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Sequence Variations in the Non-Coding Sequence of CTX Phages in Vibrio cholerae

  • Kim, Eun Jin;Yu, Hyun Jin;Kim, Dong Wook
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1473-1480
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    • 2016
  • This study focused on the variations in the non-coding sequences between ctxB and rstR of various CTX phages. The non-coding sequences of CTX-1 and CTX-cla are phage type-specific. The length of the non-coding region of CTX-1 and CTX-cla is 601 and 730 nucleotides, respectively. The non-coding sequence of CTX phage could be divided into three regions. There is a phage type-specific Variable region between two homologous Common regions (Common regions 1 and 2). The non-coding sequence of RS1 element is similar to CTX-1 except that Common region 1 is replaced by a short RS1-specific sequence. The non-coding sequences of CTX-2 and CTX-cla are homologous, indicating the non-coding sequence of CTX-2 is derived from CTX-cla. The non-coding region of CTX-O139 is similar to CTX-cla and CTX-2; however, it contains an extra phage type-specific sequence between Common region 2 and rstR. The variations in the non-coding sequences of CTX phages might be associated with the difference in the replication efficiency and the directionality in the integration into the V. cholerae chromosome.

CTX Prophages in Vibrio cholerae O1 Strains

  • Kim, Eun Jin;Lee, Dokyung;Moon, Se Hoon;Lee, Chan Hee;Kim, Dong Wook
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.725-731
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    • 2014
  • The classical biotype strains of the Vibrio cholerae O1 serogroup harbor the biotype-specific cholera-toxin encoding phage (CTX) $CTX^{cla}$, and the El Tor biotype strains contain CTX-1. Although the classical biotype strains have become extinct, a remnant of classical CTX phage is transferred to the El Tor biotype strains. The prototype El Tor strains, which produce the biotype-specific cholera toxin, are now being replaced by atypical El Tor variant strains producing classical biotype cholera toxin. The genome sequences of the CTX phages in atypical El Tor strains indicate that the CTX phages in atypical El Tor strains are a mosaic of $CTX^{cla}$ and CTX-1. Before the emergence of atypical El Tor stains in the early 1990s, unusual pre-seventh pandemic strains were isolated in the US Gulf Coast between 1973 and 1986. These strains have characteristics of atypical El Tor strains since they are El Tor biotype strains containing $CTX^{cla}$, yet the genome sequence of this CTX phage indicates that it is different from $CTX^{cla}$ and is therefore classified separately as $CTX^{US\;Gulf}$.

Molecular Characterization of Plasmids Encoding CTX-M β-Lactamases and their Associated Addiction Systems Circulating Among Escherichia coli from Retail Chickens, Chicken Farms, and Slaughterhouses in Korea

  • Jo, Su-Jin;Woo, Gun-Jo
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.270-276
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    • 2016
  • Extended-spectrum β-lactamases (ESBLs), particularly those of the CTX-M types, are the predominant resistance determinants of Escherichia coli that are rapidly spreading worldwide. To determine CTX-M types, E. coli isolates were collected from retail chickens (n = 390) and environmental samples from chicken farms (n = 32) and slaughterhouses (n = 67) in Korea. Fifteen strains harboring blaCTX-M genes were isolated from 358 E. coli isolates. The most common CTX-M type was eight of CTX-M-15, followed by six of CTX-M-1 and one of CTX-M-14. The blaCTX-M genes were identified in the isolates from retail chickens (n = 9), followed by feces, water pipes, floors, and walls. Conjugations confirmed the transferability of the plasmids carrying blaCTX-M genes to the recipient E. coli J53 strain. Furthermore, eight addiction systems carried by the replicons in CTX-M types were confirmed. The dominant system was identified as ccdAB, vagCD, and pndAC in donor strains and transconjugants. The clonal relationship between the two strains carrying blaCTX-M genes indicates that E. coli may transmit from the farm to retail chickens, suggesting a possible public health risk. Our findings demonstrate that the detection of CTX-M types in E. coli isolates is important for tracking ESBL production in animals, and suggest linkage of multiple addiction systems in plasmids bearing blaCTX-M genes.

Characterization of CTX-M-14- and CTX-M-15-Producing Escherichia coli and Klebsiella pneumoniae Isolates from Urine Specimens in a Tertiary-Care Hospital

  • Kim, Semi;Sung, Ji Youn;Cho, Hye Hyun;Kwon, Kye Chul;Koo, Sun Hoe
    • 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.

Effects of Emitted-Qi Therapy(外氣療法), Youngyeonondamtang(靈連溫膽湯) and Jeokyeonondamtang(赤連溫膽湯) on Hematopoiesis System in Mice bearing Ascites Cancer (외기요법(外氣療法)과 영연온담탕(靈連溫膽湯) 및 적연온담탕(赤連溫膽湯)이 복수암(腹水癌) 생쥐의 조혈계통(造血系統)에 미치는 영향(影響))

  • Khil Ho-Sik;Kim Kwang-Ho
    • Journal of Society of Preventive Korean Medicine
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    • v.2 no.1
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    • pp.193-208
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    • 1998
  • In order to study the effects of emitted-qi therapy, Youngyeonondamtang and Jeokyeonondamtang on the mice bearing ascites cancer caused by Sarcoma 180, the author divided the mice into normal group, control group, EQ group, YO group, JO group, CTX+QE group, CTX+YO group, CTX+JO group, and treated with emitted -qi therapy and administrated Youngyeonondamtang, Jeokyeononondaintang and Cyclophosphamide (CTX). After collecting blood from the mice, measured the values of Prothrombin Time(PT), Partial Thromboplastin Time(PTT), Fibrinogen, White Blood Cell(WBC) and Platelet. The results were as follows, 1. PT was not showed any significant change in ever)r group, being compared with the control group. 2. H increased significantly in YO group, being compared with the control group. 3. Fibrinogen decreased significantly in EQ group, YO group, CTX+EQ group, CTX+YO group, CTX+JO group, being compared with the control group. 4. WBC increased significantly in CTX+EQ group, CTX+YO group, CTX+JO group, being compared with the CTX group. 5. Platelet decreased significantly in every group, being compared with the control group. These results suggest that emitted-qi therapy, Youngyeonondamtang and Jeokyeonondamtang had the effects that could heal the hematopoiesis system in the mice bearing ascites cancer, and especially had the better effects in the case that hematopoiesis system has been impaired by CTX.

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Characteristics of Transmissible CTX-M- and CMY-Type β-Lactamase-Producing Escherichia coli Isolates Collected from Pig and Chicken Farms in South Korea

  • Shin, Seung Won;Jung, Myunghwan;Won, Ho Geun;Belaynehe, Kuastros Mekonnen;Yoon, In Joong;Yoo, Han Sang
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1716-1723
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    • 2017
  • The rapid dissemination of extended-spectrum ${\beta}$-lactamase (ESBL)-producing Escherichia coli has significantly contributed to public health hazard globally. A total of 281 E. coli strains recovered from pigs and chickens between 2009 and 2015 in South Korea were analyzed for ESBL production. ESBL phenotypes were recognized in 14 E. coli isolates; ten and three ESBL-producing isolates carried only $bla_{CTX-M}$ and $bla_{CMY}$ genes, respectively, and one isolate harbored both genes. The predominant CTX-M and CMY types were CTX-M-15 (n = 8) and CMY-2 (n = 3). We also detected ESBL-producing isolates harboring $bla_{CTX-M-65}$, $bla_{CTX-M-14}$, $bla_{CMY-6}$, $bla_{DHA-1}$, and $bla_{TEM-1}$ genes. All ESBL-producing isolates showed resistance to the extent of the fourth generation cephalosporins, along with multidrug resistance. CTX-M-15-producing isolates showed higher MIC values than CTX-M-14- and CTX-M-65-producing isolates. The $bla_{CTX-M}$ and $bla_{CMY}$ genes have the potential to be transferable. The spreading of $bla_{CMY}$ and $bla_{CTX-M}$ genes was arbitrated mainly via Frep and IncI1 plasmids. Our isolates showed clonal diversity in PFGE analysis. This is the first report of E. coli isolates carrying $bla_{CMY-6}$ in chicken from South Korea. The emergence of CMY-6 ESBLs in a population of poultry suggests that extensive screening with long-term surveillance is necessary to prevent the dissemination of ESBL from chicken to human.

CTX-M β-lactamase and plasmid-mediated quinolone resistance genes in cefotaxime-resistant gram-negative bacteria isolated from companion animals (반려동물에서 분리된 cefotaxime 내성 그람 음성균에서 CTX-M β-lactamase와 plasmid 매개 퀴놀론 내성 유전자)

  • Cho, Jae-Keun;Lee, Jung-Woo;Kim, Jeong-Mi;Park, Dae-Hyun;Jeong, Ji-yeon
    • Korean Journal of Veterinary Service
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    • v.43 no.2
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    • pp.79-88
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    • 2020
  • The aim of this study was to investigate the prevalence of CTX-M β-lactamase and plasmid-mediated quinolone resistance (PMQR) genes, and the pattern of antibiotic resistance in cefotaxime-resistant gramnegative bacteria. A total 126 gram-negative bacteria were isolated from hospitalized dogs and cats between 2018 and 2019. The most predominant isolates were E. coli (n=41), followed by Pseudomonas aeruginosa (n=25), Proteus mirabilis (n=14), Klebsiella pneumoniae (n=9), Sphingomonas paucimobilis (n=7), and Enterobacter cloacae and Serratia marcescens (respectively, n=5). Cefotaxime-resistant isolates were identified in 26.2% (33 isolates) of 126 gram-negative bacteria. CTX-M type β-lactamase were found in 15 isolates (10 E. coli, 1 Ent, cloacae and 4 K. pneumoniae, respectively). Among the CTX-M producing gram-negative bacteria, CTX-M-1 and CTX-M-9 were detected in 10 (66.7%) and 5 (33.3%) isolates, respectively. While, CTX-M-2 and CTX-M-8 were not found. PMQR genes were detected in 12 (36.4%) isolates (4 E. coli, 2 Ent, cloacae and 6 K. pneumoniae, respectively), and the predominant PMQR gene was aac(6')-lb-cr (n=9), followed by qnrB (n=8) and qnrS (n=1) alone or in combination. qnrA and qepA were not found. Additionally, 9 (60%) of 12 PMQR positive isolates were co-existence with CTX-M-1 or CTX-M-9. CTX-M or PMQR producing isolates showed highly resistance to penicillins (100%), cephalosporins (100~66.7%), monobactams (72.2%), and non-β-lactam antibiotics (94.4~61.1%) such as quinolones, trimethoprim/sulfamethoxazole, tetracycline and gentamicin. These findings showed CTX-M-1, CTX-M-9, aac(6')-lb-cr and qnrB were highly prevalent in cefotaxime-resistant Enterobacteriaceae isolates from companion animals in our region. Moreover, PMQR genes were closely associated with CTX-M type β-lactamase.

Characteristics of the Molecular Epidemiology of CTX-M-Producing Escherichia coli Isolated from a Tertiary Hospital in Daejeon, Korea

  • Kim, Semi;Sung, Ji Youn;Cho, Hye Hyun;Kwon, Kye Chul;Koo, Sun Hoe
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1643-1649
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    • 2016
  • The aims of this study were to characterize the molecular epidemiological profiles of CTX-M-producing uropathogenic Escherichia coli isolates from a tertiary hospital in Daejeon, Korea, and to investigate the genetic diversity and compare the prevalence of sequence types (STs) in different areas. Extended spectrum β-lactamase-producing E. coli strains isolated from urine were analyzed for CTX-M, integrons, and insertion sequence common regions (ISCRs) by PCR and sequencing. Multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), phylogenetic analysis, and rep-PCR were also used for molecular typing of the isolates. Of 80 CTX-M producers, 31 and 46 expressed CTX-M-15 and CTX-M-14, respectively. MLST analysis indicated that the most prevalent ST was ST131 (n = 34, 42.5%), followed by ST38 (n = 22, 27.5%), ST405 (n = 8, 10.0%), and ST69 (n = 6, 7.5%). Most CTX-M producers harbored class 1 integrons. ST131 strains belonged to phylogenetic group B2 and showed identical rep-PCR patterns, whereas ST69, ST38, and ST405 strains belonged to phylogenetic group D; the ST38 and ST405 strains displayed the same rep-PCR pattern, respectively. ST131 and ST38 isolates showed 21 and 19 distinct types, respectively, by PFGE. In Daejeon, D-ST38 CTX-M-14 producers were relatively more prevalent than in other countries and Korean cities. Our results indicate that CTX-M-producing E. coli isolates belonged mostly to ST131 or ST38 and were more related to hospital-onset than to community-onset infections and that the blaCTX-M gene may vary according to the ST.

Characterization of CTX-M-Type Extended-Spectrum Beta-Lactamase-Producing Diarrheagenic Escherichia coli Isolates in the Republic of Korea During 2008-2011

  • Kim, Jin Seok;Kim, Junyoung;Kim, Soo-Jin;Jeon, Se-Eun;Oh, Kyung Hwan;Cho, Seung-Hak;Kang, Yeon-Ho;Han, Soon Young;Chung, Gyung Tae
    • 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.

Prevalence of CTX-M-type Extended-Spectrum $\beta$-Lactamases Producing Escherichia coli and Klebsieilla pneumoniae Isolates in General Hospitals in 2005 (임상에서 분리된 CTX-M형 Extended-Spectrum $\beta$-Lactamases를 생산하는 Escherichia coli와 Klebsiella pneumoniae의 유행)

  • Kim, Yun-Tae;Kim, Tae-Un
    • 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.