• Korean Journal of Clinical Laboratory Science
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• v.43 no.2
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• pp.68-74
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• 2011

Pseudomonas aeruginosa is an aerobic, Gram-negative, glucose-nonfermenting bacterium, which has emerged as a serious opportunistic pathogen. Recently, outbreaks of carbapenem resistant P. aeruginosa give rise to significant therapeutic challenges for treating nosocomial infections. The genes of metallo-${\beta}$-lactamase (MBL), a powerful carbapenemase, are carried as a part of the mobile gene cassettes inserted into integrons playing an important role in rapid dissemination of antibiotic resistance genes among bacterial isolates. In this study, we investigated the prevalence of integron in imipenem resistant P. aeruginosa isolates. A total of 61 consecutive, non-duplicate, and imipenem resistant P. aeruginosa strains were isolated from a university hospital in the Chungcheong province of Korea. We employed repetitive extragenic palindromic sequence-based PCR (rep-PCR) method for the selection of clonally different P. aerusinosa strains. PCR and DNA sequencing were conducted for the detection of integrons. Twenty-one clonally different P. aeruginosa strains were isolated. Only one (P28) of the strains harbored $bla_{VIM-2}$ that was found as gene cassettes in class 1 integrons. Four of 21 carbapenem resistant P. aeruginosa strains harbored class 1 integron containing aminoglycoside resistance determinant. All of the integrons detected in the study contained more than one resistance gene cassette, which can mediate resistance to multiple antibiotics. To prevent further spreading of the multi-drug resistant P. aeruginosa, conseguent monitoring and clinical polices are required.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.765-770
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• 2014

This study aimed to characterize CTX-M producers of urinary E. coli and K. pneumoniae isolates and to determine the prevalence of plasmid-mediated antimicrobial resistance genes among them. Minimum inhibitory concentrations (MICs) were determined, and PCR and sequencing were performed. Among the 42 (82.3%) E. coli and 24 (77.4%) K. pneumoniae isolates containing $bla_{CTX-M}$, $bla_{CTX-M-14}$ and $bla_{CTX-M-15}$ were detected in 23 and 19 E. coli isolates, respectively, and in 7 and 17 K. pneumoniae isolates, respectively. CTX-M producers of urinary E. coli and K. pneumoniae were resistant to multiple antibiotics and contained other antimicrobial resistance genes. CTX-M-15 producers contained more antimicrobial resistance genes than did CTX-M-14 producers.

• Journal of Veterinary Clinics
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• v.25 no.4
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• pp.231-235
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• 2008

Various kinds of antibiotic intramammary infusions are used for treatment of bovine mastitis. As antibiotic-resistant bacteria are increased, the therapeutic rate for bovine mastitis is decreased. The goal of this research is to detect significant synergic effects of combination of antibiotics with sulbactam, $\beta$-lactamase inhibitor, on methicilin-resistant Staphylococcus aureus (MRSA). We used 5 strains of MRSA isolated from bovine mastitis with clinical and subclinical signs. All of the bacteria isolated had resistance to oxacillin and showed multi-resistant patterns in the antimicrobial susceptibility tests. Minimal bactericidal concentrations of ampicillin, amoxicillin, cephalexin, ampicillin/sulbactam(2:1), amoxicillin/sulbactam (2:1), and cephalexin/sulbactam (1:1) were measured according to broth microdilution method suggested by National Committee for Clinical Laboratory Standards (NCCLS, M31-A2) to compare the synergic effects of sulbactam combination with each antibiotic alone. Ampicillin and amoxicillin showed synergic antibacterial activity to 4 and 3 respectively in 5 strains of MRSA in combination with sulbactam. This study demonstrates that ampicillin/sulbactam and amoxicillin/sulbactam can be therapeutic choices for mastitis associated with MRSA.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.243-246
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• 2005

A novel esterase showing high enantioselectivity to (S)-ketoprofen ethyl ester was selected from fosmid environmental DNA library which is provided by Microbial Genomic & Applications Center. As a result of Blast search, the gene wasn't registerated in Gene Bank yet. And as we know, conserved domain region of esterase , G-X-S-X-G, wasn't discovered.$^{4)}$ And it is similar to Beta-lactamase. The DNA sequence of cloned esterase include an open reading frame consisting of 1170 bp, designated as EST-Y29, encoding a protein of 389 amino acids with a molecular mass of about 42.8 kDa. And amino acid sequence analysis revealed only a few identity (28%) to tile known esterases/lipases in the databases containing the conserved sequence motifs of esterases/lipases. when being comparison to other esterase revealed , this enzyme seems to be classified as a new member of esterase family. EST-Y29 was functionally overexpressed in a soluble form in E. coli with maximum conversion yield of (S)-ketoprofen at $65^{\circ}C$. This study demonstrates that functional screening combined with the sequential uses of restriction enzymes to exclude already known enzymes is a useful approach for isolating novel enzyme from a metagenome.

• Journal of Microbiology and Biotechnology
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• v.2 no.3
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• pp.166-173
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• 1992

The expression of Bacillus subtilis $\alpha$-amylase from the phoA-amyE fusion gene in recombinant E. coli was investigated under various environmental conditions. The overexpression of cloned $\alpha$-amylase caused retardations in cell growth and synthesis of alkaline phosphatase (AP) from the chromosomal phoA gene. The change of culture temperature from $37^\circ{C}$ to $30^\circ{C}$ increased the specific activities of both $\alpha$-amylase and $\beta$-lactamase by six and two times, respectively, whereas the AP activity remained unchanged. The experiments with chlorampenicol (a translation inhibitor) suggested the enhancement of $\alpha$-amylase activity at $30^\circ{C}$, and this was partly due to the stability of $\alpha$-amylase itself. The further decrease of the temperature to $25^\circ{C}$ slowed down both the cell growth and cloned-gene expression rate. The $\alpha$-amylase activity showed a maximum at pH of 7.4 while alkaline phosphatase was most effectively produced at pH of 8.3.

• Korean Journal of Veterinary Research
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• v.57 no.1
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• pp.9-15
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• 2017

To construct a novel vaccine candidate against bovine enterotoxigenic Escherichia coli (ETEC), FanC, the major subunit of K99 fimbriae adhesion, was inserted into secretion plasmid pYA3560 containing a ${\beta}-lactamase$ secretion system. This was then transformed into ${\Delta}asd$ ${\Delta}crp$ Salmonella (S.) Typhimurium and designated as JOL950. Secretion of recombinant fanC fimbrial antigens was confirmed by immunoblot analysis. Groups of mice were inoculated with single or double doses of JOL950. Another group was used as a negative control. Compared to control mice, all immunized mice had significantly higher levels (p < 0.05) of serum immunoglobulin (Ig)G, and secretory IgA against FanC. The IgG2a and IgG1 titer assays revealed that immunization highly induced IgG2a compared to that of IgG1, indicating that T helper-1- related cell-mediated immune responses may be elicited by JOL950. The results show that both systemic and mucosal immunities against selected fimbrial antigens of bovine ETEC expressed by a live attenuated S. Typhimurium strain are prominently produced in mice immunized with JOL950 via an oral route.

• Korean Journal of Clinical Pharmacy
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• v.23 no.1
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• pp.26-32
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• 2013

Objective: This study was to define the clinical effect on the clinical decision support system (CDSS) for prescribing antibiotics integrated with the order communication system in a National Hospital. Method: We extracted data collected before integrating the CDSS of 4,406 adult patients in 2007 and data collected after integrating the CDSS of 4,278 adult patients in 2009. These patients were 50.4% and 45.2% of all patients admitted in 2007 and 2009, respectively. The clinical effect was defined as the proportion of prescribed antibiotics, the length of antibiotics use, and the DDDs (defined daily doses) of antibiotics per 1,000 patient-days using these retrospective data. Results: There were a significant change in the proportion of patient prescribed penicillins with extended spectrum (OR=0.55, p=001), penicillins included beta-lactamase inhibitors (OR=0.75, p<.001), 3rd cephalosporin (OR=1.47, p<.001). The mean of the length of antibiotics use was decreased statistically from $6.09{\pm}5.48$ to $5.85{\pm}5.51$ days (p=.003). The DDD of glycopeptides was decreased from 24.43 DDD to 19.55 DDD per 1000 patient-days. The DDD of 3rd cephalosporins was also decreased from 15.88 to 11.65. Conclusion: Therefore, the clinical decision support system for prescribing antibiotics was effective for the clinical outcomes.

• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.465-472
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• 2019

Several reports describe antimicrobial-resistance transfer among children and the community in outbreak situations, but transfer between a child and a caregiver has not been examined in child care facilities under normal circumstances. We investigated the transfer of antimicrobial-resistance genes, resistant bacteria, or both among healthy children and teachers. From 2007 to 2009, 104 Escherichia coli isolates were obtained from four teachers and 38 children in a child care center. Twenty-six cephem-resistant isolates were obtained from children in 2007 and 2008. In 2009, cephem-resistant isolates were detected in children as well as a teacher. Nalidixic acid-resistant isolates from the same teacher for 3 years showed low similarity (<50%) to each other. However, an isolate from a teacher in 2007 and another from a child in 2008 showed high similarity (87%). Pulsed-field gel electrophoresis revealed 100% similarity for four isolates in 2007 and one isolate in 2008, and also similarity among seven isolates carrying the virulence gene (CNF1). This study yielded the following findings: (1) a gene for extended-spectrum ${\beta}$-lactamase was transferred from a child to other children and a teacher; (2) a nalidixic acid-resistant isolate was transferred from a teacher to a child; and (3) a virulent bacterium was transferred between children.

• Journal of Digital Convergence
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• v.14 no.10
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• pp.349-354
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• 2016

The emergence and dissemination of extended-spectrum ${\beta}$-lactamse (ESBL) producing Klebsiella pneumoniae isolates make it more difficult to treatment of bacterial infections. In our study, we detected ESBL genes and investigated antimicrobial susceptibility of K. pneumoniae isolates in Chungcheong province. In addition, clonality among the isolates was analyzed by repetitive element sequence (REP)-PCR. Twenty-one of 102 K. pneumoniae isolates produced CTX-M-14 and/or CTX-M-15 and showed high level (over 70%) resistance to third cephalosporins. CTX-M type ESBL producing K. pneumoniae strains isolated in our study showed diverse clonality and some of the isolates have been disseminated in the community. Enhancing infection control will be needed to prevent dissemination of the K. pneumoniae isolates. In addition, for more effective control of resistant bacteria it is considered necessary to monitor the database constructed through convergence of biological investigation and statistical analysis of antimicrobial resistance genes.

• Journal of Microbiology and Biotechnology
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• v.24 no.3
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• pp.421-426
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• 2014

To characterize the extended-spectrum beta-lactamases (ESBLs) in diarrheagenic Escherichia coli from Korea in 2008-2011, we screened seven enterotoxigenic E. coli (ETEC) and one enteroaggregative E. coli (EAEC) that produce ESBLs from a nationwide survey. All eight isolates produced CTX-M-type ESBLs, including CTX-M-12 (n = 4), CTX-M-14 (n = 2), and CTX-M-15 (n = 2). PCR-based replicon typing indicated that the $bla_{CTX-M-12}$ genes of four ETEC isolates were carried on a conjugative IncF plasmid, whereas the $bla_{CTX-M-14}$ of one EAEC was located on an IncK plasmid. This is the first report of the occurrence of $bla_{CTX-M}$ genes in clinical isolates of EAEC in Korea. The ESBL-producing isolates were shown to be different based on pulsed-field gel electrophoresis and multilocus sequence typing, whereas the four isolates with CTX-M-12 were clonally related. These observations raise an alarm for the spread of plasmid-mediated resistance to ESBL among diarrheagenic E. coli.