State, South Korea. Subjective dysphagia was assessed through self-report questionnaires using the Dysphagia Handicap Index (DHI), the Korean version of Eating Assessment Tool-10, and the Korean version of the Modified Barthel Index. In addition, the oral hypofunction assessment items included decreased chewing ability, occlusal pressure, tongue pressure, oral dryness, and oral cleanliness. RESULTS: DHI increased significantly with age, with those in their 80 s reporting the most difficulty swallowing. Oral function in terms of chewing ability (maximum occlusal pressure and number of remaining teeth), maximum occlusal pressure, and maximum tongue pressure also declined with increasing age. While there was no significant difference in oral dryness by age, those in their 80 s had dry mouth according to the criteria of the oral moisture checking device. CONCLUSIONS: In an assessment of oral function in community-dwelling, independent Korean older adults, the number of items that were assessed as oral hypofunction increased with age. The findings can be used to standardize the oral hypofunction assessment item and develop age-based individualized intervention plans for the early management of oral health and individual oral myofunctional rehabilitation in Korean community-dwelling older adults.

Validation of the Korean Academy of Geriatric Dentistry screening questionnaire and oral frailty diagnostic criteria in community-dwelling older adults.

OBJECTIVES: This study aimed to establish the validity-specifically, the sensitivity and specificity-of the screening questionnaire and diagnostic criteria for oral frailty proposed by the Korean Academy of Geriatric Dentistry (KAGD) among community-dwelling older adults. METHODS: This study enrolled 100 participants. Among various definitions of oral frailty, this study used the criteria proposed by Tanaka as the reference test. The screening questionnaire consisted of 11 items for screening physical frailty, chewing ability, swallowing difficulties, oral dryness, and tongue and lip motor function. Each question had a different scoring weight, and if the total score was 1 or higher, an oral frailty diagnostic examination proposed by the KAGD would be recommended. The diagnostic test was the oral frailty diagnostic criteria proposed by the KAGD including 6 measures: chewing ability, occlusal force, tongue pressure, oral dryness, swallowing difficulty, and oral hygiene. If a participant exhibited 2 or more positive measures, this participant was classified as "oral frail." The screening questionnaire was analyzed using a cut-off value of 1 or higher, while the diagnostic criteria utilized a cut-off of 2 or more positive measures. Sensitivity and specificity were calculated. RESULTS: The screening questionnaire showed significant power for screening oral frailty (area under the receiver operating characteristic curve, 0.783; sensitivity, 87.8%; specificity, 52.5%). The diagnostic accuracy of the newly proposed diagnostic criteria was acceptable (sensitivity, 95.1%; specificity, 42.4%). CONCLUSIONS: The newly proposed screening questionnaire and diagnostic criteria in Korea appear to be a useful tool to identify oral frailty in community-dwelling older adults.

The sesquiterpene lactone estafiatin exerts anti-inflammatory effects on macrophages and protects mice from sepsis induced by LPS and cecal ligation puncture.

BACKGROUND: Previously, we found that the water extract of Artermisia scoparia Waldst. & Kit suppressed the cytokine production of lipopolysaccharide (LPS)-stimulated macrophages and alleviated carrageenan-induced acute inflammation in mice. Artemisia contains various sesquiterpene lactones and most of them exert immunomodulatory activity. PURPOSE: In the present study, we investigated the immunomodulatory effect of estafiatin (EST), a sesquiterpene lactone derived from A. scoparia, on LPS-induced inflammation in macrophages and mouse sepsis model. STUDY DESIGN AND METHODS: Murine bone marrow-derived macrophages (BMDMs) and THP-1 cells, a human monocytic leukemia cell line, were pretreated with different doses of EST for 2 h, followed by LPS treatment. The gene and protein expression of pro-inflammatory cytokines interleukin (IL)-6, tumor necrosis factor (TNF)-α, and inducible nitric oxide synthase (iNOS) were measured by quantitative real-time polymerase chain reaction (qPCR) and Western blot analysis. The activation of nuclear factor kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) was also evaluated at the level of phosphorylation. The effect of EST on inflammatory cytokine production, lung histopathology, and survival rate was assessed in an LPS-induced mice model of septic shock. The effect of EST on the production of cytokines in LPS-stimulated peritoneal macrophages was evaluated by in vitro and ex vivo experiments and protective effect of EST on cecal ligation and puncture (CLP) mice was also assessed. RESULTS: The LPS-induced expression of IL-6, TNF-α, and iNOS was suppressed at the mRNA and protein levels in BMDMs and THP-1 cells, respectively, by pretreatment with EST. The half-maximal inhibitory concentration (IC50) of EST on IL-6 and TNF-α production were determined as 3.2 µM and 3.1 µM in BMDMs, 3 µM and 3.4 µM in THP1 cells, respectively. In addition, pretreatment with EST significantly reduced the LPS-induced phosphorylation p65, p38, JNK, and ERK in both cell types. In the LPS-induced mice model of septic shock, serum levels of IL-6, TNF-α, IL-1ß, CXCL1, and CXCL2 were lower in EST-treated mice than in the control animals. Histopathology analysis revealed that EST treatment ameliorated LPS-induced lung damage. Moreover, while 1 of 7 control mice given lethal dose of LPS survived, 3 of 7 EST-treated (1.25 mg/kg) mice and 5 of 7 EST-treated (2.5 mg/kg) mice were survived. Pretreatment of EST dose-dependently suppressed the LPS-induced production of IL-6, TNF-α and CXCL1 in peritoneal macrophages. In CLP-induced mice sepsis model, while all 6 control mice was dead at 48 h, 1 of 6 EST-treated (1.25 mg/kg) mice and 3 of 6 EST-treated (2.5 mg/kg) mice survived for 96 h. CONCLUSION: These results demonstrated that EST exerts anti-inflammatory effects on LPS-stimulated macrophages and protects mice from sepsis. Our study suggests that EST could be developed as a new therapeutic agent for sepsis and various inflammatory diseases.

Higgs inflation is still alive after the results from BICEP2.

The observed value of the Higgs boson mass indicates that the Higgs potential becomes small and flat at the scale around 10(17) GeV. Having this fact in mind, we reconsider the Higgs inflation scenario proposed by Bezrukov and Shaposhnikov. It turns out that the nonminimal coupling ξ of the Higgs squared to the Ricci scalar can be smaller than 10. For example, ξ=7 corresponds to the tensor-to-scalar ratio r≃0.2, which is consistent with the recent observation by BICEP2.

Terriglobus aquaticus sp. nov., isolated from an artificial reservoir.

A pink-pigmented, chemo-organotrophic bacterium, designated strain 03SUJ4(T), was isolated from the freshwater of Juam reservoir, Republic of Korea (35° 03' 43'' N 127° 14' 15'' E). Cells were aerobic, Gram-reaction-negative and non-motile rods. Strain 03SUJ4(T) grew at pH 6-7 (optimum, pH 6) and at 15-30 °C (optimum, 25 °C). Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolate belonged to the genus Terriglobus, showing sequence similarities of 97.09 % and 96.82 % to Terriglobus roseus DSM 18391(T) and Terriglobus saanensis SP1PR4(T), respectively. Low rpoB gene sequence similarity with members of the genus Terriglobus and different fingerprints with the repetitive primers BOX, ERIC and REP indicated that the isolate represented a novel species of the genus Terriglobus. The major cellular fatty acids were iso-C15 : 0, C16 : 0, C20 : 1ω9c, C14 : 0 and summed feature 3 (C16 : 1ω7c/C16 : 1ω6c). The DNA G+C content of strain 03SUJ4(T) was 63.2±0.1 mol% (mean±sd of three determinations). The predominant menaquinone was MK-8. The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and two unidentified phospholipids. Several phenotypic characteristics served to differentiate the novel isolate from recognized members of the genus Terriglobus. On the basis of the evidence presented in this study, a novel species, Terriglobus aquaticus sp. nov. is proposed for strain 03SUJ4(T) ( = KCTC 23332(T) = JCM 17517(T)).

Algibacter agarivorans sp. nov. and Algibacter agarilyticus sp. nov., isolated from seawater, reclassification of Marinivirga aestuarii as Algibacter aestuarii comb. nov. and emended description of the genus Algibacter.

Two yellow-pigmented, rod-shaped, non-motile, Gram-reaction-negative and aerobic bacterial strains, designated KYW560(T) and KYW563(T), were isolated from seawater collected from Gwangyang Bay, Republic of Korea. The isolates required sea salts for growth. Flexirubin-type pigments were absent. The common major cellular fatty acids (>5% of total) of the two strains were C(16:0), C(18:0), iso-C(15:0), anteiso-C(15:0), iso-C(15:1) G, iso-C(17:0) 3-OH and summed feature 3 (C(16:1)ω6c and/or C(16:1)ω7c). Strain KYW560(T) also contained iso-C(15:0) 3-OH and C(20:1)ω9c as major fatty acids. The main polar lipids were phosphatidylethanolamine, an unidentified aminolipid and two unidentified lipids. The predominant isoprenoid quinone was MK-6. The DNA G+C contents of strains KYW560(T) and KYW563(T) were 41.0 ± 0.7 and 38.3 ± 0.4 mol% (mean ± sd of three determinations), respectively. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolates belonged to the family Flavobacteriaceae, and were related to the genus Algibacter. Based on data from this taxonomic study using a polyphasic approach, it is proposed that the isolates represent novel species of the genus Algibacter, for which the names Algibacter agarivorans sp. nov. (type strain, KYW560(T) =KCTC 23855(T) =JCM 18285(T)) and Algibacter agarilyticus sp. nov. (type strain, KYW563(T) =KCTC 23857(T) =JCM 18275(T)) are proposed. Reclassification of Marinivirga aestuarii as Algibacter aestuarii comb. nov. and emended description of the genus Algibacter are also proposed.

Lutibacter agarilyticus sp. nov., a marine bacterium isolated from shallow coastal seawater.

A Gram-staining-negative, non-spore-forming, non-gliding and rod-shaped bacterial strain KYW566(T), was isolated from seawater of the Suncheon Bay, Korea, and its taxonomic position was investigated by using a polyphasic study. The cells contained MK-6 as the only respiratory quinone and contained iso-C15 : 0 (13.8 %), iso-C16 : 0 3-OH (13.1 %), anteiso-C15 : 0 (9.3 %), iso-C15 : 0 3-OH (8.7 %), iso-C15 : 1 G (6.3 %) and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) (6.0 %) as the major fatty acids. The DNA G+C content of strain KYW566(T) was 41.6 ± 0.8 mol% (mean ± sd of three determinations). A phylogenetic tree based on 16S rRNA gene sequences showed that strain KYW566(T) forms an evolutionary lineage within the radiation enclosing the members of the genus Lutibacter with Lutibacter flavus IMCC1507(T) as its nearest neighbour (96.7 % sequence similarity). A number of phenotypic characteristics distinguished strain KYW566(T) from described members of the genus Lutibacter. On the basis of the evidences presented in this study, strain KYW566(T) represents a novel species, for which the name Lutibacter agarilyticus sp. nov. is proposed. The type strain is KYW566(T) ( = KCTC 23842(T) = JCM 18281(T)).

Algibacter aquimarinus sp. nov., isolated from a marine environment, and reclassification of Pontirhabdus pectinivorans as Algibacter pectinivorans comb. nov.

An orange-coloured, rod-shaped, Gram-reaction-negative and aerobic bacterial strain, designated KYW589(T), was isolated from seawater collected from Gwangyang Bay, Republic of Korea. The isolate required sea salts for growth. Gliding motility was observed. Flexirubin-type pigments were absent. Major cellular fatty acids (>10% of the total) were iso-C15:0, iso-C17:0 3-OH, iso-C15:1 G, C16:0, iso-C15:0 3-OH and C18:0. The main polar lipids were phosphatidylethanolamine, two unidentified aminolipids and two unidentified lipids. The predominant isoprenoid quinone was MK-6. The DNA G+C content was 38.6 ± 0.7 mol% (mean ± sd of three determinations). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain KYW589(T) belongs to the family Flavobacteriaceae, and was related to the genus Algibacter. Based on data from a study using a polyphasic taxonomic approach, it is proposed that strain KYW589(T) represents a novel species belonging to the genus Algibacter, for which the name Algibacter aquimarinus sp. nov. is proposed. The type strain is KYW589(T) (=KCTC 23928(T) =JCM 18287(T)). Reclassification of Pontirhabdus pectinivorans Yi et al. 2011 to the genus Algibacter, as Algibacter pectinivorans comb. nov. (type strain JC2675(T) =KACC 14153(T) =JCM 17107(T)), is also proposed.

Sphingopyxis rigui sp. nov. and Sphingopyxis wooponensis sp. nov., isolated from wetland freshwater, and emended description of the genus Sphingopyxis.

Two yellow-pigmented, Gram-reaction-negative strains, designated 01SU5-P(T) and 03SU3-P(T), were isolated from the freshwater of Woopo wetland, Republic of Korea. Both strains were aerobic, non-motile and catalase-negative. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the two isolates belong to the genus Sphingopyxis, showing the highest level of sequence similarity with respect to Sphingopyxis witflariensis W-50(T) (95.4-95.7 %). The two novel isolates shared 99.4 % sequence similarity. DNA-DNA hybridization between the isolates and the type strain of S. witflariensis clearly suggested that strains 01SU5-P(T) and 03SU3-P(T) represent two separate novel species in the genus Sphingopyxis. The two strains displayed different fingerprints after PCR analysis using the repetitive primers BOX, ERIC and REP. Several phenotypic characteristics served to differentiate these two isolates from recognized members of the genus Sphingopyxis. The data from the polyphasic study presented here indicated that strains 01SU5-P(T) and 03SU3-P(T) should be classified as representing novel species in the genus Sphingopyxis, for which the names Sphingopyxis rigui sp. nov. and Sphingopyxis wooponensis sp. nov., respectively, are proposed. The type strain of Sphingopyxis rigui sp. nov. is 01SU5-P(T) ( = KCTC 23326(T) = JCM 17509(T)) and the type strain of Sphingopyxis wooponensis sp. nov. is 03SU3-P(T) ( = KCTC 23340(T) = JCM 17547(T)).

Aquimarina gracilis sp. nov., isolated from the gut microflora of a mussel, Mytilus coruscus, and emended description of Aquimarina spongiae.

An orange-coloured and slender rod-shaped bacterium, designated strain PSC32(T), was isolated from the gut microflora of a mussel collected from Gwangyang Bay, South Sea (Republic of Korea). Cells were Gram-reaction-negative, strictly aerobic, and catalase- and oxidase-positive. The major fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH, iso-C15 : 1 G and iso-C15 : 0 3-OH. The only isoprenoid quinone of strain PSC32(T) was MK-6 and the DNA G+C content was 36.9 mol%. Phosphatidylethanolamine, one unidentified aminolipid and two unidentified polar lipids were found as major polar lipids. A phylogenetic tree based on 16S rRNA gene sequences showed that strain PSC32(T) forms an evolutionary lineage within the genus Aquimarina and is closely related to Aquimarina spongiae A6(T) (97.0 % similarity) and to other members of the genus Aquimarina (94.4-96.5 %). Genomic DNA-DNA relatedness between strain PSC32(T) and A. spongiae A6(T) was 40.7 %. A number of phenotypic characteristics distinguished strain PSC32(T) from described members of the genus Aquimarina. On the basis of the evidence presented in this study, strain PSC32(T) represents a novel species, for which the name Aquimarina gracilis sp. nov. is proposed. The type strain is PSC32(T) ( = KCTC 23301(T) = JCM 17453(T)). An emended description of Aquimarina spongiae is given.

Marinivirga aestuarii gen. nov., sp. nov., a member of the family Flavobacteriaceae, isolated from marine environments, and emended descriptions of the genera Hyunsoonleella, Jejuia and Pontirhabdus and the species Hyunsoonleella jejuensis, Jejuia pallidilutea and Pontirhabdus pectinivorans.

Two orange, rod-shaped, Gram-reaction-negative, aerobic bacterial strains devoid of flagella and gliding motility, designated strains KYW371(T) and KS18 were isolated from a seawater sample and a shellfish Ruditapes philippinarum, respectively, collected from Gwangyang Bay, Republic of Korea. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the two strains belonged to the family Flavobacteriaceae; and that strain KYW371(T) was most closely related to Algibacter mikhailovii LMG 23988(T) (96.7 % sequence similarity), Pontirhabdus pectinivorans JC2675(T) (96.3 %), Postechiella marina M091(T) (95.6 %) and Hyunsoonleella jejuensis CNU004(T) (95.3 %). The 16S rRNA gene sequence similarity (98.8 %) and DNA-DNA relatedness (78.1 %) between strains KYW371(T) and KS18 indicated that these two strains represented a single species. The predominant cellular fatty acids of strain KYW371(T) were iso-C15 : 1 G, iso-C15 : 0, iso-C15 : 0 3-OH and iso-C17 : 0 3-OH. Flexirubin-type pigments were absent. MK-6 was the only isoprenoid quinone and the DNA G+C content was 34.8-36.6 mol%. Data from this taxonomic study employing a polyphasic approach suggested that the isolates represent a novel species in a new genus in the family Flavobacteriaceae, for which the name Marinivirga aestuarii gen. nov., sp. nov. is proposed. The type strain is KYW371(T) ( = KCTC 23449(T) = JCM 17452(T)), and an additional strain of the species is KS18 ( = KCTC 23128 = JCM 16845). Emended descriptions of the genera Hyunsoonleella, Jejuia and Pontirhabdus and the species Hyunsoonleella jejuensis, Jejuia pallidilutea and Pontirhabdus pectinivorans are also proposed.

Roseomonas riguiloci sp. nov., isolated from wetland freshwater.

A non-motile, coccobacillus-shaped and pink pigmented bacterium, designated strain 03SU10-P(T), was isolated from wetland freshwater (Woopo wetland, Republic of Korea). Cells were Gram reaction-negative and catalase- and oxidase-positive. The major fatty acids (>10% of total) were C(18:1)ω7c and summed feature 3 (iso-C(15:0) 2-OH and/or C(16:1)ω7c). The predominant respiratory lipoquinone was Q-10. The DNA G+C content was 68 mol%. The major polar lipids were phosphatidylethanolamine, phosphatidylcholine and an unknown aminolipid. Spermidine, putrescine and 1,3-diaminopropane were the major polyamines. A phylogenetic tree based on 16S rRNA gene sequence comparisons showed that strain 03SU10-P(T) formed an evolutionary lineage within the radiation enclosing the members of the genus Roseomonas. The nearest neighbour to the novel strain was Roseomonas stagni HS-69(T) (96.3% gene sequence similarity). The evidence provided by the polyphasic taxonomic approach used in this study indicated that strain 03SU10-P(T) could not be assigned to any recognized species; therefore a novel species is proposed, Roseomonas riguiloci sp. nov., with 03SU10-P(T) ( = KCTC 23339(T) = JCM 17520(T)) as the type strain.

Reclassification of Bacillus beijingensis Qiu et al. 2009 and Bacillus ginsengi Qiu et al. 2009 as Bhargavaea beijingensis comb. nov. and Bhargavaea ginsengi comb. nov. and emended description of the genus Bhargavaea.

We have carried out a polyphasic taxonomic characterization of Bacillus beijingensis DSM 19037(T) and Bacillus ginsengi DSM 19038(T), which are closely related phylogenetically to Bhargavaea cecembensis LMG 24411(T). All three strains are Gram-stain-positive, non-motile, moderately halotolerant and non-spore-forming. 16S rRNA gene sequence analyses showed that the strains constituted a coherent cluster, with sequence similarities between 99.7 and 98.7 %. The percentage similarity on the basis of amino acid sequences deduced from partial gyrB gene nucleotide sequences of these three type strains was 96.1-92.7 %. Phylogenetic trees based on the 16S rRNA gene and GyrB amino acid sequences, obtained by using three different algorithms, were consistent and showed that these three species constituted a deeply rooted cluster separated from the clades represented by the genera Bacillus, Planococcus, Planomicrobium, Sporosarcina, Lysinibacillus, Viridibacillus, Kurthia and Geobacillus, supporting their placement in the genus Bhargavaea. All three type strains have menaquinone MK-8 as the major respiratory quinone and showed similar fatty acid profiles. The main polar lipids present in the three type strains were diphosphatidylglycerol and phosphatidylglycerol, and the three strains showed peptidoglycan type A4α with L-lysine as the diagnostic diamino acid. The DNA G+C contents of Bacillus beijingensis DSM 19037(T), Bacillus ginsengi DSM 19038(T) and Bhargavaea cecembensis LMG 24411(T) were 53.1, 50.2 and 53.7 mol%, respectively. The level of DNA-DNA hybridization among the three strains was 57-39 %, indicating that they are members of different species of the genus Bhargavaea. The phenotypic data are consistent with the placement of these three species in a single genus and support their differentiation at the species level. On the basis of these data, we have emended the description of the genus Bhargavaea and propose the reclassification of Bacillus beijingensis and Bacillus ginsengi to the genus Bhargavaea, as Bhargavaea beijingensis comb. nov. (type strain ge10(T)  = DSM 19037(T)  = CGMCC 1.6762(T)) and Bhargavaea ginsengi comb. nov. (type strain ge14(T)  = DSM 19038(T)  = CGMCC 1.6763(T)).

Characterization of alginate lyase gene using a metagenomic library constructed from the gut microflora of abalone.

A metagenomic fosmid library was constructed using a genomic DNA mixture extracted from the gut microflora of abalone. The library gave an alginate lyase positive clone (AlyDW) harboring a 31.7-kbp insert. The AlyDW insert consisted of 22 open reading frames (ORFs). The deduced amino acid sequences of ORFs 11-13 were similar to those of known alginate lyase genes, which are found adjacent in the genome of Klebsiella pneumoniae subsp. aerogenes, Vibrio splendidus, and Vibrio sp. belonging to the phylum Gammaproteobacteria. Among the three recombinant proteins expressed from the three ORFs, alginate lyase activity was only observed in the recombinant protein (AlyDW11) coded by ORF 11. The expressed protein (AlyDW11) had the highest alginate lyase activity at pH 7.0 and 45°C in the presence of 1 mM AgNO(3). The alginate lyase activity of ORF 11 was confirmed to be endolytic by thin-layer chromatography. AlyDW11 preferred poly(ß-D: -mannuronate) as a substrate over poly(α-L: -guluronate). AlyDW11 contained three highly conserved regions, RSEL, QIH, and YFKAGVYNQ, which may act to stabilize the three-dimensional conformation and function of the alginate lyase.

Aquimarina mytili sp. nov., isolated from the gut microflora of a mussel, Mytilus coruscus, and emended description of Aquimarina macrocephali.

An orange, rod-shaped, gliding bacterium, designated strain PSC33(T), was isolated from the gut microflora of a mussel collected from Gwangyang Bay, South Sea (Republic of Korea). Cells were Gram-reaction-negative, strictly aerobic, and catalase- and oxidase-positive. The major fatty acids were iso-C(15:0), iso-C(17:0) 3-OH, iso-C(15:1) G, C(15:0) 3-OH and iso-C(17:1)ω9c. The only isoprenoid quinone was menaquinone-6 (MK-6). The DNA G+C content of strain PSC33(T) was 37.9 mol%. A phylogenetic tree based on 16S rRNA gene sequences showed that strain PSC33(T) formed an evolutionary lineage within the radiation encompassing members of the genus Aquimarina with Aquimarina macrocephali JAMB N27(T) as its nearest neighbour (96.3% sequence similarity). A number of phenotypic characteristics distinguished strain PSC33(T) from recognized members of the genus Aquimarina. On the basis of the data presented in this study, strain PSC33(T) is considered to represent a novel species of the genus Aquimarina, for which the name Aquimarina mytili sp. nov. is proposed. The type strain is PSC33(T) ( = KCTC 23302(T) = JCM 17454(T)). An emended description of A. macrocephali is also provided.

Gaetbulibacter aestuarii sp. nov., isolated from shallow coastal seawater, and emended description of the genus Gaetbulibacter.

A rod-shaped, yellow and strictly aerobic marine bacterium, designated KYW382(T), was isolated from seawater collected from the South Sea, Republic of Korea. Cells were Gram-negative and catalase- and oxidase-positive. The major fatty acids were iso-C(15:1) G, iso-C(15:0), iso-C(17:0) 3-OH, iso-C(15:0) 3-OH and anteiso-C(15:0). The DNA G+C content was 32.4 mol%. A phylogenetic tree based on 16S rRNA gene sequences showed that strain KYW382(T) constituted an evolutionary lineage within the radiation enclosing the members of the genus Gaetbulibacter. The closest neighbour was Gaetbulibacter saemankumensis SMK-12(T) (96.1% 16S rRNA gene sequence similarity). A number of phenotypic characteristics distinguished strain KYW382(T) from the described members of the genus Gaetbulibacter. On the basis of the data presented in this study, strain KYW382(T) represents a novel species, for which the name Gaetbulibacter aestuarii sp. nov. is proposed. The type strain is KYW382(T) (=KCTC 23303(T) =JCM 17455(T)). An emended description of the genus Gaetbulibacter is also given.

Screening and characterization of a cellulase gene from the gut microflora of abalone using metagenomic library.

A metagenomic fosmid library was constructed using genomic DNA isolated from abalone intestine. Screening of a library of 3,840 clones revealed a 36 kb insert of a cellulase positive clone (pAMHElO). A shotgun clone library was constructed using the positive clone (pAMHElO) and further screening of 3,840 shotgun clones with an approximately 5 kb insert size using a Congo red overlay revealed only one cellulase positive clone (pAMHL9). The pAMHL9 consisted of a 5,293-bp DNA sequence and three open reading frames (ORFs). Among the three ORFs, cellulase activity was only shown in the recombinant protein (CelAMll) coded by ORF3, which showed 100% identity with outer membrane protein A from Vibrio alginolyticus 12G01, but no significant sequence homology to known cellulases. The expressed protein (CelAMll) has a molecular weight of approximately 37 kDa and the highest CMC hydrolysis activity was observed at pH 7.0 and 37°C. The carboxymethyl cellulase activity was determined by zymogram active staining and different degraded product profiles for CelAMll were obtained when cellotetraose and cellopentaose were used as the substrates, while no substrate hydrolysis was observed on oligosaccharides such as cellobiose and cellotriose.

Paenibacillus wooponensis sp. nov., isolated from wetland freshwater.

A rod-shaped, endospore-forming, Gram-reaction-positive bacterium, designated strain WPCB018(T), was isolated from a fresh water sample collected from Woopo wetland, Korea. The isolate was identified as a member of the genus Paenibacillus on the basis of phenotypic characteristics and phylogenetic inference based on 16S rRNA gene sequence analysis. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and unknown aminophospholipids. The diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. The predominant menaquinone was MK-7. The major cellular fatty acids were anteiso-C(15 : 0) (32.2 %), C(16 : 0) (20.1 %) and C(18 : 0) (18.1 %). The DNA G+C content was 56.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain WPCB018(T) belongs to a cluster comprising species of the genus Paenibacillus, its closest neighbours being Paenibacillus humicus PC-147(T) (97.5 %) and Paenibcillus pasadenensis SAFN-007(T) (96.2 %). Genomic DNA-DNA hybridizations performed with strain WPCB018(T) and type strains of the species P. humicus, P. pinihumi, P. phyllosphaerae, P. pasadenensis and P. tarimensis showed relatedness values of only 10, 17, 18, 19 and 20 %, respectively. On the basis of phenotypic, molecular and genetic evidence, strain WPCB018(T) represents a novel species of the genus Paenibacillus, for which the name Paenibacillus wooponensis sp. nov. is proposed. The type strain of the novel species is WPCB018(T) ( = KCTC 13280(T) = JCM 16350(T)).

Altererythrobacter namhicola sp. nov. and Altererythrobacter aestuarii sp. nov., isolated from seawater.

Two non-motile, orange- or yellow-pigmented bacteria, designated strains KYW48(T) and KYW147(T), were isolated from seawater collected from the South Sea, Republic of Korea. Cells of both strains were Gram-reaction-negative, aerobic and catalase- and oxidase-positive. The major fatty acids of strain KYW48(T) were C(18 : 1)ω7c (35.3 %), summed feature 3 (iso-C(15 : 0) 2-OH and/or C(16 : 1)ω7c) (22.7 %), C(17 : 1)ω6c (19.8 %), C(14 : 0) 2-OH (7.4 %) and C(16 : 0) (5.9 %), and those of strain KYW147(T) were C(18 : 1)ω7c (36.0 %), summed feature 3 (18.3 %), C(16 : 0) (14.7 %), 11-methyl C(18 : 1)ω7c (10.7 %), C(16 : 0) 2-OH (9.1 %) and C(18 : 1)ω9c (8.0 %). The predominant isoprenoid quinone of both strains was ubiquinone 10 (Q-10). The DNA G+C contents of strains KYW48(T) and KYW147(T) were 63.8 and 67.2 mol%, respectively. A phylogenetic tree based on 16S rRNA gene sequences showed that strains KYW48(T) and KYW147(T) were grouped with the members of the family Erythrobacteraceae and formed a distinct clade with the members of the genus Altererythrobacter (<95.7 % sequence similarity). On the basis of the evidence presented in this study, the novel species Altererythrobacter namhicola sp. nov. (type strain KYW48(T)  = KCTC 22736(T)  = JCM 16345(T)) and Altererythrobacter aestuarii sp. nov. (type strain KYW147(T)  = KCTC 22735(T)  = JCM 16339(T)) are proposed.