Pseudomonas aeruginosa is an Effective Indicator for Screening of Quorum Sensing Inhibition by Plant Extracts
Main Article Content
Abstract
Quorum sensing is a well-known mechanism used by many bacteria to control important virulence factors through the production and subsequent response to N-acylhomoserine lactones (AHLs). Quorum sensing inhibition (QSI), targeting AHL-dependent signalling, has been reported as a strategy for the control of bacterial pathogenicity. Thus, this study aimed at investigating the capability of Pseudomonas aeruginosa as an indicator to rapidly screen for potential QSI caused by plant extracts based on pigment inhibition. Four ethanolic plant extracts including: dried flower buds of clove (Syzygium aromaticum L), bark of cinnamon (Cinnamomum cassia L), leaves of moringa (Moringa oleifera Lam), and Al-Gutub (Tribulus terrestris L) dried fruits were investigated for their QSI ability in comparison with furanone (the known QS inhibitor). Agar well diffusion assay was used, in which the indicator isolate, P. aeruginosa, was inoculated onto Pseudomonas special agar medium containing 8 mm wells to load different concentrations of the plant extracts along with furanone or phosphate buffered saline (negative control). The results showed that furanone was successful in inhibiting the pyocyanin pigment of P. aeruginosa without interfering with the bacterial growth, this verified the ability of this bacterium as a potent QS indicator organism. Importantly, clove extract at different concentrations caused complete inhibition of the pyocyanin pigment with little impact on the bacterial growth. On the use of cinnamon, only the high concentrations showed clear dye inhibition of the indicator organism. Moringa also caused certain degree of dye inhibition but only at high concentrations. Conversely, Al-Gutub neither affected on the pyocyanin production nor on the bacterial viability. In conclusion, P. aeruginosa isolate, pyocyanin producer, was successful in the screening of QS inhibitors. This simple study recommends the use of such isolate without relying on the import of certain strains of Chromobacterium violaceum, Pseudomonas aureofaciens, or Agrobacterium tumefaciens.
Received : 24 November 2023
Revised : 20 December 2023
Accepted : 01 February 2024
Published : 28 June 2024
Downloads
Article Details
How to Cite
References
Folders J, Algra J, Roelofs MS, van Loon LC, Tommassen J, Bitter W. Characterization of Pseudomonas aeruginosa chitinase, a gradually secreted protein. J Bacteriol. 2001;183:7044–7052. https://doi.org/10.1128/JB.183.24.7044-7052.2001
Dubern JF, Diggle SP. Quorum sensing by2-alkyl-4-quinolonesin Pseudomonas aeruginosa and other bacterial species. Mol Biosyst. 2008; 4:882–888. https://doi.org/10.1039/b803796p
Lee J, Zhang L. The hierarchy quorum sensing network in Pseudomonas aeruginosa. Protein Cell. 2015;6:26–41. https://doi.org/10.1007/s13238-014-0100-x
Majdura J, Jankiewicz U, Gała ̨zka A, Orzechowski S. The role of quorum sensing molecules in bacterial–plant interactions. Metabolites. 2023;13:114. https://doi.org/10.3390/metabo13010114
Li YH, Tian X. Quorum sensing and bacterial social interactions in biofilms. Sensors (Basel). 2012;12:2519–2538. https://doi.org/10.3390/s120302519
LaSarre B, Federle MJ. Exploiting quorum sensing to confuse bacterial pathogens. Microbiol Mol Biol Rev. 2013;77(1):73–111. https://doi.org/10.1128/MMBR.00046-12
Singh S, Bhatia S. Quorum sensing inhibitors: Curbing pathogenic infections through inhibition of bacterial communication. Iran J Pharm Res. 2021;20(2):486–514. 10.22037/ijpr.2020.113470.14318.
Dewick P. Medicinal Natural Products: A Biosynthetic Approach. 2nd ed. New York: John Wiley & Sons; 2002. https://doi.org/10.1002/0470846275
Asfour HZ. Anti-quorum sensing natural compounds. J Microsc Ultrastruct. 2018; 6:1-10. https://doi.org/10.4103/JMAU.JMAU_10_18
Ortega-Lozano, AJ, Hernández-Cruz EY, Gómez-Sierra, T, Pedraza-Chaverri J. Antimicrobial activity of spices popularly used in Mexico against urinary tract infections. Antibiot. 2023;12:325. https://doi.org/10.3390/antibiotics12020325
Cock E, Cheesman M. Plants of the genus Syzygium (Myrtaceae): A review on ethnobotany, medicinal properties, and phytochemistry. In: Goyal MR, editor. Bioactive Compounds of Medicinal Plants. 1st ed. Palm Bay (FL): Apple Academic Press: Palm Bay, FL, USA; 2018
Dhara L, Tripathi A. Antimicrobial activity of eugenol and cinnamaldehyde against extended spectrum beta lactamase producing Enterobacteriaceae by in vitro and molecular docking analysis. Eur J Integr Med. 2013;5(6):527–536. https://doi.org/10.1016/j.eujim.2013.08.005
Li T, Wang D, Liu N, Ma Y, Ding T, Mei Y, Li J. Inhibition of quorum sensing-controlled virulence factors and biofilm formation in Pseudomonas fluorescens by cinnamaldehyde. Int J Food Microbiol. 2018;269:98–106. https://doi.org/10.1016/j.ijfoodmicro.2018.01.023
Rao PV, Gan SH. Cinnamon: A multifaceted medicinal plant. Evidence-Based Complement Altern Med. 2014;2014,642942. https://doi.org/10.1155/2014/642942
Hussain S, Rahman R, Mushtaq A, El Zerey-Belaskri A. Clove: A review of a precious species with multiple uses. Int J Chem Biochem Sci. 2017;11:129–133. https://www.iscientific.org/wp-content/uploads/2019/10/15-IJCBS-17-11-15.pdf
Yun JW, You JR, Kim YS, Kim SH, Cho EY, Yoon JH, et al. In vitro and in vivo safety studies of cinnamon extract (Cinnamomum cassia) on general and genetic toxicology. Regul Toxicol Pharmacol. 2018;95:115–123. https://doi.org/10.1016/j.yrtph.2018.02.017
Rasool N, Saeed Z, Pervaiz M, Ali F, Younas U, Bashir R, et al. Evaluation of essential oil extracted from ginger, cinnamon and lemon for therapeutic and biological activities. Biocatal Agric Biotechnol. 2022;44:102470. https://doi.org/10.1016/j.bcab.2022.102470
Suhartono S, Ismail YS, Muhayya SR. The interference of Moringa oleifera leaf extracts to modulate quorum sensing-facilitated virulence factors. Biodiversitas. 2019;20(10):3000-3004. https://doi.org/10.13057/biodiv/d201031
Stefănescu R, Tero-Vescan A, Negroiu A, Aurică E, Vari C-E. A Comprehensive review of the phytochemical, pharmacological, and toxicological properties of Tribulus terrestris L. Biomolecules 2020;10:752. https://doi.org/10.3390/biom10050752
Mohammed MJ. Biological activity of saponins isolated from Tribulus terrestris (Fruit) on growth of some bacteria. Tikrit J Pure Sci. 2008;13(3). https://www.iasj.net/iasj/download/ce4f184455e320db
Jindal A, Kumar P, Gautam K. Evaluation of antibiotic potential of alkaloids of Tribulus terrestris L. against some pathogenic microorganisms. Int J Green Pharm. 2013;7:102–105. https://doi.org/10.4103/0973-8258.116383
Soleimanpour S, Sedighinia FS, Safipour Afshar A, Zarif R, Ghazvini K. Antibacterial activity of Tribulus terrestris and its synergistic effect with Capsella bursa-pastoris and Glycyrrhiza glabra against oral pathogens: An in-vitro study. Avicenna J phytomedicine. 2015; 5:210–217. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4469962/
Batoei S, Mahboubi M, Yari R. Antibacterial activity of Tribulus terrestris methanol extract against clinical isolates of Escherichia coli. Herba Pol. 2016;62:57–66. https://doi.org/10.1515/hepo-2016-0011
Vadakkan K, Vijayanand S, Hemapriya J, Gunasekaran R. Quorum sensing inimical activity of Tribulus terrestris against gram negative bacterial pathogens by signalling interference. 3 Biotech. 2019;9:163. https://doi.org/10.1007/s13205-019-1695-7
Ecker DJ, Crooke ST. Combinatorial drug discovery: which methods will produce the greatest value? Nat Biotechnol. 1995; 13:351–360. https://doi.org/10.1038/nbt0495-351
Ekins S, Mestres J, Testa B. In silico pharmacology for drug discovery: methods for virtual ligand screening and profiling. Br J Pharmacol. 2007;152:9–20. https://doi.org/10.1038/sj.bjp.0707305
OʼLoughlin CT, Miller LC, Siryaporn A, Drescher K, Semmelhack MF, Bassler BL. A quorum-sensing inhibitor blocks Pseudomonas aeruginosa virulence and biofilm formation. Proc Natl Acad Sci USA. 2013;110:17981–17986. https://doi.org/10.1073/pnas.1316981110
Weiland-Bräuer N, Pinnow N, Schmitz RA. Novel reporter for identification of interference with acyl homoserine lactone and autoinducer-2 quorum sensing. Appl Environ Microbiol. 2015;81:1477–1489. https://doi.org/10.1128/AEM.03290-14
Martín-Rodríguez AJ, Fernández JJ. A bioassay protocol for quorum sensing studies using Vibrio campbellii. Bio Protoc. 2016; 6: e1866. https://doi.org/10.21769/BioProtoc.1866
Noumi E, Merghni A, Alreshidi MM, Haddad O, Akmadar G, DeMartino L, et al. Chromobacterium violaceum and Pseudomonas aeruginosa PAO1: Models for evaluating anti-quorum sensing activity of Melaleuca alternifolia essential oil and its main component terpinen-4-ol. Molecules. 2018;23:2672–2688. https://doi.org/10.3390/molecules23102672
Deryabin D, Galadzhieva A, Kosyan D, Duskaev G. Plant-derived inhibitors of AHL-mediated quorum sensing in bacteria: modes of action. Int J Mol Sci. 2019;20:5588. https://doi.org/10.3390/ijms20225588
Reichling J. Anti-biofilm and virulence factor-reducing activities of essential oils and oil components as a possible option for bacterial infection control. Planta Med. 2020;86:520–537. https://doi.org/10.1055/a-1147-4671
Morohoshi T, Kato M, Fukamachi K, Kato N, Ikeda, T. N-acylhomoserine lactone regulates violacein production in Chromobacterium violaceum type strain ATCC 12472. FEMS Microbiol Lett. 2008;279:124–130. https://doi.org/10.1111/j.1574-6968.2007.01016.x
Hamad MAB, Abdulgafor AB. Bacterial and molecular detection of Pseudomonas aeruginosa in feline otitis externa in Baghdad city. J Surv Fish Sci. 2023;10(3S):728-736.
Abubakar AR, Haque M. Preparation of medicinal plants: Basic extraction and fractionation procedures for experimental purposes. J Pharm Bioallied Sci. 2008;12(1):1–10. https://doi.org/10.4103/jpbs.JPBS_175_19
Anessiny G, Perez C. Screening of plants used a green line. Folk medicine for antimicrobial activity. J Ethnopharmacol. 1993;39:119–128.https://doi.org/10.1016/0378-8741(93)90027-3
Magangana TP, Makunga NP, la Grange C, Stander MA, Fawole OA, Opara UL. Blanching pre-treatment promotes high yields, bioactive compounds, antioxidants, enzyme inactivation and antibacterial activity of ‘Wonderful’pomegranate peel extracts at three different harvest maturities. Antioxidants. 2021;10(7):1119. https://doi.org/10.3390/antiox10071119
Clinical and Laboratory Standards Institute (CLSI). Methods for dilution antimicrobial susceptibility testing for bacteria that grew aerobically. Approved Standard M7-A10, Wayne, PA Clinical and Laboratory Standards Institute. 2017.
Dulla GF, Lindow SE. Acyl-homoserine lactone-mediated cross talk among epiphytic bacteria modulates behavior of Pseudomonas syringae on leaves. ISME J. 2009; 3:825–834. https://doi.org/10.1038/ismej.2009.30
Brint JM, Ohman DE. Synthesis of multiple exoproducts in P. aeruginosa is under the control of RhlR-RhlI, another set of regulators in strain PAO1 with homology to the autoinducer-responsive LuxR-LuxI family. J Bacteriol Res. 1995; 177: 7155–7163. https://doi.org/10.1128/jb.177.24.7155-7163.1995
Pesci EC, Pearson JP, Seed PC, Iglewski BH. Regulation of las and rhl Quorum sensing in P. aeruginosa. J Bacteriol. 1997;179:3127–3132. https://doi.org/10.1128/jb.179.10.3127-3132.1997
Parsek MR, Greenberg EP. Acyl-homoserine lactone QS in gram-negative bacteria: a signaling mechanism involved in associations with higher organisms. Proc Natl Acad Sci USA. 2000; 97: 8789–8793. https://doi.org/10.1073/pnas.97.16.8789
Mohammed HA, Zgair AK. Detection of quorum sensing genes of Pseudomonas aeruginosa isolated from different areas in Iraq. Iraqi J Sci. 2022;63(11):4665–4673. https://doi.org/10.24996/ijs.2022.63.11.5
Julianti E, Rajah KK, Fidrianny I. Antibacterial activity of ethanolic extract of cinnamon bark, honey, and their combination effects against acne-causing bacteria. Sci. Pharm. 2017;85:19. https://doi.org/10.3390/scipharm85020019
Waty S, Suryanto D, Yurnaliza. Antibacterial activity of cinnamon ethanol extract (Cinnamomum burmannii) and its application as a mouthwash to inhibit streptococcus growth. IOP Conf. Series: Earth Environ Sci. 2018;130:2012049. https://doi.org/10.1088/1755-1315/130/1/012049
Tajuddin, Ahmad S, Abdul Latif, Qasmi IA. Effect of 50% ethanolic extract of Syzygium aromaticum (L.) Merr. & Perry. (clove) on sexual behaviour of normal male rats. BMC Complement and Altern Med. 2004;4:17. https://doi.org/10.1186/1472-6882-4-17
Husni E, Badriyya E, Putri L, Aldi Y. The effect of ethanol extract of Moringa leaf (Moringa oleifera Lam) against the activity and capacity of phagocytosis of macrofage cells and the percentage of leukosit cells of white mice. Pharmacog J. 2021;13(3):706-712. https://doi.org/10.5530/pj.2021.13.90
Heinz-Castro RTQ, Arredondo-Valdés R, Ordaz-Silva S, Méndez-Cortés H, Hernández-Juárez A, Chacón-Hernández JC. Evaluation of ethanol extract of Moringa oleifera Lam. as acaricide against Oligonychus punicae hirst (Trombidiformes: Tetranychidae). Insects. 2021; 12: 476. https://doi.org/10.3390/insects12050476
Hussain AA, Mohammed AA, Ibrahim HH, Abbas AH. Study the biological activities of Tribulus Terrestris extracts. World Academy of Science, Engineering and Technology. 2009; 57.
Khan, MSA, Zahin M, Hasan S, Husain FM, Ahmad I. Inhibition of quorum sensing regulated bacterial functions by plant essential oils with special reference to clove oil. Lett Appl Microbiol. 2009; 49: 354–360. https://doi.org/10.1111/j.1472-765X.2009.02666.x
Husain FM, Ahmad I, Khan MS, Ahmad E, Tahseen Q, Khan MS, et al. Sub-MICs of Mentha piperita essential oil and menthol inhibits AHL mediated quorum sensing and biofilm of Gram-negative bacteria. Front Microbiol. 2015;6:420. https://doi.org/10.3389/fmicb.2015.00420
Adhikari B, Shah PK, Karki R. Antibiogram and phytochemical analysis of cinnamon, clove, and sichuan pepper extracts. Nepal J Biotechnol. 2021;9(1):1–7. https://doi.org/10.3126/njb.v9i1.38644
Oulkheir S, Aghrouch M, El Mourabit F, Dalha F, Graich H, Amouch F, et al. Antibacterial activity of essential oils extracts from cinnamon, thyme, clove and geranium against a Gram negative and Gram positive pathogenic bacteria. J Dis Med Plants. Special Issue: New Vistas of Research in Ayurveda System of Medicine. 2017;3(2-1):1–5. https://doi.org/10.11648/j.jdmp.s.2017030201.11
Ginting EV, Retnaningrum E, Widiasih DA. Antibacterial activity of clove (Syzygium aromaticum) and cinnamon (Cinnamomum burmannii) essential oil against extended-spectrum β-lactamase-producing bacteria. Vet World. 2021;14(8):2206–2211. https://doi.org/10.14202/vetworld.2021.2206-2211
Anandhi P, Tharani M, Rajeshkumar S, Lakshmi T. Antibacterial activity of cinnamon and clove oil against wound pathogens. J Popul Ther Clin Pharmacol. 2022;28(2):e41–e46. https://doi.org/10.47750/jptcp.2022.871
Jonnalagadda S, Deshabathini UK. Anti-quorum sensing potential of Moringa oleifera seed extract. Int J Pharm Pharm Sci. 2016;8(1):76–82.
Rasamiravaka T, Labtani Q, Duez P, ElJazirihe M. Formation of biofilms by Pseudomonas aeruginosa: a review of the natural and synthetic compounds interfering with control mechanisms. Biomed Res Int. 2015a; 2015: 759348. https://doi.org/10.1155/2015/759348
Rasamiravaka T, Vandeputte OM, Pottier L, Huet J, Rabemanantsoa C, Kiendrebeogo M, Andriantsimahavandy A, Rasamindrakotroka A, Stévigny C, Duez P, El Jaziri M. Pseudomonas aeruginosa biofilm formation and persistence, along with the production of quorum sensing-dependent virulence factors, are disrupted by a triterpenoid coumarate ester isolated from Dalbergia trichocarpa, a tropical legume. PLoS One. 2015b;10:e0132791. https://doi.org/10.1371/journal.pone.0132791