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1.
Antimicrob Agents Chemother ; 67(11): e0067523, 2023 11 15.
Artículo en Inglés | MEDLINE | ID: mdl-37819082

RESUMEN

Pseudomonas aeruginosa high-risk clones pose severe threats to public health. Here, we characterize the imipenem/relebactam (IR) resistance mechanisms in P. aeruginosa high-risk clones sequence type 235 (ST235) and ST463 in China. Minimum inhibitory concentrations (MICs) were determined, and Illumina short-read sequencing was performed for 1,168 clinical carbapenem-resistant P. aeruginosa (CRPA) isolates. The gene copy number and expression level were analyzed by Illumina sequencing depth and reverse transcription-quantitative PCR, respectively. Resistance conferred by bla GES-5 was evaluated by cloning experiments. ST463 and ST235 accounted for 9.8% (115/1,168) and 4.5% (53/1,168) of total isolates, respectively, and showed high frequencies of extensively drug-resistant and difficult-to-treat resistant phenotypes. The overall IR-resistant rate in CRPA was 21.0% (245/1,168). However, the IR resistance rate was 81.7% (94/115) in ST463-PA and 52.8% (28/53) in ST235-PA. Of the ST463 isolates, 92.2% (106/115) were Klebsiella pneumoniae carbapenemase-producing P. aeruginosa (KPC-PA), and all 94 IR-resistant ST463-PA produced KPC-2. Compared to IR-susceptible ST463 KPC-2-PA, IR-resistant ST463 KPC-2-PA exhibited significantly higher bla KPC-2 copy numbers and expression levels. In ST463 KPC-2-PA, 16 mg/L relebactam resulted in additional fourfold reductions in imipenem MIC50/90 values compared to 4 mg/L relebactam. In ST235, 1.9% (1/53) carried bla IMP carbapenemase and 54.7% (29/53) carried bla GES carbapenemase. Other than the IMP producer, all 27 IR-resistant ST235-PA produced GES-5. Cloning experiments revealed that imipenem resistance in bla GES-5-carrying PAO1 transformants was generally unaffected by relebactam. In conclusion, IR-resistant CRPA isolates in China were mainly distributed in P. aeruginosa high-risk clones ST463 and ST235. The major underlying IR resistance mechanisms were bla KPC-2 overexpression and bla GES-5 carriage.


Asunto(s)
Infecciones por Pseudomonas , Pseudomonas aeruginosa , Humanos , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , beta-Lactamasas/metabolismo , Carbapenémicos/uso terapéutico , Células Clonales/metabolismo , Imipenem/farmacología , Imipenem/uso terapéutico , Pruebas de Sensibilidad Microbiana , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/metabolismo , Infecciones por Pseudomonas/tratamiento farmacológico
2.
J Hosp Infect ; 121: 128-131, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-34906601

RESUMEN

Serratia marcescens is a nosocomial pathogen with carbapenem resistance, which limits the availability of effective treatment options. In this study, molecular characterization of GES-5 carbapenemase-producing S. marcescens isolated from an outbreak in Japan was undertaken. Comparative genetic analysis revealed that the blaGES-5-encoding plasmid p2020-O-9 is a unique plasmid contributing to carbapenem resistance. Furthermore, this study highlights the need for surveillance programmes to monitor both novel and commonly occurring carbapenemases in clinical settings.


Asunto(s)
Infección Hospitalaria , Infecciones por Serratia , Antibacterianos/farmacología , Infección Hospitalaria/epidemiología , Brotes de Enfermedades , Humanos , Japón/epidemiología , Pruebas de Sensibilidad Microbiana , Plásmidos/genética , Infecciones por Serratia/epidemiología , Serratia marcescens/genética , beta-Lactamasas/genética
3.
J Glob Antimicrob Resist ; 31: 196-206, 2022 12.
Artículo en Inglés | MEDLINE | ID: mdl-36180037

RESUMEN

OBJECTIVES: This study aimed to characterize Gram negative bacteria carrying blaGES carbapenemase genes detected in wastewater from a hospital with no history of detection of clinical isolates producing GES carbapenemases. METHODS: Six hospital effluent samples were screened for carbapenemase-producing organisms (CPO) using CHROMagar mSuperCARBA and MacConkey agar with 1 µg/mL imipenem. Polymerase chain reaction (PCR) amplification and sequencing of carbapenemase genes, multilocus sequence typing, antimicrobial susceptibility testing, and whole-genome sequencing were performed. RESULTS: Among 21 CPO isolates, 11 Klebsiella spp. and 5 Enterobacter kobei isolates carried blaGES-24, and 4 E. roggenkampii and 1 Pseudomonas aeruginosa isolates carried blaGES-5. Genomic analysis of 8 representative isolates comprising 6 blaGES-24-positive and 2 blaGES-5-positive revealed that class 3 integrons with complete or defective Tn402-like transposition modules were predominantly associated with two tandem copies of blaGES-24. Furthermore, a total of 5 new class 3 integrons, In3-18 to In3-22, were identified among 5 blaGES-24 and 1 blaGES-5 plasmids. One strain each of K. pneumoniae subsp. pneumoniae and K. quasipneumoniae subsp. similipneumoniae harboring blaGES-24 plasmids also carried a rare blaVEB-1-positive class 1 integron on a non-typeable plasmid, where these blaVEB-1 plasmids had high sequence similarity. Virulence gene profiles differed between Klebsiella spp. and Enterobacter spp.; the former harbored type III fimbriae cluster, salmochelin, and T6SS type i2 gene clusters, while the latter had curli pili operon, aerobactin, T2SS gene clusters, and T6SS type i3 gene clusters. CONCLUSION: Our findings confirmed the linkage of blaGES-24 with rare Tn402-like class 3 integrons and the structural diversity of their gene cassette arrays.


Asunto(s)
Integrones , Aguas Residuales , Integrones/genética , Bacterias Gramnegativas , Hospitales , Genómica
4.
J Glob Antimicrob Resist ; 29: 116-119, 2022 06.
Artículo en Inglés | MEDLINE | ID: mdl-35231657

RESUMEN

OBJECTIVES: The aim of the study is to characterise the genomic features of three GES-producing Enterobacterales isolates from Czech hospitals. METHODS: In 2020, during a routine screening of the hospital's surfaces in Prague General Hospital, two strains (CZ862 and CZ863) that belonged to the Enterobacter cloacae complex were found to be blaGES positive. Another blaGES positive strain identified as Klebsiella oxytoca was recovered from a patient hospitalised in Pilsen. Antibiotic susceptibility profiling was done with broth microdilution assay. Conjugation/transformation experiments were performed on all three strains. Genomic DNA of the three isolates was subjected to whole genome sequencing using PacBio platform. RESULTS: Multilocus sequence types typing of CZ862 and CZ863 identified the strains as ST837 and a novel ST (ST1622). Both blaGES harbouring plasmids showed high sequence similarity and complete query coverage (100% and 99.98%) with pEcl-35771cz. Both plasmids had two copies of blaGES instead of one copy as found in pEcl-35771cz. The clinical isolate CZ598 belonged to ST180. The plasmid harboured blaGES-7 gene, cat and aac(6')-lb and the novel variant blaOXA-1011. No similar sequences were observed, suggesting a novel plasmid. CONCLUSION: The detection of the two blaGES-positive plasmids in the same hospital environment, the first report after 3 years, suggests a hidden source. This highlights the importance of the hidden sources and evolution of such plasmids on the route of spreading into clinical settings. Also, the detection of the new blaOXA-1011, which is thought in this case to be associated with carbapenem resistance, imposes a health risk if disseminated, limiting therapeutic options.


Asunto(s)
Klebsiella oxytoca , beta-Lactamasas , República Checa , Genómica , Humanos , Klebsiella oxytoca/genética , Plásmidos/genética , beta-Lactamasas/genética
5.
J Glob Antimicrob Resist ; 24: 220-227, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33385587

RESUMEN

OBJECTIVES: The spread of carbapenemase-producing Enterobacterales (CPE) with colistin resistance is a critical public health issue. We genetically characterized the clinical isolate Enterobacter roggenkampii OIPH-N260, which harboured carbapenemase genes blaIMP-1 and blaGES-5 with multiple resistance genes, including mcr-9 and blaCTX-M-9. METHODS: This isolate was characterized by whole-genome sequencing, comparative analysis of resistance plasmids, susceptibility tests, bacterial conjugation, S1-nuclease digested pulsed-field-gel electrophoresis, and Southern blot hybridization. RESULTS: The OIPH-N260 isolate exhibited resistance to most ß-lactams and colistin. It co-harboured two resistance plasmids, the blaIMP-1- and blaGES-5-encoding IncP6 plasmid pN260-3 and mcr-9- and blaCTX-M-9-encoding IncHI2 plasmid pN260-1. The comparative analysis of pN260-3 indicated that a unique blaIMP-1-surrounding region was inserted into the blaGES-5-encoding plasmid with the mobile element IS26, which plays an important role in the spread of resistance genes. pN260-1 did not possess the mcr-9 expression regulative gene qseBC. Both plasmids were transferable into other bacterial species via conjugation. CONCLUSIONS: This is the first study to report not only a blaIMP-1 and blaGES-5 co-encoding plasmid, but also the co-harbouring of another plasmid carrying mcr-9 and blaCTX-M-9 in Enterobacter cloacae complex. The development of advanced resistance via IS26-mediated insertion and the co-harbouring of resistance plasmids highlights the need to monitor for resistance genes in CPE.


Asunto(s)
Antibacterianos , Enterobacter , Antibacterianos/farmacología , Genómica , Japón , Pruebas de Sensibilidad Microbiana , Plásmidos/genética
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