Characterization of a Conjugative Multidrug Resistance IncP-2 Megaplasmid, pPAG5, from a Clinical Pseudomonas aeruginosa Isolate

Li, Meng; Guan, Congcong; Song, Gaoyu; Gao, Xiaoxi; Yang, Weina; Wang, Tietao; Zhang, Yani

doi:10.1128/spectrum.01992-21

Cited by 8 publications

(5 citation statements)

References 38 publications

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“…We found an alignment (55,243 aa) of these concatenated sequences to share between 97.4 % and 100 % identity, with the sequences of plasmids pWTJH12-KPC (CP064404) and pZPPH29-KPC (CP077978) identical to one another (they were from isolates recovered in the same hospital (Y. Li et al, 2022)). Phylogenetic analysis confirmed how closely related the newly identified megaplasmids were to pP9Me1 and the reference pBT2436-like plasmids ( Figure (6) ).…”

Section: Resultsmentioning

confidence: 94%

Phenotypic and genomic characterization ofPseudomonas aeruginosaisolates recovered from catheter-associated urinary tract infections in an Egyptian hospital

Eladawy

Thomas

Hoyles

2023

Preprint

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Catheter-associated urinary tract infections (CAUTIs) represent one of the major healthcare-associated infections, andPseudomonas aeruginosais a common Gram-negative bacterium associated with catheter infections in Egyptian clinical settings. The present study describes the phenotypic and genotypic characteristics of 31P. aeruginosaisolates recovered from CAUTIs in an Egyptian hospital over a 3-month period. Genomes of isolates were of good quality and were confirmed to beP. aeruginosaby comparison to the type strain (average nucleotide identity, phylogenetic analysis). Clonal diversity among the isolates was determined; eight different sequence types were found (STs 244, 357, 381, 621, 773, 1430, 1667 and 3765), of which 357 and 773 are considered high-risk clones. Antimicrobial resistance (AMR) testing according to EUCAST guidelines showed the isolates were highly resistant to quinolones [ciprofloxacin (12/31, 38.7 %) and levofloxacin (9/31, 29 %) followed by tobramycin (10/31, 32.5 %)], and cephalosporins (7/31, 22.5 %). Genotypic analysis of resistance determinants predicted all isolates to encode a range of AMR genes, including those conferring resistance to aminoglycosides, β-lactamases, fluoroquinolones, fosfomycin, sulfonamides, tetracyclines and chloramphenicol. One isolate was found to carry a 422,938 bp pBT2436-like megaplasmid encoding OXA-520, the first report from Egypt of this emerging family of clinically important mobile genetic elements. All isolates were able to form biofilms, and were predicted to encode virulence genes associated with adherence, antimicrobial activity, antiphagocytosis, phospholipase enzymes, iron uptake, proteases, secretion systems, and toxins. The present study shows how phenotypic analysis alongside genomic analysis may help us understand the AMR and virulence profiles ofP. aeruginosacontributing to CAUTIs in Egypt.

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Section: Resultsmentioning

confidence: 94%

Phenotypic and genomic characterization ofPseudomonas aeruginosaisolates recovered from catheter-associated urinary tract infections in an Egyptian hospital

Eladawy

Thomas

Hoyles

2023

Preprint

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“…As of 2022, the largest plasmid found to be carried by P. aeruginosa is a multidrug resistance plasmid pPAG5, which was found in a clinical strain of P. aeruginosa PAG5 isolated from a patient's urine, with a size of 513.3 kb (Li et al, 2022 ). Compared to pPAG5, pBJ86 has a larger size of 522.5 kb, and they both belong to megaplasmids.…”

Section: Discussionmentioning

confidence: 99%

“…Some pathogenic strains of Pseudomonas have already established environmental hosts, and resistance genes can be horizontally transferred from environmental organisms to human symbionts, which can bring direct public health impacts (Baker-Austin et al, 2006 ). In recent years, more and more studies have detected heavy metal resistance genes in clinical isolates, which typically coexist with various ARGs on mobile elements (Perez-Palacios et al, 2021 ; Li et al, 2022 ), and may be related to the co-selection mechanism between resistance genes. The co-selection mechanism includes co-resistance and cross-resistance.…”

Section: Discussionmentioning

confidence: 99%

Characterization of a mobilizable megaplasmid carrying multiple resistance genes from a clinical isolate of Pseudomonas aeruginosa

Mei,

Song,

Liu

et al. 2023

Front. Microbiol.

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IntroductionThe horizontal transfer of antibiotic resistance genes mediated by plasmids seriously hinders the effectiveness of modern medical treatment, and thus has attracted widespread attention. Additionally, the co-selection mechanism of antibiotic resistance genes (ARGs) and heavy metal resistance genes (MRGs) on mobile elements may further exacerbate the horizontal transfer of resistance genes.MethodsIn this study, a multidrug-resistant Pseudomonas aeruginosa strain, termed BJ86 (CHPC/NPRC1.4142), was isolated from a patient's sputum specimen. In vitro tests for antimicrobial susceptibility, conjugation, whole-genome sequencing, and bioinformatics analysis were used to explore the potential mechanisms of resistance and its spread.Results and discussionSequencing analysis indicates that P. aeruginosa BJ86 carries an amazing 522.5 kb-length megaplasmid, pBJ86, which contained a 93.5 kb-length multiple resistance region (MRR); 18 kinds of genes were identified as ARGs in this region, including tmexCD-oprJ, blaDIM−1, qnrVC6 that mediate resistance to multiple antibiotics and the operons mer that mediates heavy metal mercury resistance. In addition, there is also an 80 kb variable region (VR) on the plasmid pBJ86, and the genes encoding relaxase and type IV coupling protein (T4CP) were determined in this region, both of which are related to the conjugation and transfer ability of the plasmid. Bioinformatics analysis shows that many functional genes have insertion sequences and transposases on their flanks, which may have accumulated in the plasmid pBJ86 after multiple acquisition events. Conjugated transfer and in vitro tests for antimicrobial susceptibility verified the mobility and plasmid pBJ86-mediated resistance. To our knowledge, we are the first to report a mobilizable megaplasmid that simultaneously carried tmexCD-oprJ, blaDIM−1, qnrVC6, and the operons mer and can be transferred with frequencies of 6.24 × 10−7 transconjugants per donor cell.

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“…This is one of the reasons for the high mortality of infections caused by P. aeruginosa . Of which, the acquisition of antimicrobial in P. aeruginosa mainly through the horizontal transfer of plasmids [ 18 ]. Currently, IncP incompatibility groups have been reported in P. aeruginosa , include IncP-1 to IncP-7 and IncP-9 to IncP-14 [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

Emergence and clonal dissemination of KPC-3-producing Pseudomonas aeruginosa in China with an IncP-2 megaplasmid

Qiao

Zheng

et al. 2023

Ann Clin Microbiol Antimicrob

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Background Despite the global prevalence of Klebsiella pneumoniae Carbapenemase (KPC)-type class A β-lactamases, occurrences of KPC-3-producing isolates in China remain infrequent. This study aims to explore the emergence, antibiotic resistance profiles, and plasmid characteristics of blaKPC-3-carrying Pseudomonas aeruginosa. Methods Species identification was performed by MALDI-TOF-MS, and antimicrobial resistance genes (ARGs) were identified by polymerase chain reaction (PCR). The characteristics of the target strain were detected by whole-genome sequencing (WGS) and antimicrobial susceptibility testing (AST). Plasmids were analyzed by S1-nuclease pulsed-field gel electrophoresis(S1-PFGE), Southern blotting and transconjugation experiment. Results Five P. aeruginosa strains carrying blaKPC-3 were isolated from two Chinese patients without a history of travelling to endemic areas. All strains belonged to the novel sequence type ST1076. The blaKPC-3 was carried on a 395-kb IncP-2 megaplasmid with a conserved structure (IS6100-ISKpn27-blaKPC-3-ISKpn6-korC-klcA), and this genetic sequence was identical to many plasmid-encoded KPC of Pseudomonas species. By further analyzing the genetic context, it was supposed that the original of blaKPC-3 in our work was a series of mutation of blaKPC-2. Conclusions The emergence of a multidrug resistance IncP-2 megaplasmid and clonal transmission of blaKPC-3-producing P. aeruginosa in China underlined the crucial need for continuous monitoring of blaKPC-3 for prevention and control of its further dissemination in China.

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Characterization of a Conjugative Multidrug Resistance IncP-2 Megaplasmid, pPAG5, from a Clinical Pseudomonas aeruginosa Isolate

Abstract: The emergence and spread of plasmid-associated multidrug resistance in bacterial pathogens is a key global threat to public health. It is important to understand the mechanisms of the formation and evolution of these plasmids in patients, hospitals, and the environment.

Cited by 8 publications

References 38 publications

Phenotypic and genomic characterization ofPseudomonas aeruginosaisolates recovered from catheter-associated urinary tract infections in an Egyptian hospital

Phenotypic and genomic characterization ofPseudomonas aeruginosaisolates recovered from catheter-associated urinary tract infections in an Egyptian hospital

Characterization of a mobilizable megaplasmid carrying multiple resistance genes from a clinical isolate of Pseudomonas aeruginosa

Emergence and clonal dissemination of KPC-3-producing Pseudomonas aeruginosa in China with an IncP-2 megaplasmid

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