Birson İngti , Deepjyoti Paul ANAND PRAKASH MAURYA

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

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,

PDF

___

1. Lister PD, Wolter DJ, Hanson ND. AntibacterialResistant Pseudomonas aeruginosa: Clinical Impact and Complex Regulation of Chromosomally Encoded.Resistance Mechanisms. Clin Microbiol Rev 2009; 22 (4): 582–610.
2. Livermore DM. Multiple mechanisms of antimicrobial resistance in Pseudomonas aeruginosa: our worst nightmare? Clin Infect Dis 2002; 34: 634–640.
3. Carlos J, Moya B, Perez J, Oliver A. Stepwise upregulation of the Pseudomonas aeruginosa chromosomal cephalosporinase conferring highlevel β-lactam resistance involves three ampD homologues. Antimicrob Agents Chemother 2006; 50:1780–1787.
4. Jacoby GA. AmpC β-lactamases. Clin Microbiol Rev 2009; 22:161-182.
5. Jacobs C, Joris B, Jamin M, et al. AmpD, essential for both β-lactamase regulation and cell wall recycling, is a novel cytosolic N-acetylmuramyl-Lalanine amidase. Mol Microbiol 1995; 15: 553–9.
6. Lee S, Park YJ, Kim M, Lee HK, Han K, Kang CS. Prevalence of Ambler class A and D β- lactamases among clinical isolates of Pseudomonas aeruginosa in Korea. J Antimicrob Chemother 2005; 56: 122-127.
7. Strateva T and Yordanov D. Pseudomonas aeruginosa – a phenomenon of bacterial resistance. J Med Microbiol 2009; 58: 1133–1148.
8. Papagiannitsis CC, Tzouvelekis LS, Tzelepi E, Miriagou V. Plasmid-encoded ACC-4, an extended-spectrum cephalosporinase variant from Escherichia coli. Antimicrob Agents Chemother 2007; 51: 3763-3767.
9. Bradford PA, Urban C, Mariano N, Projan SJ, Rahal JJ, Bush K. Imipenem resistance in Klebsiella pneumoniae is associated with the combination of ACT-1, a plasmid-mediated AmpC β-lactamase, and the loss of an outer membrane protein. Antimicrob Agents Chemother 1997; 41: 563–569.
10. Chen Y, Cheng J, Wang Q, Ye Y, Li JB, Zhang XJ. ACT-3, a novel plasmid-encoded class C β- lactamase in a Klebsiella pneumoniae isolate from China. Int J Antimicrob Agents 2009; 33: 95–96.
11. Datta S, Mitra S, Viswanathan R, Saha A, Basu S. Characterization of novel plasmid-mediated β- lactamases (SHV-167 and ACT-16) associated with New Delhi metallo-β-lactamase-1 harboring isolates from neonates in India. J Med Microbiol 2014; 63: 480-482.
12. Reisbig MD, Hanson ND. The ACT-1 plasmidencoded AmpC β-lactamase is inducible: detection in a complex β-lactamase background. J Antimicrob Chemother 2002; 49:557–560.
13. Barguigua A, Otmani FE, Talmi M, et al. Prevalence and genotypic analysis of plasmidmediated β-lactamases among urinary Klebsiella pneumoniae isolates in Moroccan community. The J Antibiotics 2012; 66: 11-16.
14. Coudron PE, Hanson ND, Climo MW. Occurrence of extended-spectrum and AmpC β-lactamases in bloodstream isolates of Klebsiella pneumoniae: isolates harbor plasmid-mediated FOX-5 and ACT-1 AmpC β-lactamases. J. Clin. Microbiol 2003; 41:772–777.
15. Lorian V. Antibiotics in laboratory medicine 2005; 5th edd. Philadelphia: Lippincott Williams and Wilkins.
16. Coudron PE, Moland ES, Thomson KS. Occurrence and Detection of AmpC β-lactamases among Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis isolates at a Veterans Medical Center. J Clin Microbiol 2000; 38: 1791– 1796.
17. Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Third Informational Supplement. M100-S23. CLSI, Wayne, PA, USA, 2013.
18. Caroline D, Anaelle DC, Dominique D, Christine F, Guillaume A. Development of a set of multiplex PCR assays for the detection of genes encoding important β-lactamases in Enterobacteriaceae. J Antimicrob Chemother 2010; 65: 490–495.
19. Nass T, Cuzon G, Villegas MV, Lartigue MF, Quinn JP, Nordmann P. Genetic structures at the origin of acquisition of the β-lactamase blaKPC gene. Antimicrob Agents Chemother 2008; 52:1257–1263.
20. Rasmussen BA, Bush K, Keeney D, et al. Characterization of IMI-1 β-lactamase, a class A carbapenem-hydrolyzing enzyme from Enterobacter cloaceae. Antimicrob Agents Chemother 1996; 40: 2080–2086.
21. Nass T, Vandel L, Sougakoff W, Livermore DM, Nordmann P. Cloning and sequence analysis of the gene for a carbapenem hydrolyzing class A β- lactamase, Sme-1, from Serratia marcescens S6. Antimicrob Agents Chemother 1994; 38:1262– 1270.
22. JH Y, Yi K, Lee H, et al. Molecular characterization of metallo-β-lacatamaseproducing Acinetobacter baumannii and Acinetbacter genomospecies 3 from Korea: identification of two new integrons carrying the blaVIM-2 gene cassettes. J Antimicrob Chemother 2002; 49: 837–840.
23. Yong D, Toleman MA, Giske CG, et al. Characterization of a New Metallo-β-Lactamase Gene, blaNDM-1, and a Novel Erythromycin Esterase Gene Carried on a Unique Genetic Structure in Klebsiella pneumoniae Sequence Type 14 from India. Antimicrob Agents Chemother 2009; 53: 5046–5054.
24. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual (2nd edn). Plainview: New York: Cold Spring Harbor Laboratory Press, 1989.
25. Carattoli A, Bertini A, Villa L, Falbo V, Hopkins KL, Threlfall EJ. Identification of plasmids by PCRbased replicon typing. J Microbiol Methods 2005; 63:219-228.
26. Quale J, Bratu S, Gupta J, Landman D. Interplay of Efflux System, ampC and oprD expression in carbapenem resistance of Pseudomonas aeruginosa clinical isolates. Antimicrob Agents Chemother 2006; 50 (5): 1633-1641.
27. Versalovic J, Schneider M, de Bruijn FJ, Lupski JR. Genomic fingerprinting of bacteria using repetitive sequence based polymerase chain reaction. Methods Mol Cell Biol 1994; 5: 25–40.
28. Zhu B, zhang P, Huang Z, et al. Study on drug resistance of Pseudomonas aeruginosa plasmidmediated AmpC β-lactamase. Mol Med Reports 2013; 7: 664-668.
29. Upadhyay S, Mishra S, Sen MR, Banerjee T, Bhattacharjee A. Co-existence of Pseudomonasderived cephalosporinase among plasmid encoded CMY 2 harboring isolates of Pseudomonas aeruginosa in north India. Indian J Med Microbiol 2013; 31(3): 257-260.
30. Hanson ND, Thomson KS, Moland ES, Sanders CC, Berthold G, Penn R G. Molecular characterization of a multiply resistant Klebsiella pneumoniae encoding ESBLs and a plasmidmediated AmpC. J Antimicrob Chemother 1999; 44:377–380.
31. Wei Y, Wang J. Identification of ACT-1 PlasmidMediated AmpC β-Lactamase Producing Citrobacter freundii from a Chinese Patient. Ann Lab Med 2013; 33: 86-88.
32. Kollef MH, Ward S, Sherman G, Prentice D, Schaiff R, Huey W, Fraser VJ. Inadequate treatment of nosocomial infections is associated with certain empiric antibiotic choices. Crit Care Med 2000; 28: 3456–3464.
33. Leone M, Bourgoin A, Cambon S, Dubuc M, Albanese J, Martin C. Empirical antimicrobial therapy of septic shock patients: adequacy and impact on the outcome, Crit. Care Med 2003; 31: 462–467.
34. Pea F, Viale P. The antimicrobial therapy puzzle: could pharmacokinetic pharmacodynamic relationships be helpful in addressing the issue of appropriate pneumonia treatment in critically ill patients? Clin Infect Dis 2006; 42: 1764-1771.
35. Roberts A, Lipman J, Blot S, Rello J. Better outcomes through continuous infusion of timedependent antibiotics to critically ill patients? Curr Opin Crit Care 2008; 14: 390–396.
36. Richmond MH, Sykes RB. The β-lactamases of gram negative bacteria and their possible physiological role. Adv Microb Physiol 1973; 9: 31- 88.
37. Sykes RB, Matthew M. The β-lactamases of gram negative bacteria and their role in resistance to β- lactam antibiotics. J Antimicrob Chemother 1997; 2: 115-157.

Journal of Microbiology and Infectious Diseases-Cover

ISSN: 2146-3158
Başlangıç: 2011
Yayıncı: Sağlık Araştırmaları Derneği

Arşiv