Emergence of a multidrug-resistant Pseudomonas fulva clinical isolate co-harboring tmexCD3-toprJ3, bla (OXA-1), and bla (IMP-45) on a transferable megaplasmid

携带tmexCD3-toprJ3、bla(OXA-1)和bla(IMP-45)基因的多重耐药性黄假单胞菌临床分离株的出现,该基因位于可转移的巨型质粒上

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Abstract

INTRODUCTION: Pseudomonas fulva is a non-fermentative bacterium with both environmental adaptability and pathogenic potential that has been increasingly detected in clinical infections in recent years. This study reports the first clinical P. fulva (PF1) isolate carrying tmexCD3-toprJ3, bla (OXA-1), and bla (IMP-45) resistance genes, which was recovered from the urine of a patient with a urinary tract infection. METHODS: Species identification was performed using whole-genome sequencing and average nucleotide identity analysis, with phylogenetic placement determined by 16S rRNA. Genomic analysis identified resistance genes and plasmid structures, while plasmid transferability was assessed experimentally. Biofilm formation, stress tolerance, and virulence were evaluated using crystal violet staining, stress assays, and the Galleria mellonella model. RESULTS: PF1 exhibited resistance to carbapenems, cephalosporins, quinolones, and an elevated tigecycline MIC of 256 mg/mL, while remaining susceptible to polymyxins. The strain harbors a 449-kb transferable megaplasmid (pPF1), containing resistance genes bla (IMP-45), bla (OXA-1), and a mutated tmexCD3-toprJ3 efflux pump(T1827G and T1830C). Phylogenetic analysis showed >99% genomic similarity to clinical isolates from eastern China. PF1 demonstrated strong biofilm formation (OD620 = 3.73 ± 0.14), stress tolerance, and moderate virulence in Galleria mellonella. DISCUSSION: This study reveals the potential of P. fulva to acquire multidrug resistance and adapt to clinical environments, underscoring the need for enhanced surveillance of resistance genes in atypical Pseudomonas species.

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