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 Korea원문보기
Jo, Su-Jin
(Laboratory of Food Safety and Evaluation, Department of Biotechnology, Korea University)
,
Woo, Gun-Jo
(Laboratory of Food Safety and Evaluation, Department of Biotechnology, Korea University)
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...
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.
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.
[36]. To confirm the presence of blaCTX-M β-lactamases, the sequences of the CTX-M-1, 8, 9, and 10 families were used for specific grouping. CTX-M-type families were detected by the PCR method.
이론/모형
The antimicrobial susceptibility was determined by using the disc diffusion method, in accordance with Clinical and Laboratory Standards Institute (CLSI) guidelines [6]. The antimicrobial agents (BBL) and their corresponding concentrations were as follows: ampicillin (AM, 10 μg), piperacillin (PIP, 100 μg), amoxicillin-clavulanic acid (AmC, 20/10 μg), cephalothin (CF, 30 μg), cefazolin (CZ, 30 μg), cefoxitin (FOX, 30 μg), cefamandole (MA, 30 μg), cefotaxime (CTX, 30 μg), ceftazidime (CAZ, 30 μg), cefepime (FEP, 30 μg), imipenem (IPM, 10 μg), amikacin (AN, 30 μg), aztreonam (ATM, 30 μg), streptomycin (S, 25 μg), gentamicin (GM, 10 μg), ciprofloxacin (CIP, 30 μg), nalidixic acid (NA, 30 μg), trimethoprim-sulfamethoxazole (SXT, 1.
For comparison, a cluster analysis was performed using the Pearson correlation similarity index. The relationship was calculated by the unweighted-pair group method using average linkages.
(Seoul, Korea). The sequences were confirmed to those in the GenBank nucleotide database using the Basic Local Alignment Search Tool (BLAST) program available through the National Center for Biotechnology Information website (http://www.ncbi.nlm.nih.gov/BLAST).
성능/효과
Four of 18 plasmid replicon types, including Inc I1-Iγ, Inc FIA, IncF, and IncY, were present either alone or in combination. All the plasmids expressing blaCTX-M-1, -15 genes were either positive within two replicon types or were non-typeable, whereas plasmids harboring the blaCTX-M-14 gene showed three replicon types, Y, FIA, and F (Table 2). The predominant replicon type was confirmed as F (66.
In addition, four addiction systems in the transconjugants would imply that addiction systems might be located on a non-conjugative plasmid or in the chromosome, suggesting that those located on conjugative plasmids harbor resistance markers [32]. Furthermore, among the transconjugants, the ccdAB, vagCD, and parDE systems were associated with the IncF, IncFIA, and IncFIB replicon types, but the pndAC system was associated with the IncI1-Iγ replicon type (data not shown here). Mnif et al.
In conclusion, this study has reported the identification and characterization of CTX-M β-lactamases and their associated addiction systems. Further analysis illustrated that the detection of CTX-M-producing E.
In this study, blaCTX-M-15 was the predominant gene, followed by blaCTX-M-1 and blaCTX-M-14. The blaCTX-M genes characterized in the samples obtained from healthy chickens, chicken farms, and slaughterhouses were previously observed in animals in Korea [16,31,32].
Plasmid addiction systems can contribute to the stability and maintenance of plasmids in their hosts and may affect their distribution in the absence of antibiotic selection [8,11,24,32]. In this study, the fact that the donor strains carried blaCTX-M genes confirmed the association betweenthe eight addiction systems and the IncF plasmid replicon type. A previous study has shown that the IncF plasmid replicon type has developed the ability to diffuse in E.
1). The results revealed that 13 of 15 isolates were not genetically related, whereas the remaining two strains (13-ECO-VE-338 and 13-ECO-VE-339) isolated from the floor and chicken feces in the same farm showed high similarity to 90%. Furthermore, these two strains showed the same characteristics in CTX-M type and antimicrobial resistance pattern.
It means that the spread of blaCTX-M genes were associated with IncF plasmids [20]. Therefore, these findings suggest that all the replicon types in the strains isolated from chickens were similar to the genetic structure of the plasmids.
후속연구
coli harboring CTX-M via foodborne transmission or environmental pathways can affect human health. Therefore, further analysis is required to assess the risk posed by these microorganisms to human health, with respect to a food safety perspective.
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