Quorum sensing, a bacterial cell-to-cell communication mechanism, plays a key role in bacterial virulence and biofilm formation. Targeting quorum-sensing pathways represents a promising strategy for the development of novel antibacterial agents. This study evaluated the anti-quorum-sensing activities of 18 natural compounds, including cannabinoids, arylbenzofurans, flavonoids, caffeine, and chlorogenic acid, using the luminescent biosensor strain Vibrio harveyi MM30. V. harveyi MM30, a mutant strain deficient in the production of autoinducer-2 (AI-2) but responsive to exogenous AI-2, was used to assess the activity of test compounds on the AI-2 receptor pathway. Test compounds were incubated in AI-2-containing media, and luminescence was measured to evaluate quorum-sensing inhibition. Comparisons were made in the absence of AI-2 to determine AI-2-independent inhibitory activity. The most active compounds were further tested on methicillin-resistant Staphylococcus aureus (MRSA 7112) to determine their effects on AI-2 production in spent media. Among the tested compounds, the non-prenylated arylbenzofuran moracin M and the prenylated arylbenzofuran moracin C exhibited significant quorum-sensing inhibitory activity in the AI-2-mediated pathway. None of the test compounds significantly inhibited quorum sensing in the absence of AI-2. Five compounds (cannabigerol, cannabidiol, cannabigerolic acid, moracin M, and moracin C) were selected for further investigation in MRSA 7112 cultures. The spent media from MRSA 7112 cultures treated with moracin M (16, 32, 64 µg/mL) and cannabigerolic acid (16 µg/mL) showed significant inhibition of AI-2 production when transferred to V. harveyi MM30 cultures. Moracin M and cannabigerolic acid demonstrated potential as quorum-sensing inhibitors by targeting AI-2 production and signalling pathways in MRSA 7112 and V. harveyi. These findings suggest their potential for further development as antibacterial agents targeting quorum-sensing mechanisms.

Department of Molecular Pharmacy Faculty of Pharmacy Masaryk University 612 00 Brno Czech Republic

Department of Natural Drugs Faculty of Pharmacy Masaryk University 612 00 Brno Czech Republic

Faculty of Sciences Palacký University 779 00 Olomouc Czech Republic

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Schluter J., Schoech A.P., Foster K.R., Mitri S. The Evolution of Quorum Sensing as a Mechanism to Infer Kinship. PLoS Comput. Biol. 2016;12:e1004848. doi: 10.1371/journal.pcbi.1004848. PubMed DOI PMC

Kolenbrander P.E., Andersen R.N., Blehert D.S., Egland P.G., Foster J.S., Palmer R.J. Communication among Oral Bacteria. Microbiol. Mol. Biol. Rev. 2002;66:486–505. doi: 10.1128/MMBR.66.3.486-505.2002. PubMed DOI PMC

Fuqua W.C., Winans S.C., Greenberg E.P. Quorum Sensing in Bacteria: The LuxR-LuxI Family of Cell Density-Responsive Transcriptional Regulators. J. Bacteriol. 1994;176:269–275. doi: 10.1128/jb.176.2.269-275.1994. PubMed DOI PMC

Defoirdt T., Sorgeloos P. Monitoring of Vibrio harveyi Quorum Sensing Activity in Real Time during Infection of Brine Shrimp Larvae. ISME J. 2012;6:2314–2319. doi: 10.1038/ismej.2012.58. PubMed DOI PMC

Aharoni R., Bronstheyn M., Jabbour A., Zaks B., Srebnik M., Steinberg D. Oxazaborolidine Derivatives Inducing Autoinducer-2 Signal Transduction in Vibrio harveyi. Bioorg. Med. Chem. 2008;16:1596–1604. doi: 10.1016/j.bmc.2007.11.032. PubMed DOI

Cao J.G., Meighen E.A. Purification and Structural Identification of an Autoinducer for the Luminescence System of Vibrio harveyi. J. Biol. Chem. 1989;264:21670–21676. doi: 10.1016/S0021-9258(20)88238-6. PubMed DOI

Kadirvel M., Fanimarvasti F., Forbes S., McBain A., Gardiner J.M., Brown G.D., Freeman S. Inhibition of Quorum Sensing and Biofilm Formation in Vibrio harveyi by 4-Fluoro-DPD; a Novel Potent Inhibitor of AI-2 Signalling. Chem. Commun. 2014;50:5000–5002. doi: 10.1039/C3CC49678C. PubMed DOI

Higgins D.A., Pomianek M.E., Kraml C.M., Taylor R.K., Semmelhack M.F., Bassler B.L. The Major Vibrio Cholerae Autoinducer and Its Role in Virulence Factor Production. Nature. 2007;450:883–886. doi: 10.1038/nature06284. PubMed DOI

Henares B., Xu Y., Boon E. A Nitric Oxide-Responsive Quorum Sensing Circuit in Vibrio harveyi Regulates Flagella Production and Biofilm Formation. Int. J. Mol. Sci. 2013;14:16473–16484. doi: 10.3390/ijms140816473. PubMed DOI PMC

Keller L., Surette M.G. Communication in Bacteria: An Ecological and Evolutionary Perspective. Nat. Rev. Microbiol. 2006;4:249–258. doi: 10.1038/nrmicro1383. PubMed DOI

Surette M.G., Miller M.B., Bassler B.L. Quorum Sensing in Escherichia coli, Salmonella typhimurium, and Vibrio harveyi: A New Family of Genes Responsible for Autoinducer Production. Proc. Natl. Acad. Sci. USA. 1999;96:1639–1644. doi: 10.1073/pnas.96.4.1639. PubMed DOI PMC

Aqawi M., Gallily R., Sionov R.V., Zaks B., Friedman M., Steinberg D. Cannabigerol Prevents Quorum Sensing and Biofilm Formation of Vibrio harveyi. Front. Microbiol. 2020;11:858. doi: 10.3389/fmicb.2020.00858. PubMed DOI PMC

Kumar L., Chhibber S., Kumar R., Kumar M., Harjai K. Zingerone Silences Quorum Sensing and Attenuates Virulence of Pseudomonas Aeruginosa. Fitoterapia. 2015;102:84–95. doi: 10.1016/j.fitote.2015.02.002. PubMed DOI

Martín-Rodríguez A.J., Ticona J.C., Jiménez I.A., Flores N., Fernández J.J., Bazzocchi I.L. Flavonoids from Piper Delineatum Modulate Quorum-Sensing-Regulated Phenotypes in Vibrio harveyi. Phytochemistry. 2015;117:98–106. doi: 10.1016/j.phytochem.2015.06.006. PubMed DOI

Girennavar B., Cepeda M.L., Soni K.A., Vikram A., Jesudhasan P., Jayaprakasha G.K., Pillai S.D., Patil B.S. Grapefruit Juice and Its Furocoumarins Inhibits Autoinducer Signaling and Biofilm Formation in Bacteria. Int. J. Food Microbiol. 2008;125:204–208. doi: 10.1016/j.ijfoodmicro.2008.03.028. PubMed DOI

Vikram A., Jayaprakasha G.K., Jesudhasan P.R., Pillai S.D., Patil B.S. Suppression of Bacterial Cell–Cell Signalling, Biofilm Formation and Type III Secretion System by Citrus Flavonoids. J. Appl. Microbiol. 2010;109:515–527. doi: 10.1111/j.1365-2672.2010.04677.x. PubMed DOI

Parai D., Banerjee M., Dey P., Chakraborty A., Islam E., Mukherjee S.K. Effect of Reserpine on Pseudomonas aeruginosa Quorum Sensing Mediated Virulence Factors and Biofilm Formation. Biofouling. 2018;34:320–334. doi: 10.1080/08927014.2018.1437910. PubMed DOI

Brackman G., Defoirdt T., Miyamoto C., Bossier P., Van Calenbergh S., Nelis H., Coenye T. Cinnamaldehyde and Cinnamaldehyde Derivatives Reduce Virulence in Vibrio Spp. by Decreasing the DNA-Binding Activity of the Quorum Sensing Response Regulator LuxR. BMC Microbiol. 2008;8:149. doi: 10.1186/1471-2180-8-149. PubMed DOI PMC

Paczkowski J.E., Mukherjee S., McCready A.R., Cong J.-P., Aquino C.J., Kim H., Henke B.R., Smith C.D., Bassler B.L. Flavonoids Suppress Pseudomonas Aeruginosa Virulence through Allosteric Inhibition of Quorum-Sensing Receptors. J. Biol. Chem. 2017;292:4064–4076. doi: 10.1074/jbc.M116.770552. PubMed DOI PMC

Soni D., Smoum R., Breuer A., Mechoulam R., Steinberg D. Effect of the Synthetic Cannabinoid HU-210 on Quorum Sensing and on the Production of Quorum Sensing-Mediated Virulence Factors by Vibrio harveyi. BMC Microbiol. 2015;15:159. doi: 10.1186/s12866-015-0499-0. PubMed DOI PMC

Friedman L., Smoum R., Feldman M., Mechoulam R., Steinberg D. Does the Endocannabinoid Anandamide Affect Bacterial Quorum Sensing, Vitality, and Motility? Cannabis. Cannabinoid. Res. 2019;4:102–109. doi: 10.1089/can.2018.0055. DOI

Martinelli D., Grossmann G., Séquin U., Brandl H., Bachofen R. Effects of Natural and Chemically Synthesized Furanones on Quorum Sensing in Chromobacterium Violaceum. BMC Microbiol. 2004;4:25. doi: 10.1186/1471-2180-4-25. PubMed DOI PMC

Laganenka L., Sander T., Lagonenko A., Chen Y., Link H., Sourjik V. Quorum Sensing and Metabolic State of the Host Control Lysogeny-Lysis Switch of Bacteriophage T1. mBio. 2019;10:e01884-19. doi: 10.1128/mBio.01884-19. PubMed DOI PMC

Norizan S., Yin W.-F., Chan K.-G. Caffeine as a Potential Quorum Sensing Inhibitor. Sensors. 2013;13:5117–5129. doi: 10.3390/s130405117. PubMed DOI PMC

Castillo S., Heredia N., García S. 2(5H)-Furanone, Epigallocatechin Gallate, and a Citric-Based Disinfectant Disturb Quorum-Sensing Activity and Reduce Motility and Biofilm Formation of Campylobacter Jejuni. Folia Microbiol. 2015;60:89–95. doi: 10.1007/s12223-014-0344-0. PubMed DOI

Xu W., Zhang X., Wang L., Zeng W., Sun Y., Zhou C., Zhou T., Shen M. Effect of Chlorogenic Acid on the Quorum-sensing System of Clinically Isolated Multidrug-resistant Pseudomonas aeruginosa. J. Appl. Microbiol. 2022;132:1008–1017. doi: 10.1111/jam.15275. PubMed DOI

Čulenová M., Sychrová A., Hassan S.T.S., Berchová-Bímová K., Svobodová P., Helclová A., Michnová H., Hošek J., Vasilev H., Suchý P., et al. Multiple In Vitro Biological Effects of Phenolic Compounds from Morus Alba Root Bark. J. Ethnopharmacol. 2020;248:112296. doi: 10.1016/j.jep.2019.112296. PubMed DOI

Ramić D., Klančnik A., Možina S.S., Dogsa I. Elucidation of the AI-2 Communication System in the Food-Borne Pathogen Campylobacter Jejuni by Whole-Cell-Based Biosensor Quantification. Biosens. Bioelectron. 2022;212:114439. doi: 10.1016/j.bios.2022.114439. PubMed DOI

Hong N.T.X., Baruah K., Vanrompay D., Bossier P. Characterization of Phenotype Variations of Luminescent and Non-luminescent Variants of Vibrio harveyi Wild Type and Quorum Sensing Mutants. J. Fish Dis. 2016;39:317–327. doi: 10.1111/jfd.12365. PubMed DOI

Mok K.C. Vibrio harveyi Quorum Sensing: A Coincidence Detector for Two Autoinducers Controls Gene Expression. EMBO J. 2003;22:870–881. doi: 10.1093/emboj/cdg085. PubMed DOI PMC

Cluzel M.-E., Zanella-Cléon I., Cozzone A.J., Fütterer K., Duclos B., Molle V. The Staphylococcus Aureus Autoinducer-2 Synthase LuxS Is Regulated by Ser/Thr Phosphorylation. J. Bacteriol. 2010;192:6295–6301. doi: 10.1128/JB.00853-10. PubMed DOI PMC

Appendino G., Giana A., Gibbons S., Maffei M., Gnavi G., Grassi G., Sterner O. A Polar Cannabinoid from Cannabis sativa Var. Carma. Nat. Prod. Commun. 2008;3:1934578X0800301. doi: 10.1177/1934578X0800301207. DOI

Appendino G., Gibbons S., Giana A., Pagani A., Grassi G., Stavri M., Smith E., Rahman M.M. Antibacterial Cannabinoids from Cannabis sativa: A Structure−Activity Study. J. Nat. Prod. 2008;71:1427–1430. doi: 10.1021/np8002673. PubMed DOI

DeKeersmaecker S.C.J., Vanderleyden J. Constraints on Detection of Autoinducer-2 (AI-2) Signalling Molecules Using Vibrio harveyi as a Reporter. Microbiology. 2003;149:1953–1956. doi: 10.1099/mic.0.C0117-0. PubMed DOI

Filer C.N. Acidic Cannabinoid Decarboxylation. Cannabis. Cannabinoid. Res. 2022;7:262–273. doi: 10.1089/can.2021.0072. PubMed DOI PMC

Naik R., Harmalkar D.S., Xu X., Jang K., Lee K. Bioactive Benzofuran Derivatives: Moracins A–Z in Medicinal Chemistry. Eur. J. Med. Chem. 2015;90:379–393. doi: 10.1016/j.ejmech.2014.11.047. PubMed DOI

Škovranová G., Čulenová M., Treml J., Dzurická L., Marova I., Sychrová A. Prenylated Phenolics from Morus Alba against MRSA Infections as a Strategy for Wound Healing. Front. Pharmacol. 2022;13:1068371. doi: 10.3389/fphar.2022.1068371. PubMed DOI PMC

Rasmussen T.B., Manefield M., Andersen J.B., Eberl L., Anthoni U., Christophersen C., Steinberg P., Kjelleberg S., Givskov M. How Delisea Pulchra Furanones Affect Quorum Sensing and Swarming Motility in Serratia Liquefaciens MG1. Microbiology. 2000;146:3237–3244. doi: 10.1099/00221287-146-12-3237. PubMed DOI

Ahmad A., Viljoen A.M., Chenia H.Y. The Impact of Plant Volatiles on Bacterial Quorum Sensing. Lett. Appl. Microbiol. 2015;60:8–19. doi: 10.1111/lam.12343. PubMed DOI

Yue J., Yang H., Liu S., Song F., Guo J., Huang C. Influence of Naringenin on the Biofilm Formation of Streptococcus Mutans. J. Dent. 2018;76:24–31. doi: 10.1016/j.jdent.2018.04.013. PubMed DOI