Biofilm formation is a virulence factor of bacteria. The goal of this study was to understand the mechanisms of biofilm formation by methicillin-resistant Staphylococcus aureus (MRSA). Whole-genome sequencing of eight MRSA strains was performed to identify sequence variations in genes related to biofilm formation. Thirty-one genes involved in MRSA biofilm formation were analyzed and 11 amino acid sequence variations in four genes related to autolysis were found. These variations include E121D and H387 N in ArlS; Q117K, T424S, K428T, A509S, V752E, A754V, and T771A in Atl; T184K in CidC; and D251N in CidR. Among the 26 clinical MRSA isolates studied, 13 isolates were nonbiofilm producers and were found to harbor these mutations. Furthermore, all of these 13 isolates belonged to ST59. Ten of these 13 ST59 isolates became able to produce biofilms when they were incubated with extracellular DNA from MRSA N315. Results of this study suggest that sequence variations in arlS, atl, cidC, and cidR genes may render MRSA unable to produce biofilms. Further investigations are needed to correlate these sequence variations with the biofilm-forming ability of MRSA isolates.
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References
1.
GouldI.M., DavidM.Z., EspositoS., GarauJ., LinaG., MazzeiT., and PetersG.. 2012. New insights into meticillin-resistant Staphylococcus aureus (MRSA) pathogenesis, treatment and resistance. Int. J. Antimicrob. Agents, 39:96–104.
2.
CeriH., OlsonM.E., StremickC., ReadR.R., MorckD., and ABuret. 1999. The Calgary Biofilm Device: new technology for rapid determination of antibiotic susceptibilities of bacterial biofilms. J. Clin. Microbiol., 37:1771–1776.
3.
CostertonW., VeehR., ShirtliffM., PasmoreM., PostC., and GEhrlich. 2003. The application of biofilm science to the study and control of chronic bacterial infections. J. Clin. Invest., 112:1466–1477.
4.
O'GaraJ.P.2007. ica and beyond: biofilm mechanisms and regulation in Staphylococcus epidermidis and Staphylococcus aureus. FEMS Microbiol. Lett., 270:179–188.
5.
ArciolaC.R., CampocciaD., SpezialeP., MontanaroL., and CostertonJ.W.. 2012. Biofilm formation in Staphylococcus implant infections. A review of molecular mechanisms and implications for biofilm-resistant materials. Biomaterials., 33:5967–5982.
GeogheganJ.A., CorriganR.M., GruszkaD.T., SpezialeP., O'GaraJ.P., PottsJ.R., and FosterT.J.. 2010. Role of surface protein SasG in biofilm formation by Staphylococcus aureus. J. Bacteriol., 192:5663–5673.
8.
QinZ., OuY., YangL., ZhuY., Tolker-NielsenT., MolinS., and QuD.. 2007. Role of autolysin-mediated DNA release in biofilm formation of Staphylococcus epidermidis. Microbiology, 153:2083–2092.
9.
Toledo-AranaA., MerinoN., Vergara-IrigarayM., DebarbouilleM., PenadesJ.R., and LasaI.. 2005. Staphylococcus aureus develops an alternative, ica-independent biofilm in the absence of the arlRS two-component system. J. Bacteriol., 187:5318–5329.
10.
BiswasR., VogguL., SimonU.K., HentschelP., ThummG., and GotzF.. 2006. Activity of the major staphylococcal autolysin Atl. FEMS Microbiol. Lett., 259:260–268.
11.
VanhommerigE., MoonsP., PiriciD., LammensC., HernalsteensJ.P., De GreveH., Kumar-SinghS., GoossensH., and Malhotra-KumarS.. 2014. Comparison of biofilm formation between major clonal lineages of methicillin resistant Staphylococcus aureus. PLoS One, 9:e104561.
12.
Aires de Sousa, CrisostomoM., M.I., SanchesI.S., WuJ.S., FuzhongJ., TomaszA., and de LencastreH.. 2003. Frequent recovery of a single clonal type of multidrug-resistant Staphylococcus aureus from patients in two hospitals in Taiwan and China. J. Clin. Microbiol., 41:159–163.
13.
WangC.C., LoW.T., ChuM.L., and SiuL.K.. 2004. Epidemiological typing of community-acquired methicillin-resistant Staphylococcus aureus isolates from children in Taiwan. Clin. Infect. Dis., 39:481–487.
14.
HuangY.H., TsengS.P., HuJ.M., TsaiJ.C., HsuehP.R., and TengL.J.. 2007. Clonal spread of SCCmec type IV methicillin-resistant Staphylococcus aureus between community and hospital. Clin. Microbiol. Infect., 13:717–724.
15.
WangW.Y., ChiuehT.S., SunJ.R., TsaoS.M., and LuJ.J.. 2012. Molecular typing and phenotype characterization of methicillin-resistant Staphylococcus aureus isolates from blood in Taiwan. PLoS One, 7:e30394.
16.
ChenC.J., and HuangY.C.. 2014. New epidemiology of Staphylococcus aureus infection in Asia. Clin. Microbiol. Infect., 20:605–623.
17.
LuJ.J., LeeS.Y., HwaS.Y., and YangA.H.. 2005. Septic arthritis caused by vancomycin-intermediate Staphylococcus aureus. J. Clin. Microbiol., 43:4156–4158.
18.
WangW.Y., LeeS.Y., ChiuehT.S., and LuJ.J.. 2009. Molecular and phenotypic characteristics of methicillin-resistant and vancomycin-intermediate staphylococcus aureus isolates from patients with septic arthritis. J. Clin. Microbiol., 47:3617–3623.
19.
HoC.M., HsuehP.R., LiuC.Y., LeeS.Y., ChiuehT.S., ShyrJ.M., TsaoS.M., ChuangY.C., YanJ.J., WangL.S., et al.2010. Prevalence and accessory gene regulator (agr) analysis of vancomycin-intermediate Staphylococcus aureus among methicillin-resistant isolates in Taiwan—SMART program, 2003. Eur. J. Clin. Microbiol. Infect. Dis., 29:383–389.
20.
ChenC.J., HuangY.C., SuL.H., WuT.L., HuangS.H., ChienC.C., ChenP.Y., LuM.C., and KoW.C.. 2014. Molecular epidemiology and antimicrobial resistance of methicillin-resistant Staphylococcus aureus bloodstream isolates in Taiwan, 2010. PLoS One, 9:e101184.
21.
BoseJ.L., LehmanM.K., FeyP.D., and BaylesK.W.. 2012. Contribution of the Staphylococcus aureus Atl AM and GL murein hydrolase activities in cell division, autolysis, and biofilm formation. PLoS One, 7:e42244.
22.
ChenC.J., LinM.H., ShuJ.C., and LuJ.J.. 2014. Reduced susceptibility to vancomycin in isogenic Staphylococcus aureus strains of sequence type 59: tracking evolution and identifying mutations by whole-genome sequencing. J. Antimicrob. Chemother., 69:349–354.
BeenkenK.E., DunmanP.M., McAleeseF., MacapagalD., MurphyE., ProjanS.J., BlevinsJ.S., and SmeltzerM.S.. 2004. Global gene expression in Staphylococcus aureus biofilms. J. Bacteriol., 186:4665–4684.
25.
Parra-RuizJ., Bravo-MolinaA., Pena-MonjeA., and Hernandez-QueroJ.. 2012. Activity of linezolid and high-dose daptomycin, alone or in combination, in an in vitro model of Staphylococcus aureus biofilm. J. Antimicrob. Chemother., 67:2682–2685.
26.
SchlagS., NerzC., BirkenstockT.A., AltenberendF., and GotzF.. 2007. Inhibition of staphylococcal biofilm formation by nitrite. J. Bacteriol., 189:7911–7919.
27.
EnrightM.C., DayN.P., DaviesC.E., PeacockS.J., and SprattB.G.. 2000. Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus. J. Clin. Microbiol., 38:1008–1015.
28.
FournierB., and HooperD.C.. 2000. A new two-component regulatory system involved in adhesion, autolysis, and extracellular proteolytic activity of Staphylococcus aureus. J. Bacteriol., 182:3955–3964.
29.
RenzoniA., BarrasC., FrancoisP., CharbonnierY., HugglerE., GarzoniC., KelleyW.L., MajcherczykP., SchrenzelJ., LewD.P., et al.2006. Transcriptomic and functional analysis of an autolysis-deficient, teicoplanin-resistant derivative of methicillin-resistant Staphylococcus aureus. Antimicrob. Agents Chemother., 50:3048–3061.
30.
DwyerB.E., NewtonK.L., KisielaD., SokurenkoE.V., and CleggS.. 2011. Single nucleotide polypmorphisms of fimH associated with adherence and biofilm formation by serovars of Salmonella enterica. Microbiology, 157:3162–3171.
31.
PozziC., WatersE.M., RudkinJ.K., SchaefferC.R., LohanA.J., TongP., LoftusB.J., PierG.B., FeyP.D., MasseyR.C., et al.2012. Methicillin resistance alters the biofilm phenotype and attenuates virulence in Staphylococcus aureus device-associated infections. PLoS Pathog., 8:e1002626.
32.
WangD., JinQ., XiangH., WangW., GuoN., ZhangK., TangX., MengR., FengH., LiuL., et al.2011. Transcriptional and functional analysis of the effects of magnolol: inhibition of autolysis and biofilms in Staphylococcus aureus. PLoS One, 6:e26833.
33.
MerckollP., JonassenT.O., VadM.E., JeanssonS.L., and MelbyK.K.. 2009. Bacteria, biofilm and honey: a study of the effects of honey on ‘planktonic’ and biofilm-embedded chronic wound bacteria. Scand. J. Infect. Dis., 41:341–347.
34.
RiceK.C., MannE.E., EndresJ.L., WeissE.C., CassatJ.E., SmeltzerM.S., and BaylesK.W.. 2007. The cidA murein hydrolase regulator contributes to DNA release and biofilm development in Staphylococcus aureus. Proc. Natl. Acad. Sci. U. S. A., 104:8113–8118.
35.
HsuC.Y., LinM.H., ChenC.C., ChienS.C., ChengY.H., SuI.N., and ShuJ.C.. 2011. Vancomycin promotes the bacterial autolysis, release of extracellular DNA, and biofilm formation in vancomycin-non-susceptible Staphylococcus aureus. FEMS Immunol. Med. Microbiol., 63:236–247.
36.
PattonT.G., RiceK.C., FosterM.K., and BaylesK.W.. 2005. The Staphylococcus aureus cidC gene encodes a pyruvate oxidase that affects acetate metabolism and cell death in stationary phase. Mol. Microbiol., 56:1664–1674.
37.
YangS.J., DunmanP.M., ProjanS.J., and BaylesK.W.. 2006. Characterization of the Staphylococcus aureus CidR regulon: elucidation of a novel role for acetoin metabolism in cell death and lysis. Mol. Microbiol., 60:458–468.
38.
HellW., ReichlS., AndersA., and GatermannS.. 2003. The autolytic activity of the recombinant amidase of Staphylococcus saprophyticus is inhibited by its own recombinant GW repeats. FEMS Microbiol. Lett., 227:47–51.
39.
MemmiG., NairD.R., and Cheung AA.. 2012. Role of ArlRS in autolysis in methicillin-sensitive and methicillin-resistant Staphylococcus aureus strains. J. Bacteriol., 194:759–767.
40.
HuangY.C., and ChenC.J.. 2011. Community-associated meticillin-resistant Staphylococcus aureus in children in Taiwan, 2000s. Int. J. Antimicrob. Agents, 38:2–8.
41.
ChenC.J., HsuehP.R., SuL.H., ChiuC.H., LinT.Y., and HuangY.C.. 2009. Change in the molecular epidemiology of methicillin-resistant Staphylococcus aureus bloodstream infections in Taiwan. Diagn. Microbiol. Infect. Dis., 65:199–201.
42.
KudirkieneE., CohnM.T., StablerR.A., StrongP.C., SernieneL., WrenB.W., NielsenE.M., MalakauskasM., and BrondstedL.. 2012. Phenotypic and genotypic characterizations of Campylobacter jejuni isolated from the broiler meat production process. Curr. Microbiol., 65:398–406.
43.
NaparstekL., CarmeliY., Navon-VeneziaS., and BaninE.. 2014. Biofilm formation and susceptibility to gentamicin and colistin of extremely drug-resistant KPC-producing Klebsiella pneumoniae. J. Antimicrob. Chemother., 69:1027–1034.
44.
LeeS.C., LeeC.W., ShihH.J., ChiouM.J., SeeL.C., and SiuL.K.. 2013. Clinical features and risk factors of mortality for bacteremia due to community-onset healthcare-associated methicillin-resistant S. aureus. Diagn. Microbiol. Infect. Dis., 76:86–92.
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