Abstract
The aim of this study is to evaluate the antibiofilm and antibacterial effects of auranofin against WT-ETBF, rETBF, WT-NTBF and clinically isolated Bacteroides fragilis strains. The minimum inhibitory concentration and biofilm inhibitory concentration of 0.25 μg/ml for auranofin against B. fragilis were determined, and the biofilm eradication concentration was 1 μg/ml. At an auranofin concentration of 0.5 μg/ml, little cellular metabolic activity was found. Confocal laser scanning microcopy results confirmed the inhibition of biofilm by auranofin. The effects of auranofin on the outer membrane protein (ompA) gene and the RND-type efflux pump (bmeB3) gene were investigated using quantitative real-time polymerase chain reaction (qRT-PCR). The qRT-PCR results showed that treatment with auranofin significantly reduced the gene expression compared to controls without auranofin. These data indicate the applicability of auranofin as a repurposed drug due to its inhibitory effect against biofilm formation of B. fragilis. Therefore, our study demonstrates that auranofin, already approved for human use, is a promising drug that has strong antibiofilm and antibacterial activity against B. fragilis.
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References
AbdelKhalek A, Abutaleb NS, Elmagarmid KA, Seleem MN (2018) Repurposing auranofin as an intestinal decolonizing agent for vancomycin-resistant enterococci. Sci Rep 8:8353. https://doi.org/10.1038/s41598-018-26674-0
Abouelhassan Y et al (2017) Nitroxoline: a broad-spectrum biofilm-eradicating agent against pathogenic bacteria. Int J Antimicrob Agents 49:247–251. https://doi.org/10.1016/j.ijantimicag.2016.10.017
Alav I, Sutton JM, Rahman KM (2018) Role of bacterial efflux pumps in biofilm formation. J Antimicrob Chemother 73:2003–2020. https://doi.org/10.1093/jac/dky042
Boleij A et al (2015) The Bacteroides fragilis toxin gene is prevalent in the colon mucosa of colorectal cancer patients. Clin Infect Dis 60:208–215. https://doi.org/10.1093/cid/ciu787
Bundgaard-Nielsen C, Baandrup UT, Nielsen LP, Sorensen S (2019) The presence of bacteria varies between colorectal adenocarcinomas, precursor lesions and non-malignant tissue. BMC Cancer 19:399. https://doi.org/10.1186/s12885-019-5571-y
Chen H, Wubbolts RW, Haagsman HP, Veldhuizen EJA (2018) Inhibition and eradication of Pseudomonas aeruginosa biofilms by host defence peptides. Sci Rep 8:10446. https://doi.org/10.1038/s41598-018-28842-8
Clinical and Laboratory Standards Institute (CLSI) (2018) Methods for antimicrobial susceptibility testing of anaerobic bacteria, 9th edn. CLSI, Wayne, p M11
Costa OYA, Raaijmakers JM, Kuramae EE (2018) Microbial extracellular polymeric substances: ecological function and impact on soil aggregation. Front Microbiol 9:1636. https://doi.org/10.3389/fmicb.2018.01636
Debnath A et al (2012) A high-throughput drug screen for Entamoeba histolytica identifies a new lead and target. Nat Med 18:956–960. https://doi.org/10.1038/nm.2758
de Vos WM (2015) Microbial biofilms and the human intestinal microbiome. NPJ Biofilms Microbiomes 1:15005. https://doi.org/10.1038/npjbiofilms.2015.5
Donelli G, Vuotto C, Cardines R, Mastrantonio P (2012) Biofilm-growing intestinal anaerobic bacteria. FEMS Immunol Med Microbiol 65:318–325. https://doi.org/10.1111/j.1574-695X.2012.00962.x
Felipe V et al (2019) Chitosan disrupts biofilm formation and promotes biofilm eradication in Staphylococcus species isolated from bovine mastitis. Int J Biol Macromol 126:60–67. https://doi.org/10.1016/j.ijbiomac.2018.12.159
Flemming HC, Neu TR, Wozniak DJ (2007) The EPS matrix: the “house of biofilm cells”. J Bacteriol 189:7945–7947. https://doi.org/10.1128/JB.00858-07
Goel S, Mishra P (2018) Thymoquinone inhibits biofilm formation and has selective antibacterial activity due to ROS generation. Appl Microbiol Biotechnol 102:1955–1967. https://doi.org/10.1007/s00253-018-8736-8
Harbut MB et al (2015) Auranofin exerts broad-spectrum bactericidal activities by targeting thiol-redox homeostasis. Proc Natl Acad Sci USA 112:4453–4458. https://doi.org/10.1073/pnas.1504022112
Holmgren A, Lu J (2010) Thioredoxin and thioredoxin reductase: current research with special reference to human disease. Biochem Biophys Res Commun 396:120–124. https://doi.org/10.1016/j.bbrc.2010.03.083
Hwang-Bo H et al (2017) Auranofin, an inhibitor of thioredoxin reductase, induces apoptosis in hepatocellular carcinoma Hep3B cells by generation of reactive oxygen species. Gen Physiol Biophys 36:117–128. https://doi.org/10.4149/gpb_2016043
Joo HS, Otto M (2012) Molecular basis of in vivo biofilm formation by bacterial pathogens. Chem Biol 19:1503–1513. https://doi.org/10.1016/j.chembiol.2012.10.022
Kato N et al (2000) A new subtype of the metalloprotease toxin gene and the incidence of the three bft subtypes among Bacteroides fragilis isolates in Japan. FEMS Microbiol Lett 182:171–176. https://doi.org/10.1111/j.1574-6968.2000.tb08892.x
Kim HR, Rhee KJ, Eom YB (2019) Anti-biofilm and antimicrobial effects of zerumbone against Bacteroides fragilis. Anaerobe 57:99–106. https://doi.org/10.1016/j.anaerobe.2019.04.001
Kowalski RP, Yates KA, Romanowski EG, Karenchak LM, Mah FS, Gordon YJ (2005) An ophthalmologist’s guide to understanding antibiotic susceptibility and minimum inhibitory concentration data. Ophthalmology 112:1987. https://doi.org/10.1016/j.ophtha.2005.06.025
Mendes RF et al (2017) The essential oil from the fruits of the Brazilian spice Xylopia sericea A. St.-Hil. presents expressive in vitro antibacterial and antioxidant activity. J Pharm Pharmacol 69:341–348. https://doi.org/10.1111/jphp.12698
Molina-Manso D et al (2013) In vitro susceptibility to antibiotics of staphylococci in biofilms isolated from orthopaedic infections. Int J Antimicrob Agents 41:521–523. https://doi.org/10.1016/j.ijantimicag.2013.02.018
Myers LL, Shoop DS, Collins JE, Bradbury WC (1989) Diarrheal disease caused by enterotoxigenic Bacteroides fragilis in infant rabbits. J Clin Microbiol 27:2025–2030
Park SH, Lee JH, Berek JS, Hu MC (2014) Auranofin displays anticancer activity against ovarian cancer cells through FOXO3 activation independent of p53. Int J Oncol 45:1691–1698. https://doi.org/10.3892/ijo.2014.2579
Pierce JV, Bernstein HD (2016) Genomic diversity of enterotoxigenic strains of Bacteroides fragilis. PLoS ONE 11:e0158171. https://doi.org/10.1371/journal.pone.0158171
Poole K, Srikumar R (2001) Multidrug efflux in Pseudomonas aeruginosa: components, mechanisms and clinical significance. Curr Top Med Chem 1:59–71
Pumbwe L, Ueda O, Yoshimura F, Chang A, Smith RL, Wexler HM (2006) Bacteroides fragilis BmeABC efflux systems additively confer intrinsic antimicrobial resistance. J Antimicrob Chemother 58:37–46. https://doi.org/10.1093/jac/dkl202
Pumbwe L, Skilbeck CA, Wexler HM (2008) Presence of quorum-sensing systems associated with multidrug resistance and biofilm formation in Bacteroides fragilis. Microb Ecol 56:412–419. https://doi.org/10.1007/s00248-007-9358-3
Reichhardt C, Parsek MR (2019) Confocal laser scanning microscopy for analysis of Pseudomonas aeruginosa biofilm architecture and matrix localization. Front Microbiol 10:677. https://doi.org/10.3389/fmicb.2019.00677
Rhee KJ et al (2009) Induction of persistent colitis by a human commensal, enterotoxigenic Bacteroides fragilis, in wild-type C57BL/6 mice. Infect Immun 77:1708–1718. https://doi.org/10.1128/IAI.00814-08
Rocha ER, Tzianabos AO, Smith CJ (2007) Thioredoxin reductase is essential for thiol/disulfide redox control and oxidative stress survival of the anaerobe Bacteroides fragilis. J Bacteriol 189:8015–8023. https://doi.org/10.1128/JB.00714-07
Roder C, Thomson MJ (2015) Auranofin: repurposing an old drug for a golden new age. Drugs R&D 15:13–20. https://doi.org/10.1007/s40268-015-0083-y
Rosenblatt JE, Stewart PR (1975) Anaerobic bag culture method. J Clin Microbiol 1:527–530
Schwarz S et al (2010) Editorial: assessing the antimicrobial susceptibility of bacteria obtained from animals. J Antimicrob Chemother 65:601–604. https://doi.org/10.1093/jac/dkq037
Smith SG, Mahon V, Lambert MA, Fagan RP (2007) A molecular Swiss army knife: OmpA structure, function and expression. FEMS Microbiol Lett 273:1–11. https://doi.org/10.1111/j.1574-6968.2007.00778.x
Swidsinski A, Weber J, Loening-Baucke V, Hale LP, Lochs H (2005) Spatial organization and composition of the mucosal flora in patients with inflammatory bowel disease. J Clin Microbiol 43:3380–3389. https://doi.org/10.1128/JCM.43.7.3380-3389.2005
Thangamani S et al (2016a) Antibacterial activity and mechanism of action of auranofin against multi-drug resistant bacterial pathogens. Sci Rep 6:22571. https://doi.org/10.1038/srep22571
Thangamani S, Mohammad H, Abushahba MF, Sobreira TJ, Seleem MN (2016b) Repurposing auranofin for the treatment of cutaneous staphylococcal infections. Int J Antimicrob Agents 47:195–201. https://doi.org/10.1016/j.ijantimicag.2015.12.016
Wexler HM (2007) Bacteroides: the good, the bad, and the nitty-gritty. Clin Microbiol Rev 20:593–621. https://doi.org/10.1128/CMR.00008-07
Wexler HM, Tenorio E, Pumbwe L (2009) Characteristics of Bacteroides fragilis lacking the major outer membrane protein, OmpA. Microbiology 155:2694–2706. https://doi.org/10.1099/mic.0.025858-0
Woo SG et al (2017) The effectiveness of anti-biofilm and anti-virulence properties of dihydrocelastrol and dihydrocelastryl diacetate in fighting against methicillin-resistant Staphylococcus aureus. Arch Microbiol 199:1151–1163. https://doi.org/10.1007/s00203-017-1386-x
Acknowledgements
This research was supported by Soonchunhyang University Research Fund (Grant no. 20190911) and a Grant (NRF-2017R1D1A1B03032960) of the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education.
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Jang, HI., Eom, YB. Antibiofilm and antibacterial activities of repurposing auranofin against Bacteroides fragilis. Arch Microbiol 202, 473–482 (2020). https://doi.org/10.1007/s00203-019-01764-3
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DOI: https://doi.org/10.1007/s00203-019-01764-3