title: Efflux as a glutaraldehyde resistance mechanism in Pseudomonas fluorescens and Pseudomonas aeruginosa biofilms.
abstract: A major challenge in microbial biofilm control is biocide resistance. Phenotypic adaptations and physical protective effects have been historically thought to be the primary mechanisms for glutaraldehyde resistance in bacterial biofilms. Recent studies indicate the presence of genetic mechanisms for glutaraldehyde resistance, but very little is known about the contributory genetic factors. Here, we demonstrate that efflux pumps contribute to glutaraldehyde resistance in Pseudomonas fluorescens and Pseudomonas aeruginosa biofilms. The RNA-seq data show that efflux pumps and phosphonate degradation, lipid biosynthesis, and polyamine biosynthesis metabolic pathways were induced upon glutaraldehyde exposure. Furthermore, chemical inhibition of efflux pumps potentiates glutaraldehyde activity, suggesting that efflux activity contributes to glutaraldehyde resistance. Additionally, induction of known modulators of biofilm formation, including phosphonate degradation, lipid biosynthesis, and polyamine biosynthesis, may contribute to biofilm resistance and resilience. Fundamental understanding of the genetic mechanism of biocide resistance is critical for the optimization of biocide use and development of novel disinfection strategies. Our results reveal genetic components involved in glutaraldehyde resistance and a potential strategy for improved control of biofilms.
authors.affiliation: Department of Civil and Environmental Engineering, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: Amit
authors.initials: A
authors.lastname: Vikram
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Bomberger
authors.affiliation: Department of Civil and Environmental Engineering, University of Pittsburgh, Pittsburgh, Pennsylvania, USA Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA BibbyKJ@pitt.edu.
authors.forename: Kyle J
authors.initials: KJ
authors.lastname: Bibby
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects*; D004202:Disinfectants / Q000494:pharmacology; D024881:Drug Resistance, Bacterial; D005976:Glutaral / Q000494:pharmacology*; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000378:metabolism*; D011551:Pseudomonas fluorescens / Q000187:drug effects* / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D004202:Disinfectants; D005976:Glutaral
references.citation: J Antimicrob Chemother. 2002 Apr;49(4):597-9
references.pmid: 11909832
references.citation: Environ Sci Technol. 2014 Nov 4;48(21):13001-9
references.pmid: 25279933
references.citation: Antimicrob Agents Chemother. 1992 Jun;36(6):1208-14
references.pmid: 1416820
references.citation: Biofouling. 2011 Mar;27(3):337-46
references.pmid: 21512918
references.citation: Antimicrob Agents Chemother. 2014 Nov;58(11):6424-31
references.pmid: 25136007
references.citation: Bioinformatics. 2003 Aug 12;19(12):1477-83
references.pmid: 12912827
references.citation: J Appl Microbiol. 2002;92 Suppl:55S-64S
references.pmid: 12000613
references.citation: Antimicrob Agents Chemother. 2005 Feb;49(2):849-52
references.pmid: 15673787
references.citation: Antimicrob Agents Chemother. 2009 Dec;53(12):5230-5
references.pmid: 19752278
references.citation: Antimicrob Agents Chemother. 2002 May;46(5):1469-74
references.pmid: 11959584
references.citation: Antimicrob Agents Chemother. 2009 Sep;53(9):4015-8
references.pmid: 19581465
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: J Bacteriol. 2006 Apr;188(7):2355-63
references.pmid: 16547021
references.citation: Nat Methods. 2008 Jul;5(7):621-8
references.pmid: 18516045
references.citation: J Hosp Infect. 2010 Nov;76(3):200-5
references.pmid: 20638752
references.citation: J Bacteriol. 2002 Feb;184(4):1140-54
references.pmid: 11807075
references.citation: Antimicrob Agents Chemother. 2011 Jun;55(6):2648-54
references.pmid: 21422224
references.citation: J Ind Microbiol Biotechnol. 2002 Jul;29(1):10-5
references.pmid: 12080421
references.citation: Microbiology. 2003 Mar;149(Pt 3):707-14
references.pmid: 12634339
references.citation: J Hosp Infect. 2013 Aug;84(4):316-8
references.pmid: 23831280
references.citation: Biotechnol Bioeng. 2008 May 1;100(1):1-18
references.pmid: 18366134
references.citation: Proc Natl Acad Sci U S A. 2011 Jul 12;108(28):11393-8
references.pmid: 21705661
references.citation: J Biol Chem. 2010 Dec 10;285(50):39224-38
references.pmid: 20876533
references.citation: J Biol Chem. 2009 Apr 10;284(15):9899-907
references.pmid: 19196710
references.citation: Curr Med Chem. 2011;18(19):2969-80
references.pmid: 21651484
references.citation: BMC Genomics. 2013;14:350
references.pmid: 23705625
references.citation: J Hosp Infect. 2011 Jul;78(3):163-70
references.pmid: 21664533
references.citation: J Med Microbiol. 2008 Aug;57(Pt 8):966-73
references.pmid: 18628497
references.citation: Infect Immun. 2014 Sep;82(9):3764-74
references.pmid: 24958708
references.citation: Genome Biol. 2010;11(3):R25
references.pmid: 20196867
references.citation: J Bacteriol. 2001 Jun;183(11):3268-75
references.pmid: 11344133
references.citation: J Ind Microbiol Biotechnol. 2006 Sep;33(9):741-9
references.pmid: 16609854
references.citation: Infect Control Hosp Epidemiol. 2011 Dec;32(12):1173-8
references.pmid: 22080655
references.citation: Antimicrob Agents Chemother. 2010 Jul;54(7):2920-7
references.pmid: 20457816
references.citation: Scientifica (Cairo). 2014;2014:231083
references.pmid: 24834360
references.citation: Antimicrob Agents Chemother. 2008 Dec;52(12):4478-82
references.pmid: 18838593
references.citation: J Bacteriol. 2012 Feb;194(4):813-26
references.pmid: 22155771
references.citation: J Hosp Infect. 2007 Jun;65 Suppl 2:60-72
references.pmid: 17540245
references.citation: Environ Microbiol. 2011 Apr;13(4):1078-90
references.pmid: 21261797
references.citation: Biofouling. 2011 Oct;27(9):1017-32
references.pmid: 22011093
references.citation: Methods Mol Biol. 2000;132:365-86
references.pmid: 10547847
references.citation: Clin Microbiol Rev. 1999 Jan;12(1):147-79
references.pmid: 9880479
references.citation: Environ Microbiol. 2010 Jul;12(7):2034-47
references.pmid: 20406298
references.citation: J Mol Microbiol Biotechnol. 2001 Apr;3(2):255-64
references.pmid: 11321581
references.citation: Int J Food Microbiol. 2010 Jun 15;140(2-3):109-16
references.pmid: 20471125
references.citation: BMC Microbiol. 2012;12:261
references.pmid: 23153211
references.citation: J Antimicrob Chemother. 2011 May;66(5):1036-43
references.pmid: 21393196
references.citation: Antimicrob Agents Chemother. 2001 Jan;45(1):105-16
references.pmid: 11120952
references.citation: Expert Opin Investig Drugs. 2013 Feb;22(2):191-206
references.pmid: 23215733
references.citation: Trends Microbiol. 2014 Jun;22(6):326-33
references.pmid: 24598086
references.citation: Mol Microbiol. 2011 Apr;80(1):195-218
references.pmid: 21276098
references.citation: Nat Protoc. 2009;4(1):44-57
references.pmid: 19131956
references.citation: N Engl J Med. 2003 Jan 16;348(3):214-20
references.pmid: 12529461
references.citation: Antimicrob Agents Chemother. 2005 Feb;49(2):781-2
references.pmid: 15673767
references.citation: J Vis Exp. 2011;(47). pii: 2437. doi: 10.3791/2437
references.pmid: 21307833
references.citation: Antimicrob Agents Chemother. 2001 Feb;45(2):428-32
references.pmid: 11158736
references.citation: In Vivo. 2006 May-Jun;20(3):361-6
references.pmid: 16724671