• Korean Journal of Microbiology
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• v.37 no.4
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• pp.277-283
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• 2001

The emergence of extended spectrum beta-lactamase(ESBL) producing bacteria is causing very serious problems in Korea. Although there have been many reports about these bacteria isolated from patients and clinical specimens, there is no report of ESBL-producing organisms isolated from natural evironment in Korea. This is the first study on the ESBL producing bacteria out of the medical system in Korea. Twenty-six ESBL producing bacteria were isolated only from sewerage plant drain water at Kwang-an beach among the sampling collected sites including snakehead fish plants in Myungi, Aquaculture Engineering Lab. in Pukyong National University and two public-bathrooms in Pusan, Korea. ESBL producing bacteria were identified by double-disk synergy test, conjugation, isoelectric focusing values and PCR. The species of ESBL producing bacteria were Enterobacter cloacae(4 strains), E. sakazakii(8 strains), Klebsiella pneumoniae subsp. pneumoniae(8 strains) and K. pneumoniae subsp. ozaenae(6 strains). TEM and SHV specific PCR products were detected from all the ESBL strains produced TEM+SHV products on the PCR plates. The pI values of ESBL produced by Klebsiella pneumoniae subsp. pneumoniae, K. pneumoniae subsp. ozaenae, Enterobacter cloacae, and E. sakazakii were 5.9, 5.9+5.4; 5.9, $5.9+5.4;{\ge}8.5$, 8.0+5.4, and 8.0+5.4, respectively on the IEF. Seven strains of the isolates were transfered their genes to E. coli RG488 $Rif^r$ by conjugation.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.10 no.9
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• pp.2516-2522
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• 2009

The data which analyze the results of blood cultures are crucial basic information of the empirical therapy for patients with infection since the patterns of the species of microorganism isolated from blood and the results of antibiotic susceptibility test vary depending upon patients general features. Especially, in case of ESBL-producing bacteria, there is a close relation with use of antibiotics. Therefore, we carried out the research with the results of blood culture and antibiotic. 1. Total 39,305 cases of blood culture samples were investigated and positive patients of 2,216 (20.0%) were found. Among those, there were 40 patients with ESBL positive, and blood culture positive samples were 4,798 (12.2%). ESBL positive bacteria were found in 86 samples (including double checked culture bacteria). 2. The majority of ESBL producing bacteria were E. coli, K. pneumoniae and K. oxitoca as ordering based on the number. 3. The research showed the results that there were more females than male with the bacterias, more E. coli in over 50 years old aged group than other bacterias, more K. pneumoniae and K.oxitoca in less 1 year old aged group than other bacterias and largest numbers of patients with 13 patients (32.5%) in Chungcheongnam-do province were found. 4. The most common ESBL producing bacteria were E. coli throughout 3 years, but K, pneumoniae and K. oxitoca were also fairly found. Interestingly, E. coli was highly found in over 50 years old patients.

• Pediatric Infection and Vaccine
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• v.22 no.1
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• pp.29-35
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• 2015

Purpose: The purpose of this study was to investigate the clinical characteristics and outcome of febrile urinary tract infections (UTIs) caused by community-acquired extended-spectrum ${\beta}$-lactamase (CA-ESBL)-producing and -nonproducing bacteria. Methods: We analyzed febrile UTIs in children hospitalized at Gachon University Gil Medical Center from January 2011 to December 2013 through retrospective data collection from their medical records. Results: Among pathogens causing 374 episodes of UTIs, the proportion of ESBL-producing bacteria was 13.1% (49/374). The proportion of ESBL-producing Escherichia coli and Klebsiella spp. was 13.6% (48/354) and 5.0% (1/20), respectively. There was no significant difference between the CA-ESBL and CA non-ESBL groups in duration of fever ($4.2{\pm}2.7$ vs. $3.7{\pm}2.1$ days, P=0.10) and bacterial eradication rate with empirical antibiotics (100% vs. 100%). The risk of cortical defects on renal scan significantly depended on existence of vesicoureteral reflux rather than ESBL production of pathogen. Conclusions: There was no significant difference between the CA-ESBL and CA non-ESBL groups in renal cortical defects and clinical outcome. Careful choice of antibiotics is important for treatment of community-acquired UTI in children.

• Biomedical Science Letters
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• v.13 no.4
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• pp.293-304
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• 2007

The emergence of extended spectrum $\beta$-lactamase (ESBL) producing bacteria is worldwide concern. Until recently, the most frequently identified strains in the Republic of Korea were E. coli and Klebsiella spp. The incidence of resistance to extended spectrum $\beta$-lactam antibiotics is increasing in Wonju city, Korea. Total 57 strains of ESBL producing E. coli and Klebsiella species were isolated from Wonju Christian Hospital during a 9 month-period from April to December, 2003. To determine the prevalence and genotypes of the ESBL producing clinical isolates, antibiotic susceptibility and ESBL activity test by VITEK system and double disk synergy (DDS) test, and PCR based genotyping were performed. Fourteen (82%) isolates of 17 ESBL producing E. coli were found to have $bla_{TEM}$ gene and 5 (29%) isolates were found to have $bla_{CTX-M}$ gene by polymerase chain reaction (PCR). Thirty (75%) isolates of 40 ESBL producing Klebsiella species with $bla_{TEM}$ gene, 38 (95%) isolates with $bla_{SHV}$ gene, and 7 (20%) isolates with $bla_{CTX-M}$ type gene were also identified. Enterobacterial repetitive intergenic consensus (ERIC) PCR and similarity index by dendrogram for genetical similarity to band pattern of each clinical isolates were examined. ESBL producing E. coli were grouped into 6 clusters up to 84% of similarity index and Klebsiella species were grouped into 12 clusters up to 76% of similarity index. In conclusion, ESBL producing clinical isolates were characterized with the results from antimicrobial resistance pattern and genetical similarity using ERiC PCR.

• Journal of Microbiology and Biotechnology
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• v.28 no.9
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• pp.1563-1572
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• 2018

Gold nanoparticles (AuNP) and their conjugates have been gaining a great deal of recognition in the medical field. Meanwhile, extended-spectrum ${\beta}$-lactamases (ESBL)-producing bacteria are also demonstrating a challenging problem for health care. The aim of this study was the biosynthesis of AuNP using Rosa damascenes petal extract and conjugation of ceftriaxone antibiotic (Cef-AuNP) in inhibiting ESBL-producing bacteria and study of in vitro anticancer activity. Characterization of the synthesized AuNP and Cef-AuNP was studied. ESBL-producing strains, Acinetobacter baumannii ACI1 and Pseudomonas aeruginosa PSE4 were used for testing the efficacy of Cef-AuNP. The cells of MCF-7 breast cancer were treated with previous AuNP and Cef-AuNP at different time intervals. Cytotoxicity effects of apoptosis and its molecular mechanism were evaluated. Ultraviolet-visible spectroscopy and Fourier transform infrared spectroscopy established the formation of AuNP and Cef-AuNP. Transmission electron microscope demonstrated that the formed nanoparticles were of different shapes with sizes of 15~35 nm and conjugation was established by a slight increase in size. Minimum inhibitory concentration (MIC) values of Cef-AuNP against tested strains were obtained as 3.6 and $4{\mu}g/ml$, respectively. Cef-AuNP demonstrated a decrease in the MIC of ceftriaxone down to more than 27 folds on the studied strains. The biosynthesized AuNP displayed apoptotic and time-dependent cytotoxic effects in the cells of MCF-7 at a concentration of $0.1{\mu}g/ml$ medium. The Cef-AuNP have low significant effects on MCF-7 cells. These results enhance the conjugating utility in old unresponsive ceftriaxone with AuNP to restore its efficiency against otherwise resistant bacterial pathogens. Additionally, AuNP may be used as an alternative chemotherapeutic treatment of MCF-7 cancer cells.

• Microbiology and Biotechnology Letters
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• v.46 no.2
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• pp.135-144
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• 2018

The rapid increase in number and diversity of Extended Spectrum ${\beta}$-Lactamases (ESBLs) producing Enterobacteriaceae in natural aquatic environment is a major health concern worldwide. This study investigates abundance and distribution of ESBL producing multidrug resistant Enterobacteriaceae and molecular characterization of ESBL genes among isolates from highly polluted stretch of river Yamuna, India. Water samples were collected from ten different sites distributed across Delhi stretch of river Yamuna, during 2014-15. A total of 506 non duplicate Enterobacteriaceae isolates were obtained. Phenotypic detection of ESBL production and antibiotic sensitivity for 15 different antibiotics were performed according to CLSI guidelines (Clinical and Laboratory Standard Institute, 2015). A subset of ESBL positive Enterobacteriaceae isolates were identified by 16S rRNA gene and screened for ESBL genes, such as $bla_{CTX-M}$, $bla_{TEM}$ and $bla_{OXA}$. Out of 506 non-duplicate bacterial isolates obtained, 175 (34.58%) were positive for ESBL production. Susceptibility pattern for fifteen antibiotics used in this study revealed higher resistance to cefazolin, rifampicin and ampicillin. A high proportion (76.57%) of ESBL positive isolates showed multidrug resistance phenotype, with MAR index of 0.39 at Buddha Vihar and Old Delhi Railway bridge sampling site. Identification and PCR based characterization of ESBL genes revealed the prevalence of $bla_{CTX-M}$ and $bla_{TEM}$ genes to be 88.33% and 61.66%, respectively. Co-occurrence of $bla_{CTX-M}$ and $bla_{TEM}$ genes was detected in 58.33% of the resistant bacteria. The $bla_{OXA}$ gene was not detected in any isolates. This study highlights deteriorating condition of urban aquatic environment due to rising level of ESBL producing Enterobacteriaceae with multidrug resistance phenotype.

• Korean Journal of Microbiology
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• v.50 no.2
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• pp.114-118
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• 2014

A total of 122 ESBL-producing intestinal bacteria were collected from regional hospitals in the Chungcheong area. Combination disk test (CDT) was performed for antimaicrobial susceptability using cefotaxime and cefotaxime/clavulanate according to Clinical Laboratory Standard Institute (CLSI). Mutiplex PCR using specific primers was performed for a detection of ESBL-genotypes and enterobacterial repetitive intergenic consensus (ERIC)-PCR was carried out for the tracking of molecular epidemiology. In the confirmation test using CDT, 73 out of 76 (96.1%) ESBL-producing Escherichia coli and 43 out of 46 (93.4%) ESBL-producing Klebsiella pnemoniae were positive. In the multiplex PCR, 60.5% of E. coil were positive for CTX-M-2 type gene and 56.5% of K. pneumoniae were positive for VEB -1 type gene. In the ERIC-PCR, E. coil isolates formed 5 clusters and K. pneumoniae isolates were grouped into 4 clusters depending on region. Genotypes of clinical isolates are useful for detection and differentiation of ESBL producing intestinal bacteria. The ERIC-PCR method is thought to be helpful for establishing a regional surveillance system for infection due to its formation of different clusters depending on region.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.342-351
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• 2006

The aim of this study was to survey susceptibilities of Escherichia coli and Klebsiella pneumoniae isolates against cefotaxime and to determine the prevalences of CTX-M type extended-spectrum $\beta$-lactamases (ESBLs) producing E. coli and K. pneumoniae in Korea. During the period of February to July, 2005, 153 E. coli and 52 K. neumoniae isolates were collected from 2 hospitals in Busan. Antimicrobial susceptibilities to cefotaxime were tested by the disk diffusion method. ESBL production of E. coli and K. pneumoniae was determined by the double disk synergy test. MICs of $\beta$-lactam antibiotics were determined by the agar dilution method. Blac$_{CTX-M}$ genes of the organism were detected by PCR. Among 153 isolates of E. coli and 52 isolates of K. neumoniae, 27 (17.6%) and 25 (48.0%) were intermediate or resistant to cefotaxime, respectively. Twenty-three (15.0%) isolates out of 153 E. coli and 13 (25.0%) out of 52 K. neumoniae isolates showed positive results for ESBL by the double disk synergy test. Twenty isolates out of 23 ESBL producing E. coli and 12 out of 13 ESBL producing K. neumoniae isolates harbored biacTx-M gene,11 of ESBL producing E. coli and 12 of ESBL producing K. neuinoniae isolates harbored bla$_{CTX-M}$ gene, 11 of the ESBL producing E. coli and 2 of ESBL producing K. neumoniae isolates harbored bla$_{TEM}$ gene, and 1 of the ESBL producing E. coli and 12 of ESBL producing K. neumoniae isolates harbored bla$_{SHV}$ gene. E. coli and K. neumoniae isolates producing CTX-M-type ESBLs were not uncommon in Korea. It is thought that continuous survey are necessary for inspecting the spread and novel variants of CTX-M-type ESBL genes. Further me]'e investigation and research on ESBL producing strains are needed in order to prevent the spread of resistant bacteria.

• Journal of Veterinary Clinics
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• v.27 no.2
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• pp.125-129
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• 2010

A 2-year-old female Pomeranian dog was referred with multiple pelvic fractures. The surgical correction was performed for the fractures. However, after the surgery, purulent exudation was occurred in the surgical site. Antibiotic susceptibility test revealed that the isolated bacteria are resistant to penicillins, cephalosporins, aminoglycosides, quinolones, and trimethoprim/sulfamethoxazole. Bacterial identification and extended-spectrum $\beta$-lactamase (ESBL) confirming test indicated that the isolated bacteriae is ESBL-producing Klebsiella pneumoniae. Minimum inhibitory concentration (MIC) and maximum bactericidal concentration (MBC) tests revealed that meropenem, one of carbapenems, is the only effective antibiotic. The patient was treated with meropenem for 5 days. After 10 days, the exudation was disappeared and the infection was cured. The molecular typing of the ESBL revealed that TEM-1 ESBL is present in the bacteria isolated from the patient. The bacteria isolated from the owner's palm also revealed that TEM-1 and SHV-1 ESBLs are present.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.394-401
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• 2016

Recently, the clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (CRISPR/Cas9) system, a genome editing technology, was shown to be versatile in treating several antibiotic-resistant bacteria. In the present study, we applied the CRISPR/Cas9 technology to kill extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli. ESBL bacteria are mostly multidrug resistant (MDR), and have plasmid-mediated antibiotic resistance genes that can be easily transferred to other members of the bacterial community by horizontal gene transfer. To restore sensitivity to antibiotics in these bacteria, we searched for a CRISPR/Cas9 target sequence that was conserved among >1,000 ESBL mutants. There was only one target sequence for each TEM- and SHV-type ESBL, with each of these sequences found in ~200 ESBL strains of each type. Furthermore, we showed that these target sequences can be exploited to re-sensitize MDR cells in which resistance is mediated by genes that are not the target of the CRISPR/Cas9 system, but by genes that are present on the same plasmid as target genes. We believe our Re-Sensitization to Antibiotics from Resistance (ReSAFR) technology, which enhances the practical value of the CRISPR/Cas9 system, will be an effective method of treatment against plasmid-carrying MDR bacteria.