Streptococcus taonis sp. nov., a novel bacterial species isolated from a blood culture of a patient.

One Gram stain-positive, catalase-negative, α-hemolytic, chain-forming or paired cocci, designated ST22-14T, was isolated from a blood culture of a child with suspected infection. The results of 16S rRNA gene sequences analyses showed that the most closely related species to strain ST22-14T were "Streptococcus vulneris" DM3B3T (99.2%), Streptococcus mitis NCTC 12261T (99.0%), "Streptococcus gwangjuense" ChDC B345T, (99.0%), Streptococcus oralis subsp. dentisani 7747T (99.0%), Streptococcus downii CECT 9732T (99.0%), and Streptococcus infantis ATCC 700779T (98.9%). The genome of strain ST22-14T consists of 2,053,261 bp with a G + C content of 39.4%. Average nucleotide identity values between strain ST22-14T and Streptococcus mitis NCTC 12261T or other five species were from 82.2 to 88.0%. In silico DNA-DNA hybridization of ST22-14T showed an estimated DNA reassociation value of 34.6% with the closest species. The main cellular fatty acids of strain ST22-14T were 16:0, 18:0, 14:0, 18:1ω7c and 18:1ω6c. Based on these results, strain ST22-14T should be classified as a novel species of genus Streptococcus, for which the name Streptococcus taonis sp. nov. is proposed (type strain ST22-14T = NBRC 116002T = BCRC 81402T).

Streptococcus vulneris sp. nov., isolated from wound of patient with diabetic foot ulcer (DFU).

A new α-haemolytic streptococcal strain, designated DM3B3T, was isolated from the wound of a diabetic foot ulcer (DFU) patient. Phylogenetic analysis based on 16S rRNA full-gene sequencing (1563 bp) revealed highest sequence similarity to Streptococcus mitis (99.7%), followed by "Streptococcus gwangjuense" (99.6%), and Streptococcus pseudopneumoniae (99.5%). Comparison of five housekeeping genes, groEL, rpoB, sodA, recA and pheS, revealed that strain DM3B3T was well separated from the Streptococcus reference strains. The complete genome of strain DM3B3T consisted of 1,963,039 bp with a G + C content of 41.0 mol%. Average nucleotide identity values between strain DM3B3T and Streptococcus mitis NCTC 12261T, "Streptococcus gwangjuense" ChDC B345T, and Streptococcus pseudopneumoniae ATCC BAA-960T were 93.8%, 94.4%, and 92.2%, respectively. The highest digital DNA-DNA hybridization value with respect to the closest species was 57.5%, i.e., below the species cut-off of 70% hybridization. The main cellular fatty acids of strain DM3B3T were 16:0, 18:1ω7c, 18:1ω9c and 18:0. On the basis of phylogenetic, genotypic and phenotypic data, we propose to classify this isolate as representative of a novel species of the genus Streptococcus, Streptococcus vulneris sp. nov., in reference to its isolation from wound, with strain DM3B3T (= NBRC 114638T = BCRC 81288T) as the type strain.

Enterococcus alishanensis sp. nov., a novel lactic acid bacterium isolated from fresh coffee beans.

A coccus-shaped organism, designated ALS3T, was isolated from fresh coffee cherries collected at a farm located in the Ali Mountain region of Taiwan. Sequence analysis of its 16S rRNA gene indicated that strain ALS3T belongs to the genus Enterococcus and has more than 98.5 % sequence similarity to Enterococcus pallens and Enterococcus hermanniensis. When comparing the ALS3T genome with these two type strains, the average nucleotide identity values and digital DNA-DNA hybridization values were 72.6-73.3 and 19.2 %, respectively. The G+C content of the genomic DNA from strain ALS3T was 35.6 mol%. Results of sequence analysis, together with enzymatic activities and characteristics of carbohydrate metabolism, indicated that strain ALS3T is distinct and represents a novel species, for which the name Enterococcus alishanensis sp. nov. is proposed. The type strain is ALS3T (=NBRC 109593T=BCRC 80605T).

Fructobacillus apis sp. nov., isolated from the gut of honeybee (Apis mellifera).

A Gram-positive, facultatively anaerobic, catalase-negative, fructose-dependent strain (W13T) was isolated from the gut of honeybee (Apis mellifera). Phylogenetic analysis based on 16S rRNA gene sequencing indicated that strain W13T represents a distinct line of descent within the genus Fructobacillus, with the closest neighbours being Fructobacillus broussonetiae BCRC 81240T (98.9 % sequence similarity) and Fructobacillus durionis DSM 19113T (96.8 % sequence similarity). Comparative sequencing of the additional phylogenetic markers rpoC and recA confirmed the 16S rRNA gene tree topology. The complete genome of strain W13T consisted of 1 292 712 bp with a G+C content of 48.3 mol%. Pairwise comparisons of the average nucleotide identity values and digital DNA-DNA hybridization values between the genomes of W13T and its close phylogenetic neighbours, F. broussonetiae BCRC 81240T and F. durionis DSM 19113T, resulted in 76.2-84.1 % and 20.2-27.6 %, respectively. The main cellular fatty acids of strain W13T were C16 : 0, C18 : 1 ω9c and C18 : 1 ω7c. Thus, we propose a novel species within the genus Fructobacillus, with the name Fructobacillus apis sp. nov. and the type strain is W13T (= NBRC 115637T=BCRC 81365T).

Streptococcus vaginalis sp. nov., a novel bacterial species isolated from vaginal swabs of a pregnant woman with diabetes.

Sequences targeted at the V3 and V4 16S rRNA hypervariable regions of a streptococcal strain (P1L01T) isolated from vaginal swabs of a pregnant woman with diabetes were 100% similar to those of Streptococcus anginosus subsp. whileyi. However, phylogenetic analysis based on 16S rRNA full-gene sequencing (1562 bp) revealed highest sequence similarity to Streptococcus periodonticum (98.7%), followed by Streptococcus anginosus subsp. whileyi (98.7%), and Streptococcus anginosus subsp. anginosus (98.4%). Phylogenies of housekeeping genes rpoB and groEL were compared to improve classification, and the results showed a clear separation between strain P1L01T and closely related Streptococcus type strains. The complete genome of strain P1L01T consisted of 2,108,769 bp with a G + C content of 38.5 mol%. Average nucleotide identity values, based on genome sequencing, between strain P1L01T and Streptococcus periodonticum KCOM 2412T, Streptococcus anginosus subsp. whileyi CCUG 39159T, and Streptococcus anginosus subsp. anginosus NCTC 10713T were 95.5%, 94.3%, and 95.3%, respectively. The highest in silico DNA-DNA hybridization value with respect to the closest species was 66.2%, i.e., below the species cutoff of 70% hybridization. The main cellular fatty acids of strain P1L01T were 16:0, 18:1ω7c, and 14:0. On the basis of phylogenetic, genotypic and phenotypic data, we propose to classify this isolate as representative of a novel species of the genus Streptococcus, Streptococcus vaginalis sp. nov., in reference to its isolation from vaginal swabs, with strain P1L01T (= NBRC 114754T = BCRC 81289T) as the type strain.

Leuconostoc litchii sp. nov., a novel lactic acid bacterium isolated from lychee.

A novel lactic acid bacterium, strain MB7T, was isolated from lychee in Taiwan. MB7T is Gram-staining-positive, catalase-negative, non-motile, non-haemolytic, facultatively anaerobic, coccoid-shaped, heterofermentative and mainly produces d-lactic acid from glucose. Comparative analysis of 16S rRNA, pheS and rpoA gene sequences has demonstrated that the novel strain represented a member of the genus Leuconostoc. 16S rRNA gene sequencing results indicated that MB7T had the same sequence similarity of 99.25 % to four type strains of members of the genus Leuconostoc: Leuconostoc mesenteroides subsp. dextranicum DSM 20484T, Leuconostoc mesenteroides subsp. jonggajibkimchii DRC 1506T, Leuconostoc mesenteroides subsp. mesenteroides ATCC 8293T and Leuconostoc suionicum DSM 20241T. Additionally, high 16S rRNA sequence similarities were also observed with Leuconostoc mesenteroides subsp. cremoris ATCC 19254T (99.12 %) and Leuconostoc pseudomesenteroides NRIC 1777T (98.69 %). When comparing the genomes of these type strains, the average nucleotide identity values and digital DNA-DNA hybridization values of MB7T with these type strains were 76.57-80.53 and 22.0-22.6 %, respectively. MB7T also showed different phenotypic characteristics to other most closely related species of the genus Leuconostoc, such as carbohydrate metabolizing ability, halotolerance and growth at various pHs. On the basis of phenotypic and genotypic properties, strain MB7T represents a novel species belonging to the genus Leuconostoc, for which the name Leuconostoc litchii sp. nov. is proposed. The type strain is MB7T (=BCRC 81077T=NBRC 113542T).

Lactobacillus musae sp. nov., a novel lactic acid bacterium isolated from banana fruits.

Two Gram-stain-positive, catalase-negative, rod-shaped, bacterial strains (313T and 311) were isolated from banana fruits in Taiwan. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the highest similarity to both strains corresponded to the type strain of Lactobacillus nantensis (99.19 %), followed by Lactobacillus crustorum (98.99 %), Lactobacillus heilongjiangensis (98.59 %) and Lactobacillus farciminis (98.52 %). Phylogenetic analysis based on the sequences of two housekeeping genes, pheS and rpoA, revealed that these two strains were well separated from the Lactobacillus reference strains. DNA-DNA relatedness values revealed genotype separation of the two strains from the above four species. The DNA G+C content of strain 313T was 35.5 mol%. The strains were homofermentative and mainly produced l-lactic acid from glucose. The major cellular fatty acids of strain 313T were 18 : 1ω6c and/or 18 : 1ω7c, 16 : 0, and 19 : 1ω6c and/or 19 : 0 cyclo ω10c. Based on their physiological and genotypic characteristics, the isolates represent a novel species of the genus Lactobacillus, for which the name Lactobacillusmusae sp. nov. is proposed. The type strain is 313T=NBRC 112868T=BCRC 81020T).

Comparative genomic analysis of bacteriocin-producing Weissella cibaria 110.

Weissella cibaria 110 was isolated from plaa-som, a Thai fermented fish product, and known to produce the weissellicin 110 bacteriocin. We carried out comprehensive comparative genomic analysis of W. cibaria 110 with four other non-bacteriocin-producing W. cibaria strains and identified potential antibiotic-resistant genes. We further identified a type III restriction-modification system, a TA system, and a bacteriocin gene cluster that are unique in W. cibaria 110. Genes related to bacteriocin biosynthesis are organized in clusters and are encoded with minimum genetic machinery consisting of structural cognate immunity genes, including ABC transporter and immunity protein. Finally, we predicted W. cibaria 110 to produce a class IId bacteriocin, weissellicin 110, which is 31 amino acids in length and contains a 21-amino-acid N-terminal leader peptide. This is the first bacteriocin-producing sequencing genome in W. cibaria, and we describe the difference between the bacteriocin-producing and non bacteriocin-producing strains from genome point of view.

Diversity of Lactic Acid Bacteria Associated with Banana Fruits in Taiwan.

Banana is a popular fruit worldwide. The lactic acid bacteria (LAB) microflora in banana fruits has not been studied in detail. A total of 164 LAB were isolated from banana fruits in Taiwan. These isolates were initially divided into nine groups (r1 to r9) using restriction fragment length polymorphism analysis and 16S ribosomal DNA sequencing. Isolates belonging to Lactobacillus plantarum group were further divided into three additional groups using multiplex PCR assay targeting the recA gene. The most common bacterial genera found in banana fruits were Lactobacillus and Weissella. The distribution of LAB indicated that, in most cases, neighboring regions shared common strains, but there were still some differences between regions. On the basis of phylogenetic analysis of 16S rRNA, rpoA, and pheS gene sequences, two strains included in the genera Lactobacillus were identified as potential novel species or subspecies. In addition, a total 36 isolates were found to have bacteriocin-producing abilities. These results suggest that various LAB are associated with banana fruits in Taiwan. This is the first report describing the distribution and varieties of LAB associated with banana fruits. In addition, one potential novel LAB species was also found in this study.

Secretion of one adipokine Nampt/Visfatin suppresses the inflammatory stress-induced NF-κB activity and affects Nampt-dependent cell viability in Huh-7 cells.

Nampt/visfatin acts in both intracellular and extracellular compartments to regulate multiple biological roles, including NAD metabolism, cancer, inflammation, and senescence. However, its function in chronic inflammation and carcinogenesis in hepatocellular carcinoma (HCC) has not been well-defined. Here we use Huh-7 hepatoma cells as a model to determine how Nampt/visfatin affects cellular survival under oxidative stress. We found that the transition of Nampt/visfatin from intracellular into extracellular form was induced by H2O2 treatment in 293T cells and confirmed that this phenomenon was not due to cell death but through the secretion of Nampt/visfatin. In addition, Nampt/visfatin suppressed cell viability in oxidative treatment in Huh-7 cells and acted on the inhibition of hepatoma cell growth. Oxidative stress also reduced the Nampt-mediated activation of NF-κB gene expression. In this study, we identify a novel feature of Nampt/visfatin which functions as an adipokine that can be secreted upon cellular stress. Our results provide an example to understand how adipokine interacts with chemotherapeutic treatment by oxidative stress in HCC.

Lactococcus formosensis sp. nov., a lactic acid bacterium isolated from yan-tsai-shin (fermented broccoli stems).

A coccal-shaped organism, designated 516(T), was isolated from yan-tsai-shin (fermented broccoli stems), a traditional fermented food in Taiwan. 16S rRNA gene sequencing results showed that strain 516(T) had 98.9 % sequence similarity to that of the type strain Lactococcus garvieae NBRC 100934(T). Comparison of three housekeeping genes, rpoA, rpoB and pheS, revealed that strain 516(T) was well separated from Lactococcus garvieae NBRC 100934(T). DNA-DNA hybridization studies indicated that strain 516(T) had low DNA relatedness with Lactococcus garvieae NBRC 100934(T) (46.1 %). The DNA G+C content of strain 516(T) was 38.1 mol% and the major fatty acids were C16 : 0 (22.7 %), C19 : 0 cyclo ω8c (17.9 %) and summed feature 7 (29.0 %). Based on the evidence, strain 516(T) represents a novel species of the genus Lactococcus, for which the name Lactococcus formosensis sp. nov. is proposed. The type strain is 516(T) ( = NBRC 109475(T) = BCRC 80576(T)).

Diversity of lactic acid bacteria associated with fresh coffee cherries in Taiwan.

A total of 102 lactic acid bacteria (LAB) were isolated from three different coffee farms in Taiwan. These isolates were classified and identified by the restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Heterofermentative Leuconostoc, and Weissella species were the most common LAB found in two farms located at an approximate altitude of 800 m. Lactococcus lactis subsp. lactis was the most common LAB found in the remaining farm was located at an approximate altitude of 1,200 m. It is therefore suggested that the altitude and climate may affect the distribution of LAB. On the basis of phylogenetic analysis, two strains included in the genera Enterococcus were considered as two potential novel species or subspecies. In addition, a total of 34 isolates showed the antifungal activity against Aspergillus flavus. Moreover, seven Lactococcus lactis subsp. lactis strains and one Enterococcus faecalis strain were found to have bacteriocin-like inhibitory substance-producing capability. These results suggest that various LAB are associated with fresh coffee cherries in Taiwan. Some of the isolates found in this study showed potential as antifungal agents.

Enterocin T, a novel class IIa bacteriocin produced by Enterococcus sp. 812.

Enterococcus sp. 812, isolated from fresh broccoli, was previously found to produce a bacteriocin active against a number of Gram-positive bacteria, including Listeria monocytogenes. Bacteriocin activity decreased slightly after autoclaving (121 °C for 15 min), but was inactivated by protease K. Mass spectrometry analysis revealed the bacteriocin mass to be approximately 4,521.34 Da. N-terminal amino acid sequencing yielded a partial sequence, NH2-ATYYGNGVYXDKKKXWVEWGQA, by Edman degradation, which contained the consensus class IIa bacteriocin motif YGNGV in the N-terminal region. The obtained partial sequence showed high homology with some enterococcal bacteriocins; however, no identical peptide or protein was found. This peptide was therefore considered to be a novel bacteriocin produced by Enterococcus sp. 812 and was termed enterocin T.

Lactococcus taiwanensis sp. nov., a lactic acid bacterium isolated from fresh cummingcordia.

One coccal strain, designated 0905C15(T), was isolated from fresh cummingcordia, which is the main ingredient of pobuzihi (fermented cummingcordia), a traditional fermented food in Taiwan. 16S rRNA gene sequencing results showed that strain 0905C15(T) had 98.22-98.82 % sequence similarity to that of the type strains of four Lactococcus lactis subspecies (L. lactis subsp. lactis BCRC 12312(T), L. lactis subsp. cremoris BCRC 12586(T), L. lactis subsp. hordniae BCRC 80474(T) and L. lactis subsp. tructae BCRC 80475(T)). Comparison of two housekeeping genes, recA and rpoB, revealed that strain 0905C15(T) was well separated from the reference strains of the genus Lactococcus. DNA-DNA hybridization studies indicated that strain 0905C15(T) had low DNA relatedness to the four Lactococcus lactis subspecies (9.7-15.24 %). The DNA G+C content of strain 0905C15(T) was 39.6 mol %. Based on the evidence, strain 0905C15(T) represents a novel species of the genus Lactococcus, for which the name Lactococcus taiwanensis sp. nov. is proposed. The type strain is 0905C15(T) ( = NBRC 109049(T) = BCRC 80460(T)).

Enterococcus saccharolyticus subsp. taiwanensis subsp. nov., isolated from broccoli.

A coccal strain isolated from fresh broccoli was initially identified as Enterococcus saccharolyticus; however, molecular identification and phenotypic traits did not support this identification. DNA-DNA hybridization with the type strain of E. saccharolyticus (76.4 % relatedness), DNA G+C content (35.7 mol%), phylogenetic analysis based on 16S rRNA, pheS and rpoA gene sequences, rep-PCR fingerprinting and profiles of cellular fatty acids, whole-cell proteins and enzyme activities, together with carbohydrate metabolism characteristics, indicated that this strain is distinct and represents a novel subspecies, for which the name Enterococcus saccharolyticus subsp. taiwanensis subsp. nov. is proposed. The type strain is 812(T) ( = NBRC 109476(T) = BCRC 80575(T)). Furthermore, we present an emended description of Enterococcus saccharolyticus and proposal of Enterococcus saccharolyticus subsp. saccharolyticus subsp. nov. (type strain ATCC 43076(T) = CCUG 27643(T) = CCUG 33311(T) = CIP 103246(T) = DSM 20726(T) = JCM 8734(T) = LMG 11427(T) = NBRC 100493(T) = NCIMB 702594(T)).

Surgical Management for Complications during Closure of Atrial Septal Defect with Amplatzer Device.

BACKGROUND: Transcatheter closure of secundum type atrial septal defect (ASD) has become a standard procedure in most medical centers. Although the procedure is invasive and has a shorter recovery duration and a lower complication rate compared with surgery, it is not risk-free. In this retrospective chart review case series, we report our experience of management of complications after placement of an ASD occluder. METHODS: Between January 2000 and December 2010, a total of 508 patients in our hospital underwent closure of secundum type ASD using an Amplatzer septal occluder (ASO). Six of the patients (1.2%) had device embolization or migration warranting surgical retrieval and repair. RESULTS: All the devices were removed and the defects were repaired successfully without any mortality. CONCLUSIONS: Surgical intervention for complications of ASO placement in patients who underwent closure of secundum type ASD is safe and effective. KEY WORDS: Amplatzer occluder; Atrial septal defect; Surgical management.

Diversity of lactic acid bacteria in sian-sianzih (fermented clams), a traditional fermented food in Taiwan.

BACKGROUND: Sian-sianzih (fermented clams) is a popular traditional fermented food in Taiwan. The lactic acid bacteria (LAB) microflora in sian-sianzih have not been studied in detail. In this study, LAB from sian-sianzih were isolated, characterized and identified. RESULTS: A total of 186 cultures of LAB were isolated from seven sian-sianzih samples and 29 cultures were isolated from its main raw substrate: clams. The identification results revealed up to 11 distinct bacterial species belonging to five genera in sian-sianzih, and three species belonging to two genera in clams. The most common bacterial genera in sian-sianzih were Lactobacillus and Weissella, followed by Leuconostoc, Pediococcus and Lactococcus. A regional similarity in LAB, with differences in diversity, was observed in the current study. On the other hand, Lactococcus lactis subsp. lactis was the most common species found in raw clam samples. The results also suggested that greater LAB diversity could be observed in wild clams than in cultured ones. Furthermore, antibacterial activities of the isolates were determined, and one Weisella hellenica strain showed inhibitory activity against the indicator strain Lactobacilluas sakei JCM 1157(T) . A sensory assessment of seven sian-sianzih samples was also performed and the results indicated that diversity of LAB has a great effect on its aroma and taste formation. CONCLUSION: The results demonstrate that various LAB species are distributed in sian-sianzih and have a great effect on the flavor of sian-sianzih.

Isolation and characterisation of lactic acid bacteria from jiang-gua (fermented cucumbers), a traditional fermented food in Taiwan.

BACKGROUND: Jiang-gua (fermented cucumbers) is a popular traditional fermented food in Taiwan. The microflora of lactic acid bacteria (LAB) in jiang-gua have not been investigated in detail. In this study, LAB from jiang-gua were isolated, characterised and identified. RESULTS: A total of 103 LAB were isolated; 70 cultures were isolated from jiang-gua samples and 33 cultures were isolated from its raw substrate, cucumber. These isolates were mainly characterised phenotypically and then divided into seven groups (A-G) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. The isolates were identified as Enterococcus casseliflavus, Leuconostoc lactis, Leuconostoc mesenteroides, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paraplantarum, Lactococcus lactis subsp. lactis, Weissella cibaria and Weissella hellenica. The antibacterial activities of the isolates were determined and 11 Lc. lactis subsp. lactis strains showed inhibitory activity against the indicator strain Lactobacillus sakei JCM 1157(T) . CONCLUSION: Heterofermentative W. cibaria and Leu. lactis were the major LAB found in jiang-gua samples without soy sauce. In soy sauce-added samples, homofermentative L. pentosus and L. plantarum were the most abundant LAB. In addition, the results also suggested that HhaI and RsaI restriction enzymes could be applied to distinguish W. hellenica and Weissella paramesenteroides.

Isoelectric focusing management: an investigation for salt interference and an algorithm for optimization.

Two-dimensional electrophoresis (2-DE) has evolved into a robust separation technique in proteomic research. However, one of the major challenges in 2-DE experiments, the reproducibility of the first dimensional electrophoresis (IEF), has remained unsolved. It is well-known that the quality of IEF experiments is significantly affected by the salt interference. Nevertheless, the interference mechanisms of salts in IEF have never been systematically investigated. In this study, we comprehensively investigated the interference effects in IEF due to various kinds of simple and buffer salts in protein samples. Two interference schemes were proposed accordingly to elucidate the interference mechanisms of salts in IEF. Furthermore, to increase the reproducibility of IEF, we proposed that conductivity measurement is a feasible method to assess the salt content of 2-DE samples and developed an algorithm to predict the optimal total volt-hours (Vh) required for protein focusing in IEF. The developed algorithm had been evaluated under various IEF conditions for a variety of 2-DE samples and proven to be a reliable guide. In sum, information disclosed in this study should be of use for increasing the reproducibility and thus the applicability of 2-DE in current proteomics.

A red line not to cross: evaluating the limitation and properness of gel image tuning procedures.

Currently, results of gel electrophoresis are commonly documented in digital formats by image acquisition instruments. In this study, gel images tuned by a common image processing software package, Photoshop, were assessed to understand the transforming algorithms and their impacts on quantitative analysis. TotalLab 100, an electrophoresis gel image analysis software package, was applied for image quantitation and evaluation. The three most frequently used image tuning functions-adjustments of the brightness, contrast, and grayscale span (level) of images-were investigated using both data generated from a standard grayscale tablet and an actual electrophoresis gel image. The influences of these procedures were analyzed for the grayscale transformation between the input and output images. Although all three procedures differentially improved the visualization of the input image, adjusting the contrast of images disrupted the quantitative information because of its nonlinear transforming algorithm. Under certain conditions, adjusting the brightness or the level of images could preserve the quantitative information because of the linear transforming algorithms. It was found that when the minimum and maximum grayscales of a gel image were recognized, using a commercial software package to maximally stretch the level may significantly improve the quality of a gel image without jeopardizing quantitative analysis.