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mSphere ; 4(5)2019 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-31554719


The continuous emergence of carbapenemase-producing Enterobacteriaceae (CPE) presents a great public health challenge. Mitigation of CPE spread in the environment is crucial, particularly from a One Health perspective. Here we describe the isolation of CPE strain SNI47 from influent water of a sewage treatment plant in Japan. SNI47 was identified as Klebsiella quasipneumoniae subsp. quasipneumoniae by phylogenetic analysis and was resistant to ß-lactams, including carbapenems. Of four plasmids detected from SNI47, the 185,311-bp IncA/C2 plasmid (pTMSNI47-1), which carried 10 drug resistance genes, including genes for four ß-lactamases (bla CTX-M-2, bla DHA-1, bla KHM-1, and bla OXA-10), was transferred to Escherichia coli J53 via conjugation. The MICs of all tested ß-lactams for the transconjugant were higher than for the recipient. We constructed recombinant plasmids, into which each ß-lactamase gene was inserted, and used them to transform E. coli DH5α cells, demonstrating that KHM-1 enhanced carbapenem resistance. In addition, these ß-lactamases were responsible for a wide-spectrum ß-lactam resistance acquisition with mutual compensation. KHM-1, recognized as a rare type of metallo-ß-lactamase, was detected in a transferable plasmid, from a sewage treatment plant, involved in horizontal gene transfer. The detection of such plasmids raises a health risk alarm for CPE dissemination.IMPORTANCE In our investigation of urban wastewater in Japan, carbapenem-resistant Klebsiella quasipneumoniae subsp. quasipneumoniae was isolated that carried the pTMSNI47-1 plasmid, which carries four ß-lactamase genes and has transferability among Enterobacteriaceae pTMSNI47-1 was found to encode a rarely reported carbapenemase, KHM-1. Cooperative effects of ß-lactamases encoded by pTMSNI47-1 appeared to have broad-spectrum resistance to ß-lactams. The detection of the KHM-1 gene in urban wastewater suggests that such a rare antimicrobial resistance (AMR) gene can be pooled in the environment, potentially emerging as an AMR determinant in a pathogen. When the number of ß-lactamase resistance genes is increased in one plasmid, the transfer of this plasmid can confer broad-spectrum resistance to ß-lactams, even if the individual gene confers narrow-spectrum resistance. The present study adds important information about the potential risk of sewage treatment plants as reservoirs and environmental suppliers of AMR genes, contributing to the public health from a One Health perspective.

Shokuhin Eiseigaku Zasshi ; 60(3): 45-51, 2019.
Artigo em Japonês | MEDLINE | ID: mdl-31391410


This study aimed to survey the trend of antimicrobial resistance in Escherichia coli obtained from retail meat. We examined the susceptibilities of 1,115 E. coli isolates obtained from chicken, beef, pork, venison, and wild boar meat from 2011 to 2017 in Tokyo to 14 antimicrobials (ampicillin, cefotaxime (CTX), streptomycin, gentamicin, kanamycin, tetracycline (TC), chloramphenicol, nalidixic acid, ciprofloxacin, sulfamethoxazole-trimethoprim, fosfomycin, amikacin, imipenem, and meropenem). Of all the tested isolates, 18.7% (135/721) isolates from chicken, 77.0% (117/152) from beef, 46.6% (89/187) from pork, 100% (28/28) from venison, and 92.6% (25/27) from wild boar meat were susceptible to all tested antimicrobials. Furthermore, TC resistance was the most common, with rates as high as 56.7% (409/721) and 40.6% (76/187) in the isolates from chicken and pork, respectively. CTX resistance was detected in 4.9% (25/506) of the isolates from domestic chicken and 23.7% (51/215) of the isolates from imported chicken. Moreover, CTX resistance rate in isolates from domestic chicken was significantly lower in 2016 (0.9%, 1/111) and in 2017 (0.8%, 1/121) than in 2012 (10.6%, 17/161). In conclusion, E. coli isolates from retail meat were most commonly resistant to TC, and CTX resistance was higher in E. coli isolates from imported chicken than in E. coli isolates from domestic chicken.

Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Escherichia coli/efeitos dos fármacos , Microbiologia de Alimentos , Carne/microbiologia , Animais , Bovinos , Galinhas , Cervos , Testes de Sensibilidade Microbiana , Prevalência , Suínos , Tóquio
Microbiol Immunol ; 61(12): 554-557, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29052266


In this study, the presence of the mcr-1 gene in Escherichia coli from retail meat in Japan was investigated. Nine E. coli isolates (eight from chickens and one from pork) carried the mcr-1 gene on the plasmid. In six isolates from domestic chickens, mcr-1 was located on the IncI2 plasmid, which is approximately 60 kb in size. In the remaining three isolates from imported chicken and pork, mcr-1 was located on the IncX4 plasmid (30 kb).

Antibacterianos/farmacologia , Colistina/farmacologia , Farmacorresistência Bacteriana , Proteínas de Escherichia coli/genética , Escherichia coli/efeitos dos fármacos , Escherichia coli/isolamento & purificação , Carne/microbiologia , Animais , Galinhas , Escherichia coli/classificação , Proteínas de Escherichia coli/metabolismo , Contaminação de Alimentos/análise , Contaminação de Alimentos/economia , Contaminação de Alimentos/estatística & dados numéricos , Japão , Carne/economia , Plasmídeos/genética , Plasmídeos/metabolismo , Suínos
J Vet Med Sci ; 78(7): 1183-7, 2016 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-27000951


We surveyed prevalence and contamination levels of Listeria monocytogenes in ready-to-eat foods between 2000 and 2012 in Tokyo. L. monocytogenes was isolated from 52 (1.7%) out of 2,980 samples. Comparing the prevalence in the study period, 2.2% were positive in the former period (2000-2005) and 1.2% in the latter (2006-2012). Using the most probable number (MPN) technique, 32 samples were contaminated with fewer than 0.3 L. monocytogenes/g, 10 samples with 0.3-1.0/g and 4 samples with more than 1.0/g (the maximum was 2.3/g). The most common serovar was 1/2a, followed by 1/2b, 4b and 1/2c. We revealed that ready-to-eat foods in Tokyo were contaminated with L. monocytogenes, although the contamination levels were low.

Fast Foods/microbiologia , Contaminação de Alimentos , Microbiologia de Alimentos , Listeria monocytogenes/isolamento & purificação , Contaminação de Alimentos/estatística & dados numéricos , Japão , Prevalência
J Vet Med Sci ; 78(3): 477-9, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26537550


PCR serogrouping methods were used to examine strains of L. monocytogenes isolated in Japan. Among 187 strains, 99.5% were classified into 4 PCR serogroups corresponding to conventional serotypes. Only one isolate had a new PCR profile, which may be a variant of serogroup IVb.

Listeria monocytogenes/classificação , Japão , Listeria monocytogenes/isolamento & purificação , Tipagem Molecular/métodos , Reação em Cadeia da Polimerase Multiplex , Sorotipagem/métodos