Characteristics of Transmissible CTX-M- and CMY-Type β-Lactamase-Producing Escherichia coli Isolates Collected from Pig and Chicken Farms in South Korea원문보기
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 pr...
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.
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.
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문제 정의
The finding of phylogenetic typing was applied to determine the pattern of antimicrobial resistance and the β-lactamase gene distributions among the ESBL-producing E. coli isolates investigated in this work.
However, much of them focused on the prevalence of resistant strains and resistance gene. Therefore, this work aimed to study the prevalence of ESBL-producing E. coli isolates recovered from pigs and chickens that had been collected in South Korea, and also to analyze the transfer of ESBL-associated genes and resistance among E. coli strains.
제안 방법
In this study we investigated ESBL-producing E. coli isolates harboring various types of β-lactamase genes.
대상 데이터
DNA templates f or PCR were obtained by centrifugation of bacterial colony suspensions grown on tryptic soy broth in 500 μl of distilled water, following boiling for 10 min at 100℃. Sequence alignment and comparison with known sequences were carried out by searching the GenBank database using the BLAST search tool available at the National Center for Biotechnology Information Website (http://www.ncbi.nlm.nih.gov/BLAST).
이론/모형
All isolates producing ESBL were subjected to phylogenetic grouping to assign into one of the four phylogenetic groups (A, B1, B2, and D) using a multiplex PCR method [23]. This was performed by targeting two genetic determinants (chuA and yjaA) and an anonymous DNA fragment (TSPE4.
Bacterial strains were assessed for antimicrobial susceptibility using a disc diffusion assay, according to the Clinical and Laboratory Standards Institute (CLSI) procedure [18]. The antimicrobial agents (Oxoid, UK) were as follows: ampicillin, 10 μg; ceftazidime, 30 μg; cefotaxime, 30 μg; ciprofloxacin, 5 μg; gentamicin, 10 μg; nalidixic acid, 30 μg; tetracycline, 30 μg; and trimethoprim-sulfamethoxazole, 1.
Conjugation assays were performed to determine the transferability of β-lactamase resistance genes on ESBL-producing isolates by using the broth mating method.
Similarity analysis was performed by using the Dice coefficient, and clustering was done by the unweighted-pair group method using average linkages. For 14 ESBL-producing E.
Analysis of gel images were performed using GelCompar II software (Applied Maths, Belgium). Unweighted pair group method with arithmetic means analysis was employed to construct clustering based on the Dice similarity index.
성능/효과
In this study, the MIC test for five β-lactam agents showed that the average MIC of each agent for isolates that produced CTX-M-15 was higher than that produced CTX-M-65 and CTX-M-14, especially for cefotaxime and ceftriaxone (data not shown).
In this study, three of the E. coli strains carrying blaCTX-M genes and one isolate carrying blaCMY genes belonged to phylogenetic groups B2 and D, known for their virulence. These results may raise interest concerning the transfer of CTX-M- or CMY-producing E.
Three different types of blaCTX-M, namely, blaCTX-M-65, blaCTX-M-15, and blaCTX-M-14 and two different CMY-type (blaCMY-2 and blaCMY-6) β-lactamase genes were observed.
후속연구
More studies are needed with a better long-term surveillance to trace the evolution and dissemination of CTX-M- and CMY-type β-lactamases between different food-producing animals.
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