IncFIIk plasmid harbouring an amplification of 16S rRNA methyltransferase-encoding gene rmtH associated with mobile element ISCR2
Objectives To investigate the resistance mechanisms and genetic support underlying the high resistance level of the Klebsiella pneumoniae strain CMUL78 to aminoglycoside and β-lactam antibiotics. Methods Antibiotic susceptibility was assessed by the disc diffusion method and MICs were determined by the microdilution method. Antibiotic resistance genes and their genetic environment were characterized by PCR and Sanger sequencing. Plasmid contents were analysed in the clinical strain and transconjugants obtained by mating-out assays. Complete plasmid sequencing was performed with PacBio and Illumina technology. Results Strain CMUL78 co-produced the 16S rRNA methyltransferase (RMTase) RmtH, carbapenemase OXA-48 and ESBL SHV-12. The rmtH - and bla SHV-12 -encoding genes were harboured by a novel ∼115 kb IncFII k plasmid designated pRmtH, and bla OXA-48 by a ∼62 kb IncL/M plasmid related to pOXA-48a. pRmtH plasmid possessed seven different stability modules, one of which is a novel hybrid toxin–antitoxin system. Interestingly, pRmtH plasmid harboured a 4-fold amplification of an rmtH -IS CR2 unit arranged in tandem and inserted within a novel IS 26 -based composite transposon designated Tn 6329 . Conclusions This is the first known report of the 16S RMTase-encoding gene rmtH in a plasmid. The rmtH -IS CR2 unit was inserted in a composite transposon as a 4-fold tandem repeat, a scarcely reported organization.
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