Anti-Virulence and Antibiofilm Activities of Stenochlaena palustris Extract Against Multidrug-Resistant Pseudomonas aeruginosa
DOI:
https://doi.org/10.65761/jbs.2026.45Keywords:
Stenochlaena palustris; Pseudomonas aeruginosa; Anti-virulence; Biofilm; Quorum sensing.Abstract
Background: The increasing prevalence of multidrug-resistant (MDR) Pseudomonas aeruginosa has created an urgent need for alternative strategies that target bacterial virulence and biofilm formation. Stenochlaena palustris is a medicinal and edible fern containing diverse bioactive phytochemicals with potential antimicrobial properties.
This study evaluated the anti-virulence and antibiofilm activities of S. palustris leaf ethanol extract (KLEE) against P. aeruginosa.
Methods: KLEE was prepared using 70% ethanol and evaluated for effects on bacterial motility, pyocyanin production, LasA protease activity, planktonic growth, biofilm formation, mature biofilm disruption, and extracellular polysaccharide production. Biofilm biomass was quantified using crystal violet staining. LC-HRMS was used to characterize phytochemical constituents, while acute toxicity was assessed using Rasbora lateristriata embryos.
Results: KLEE demonstrated concentration-dependent inhibition of important virulence-associated traits, including swarming and twitching motility, pyocyanin production, LasA activity, biofilm formation, mature biofilm biomass, and EPS production. LC-HRMS analysis indicated the presence of multiple potentially bioactive compounds. The toxicity assay provided preliminary evidence regarding the extract's biological safety.
Conclusion: S. palustris leaf extract shows promising anti-virulence and antibiofilm activity against P. aeruginosa and may serve as a potential source of natural compounds for combating MDR bacterial infections.
References
1. Artini M, Papa R, Sapienza F, Božović M, Vrenna G, Tuccio Guarna Assanti V, Sabatino M, Garzoli S, Fiscarelli EV, Ragno R, Selan L. Essential oils biofilm modulation activity and machine learning analysis on Pseudomonas aeruginosa isolates from cystic fibrosis patients. Microorganisms. 2022 Apr 24;10(5):887. https://doi.org/10.3390/microorganisms10050887
2. Chadha J, Khullar L, Gulati P, Chhibber S, Harjai K. Repurposing albendazole as a potent inhibitor of quorum sensing-regulated virulence factors in Pseudomonas aeruginosa: novel prospects of a classical drug. Microbial Pathogenesis. 2024 Jan 1;186:106468. https://doi.org/10.1016/j.micpath.2023.106468
3. Cheng X, Lu M, Qiu H, Li Y, Huang L, Dai W. Spontaneous quorum-sensing hierarchy reprogramming in Pseudomonas aeruginosa laboratory strain PAO1. AMB Express. 2022 Jan 26;12(1):6. https://doi.org/10.1186/s13568-022-01344-7
4. De Sousa T, Hébraud M, Dapkevicius ML, Maltez L, Pereira JE, Capita R, Alonso-Calleja C, Igrejas G, Poeta P. Genomic and Metabolic Characteristics of the Pathogenicity in Pseudomonas aeruginosa. International journal of molecular sciences. 2021 Nov 29;22(23):12892. https://doi.org/10.3390/ijms222312892
5. Eltayb EK, Alqahtani FY, Alkahtani HM, Alsarra IA, Alfaraj R, Aleanizy FS. Attenuation of Pseudomonas aeruginosa quorum sensing virulence of biofilm and pyocyanin by mBTL-loaded calcium alginate nanoparticles. Polymers. 2022 Sep 2;14(17):3655. https://doi.org/10.3390/polym14173655
6. Gerner E, Almqvist S, Thomsen P, Werthen M, Trobos M. Sodium salicylate influences the Pseudomonas aeruginosa biofilm structure and susceptibility towards silver. International Journal of Molecular Sciences. 2021 Jan 21;22(3):1060. https://doi.org/10.3390/ijms22031060
7. Gervasoni LF, Peixoto IC, Imperador AC, De Oliveira LB, Correia LF, de Oliveira Vieira KC, Saeki EK, da Silva Lima PE, Mareco EA, Pereira VC, Eller LK. Relationship between antibiotic resistance, biofilm formation, virulence factors and source of origin of Pseudomonas aeruginosa environmental isolates with regard to the presence of metallo-β-lactamase-encoding genes. Microbial Pathogenesis. 2023 Sep 1;182:106223. https://doi.org/10.1016/j.micpath.2023.106223
8. Hemmati J, Nazari M, Abolhasani FS, Ahmadi A, Asghari B. In vitro investigation of relationship between quorum-sensing system genes, biofilm forming ability, and drug resistance in clinical isolates of Pseudomonas aeruginosa. BMC microbiology. 2024 Mar 25;24(1):99. https://doi.org/10.1186/s12866-024-03249-w
9. Hendra R, Army MK, Frimayanti N, Teruna HY, Abdulah R, Nugraha AS. α-glucosidase and α-amylase inhibitory activity of flavonols from Stenochlaena palustris (Burm. f.) Bedd. Saudi Pharmaceutical Journal. 2024 Feb;32(2):1-8. https://doi.org/10.1016/j.jsps.2023.101940
10. Hendra R, Khodijah R, Almurdani M, Haryani Y, Nugraha AS, Frimayanti N, Teruna HY, Abdulah R. Free Radical Scavenging, Anti‐Infectious, and Toxicity Activities from Stenochlaena palustris (Burm. f.) Bedd. Extracts. Advances in Pharmacological and Pharmaceutical Sciences. 2022;2022(1):5729217. https://doi.org/10.1155/2022/5729217
11. Jurado-Martín I, Sainz-Mejías M, McClean S. Pseudomonas aeruginosa: an audacious pathogen with an adaptable arsenal of virulence factors. International journal of molecular sciences. 2021 Mar 18;22(6):3128. https://doi.org/10.3390/ijms22063128
12. Kalgudi R, Tamimi R, Kyazze G, Keshavarz T. Effect of quorum quenchers on virulence factors production and quorum sensing signalling pathway of non-mucoid, mucoid, and heavily mucoid Pseudomonas aeruginosa. World Journal of Microbiology and Biotechnology. 2022 Sep;38(9):163. https://doi.org/10.1007/s11274-022-03339-9
13. Li Q, Mao S, Wang H, Ye X. The molecular architecture of Pseudomonas aeruginosa quorum-sensing inhibitors. Marine Drugs. 2022 Jul 28;20(8):488. https://doi.org/10.3390/md20080488
14. Mohan MS, Salim SA, Ranganathan S, Parasuraman P, Anju VT, Ampasala DR, Dyavaiah M, Lee JK, Busi S. Attenuation of Las/Rhl quorum sensing regulated virulence and biofilm formation in Pseudomonas aeruginosa PAO1 by Artocarpesin. Microbial Pathogenesis. 2024 Apr 1;189:106609. https://doi.org/10.1016/j.micpath.2024.106609
15. Oluwabusola ET, Katermeran NP, Poh WH, Goh TM, Tan LT, Diyaolu O, Tabudravu J, Ebel R, Rice SA, Jaspars M. Inhibition of the quorum sensing system, elastase production and biofilm formation in Pseudomonas aeruginosa by psammaplin A and bisaprasin. Molecules. 2022 Mar 6;27(5):1721. https://doi.org/10.3390/molecules27051721
16. Patel M, Siddiqui AJ, Ashraf SA, Surti M, Awadelkareem AM, Snoussi M, Hamadou WS, Bardakci F, Jamal A, Jahan S, Sachidanandan M. Lactiplantibacillus plantarum-derived biosurfactant attenuates quorum sensing-mediated virulence and biofilm formation in Pseudomonas aeruginosa and Chromobacterium violaceum. Microorganisms. 2022 May 13;10(5):1026. https://doi.org/10.3390/microorganisms10051026
17. Rather MA, Mandal M. Attenuation of biofilm and quorum sensing regulated virulence factors of an opportunistic pathogen Pseudomonas aeruginosa by phytofabricated silver nanoparticles. Microbial Pathogenesis. 2023 Dec 1;185:106433. https://doi.org/10.1016/j.micpath.2023.106433
18. Reynolds D, Kollef M. The epidemiology and pathogenesis and treatment of Pseudomonas aeruginosa infections: an update. Drugs. 2021 Dec;81(18):2117-31. https://doi.org/10.1007/s40265-021-01635-6
19. Rohatgi N, Ganapathy D, Sathishkumar P. Eradication of Pseudomonas aeruginosa biofilm using quercetin-mediated copper oxide nanoparticles incorporated in the electrospun polycaprolactone nanofibrous scaffold. Microbial Pathogenesis. 2023 Dec 1;185:106453. https://doi.org/10.1016/j.micpath.2023.106453
20. Sendra E, Fernández-Muñoz A, Zamorano L, Oliver A, Horcajada JP, Juan C, Gómez-Zorrilla S. Impact of multidrug resistance on the virulence and fitness of Pseudomonas aeruginosa: a microbiological and clinical perspective. Infection. 2024 Aug;52(4):1235-68. https://doi.org/10.1007/s15010-024-02313-x
21. Sultan M, Arya R, Kim KK. Roles of two-component systems in Pseudomonas aeruginosa virulence. International journal of molecular sciences. 2021 Nov 10;22(22):12152. https://doi.org/10.3390/ijms222212152
22. Thakur M, Kumar Y, Yadav V, Pramanik A, Dubey KK. Understanding resistance acquisition by Pseudomonas aeruginosa and possible pharmacological approaches in palliating its pathogenesis. Biochemical Pharmacology. 2023 Sep 1;215:115689. https://doi.org/10.1016/j.bcp.2023.115689
23. Tuon FF, Dantas LR, Suss PH, Tasca Ribeiro VS. Pathogenesis of the Pseudomonas aeruginosa biofilm: a review. Pathogens. 2022 Feb 27;11(3):300. https://doi.org/10.3390/pathogens11030300
24. Vetrivel A, Ramasamy M, Vetrivel P, Natchimuthu S, Arunachalam S, Kim GS, Murugesan R. Pseudomonas aeruginosa biofilm formation and its control. Biologics. 2021 Oct 15;1(3):312-36. https://doi.org/10.3390/biologics1030019
25. Wang S, Feng Y, Han X, Cai X, Yang L, Liu C, Shen L. Inhibition of virulence factors and biofilm formation by wogonin attenuates pathogenicity of Pseudomonas aeruginosa PAO1 via targeting pqs quorum-sensing system. International Journal of Molecular Sciences. 2021 Nov 24;22(23):12699. https://doi.org/10.3390/ijms222312699
26. Ye Z, Silva DM, Traini D, Young P, Cheng S, Ong HX. An adaptable microreactor to investigate the influence of interfaces on Pseudomonas aeruginosa biofilm growth. Applied microbiology and biotechnology. 2022 Feb;106(3):1067-77. https://doi.org/10.1007/s00253-021-11746-5
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