• Journal of the Korean Applied Science and Technology
• /
• v.39 no.6
• /
• pp.916-923
• /
• 2022

Water samples were collected from three spots(Namcheon, Changwoncheon and Cheongwoonji) in Changwon and genomic DNA was isolated from them. Quantitative PCR was performed with the isolated DNA as template and primers targeting five different class A extended-spectrum beta-lactamase(ESBL) genes(bla_OXA-1, bla_SHV, bla_TEM, bla_CTX-M-1, bla_CTX-M-9). The number of total ESBL genes from each sample showed large variations between each sample. Thirty nanograms of DNA from Namcheon contained 1.93×10⁶ copies of ESBL genes whereas the same amount of DNA from Changwoncheon contained 1.47×10⁵ copies of ESBL genes. However, the same amount of DNA from Cheongwoonji pond contained only 9.5×10³ copies of ESBL genes. The ratio of each ESBL genes showed little difference between Namcheon river and Changwoncheon river, but DNA from Cheongwoonji pond showed a large difference from the rest. bla_OXA-1 gene was present at 65.3%, and bla_CTX-M-1 gene 33.6% for Namcheon comprising together almost 99%. bla_OXA-1 gene was present at 64.1%, and bla_CTX-M-1 gene 19.1% for Changwoncheon comprising together over 83%. bla_CTX-M-1 gene was present at 87.5% and bla_CTX-M-9 genes 9.8% for Cheongwoonji, a pond which is a closed and isolated environment.

• Fisheries and Aquatic Sciences
• /
• v.16 no.4
• /
• pp.221-227
• /
• 2013

In total, 131 Escherichia coli isolates from surface seawater of the Gomso Bay, of Korea, were analyzed for their susceptibility to 22 different antimicrobials and for genes associated with antimicrobial resistance and virulence. According to the disk diffusion susceptibility test, the resistance to tetracycline was most prevalent (33.6%), followed by that to ampicillin (22.1%), ticarcillin (22.1%), and trimethoprim (16.8%). More than 46.6% of the isolates were resistant to at least one antimicrobial, and 22.9% were resistant to three or more classes of antimicrobials; these were consequently defined as multidrug resistant. We further found that 29 ampicillin-resistant isolates possessed genes encoding TEM-type (93.1%) and SHV-type (6.9%) ${\beta}$-lactamases. Among the 44 tetracycline-resistant isolates, tetA and tetC were found in 35 (79.5%) and 19 (43.2%), respectively, whereas tetB was detected in only three isolates (6.8%). With regard to virulence genes, merely 0.8% (n = 1) and 2.3% (n = 3) of the isolates were positive for the enteroaggregative E. coli-associated plasmid (pCVD432) gene and the enteropathogenic E. coli-specific attaching and effacing (eae) gene, respectively. Overall, these results not only provide novel insight into the necessity for seawater sanitation in Gomso Bay, but they help reduce the risk of contamination of antimicrobial-resistant bacteria.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
• /
• 1996.11a
• /
• pp.131-139
• /
• 1996

To quantitatively investigate the effects of manure compost on the soil and water environment including ground water the elution patterns of anions and hydraulic conductivity wore estimated with four different depth(15, 30, 45 60cm) and four variable ratio of compost treatment(0, 2, 4, 6%) through soil column test. 1. There were over 95% of elution of chloride and nitrate within 0.1 pore volume(PV), and sulfate within 0.2 PV. With 2 ton/10a of cow manure compost treatment recommended total 40 kg/10a of anions added was recovered as effluent at the amount of 17kg chloride, 5.4kg nitrate, and 13.2kg sulfate, respectively However, phosphate rarely recovered in the effluent due to the strong affinity for sorption sites in soils. 2. In multi-layered soil column the maximum peaks of each anion eluted were retardated with increasing soil depth and the amount of organic matter(OM) treatment. 3. With increasing OM up to 2% the saturated hydraulic conductivity(SHC) was greatly decreased, but the slight decrease in SHC was found by addition of OM greater than 4％. In multi-layered soil column SHC was more effected by the lower SHV layer than by that of the higher.

• Korean Journal of Clinical Laboratory Science
• /
• v.38 no.3
• /
• pp.173-178
• /
• 2006

The production of extended-spectrum ${\beta}$-lactamases ($ESBL_S$) is the main mechanism of bacterial resistance to third-generation cephalosporins and monobactams, whose prevalence varies depending on the different geographical areas. In the last years it has increased notably to the point of being considered a health problem of great importance. The characterization of the ESBLs producing Klebsiella penumoniae strains present in clinical isolates is time-consuming. I describe here the development of a new system, which consists of a multiplex PCR. I found 51 K. pneumoniae strains to be presumptive strains ESBLs producers by clinical and laboratory standards institute (CLSI) guidelines. The double disc synergy test showed 47 positive K. pneumoniae, which were K. pneumoniae isolates. All ESBLs producing K. pneumoniae strains were resistant to antibiotic amikacin, gentamicin and ciprofloxacin. By multiplex PCR analysis, $bla_{TEM}$ gene in 17 strains 44 $bla_{SHV}$ genes and $bla_{CTX}$ genes in 33 strains were identified. In this study, the multiplex polymerase chain reaction (PCR) assay was a good method to detect and differentiate ESBLs producing K. penumoniae strains in clinical isolates.

• Journal of Microbiology and Biotechnology
• /
• v.30 no.7
• /
• pp.974-981
• /
• 2020

Sequence type 410 (ST410) of Escherichia coli is an extraintestinal pathogen associated with multi drug resistance. In this study, we aimed to investigate the horizontal propagation pathway of a high-risk clone of E. coli ST410 that produces Klebsiella pneumoniae carbapenemase (KPC). blaKPC-encoding E. coli and K. pneumoniae isolates were evaluated, and complete sequencing and comparative analysis of blaKPC-encoding plasmids from E. coli and K. pneumoniae, antimicrobial susceptibility tests, polymerase chain reaction, multilocus sequence typing, and conjugal transfer of plasmids were performed. Whole-genome sequencing was performed for plasmids mediating KPC-2 production in E. coli and K. pneumoniae clinical isolates. Strains E. coli CPEc171209 and K. pneumoniae CPKp171210 were identified as ST410 and ST307, respectively. CPEc171209 harbored five plasmids belonging to serotype O8:H21, which is in the antimicrobial-resistant clade C4/H24. The CPKp171210 isolate harbored three plasmids. Both strains harbored various additional antimicrobial resistance genes. The IncX3 plasmid pECBHS_9_5 harbored blaKPC-2 within a truncated Tn4401a transposon, which also contains blaSHV-182 with duplicated conjugative elements. This plasmid displayed 100% identity with the IncX3 plasmid pKPBHS_10_3 from the K. pneumoniae CPKp171210 ST307 strain. The genes responsible for the conjugal transfer of the IncX3 plasmid included tra/trb clusters and pil genes coding the type IV pilus. ST410 can be transmitted between patients, posing an elevated risk in clinical settings. The emergence of a KPC-producing E. coli strain (ST410) is concerning because the blaKPC-2-bearing plasmids may carry treatment resistance across species barriers. Transgenic translocation occurs among carbapenem-resistant bacteria, which may spread rapidly via horizontal migration.

• Biomedical Science Letters
• /
• v.13 no.4
• /
• pp.293-304
• /
• 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
• /
• v.28 no.7
• /
• pp.1178-1184
• /
• 2018

The spread of antibiotic-resistant Enterobacteriaceae in the community is one of the main challenges for antibiotic treatment of community-onset infections. We evaluated the microbiological and molecular characteristics of stool samples from adults with comprehensive health examinations. Of 109 fecal samples, bacterial growth was observed in 86 samples and 61 gram-negative bacterial isolates were identified, of which 45 were Escherichia coli isolates (73.8%). Two isolates of Raoultella showed imipenem resistance, and both E. coli and Citrobacter freundii showed intermediate resistance to imipenem. Colistin resistance was identified in isolates of Klebsiella variicola and Salmonella subterranean, but no isolates carried mcr-1. As for E. coli genotypes, 35 sequence types were identified. $bla_{TEM-1}$, $bla_{TEM-30}$, and $bla_{CTX-M}$ were identified in 15, 1, and 4 E. coli isolates, respectively. In addition, all four Klebsiella pneumoniae isolates carried $bla_{SHV}$. Many genotypes that have been identified in isolates causing human infections were found in isolates in this study. There is a need to control the rise and spread of antibiotic-resistant pathogens by fecal carriage.

• Korean Journal of Veterinary Service
• /
• v.40 no.3
• /
• pp.193-200
• /
• 2017

This study was carried out to investigate the antimicrobial resistance profiles and resistance genes in 62 Escherichia coli isolated from dogs and cats hospitalized at animal hospitals in Daegu. E. coli isolates showed high resistance to nalidixic acid (46.8%) and ampicillin (45.2%). Resistance to the other antimicrobial agents was less than 30%, and no resistant isolates were detected for imipenem and amikacin. Of the 28 ampicillin-resistant isolates, TEM and CTX-M genes were detected in 16 (57.1%) and 11 (39.3%), respectively. The aadA gene was found in 4 (26.7%) of 15 gentamicin-resistant isolates, and strA-strB gene was found in 10 (66.7%) isolates. The sul I and sul II genes were detected in 11 (61.1%) and 14 (77.8%) of 18 trimethoprim/sulfamethoxazole-resistant isolates, and tetB gene in 9 (81.8%) of 11 minocycline-resistant isolates, and cmlA gene in 2 (22.2%) of 8 chloramphenicol-resistant isolates. The qnrB and qnrS genes were found in 3 (10.3%) and 1 (3.4%) of 28 nalidixic acid-resistant isolates, respectively. Whereas, none of the SHV, CMY-2, tetA, dfr Ia and dfr VII, and qnrA genes were found. Our results show a wide variety of resistance genes in E. coli isolates from dogs and cats. This study also represents the first report of qnrB and qnrS gene producing E. coli isolates from dogs in republic of Korea.

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

• Korean Journal of Veterinary Service
• /
• v.37 no.1
• /
• pp.19-27
• /
• 2014

This study was conducted to investigate the antimicrobial resistance, presence of ${\beta}$-lactamase genes and integrons in 83 ESBL-producing Escherichia coli isolated from Nakdong river and Geumho river in Daegu. Among the ${\beta}$-lactam antimicrobials, all isolates were resistant to ampicillin, cephalothin, cefamandole and cefotaxime, followed by piperacillin (98.8%), ampicillin/sulbactam (86.7%), aztreonam (60.2%) and cefepime (59.0%), whereas resistance to piperacillin/tazobacram, ticarcillin/clavulanic acid and cefoxitin was less than 30%. Many of the ESBL-producing Escherichia coli were also resistant to non-${\beta}$-lactams antimicrobials such as nalidixic acid (83.1%), sulfonamides (72.3%), ciprofloxacin (62.7%) and gentamicin (38.6%). All isolates showed resistance to seven or more antimicrobial agents. The most frequently detected gene was $bla_{TEM+CTX-M}$ (49.4%), followed by $bla_{CTX-M}$ (27.7%), $bla_{TEM}$ (6.0%) and $bla_{OXA}$ (1.2%). But $bla_{SHV}$ was not found. Class 1 integrons were found in 61.4% (51 isolates) of isolates, however, class 2 and 3 integrons were not detected. The results showed water from Nakdong river and Geumho river is contaminated with ESBL-producing E. coli isolates. These results suggest the need for further investigation of antibiotic resistant bacteria to prevent public health impacts in the water environment.