Expansion of plasmid mediated blaACT-2 among Pseudomonas aeruginosa associated with postoperative infection and its transcriptional response under cephalosporin stress

Year 2017, Volume: 07 Issue: 02, 75 - 82, 01.06.2017

Birson İngti , Deepjyoti Paul Anand Prakash Maurya

https://doi.org/10.5799/jmid.vi.328788

Abstract

Objectives: Organisms harboring multiple plasmid mediated β-lactamases are major concerns in nosocomial
infections. Among these plasmid mediated β-lactamases, ACT (EBC family) is a clinically important enzyme capable
of hydrolyzing broad spectrum cephalosporins. Therefore, the present study was undertaken to determine the
prevalence of ACT determinant along with other co-existing β-lactamase genes in P. aeruginosa strains.
Methods: A total of 176 Pseudomonas isolates were phenotypically screened for the presence of AmpC β-lactamase
by M3DET Method followed by Molecular detection using PCR assay. Transcriptional evaluation of blaACT-2 gene was
analyzed by RT-PCR and its transferability was performed by transformation and conjugation.
Results: Present study demonstrates the presence of ACT-2 allele among 12 strains of P. aeruginosa. Co-existence
of other β-lactamase genes were encountered among ACT-2 harboring strains which includes CTX-M (n=2), SHV
(n=3), TEM (n=2), VEB (n=2), OXA-10 (n=1), CIT (n=2) and DHA (n=3). Fingerprinting by REP PCR revealed the
isolates harboring ACT-2 to be distinct and these isolates showed high resistance to expanded-spectrum
cephalosporins and even to carbapenem group of drugs. This ACT-2 allele was encoded in the plasmid (L/M, FIA,
FIB Inc. Group) and conjugatively transferable. Transcriptional analysis revealed a significant increase in ACT-2
expression (483 fold) when induced by ceftriaxone at 4 µg/ml followed by ceftazidime at 8 µg/ml (31 fold) and
cefotaxime 4 µg/ml (8 fold).
Conclusion: In this study detection of ACT-2 plasmid mediated AmpC β-lactamase along with other β-lactamase
genes in clinical isolates of P. aeruginosa represents a serious therapeutic challenge. Therefore, revision in
antimicrobial policy is required for effective treatment of patients infected with pathogen expressing this mechanism.
J Microbiol Infect Dis 2017; 7(2): 75-82

Keywords

Plasmid mediated, ACT-2 β-lactamase, P. aeruginosa

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