Restricted accessResearch articleFirst published online 2020-10
Exploring the Interplay of Resistance Nodulation Division Efflux Pumps,Amp C and Opr D in Antimicrobial Resistance of Burkholderia cepacia Complex in Clinical Isolates
This study aimed at investigating the association of gene expression of multidrug efflux pumps (MexA, MexC, MexE, and MexX), the outer membrane porin OprD, and the β-lactamase AmpC with the antimicrobial susceptibility among 44 clinical isolates of Burkholderia cepacia complex (Bcc).
Results:
Increased expression of ampC gene showed significant association with reduced susceptibility to chloramphenicol. In fact, reduced susceptibility to chloramphenicol was correlated with overexpression of most genes (ampC, mexC, mexE, and mexX) studied here in majority (>95%) of the Bcc isolates. Increased mexA expression showed significant association with reduced susceptibility to β-lactam antimicrobials (ceftazidime, piperacillin-tazobactam, and meropenem) and co-trimoxazole. Reduced susceptibility to meropenem also showed significant correlation with overexpression of mexC and mexX, whereas reduced susceptibility to ceftazidime was also associated with mexE overexpression. Reduced susceptibility to levofloxacin was significantly associated with overexpression of mexX. The involvement of the efflux pumps in levofloxacin and ceftazidime resistance was further inferred from the finding that the efflux pump inhibitor, carbonyl cyanide m-chlorophenylhydrazone reduced minimum inhibitory concentrations for both the antimicrobials.
Conclusions:
To conclude, this study explored the high-level expression of mexC, mexE, and mexX efflux pumps genes and ampC in the clinical isolates of Bcc, which can be targeted at treating infections caused by Bcc.
Get full access to this article
View all access options for this article.
References
1.
VanlaereE., BaldwinA., GeversD., HenryD., De BrandtE., LiPumaJ.J, MahenthiralingamE., SpeertD.P, DowsonC., and VandammeP.. 2009. K. Taxon, a complex within the Burkholderia cepacia complex, comprises at least two novel species, Burkholderia contaminans sp. nov. and Burkholderia lata sp. nov. Int. J. Syst. Evol. Microbiol. 59,(Pt. 1)::102–111.
2.
SmetB., MayoM., PeetersC., ZlosnikJ.E, SpilkerT., HirdT.J, LiPumaJ.J, KiddT.J, KaestliM., GintherJ.L, WagnerD.M, KeimP., BellS.C, JacobsJ.A, CurrieB.J, and VandammeP.A.. 2015. Burkholderia stagnalis sp. nov. and Burkholderia territorii sp. nov., two novel Burkholderia cepacia complex species from environmental and human sources. Int. J. Syst. Evol. Microbiol. 65:2265–2271.
3.
KrishnamoorthyG., LeusI.V, WeeksJ.W, WolloscheckD., RybenkovV.V, and ZgurskayaH.I.. 2017. Synergy between active efflux and outer membrane diffusion defines rules of antibiotic permeation into Gram-negative bacteria. MBio. 8:1–16.
4.
TomasM., DoumithM., WarnerM., TurtonJ.F, BeceiroA., BouG., LivermoreD.M, and WoodfordN.. 2010. Efflux pumps, OprD porin, AmpC beta-lactamase, and multiresistance in Pseudomonas aeruginosa isolates from cystic fibrosis patients. Antimicrob. Agents Chemother. 54:2219–2224.
5.
QualeJ., BratuS., GuptaJ., and LandmanD.. 2006. Interplay of efflux system, ampC, and oprD expression in carbapenem resistance of Pseudomonas aeruginosa clinical isolates. Antimicrob. Agents Chemother. 50:1633–1641.
6.
PodneckyN.L., RhodesK.A, and SchweizerH.P.. 2015. Efflux pump-mediated drug resistance in Burkholderia. Front Microbiol. 6:305.
7.
TsengS.P., TsaiW.C, LiangC.Y, LinY.S, HuangJ.W, ChangC.Y, TyanY.C, and LuP.L.. 2014. The contribution of antibiotic resistance mechanisms in clinical Burkholderia cepacia complex isolates: An emphasis on efflux pump activity. PLoS One, 9:e104986.
8.
GautamV., ShafiqN., SinghM., RayP., SinghalL., JaiswalN.P, PrasadA., SinghS., and AgarwalA.. 2015. Clinical and in vitro evidence for the antimicrobial therapy in Burkholderia cepacia complex infections. Expert. Rev. Anti. Infect. Ther. 13:629–663.
9.
BonacorsiS., FitoussiF., LhopitalS., and BingenE.. 1999. Comparative in vitro activities of meropenem, imipenem, temocillin, piperacillin, and ceftazidime in combination with tobramycin, rifampin, or ciprofloxacin against Burkholderia cepacia isolates from patients with cystic fibrosis. Antimicrob. Agents. Chemother. 43:213–217.
10.
ZhouJ., ChenY., TabibiS., AlbaL., GarberE., and SaimanL.. 2007. Antimicrobial susceptibility and synergy studies of Burkholderia cepacia complex isolated from patients with cystic fibrosis. Antimicrob. Agents Chemother. 51:1085–1088.
11.
RhodesK.A., and SchweizerH.P.. 2016. Antibiotic resistance in Burkholderia species. Drug Resist. Updat. 28:82–90.
12.
LandmanD., BratuS., KocharS., PanwarM., TrehanM., DoymazM., and QualeJ.. 2007. Evolution of antimicrobial resistance among Pseudomonas aeruginosa, Acinetobacter baumannii and Klebsiella pneumoniae in Brooklyn, NY. J. Antimicrob. Chemother. 60:78–82.
13.
CastanheiraM., DeshpandeL.M, MathaiD., BellJ.M, JonesR.N, and MendesR.E.. 2010. Early dissemination of NDM-1- and OXA-181-producing Enterobacteriaceae in Indian Hospitals: Report from the SENTRY Antimicrobial Surveillance Program (2006–2007). Antimicrob. Agents Chemother. 55:1274–1278.
14.
FurlanJ.P.R., SanchezD.G, FachinA.L, and StehlingE.G.. 2018. Presence of beta-lactamase encoding genes in Burkholderia cepacia complex isolated from soil. Microb. Drug Resist. 24:347–352.
15.
GuglierameP., PascaM.R, De RossiE., BuroniS., ArrigoP., ManinaG., and RiccardiG.. 2006. Efflux pump genes of the resistance-nodulation-division family in Burkholderia cenocepacia genome. BMC Microbiol. 6:66.
16.
BuroniS., PascaM.R, FlannaganR.S, BazziniS., MilanoA., BertaniI., VenturiV., ValvanoM.A, and RiccardiG.. 2009. Assessment of three resistance-nodulation-cell division drug efflux transporters of Burkholderia cenocepacia in intrinsic antibiotic resistance. BMC Microbiol. 9:200.
17.
GautamV., RayP., VandammeP., ChatterjeeS.S, DasA., SharmaK., RanaS., GargR.K, MadhupS.K, MahajanM., and SharmaM.. 2009. Identification of lysine positive non-fermenting gram negative bacilli (Stenotrophomonas maltophilia and Burkholderia cepacia complex). J. Indian. Med. Microbiol. 27:128–133.
18.
MahenthiralingamE., BischofJ., ByrneS.K, RadomskiC., DaviesJ.E, Av-GayY., and VandammeP.. 2000. DNA-Based diagnostic approaches for identification of Burkholderia cepacia complex, Burkholderia vietnamiensis, Burkholderia multivorans, Burkholderia stabilis, and Burkholderia cepacia genomovars I and III. J. Clin. Microbiol. 38:3165–3173.
19.
SpilkerT., BaldwinA., BumfordA., DowsonC.G, MahenthiralingamE., and LiPumaJ.J.. 2009. Expanded multilocus sequence typing for Burkholderia species. J. Clin. Microbiol. 47:2607–2610.
20.
Clinical and Laboratory Standards Institute (CLSI). 2018. Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Fifth Informational Supplement. M100-S25, Vol. 35. CLSI, Wayne, PA.
21.
YigitH., QueenanA.M, AndersonG.J, Domenech-SanchezA., BiddleJ.W, StewardC.D, AlbertiS., BushK., and TenoverF.C.. 2001. Novel carbapenem-hydrolyzing beta-lactamase, KPC-1, from a carbapenem-resistant strain of Klebsiella pneumoniae. Antimicrob. Agents Chemother. 45:1151–1161.
22.
Afzal-ShahM., WoodfordN., and LivermoreD.M.. 2001. Characterization of OXA-25, OXA-26, and OXA-27, molecular class D beta-lactamases associated with carbapenem resistance in clinical isolates of Acinetobacter baumannii. Antimicrob. Agents Chemother. 45:583–588.
23.
DeshpandeP., RodriguesC., ShettyA., KapadiaF., HedgeA., and SomanR.. 2010. New Delhi Metallo-beta lactamase (NDM-1) in Enterobacteriaceae: treatment options with carbapenems compromised. J. Assoc. Physicians India, 58:147–149.
24.
NamS.W., ChenX., LimJ., KimS.H, KimS.T, ChoY.H, YoonJ., and ParkS.. 2011. In vivo fluorescence imaging of bacteriogenic cyanide in the lungs of live mice infected with cystic fibrosis pathogens. PLoS One, 6:e21387.
25.
MeletiadisJ., PournarasS., RoilidesE., and WalshT.J.. 2010. Defining fractional inhibitory concentration index cutoffs for additive interactions based on self-drug additive combinations, Monte Carlo simulation analysis, and in vitro-in vivo correlation data for antifungal drug combinations against Aspergillus fumigatus. Antimicrob. Agents Chemother. 54:602–609.
26.
ListerP.D., WolterD.J, and HansonN.D.. 2009. Antibacterial-resistant Pseudomonas aeruginosa: clinical impact and complex regulation of chromosomally encoded resistance mechanisms. Clin. Microbiol. Rev. 22:582–610.
27.
Livermore, D.M. 1992. Interplay of impermeability and chromosomal beta-lactamase activity in imipenem-resistant Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 36:2046–2048.
28.
StratevaT., and YordanovD.. 2009. Pseudomonas aeruginosa—a phenomenon of bacterial resistance. J. Med. Microbiol. 58, (Pt. 9)::1133–1148.
29.
Jacoby, G.A. 2009. AmpC beta-lactamases. Clin. Microbiol. Rev. 22:161–182.
30.
XavierD.E., PicaoR.C, GirardelloR., FehlbergL.C, and GalesA.C.. 2010. Efflux pumps expression and its association with porin down-regulation and beta-lactamase production among Pseudomonas aeruginosa causing bloodstream infections in Brazil. BMC Microbiol. 10:217.
31.
NikaidoH.1996. Multidrug efflux pumps of gram-negative bacteria. J. Bacteriol. 178:5853–5859.
LiX.Z., and NikaidoH.. 2009. Efflux-mediated drug resistance in bacteria: an update. Drugs, 69:1555–1623.
34.
BurnsJ.L., WadsworthC.D, BarryJ.J, and GoodallC.P. 1996. Nucleotide sequence analysis of a gene from Burkholderia (Pseudomonas) cepacia encoding an outer membrane lipoprotein involved in multiple antibiotic resistance. Antimicrob. Agents Chemother. 40:307–313.
35.
KohlerT., Michea-HamzehpourM., HenzeU., GotohN., CurtyL.K, and PechereJ.C.. 1997. Characterization of MexE-MexF-OprN, a positively regulated multidrug efflux system of Pseudomonas aeruginosa. Mol. Microbiol. 23:345–354.
36.
NairB.M., CheungK.JJr., GriffithA., and BurnsJ.L. 2004. Salicylate induces an antibiotic efflux pump in Burkholderia cepacia complex genomovar III (B. cenocepacia). J. Clin. Invest. 113:464–473.
37.
HoldenM.T., Seth-SmithH.M, CrossmanL.C, SebaihiaM., BentleyS.D, Cerdeno-TarragaA.M, ThomsonN.R, BasonN., QuailM.A, SharpS., CherevachI., ChurcherC., GoodheadI., HauserH., HolroydN., MungallK., ScottP., WalkerD., WhiteB., RoseH., IversenP., Mil-HomensD., RochaE.P, FialhoA.M, BaldwinA., DowsonC., BarrellB.G, GovanJ.R, VandammeP., HartC.A, MahenthiralingamE., and ParkhillJ.. 2009. The genome of Burkholderia cenocepacia J2315, an epidemic pathogen of cystic fibrosis patients. J. Bacteriol. 191:261–277.
38.
DrevinekP., and MahenthiralingamE.. 2010. Burkholderia cenocepacia in cystic fibrosis: epidemiology and molecular mechanisms of virulence. Clin. Microbiol. Infect. 16:821–830.
39.
CoenyeT., Van AckerH., PeetersE., SassA., BuroniS., RiccardiG., and MahenthiralingamE.. 2011. Molecular mechanisms of chlorhexidine tolerance in Burkholderia cenocepacia biofilms. Antimicrob. Agents Chemother. 55:1912–1919.
40.
BazziniS., UdineC., SassA., PascaM.R, LongoF., EmilianiG., FondiM., PerrinE., DecorosiF., VitiC., GiovannettiL., LeoniL., FaniR., RiccardiG., MahenthiralingamE., and BuroniS.. 2011. Deciphering the role of RND efflux transporters in Burkholderia cenocepacia. PLoS One, 6:e18902.
41.
RushtonL., SassA., BaldwinA., DowsonC.G, DonoghueD., and MahenthiralingamE.. 2013. A key role for efflux in the preservative susceptibility and adaptive resistance of Burkholderia cepacia complex bacteria. Antimicrob. Agents Chemother. 57:2972–2980.
42.
ParrT.R.Jr., MooreR.A, MooreL.V, and HancockR.E.. 1987. Role of porins in intrinsic antibiotic resistance of Pseudomonas cepacia. Antimicrob. Agents Chemother. 31:121–123.
43.
ChopraI., and RobertsM.. 2001. Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. Microbiol. Mol. Biol. Rev. 65:232–260.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
