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        Characterization of antibiotic-resistant, coagulase-negative staphylococci from fresh produce and description of Staphylococcus shinii sp. nov. isolated from chives

        Cho Gyu-Sung,Li Bo,Brinks Erik,Franz Charles M. A. P. 한국미생물학회 2022 The journal of microbiology Vol.60 No.9

        Coagulase-negative Staphylococcus (CoNS) species may possess antibiotic resistance genes and have been associated with nosocomial infections. In this study, 91 CoNS with decreased susceptibility to oxacillin were isolated from fresh produce using oxacillin containing agar plates. Their antibiotic resistances were determined phenotypically and all isolates were identified by rep-PCR, 16S rRNA and rpoB gene sequencing. Furthermore, the genomes of representative strains were sequenced in order to confirm species identification by phylogenomics. The majority (64 of 91) of the CoNS strains could be identified as Mammaliicoccus (M.) fleurettii, while 13 were identified as M. sciuri, 8 as M. vitulinus, 2 as Staphylococcus (S.) epidermidis and single strains each as S. warneri, S. xylosus, Staphylococcus spp. and S. casei. Most of the strains were generally susceptible to clinically-relevant antibiotics, but only few (< 7%) strains possessed multiple resistances. Both oxacillin and cefoxitin resistant isolates were considered to be presumptive methicillin-resistant CoNS. From whole genome sequencing data of 6 representative strains, the mecA gene, accessory genes and the SCC loci were compared, which revealed high variability between some of the strains. The major fatty acids of K22-5MT strain included anteiso-C15:0, iso-C15:0, iso-C17:0, anteiso-C17:0, C18:0, and C20:0. Average nucleotide identity and digital DNA-DNA hybridization values indicated that Staphylococcus strain K22-5MT was below the species delineation cutoff values for ANI (less than 91%) and DDH (less than 44.4%), with the most closely related species being the S. pseudoxylosus S04009T type strain. Thus, strain K22- 5MT (=DSM 112532T, =LMG 32324T) represents a novel species, for which the name Staphylococcus shinii sp. nov. is proposed.

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        Comparative Study of Fecal Microbiota in Patients with Type II Diabetes after Consumption of Apple Juice for 4 Weeks

        조규성,Anja König,Stephanie Seifert,Alexander Hanak,Alexander Roth,Melanie Huch,Achim Bub,Bernhard Watzl,Charles M.A.P. Franz 한국식품과학회 2015 Food Science and Biotechnology Vol.24 No.6

        The effect of cloudy apple juice on fecal microbiota of type 2 diabetics was studied. Five volunteers consumed apple juice while 5 control volunteers received an isocaloric control beverage daily for 4 weeks. DGGE profile analysis showed high diversity between volunteers that did not change over the intervention period using primers for Firmicutes, Bacteroidetes, bifidobacteria, enterococci, and enterobacteria. An exception was observed using lactobacilli primers, perhaps as the result of the dietary influence. Consumption of apple juice was not correlated with changes in DGGE profiles. Quantitative PCR was used to investigate the effect of apple juice on bacterial counts in different subgroups. Apple juice did not lead to significantly (p>0.05) different numbers of total bacteria, enterobacteria, bifidobacteria, lactobacilli, or Bacteroidetes, but caused a significant (p<0.05) decrease in numbers of enterococci, and a smaller but also significant decrease in numbers of Firmicutes, when comparing before and after intervention with apple juice.

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        Development of a Quantitative PCR for Detection of Lactobacillus plantarum Starters During Wine Malolactic Fermentation

        ( Gyu Sung Cho ),( Sabrina Krauß ),( Melanie Huch ),( Maret Du Toit ),( Charles M. A. P. Franz ) 한국미생물 · 생명공학회 2011 Journal of microbiology and biotechnology Vol.21 No.12

        A quantitative, real-time PCR method was developed to enumerate Lactobacillus plantarum IWBT B 188 during the malolactic fermentation (MLF) in Grauburgunder wine. The qRT-PCR was strain-specific, as it was based on primers targeting a plasmid DNA sequence, or it was L. plantarum-specific, as it targeted a chromosomally located plantaricin gene sequence. Two 50 l wine fermentations were prepared. One was inoculated with 15 g/hl Saccharomyces cerevisiae, followed by L. plantarum IWBT B 188 at 3.6 × 106 CFU/ml, whereas the other was not inoculated (control). Viable cell counts were performed for up to 25 days on MRS agar, and the same cells were enumerated by qRT-PCR with both the plasmid or chromosomally encoded gene primers. The L. plantarum strain survived under the harsh conditions in the wine fermentation at levels above 105/ml for approx. 10 days, after which cell numbers decreased to levels of 103 CFU/ml at day 25, and to below the detection limit after day 25. In the control, no lactic acid bacteria could be detected throughout the fermentation, with the exception of two sampling points where ca. 1 × 102 CFU/ml was detected. The minimum detection level for quantitative PCR in this study was 1 × 102 to 1 × 103 CFU/ml. The qRT-PCR results determined generally overestimated the plate count results by about 1 log unit, probably as a result of the presence of DNA from dead cells. Overall, qRT-PCR appeared to be well suited for specifically enumerating Lactobacillus plantarum starter cultures in the MLF in wine.

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