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Almost all cases of carbapenemase-producing Enterobacteriaceae infections in Japan are caused by  bla  IMP -positive Enterobacteriaceae (especially  bla  IMP-6 ) and infections caused by other types of carbapenemase-producing Enterobacteriaceae are quite rare. We examined drug resistance genes co-harboring with  bla  IMP-6  and their inoculum size effects. We screened β-lactamase genes, plasmid-mediated quinolone resistance (PMQR) genes, and aminoglycoside-modifying enzyme genes by PCR and performed sequencing for 14  bla  IMP-6 -positive Enterobacteriaceae. Further, all PMQR-positive isolates were submitted to conjugation and inoculum effect evaluation. Our data showed that 13 of the 14 isolates harbored CTX-M-2 and one co-harbored CTX-M-2 and CTX-M-1 as extended-spectrum β-lactamases. All isolates carried one or more PMQRs;  aac(6’)-Ib-cr  was the most prevalent (92.8%), and was followed by  oqxA  (64.3%),  qnrS  (50%),  oqxAB  (21.4%), and  qnrB  (14.3%). However,  Klebsiella pneumoniae  contains chromosomal OqxAB. Inoculum size effects were significant in all strains for meropenem, 13 strains for imipenem, 7 for levofloxacin, and 3 for amikacin. We observed that 11 of the experimental strains (100%), 8 strains (72.7%), and 1 strain showed inoculum size effects for meropenem, imipenem, and amikacin, respectively. However, four strains harbored  qnr  genes and two strains harbored  qnr  genes and QRDR mutations concurrently; no inoculum size effect was seen for levofloxacin. The  bla  IMP-6 -positive Enterobacteriaceae that we studied was found to harbor at least one plasmid-mediated drug resistance gene. The inoculum size effect for carbapenems was thought to be mainly due to IMP-6-type metallo-β-lactamase; however  qnrB  and  qnrS  also had a minimal impact on the inoculum size effect for levofloxacin.

plos one

Comparison of the inoculum size effects of antibiotics on IMP-6 β-lactamase-producing Enterobacteriaceae co-harboring plasmid-mediated quinolone resistance genes