Green Synthesis of Silver Nanoparticles Using Glycyrrhiza glabra Root Extract for Antibacterial and Antibiofilm Activity Against Multidrug-Resistant Pseudomonas aeruginosa Clinical Isolates
DOI:
https://doi.org/10.21271/ZJPAS.38.2.5Keywords:
Multidrug resistant, Pseudomonas aeruginosa, Green silver nanoparticle, Glycyrrhiza glabra root, Antibiofilm.Abstract
Background: Pseudomonas aeruginosa is an opportunistic pathogen that contributes to treatment failure due to its strong biofilm-forming ability and multidrug resistance (MDR). This study intended to assess the antibacterial and antibiofilm properties of silver nanoparticles (AgNPs) synthesized from Glycyrrhiza glabra root extract against MDR-P. aeruginosa clinical isolates.
Methods: 128 clinical specimens were collected from patients who were receiving medical care in Erbil, Kurdistan Region, Iraq. Clinical specimens were cultured and analyzed using standard biochemical tests to identify P. aeruginosa. Antibiotic susceptibility testing was performed for identify MDR isolate. Biofilm production was evaluated with a microtiter plate technique through safranin staining. AgNPs were synthesized from an aqueous extract of G. glabra root. The antibacterial activity was assessed by a well diffusion test, and the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined using microdilution in 96-well plates. AgNPs were tested for their capacity to inhibit and eradicate biofilms.
Results: Thirty-four (26.6%) isolates were identified as P. aeruginosa, of which 26 (76.5%) exhibited MDR. Among these MDR isolates, 80.7% exhibited robust biofilm formation, 11.5% moderate, 3.8% weak, and 3.8% produced no biofilm. AgNPs demonstrated inhibition zones measuring 17 mm in 1500 µg/mL, and MIC values between 62.5 and 500 µg/mL. At a concentration of 500 µg/mL, AgNPs exhibited 97% biofilm inhibition and 85% eradication.
Conclusion: AgNPs derived from G. glabra root extract shown significant antibacterial and antibiofilm properties against MDR-P. aeruginosa isolates. These findings demonstrate their potential as alternative therapeutic agents for the management of MDR-P. aeruginosa infections.
References
Abdulhaq, N., Nawaz, Z., Zahoor, M. A. & Siddique, A. B. 2020. Association of biofilm formation with multi drug resistance in clinical isolates of Pseudomonas aeruginosa. Excli j, 19, 201-208.
Aflakian, F. & Hashemitabar, G. 2025. Biosynthesized silver nanoparticles at subinhibitory concentrations as inhibitors of quorum sensing, pathogenicity, and biofilm formation in Pseudomonas aeruginosa PAO1. Heliyon, 11.
Afrasiabi, S. & Partoazar, A. 2024. Targeting bacterial biofilm-related genes with nanoparticle-based strategies. Front Microbiol, 15, 1387114.
Al-Momani, H., Almasri, M., Al Balawi, D. A., Hamed, S., Albiss, B. A., Aldabaibeh, N., Ibrahim, L., Albalawi, H., Al Haj Mahmoud, S., Khasawneh, A. I., Kilani, M., Aldhafeeri, M., Bani-Hani, M., Wilcox, M., Pearson, J. & Ward, C. 2023. The efficacy of biosynthesized silver nanoparticles against Pseudomonas aeruginosa isolates from cystic fibrosis patients. Scientific Reports, 13, 8876.
Alobaid, H. M., Daghestani, M. H., AL-Malahi, N. M., Alzahrani, S. A., Hassen, L. M. & Metwally, D. M. 2022. Exploring the effect of silver nanoparticles on gene expression in colon cancer cell line HCT116. Green Processing and Synthesis, 11, 1108-1117.
Anees Ahmad, S., Sachi Das, S., Khatoon, A., Tahir Ansari, M., Afzal, M., Saquib Hasnain, M. & Kumar Nayak, A. 2020. Bactericidal activity of silver nanoparticles: A mechanistic review. Materials Science for Energy Technologies, 3, 756-769.
Ansari, M. A., Kalam, A., Al-Sehemi, A. G., Alomary, M. N., AlYahya, S., Aziz, M. K., Srivastava, S., Alghamdi, S., Akhtar, S., Almalki, H. D., Adil, S. F., Khan, M. & Hatshan, M. R. 2021. Counteraction of Biofilm Formation and Antimicrobial Potential of Terminalia catappa Functionalized Silver Nanoparticles against Candida albicans and Multidrug-Resistant Gram-Negative and Gram-Positive Bacteria. Antibiotics, 10, 725.
Antunes, L. C. M., Ferreira, R. B. R., Buckner, M. M. C. & Finlay, B. B. 2010. Quorum sensing in bacterial virulence. Microbiology, 156, 2271-2282.
Azam, M. W. & Khan, A. U. 2019. Updates on the pathogenicity status of Pseudomonas aeruginosa. Drug Discovery Today, 24, 350-359.
Bala Subramaniyan, S., Senthilnathan, R., Arunachalam, J. & Anbazhagan, V. 2020. Revealing the Significance of the Glycan Binding Property of Butea monosperma Seed Lectin for Enhancing the Antibiofilm Activity of Silver Nanoparticles against Uropathogenic Escherichia coli. Bioconjugate Chemistry, 31, 139-148.
Bauer, A. W., Kirby, W. M. M., Sherris, J. C. & Turck, M. 1966. Antibiotic Susceptibility Testing by a Standardized Single Disk Method. American Journal of Clinical Pathology, 45, 493-496.
Bhuiya, M., Sarkar, M. K. I., Sohag, M. H., Ali, H., Roy, C. K., Akther, L. & Sarker, A. F. 2018. Enumerating Antibiotic Susceptibility Patterns of Pseudomonas aeruginosa Isolated from Different Sources in Dhaka City. The Open Microbiology Journal, 12, 172-180.
Brindhadevi, K., LewisOscar, F., Mylonakis, E., Shanmugam, S., Verma, T. N. & Pugazhendhi, A. 2020. Biofilm and Quorum sensing mediated pathogenicity in Pseudomonas aeruginosa. Process Biochemistry, 96, 49-57.
Chegini, Z., Shariati, A., Alikhani, M. Y., Safaiee, M., Rajaeih, S., Arabestani, M. & Azizi, M. 2024. Antibacterial and antibiofilm activity of silver nanoparticles stabilized with C-phycocyanin against drug-resistant Pseudomonas aeruginosa and Staphylococcus aureus. Front Bioeng Biotechnol, 12, 1455385.
Chimi, L. Y., Noubom, M., Bisso, B. N., Singor Njateng, G. S. & Dzoyem, J. P. 2024. Biofilm Formation, Pyocyanin Production, and Antibiotic Resistance Profile of Pseudomonas aeruginosa Isolates from Wounds. International Journal of Microbiology, 2024, 1207536.
Cifuentes-Jiménez, C., Bolaños-Carmona, M. V., Enrich-Essvein, T., Rodríguez-Navarro, A. B., González-López, S., Yamauti, M. & Álvarez-Lloret, P. 2024. Green synthesis of chitosan- and fluoride-functionalized silver nanoparticles using Camellia sinensis: Characterization and dental applications. International Journal of Biological Macromolecules, 268, 131676.
Clinical and Laboratory Standards Institute 2020. Performance Standards for Antimicrobial Susceptibility Testing. 30th ed. Wayne, PA: CLSI.
Das, M. C., Sandhu, P., Gupta, P., Rudrapaul, P., De, U. C., Tribedi, P., Akhter, Y. & Bhattacharjee, S. 2016. Attenuation of Pseudomonas aeruginosa biofilm formation by Vitexin: A combinatorial study with azithromycin and gentamicin. Scientific Reports, 6, 23347.
Dybkova, S., Goncharuk, O., Rieznichenko, L., Terpiłowski, K., Borysenko, L., Gruzina, T., Dybkova, K. & Szewczuk-Karpisz, K. 2024. Aqueous dispersions of ‘green’ silver nanoparticles for eco-applications: Synthesis, structure and biosafety. Journal of Molecular Liquids, 415, 126319.
El-Telbany, M. & El-Sharaki, A. 2022. Antibacterial and anti-biofilm activity of silver nanoparticles on multi-drug resistance pseudomonas aeruginosa isolated from dental-implant. Journal of Oral Biology and Craniofacial Research, 12, 199-203.
Fernebro, J. 2011. Fighting bacterial infections—Future treatment options. Drug Resistance Updates, 14, 125-139.
Ghafoor, A., Hay, I. D. & Rehm, B. H. 2011. Role of exopolysaccharides in Pseudomonas aeruginosa biofilm formation and architecture. Appl Environ Microbiol, 77, 5238-46.
Goda, R. M., El-Baz, A. M., Khalaf, E. M., Alharbi, N. K., Elkhooly, T. A. & Shohayeb, M. M. 2022. Combating Bacterial Biofilm Formation in Urinary Catheter by Green Silver Nanoparticle. Antibiotics 11, 495.
Gondil, V. S., Kalaiyarasan, T., Bharti, V. K. & Chhibber, S. 2019. Antibiofilm potential of Seabuckthorn silver nanoparticles (SBT@AgNPs) against Pseudomonas aeruginosa. 3 Biotech, 9, 402.
Gusso, B. P., Almeida, A. R., Nunes, M. R., Becker, D., Hotza, D., da Rosa, C. G., dos Santos, V. V. & da Silva, B. F. 2025. The Antimicrobial Activity of Silver Nanoparticles Biosynthesized Using Cymbopogon citratus Against Multidrug-Resistant Bacteria Isolated from an Intensive Care Unit. Pharmaceuticals, 18, 1120.
Haji, S. H., Ganjo, A. R., Faraj, T. A., Fatah, M. H. & Smail, S. B. 2024. The enhanced antibacterial and antibiofilm properties of titanium dioxide nanoparticles biosynthesized by multidrug-resistant Pseudomonas aeruginosa. BMC Microbiology, 24, 379.
Hetta, H. F., Ramadan, Y. N., Al-Harbi, A. I., A. Ahmed, E., Battah, B., Abd Ellah, N. H., Zanetti, S. & Donadu, M. G. 2023. Nanotechnology as a Promising Approach to Combat Multidrug Resistant Bacteria: A Comprehensive Review and Future Perspectives. Biomedicines, 11, 413.
Huq, M. A. & Akter, S. 2021. Bacterial Mediated Rapid and Facile Synthesis of Silver Nanoparticles and Their Antimicrobial Efficacy against Pathogenic Microorganisms. Materials (Basel), 14.
Juzer, T., Soundharajan, R. & Srinivasan, H. 2025. Camellia sinensis mediated silver nanoparticles: eco-friendly antimicrobial agent to control multidrug resistant Gram-positive Staphylococcus aureus. Discover Nano, 20, 92.
Kambale, E. K., Nkanga, C. I., Mutonkole, B.-P. I., Bapolisi, A. M., Tassa, D. O., Liesse, J.-M. I., Krause, R. W. M. & Memvanga, P. B. 2020. Green synthesis of antimicrobial silver nanoparticles using aqueous leaf extracts from three Congolese plant species (Brillantaisia patula, Crossopteryx febrifuga and Senna siamea). Heliyon, 6.
Kamer, A. M. A., El Maghraby, G. M., Shafik, M. M. & Al-Madboly, L. A. 2024. Silver nanoparticle with potential antimicrobial and antibiofilm efficiency against multiple drug resistant, extensive drug resistant Pseudomonas aeruginosa clinical isolates. BMC Microbiology, 24, 277.
Khan, J., Tarar, S. M., Gul, I., Nawaz, U. & Arshad, M. 2021. Challenges of antibiotic resistance biofilms and potential combating strategies: a review. 3 Biotech, 11, 169.
Magiorakos, A. P., Srinivasan, A., Carey, R. B., Carmeli, Y., Falagas, M. E., Giske, C. G., Harbarth, S., Hindler, J. F., Kahlmeter, G., Olsson-Liljequist, B., Paterson, D. L., Rice, L. B., Stelling, J., Struelens, M. J., Vatopoulos, A., Weber, J. T. & Monnet, D. L. 2012. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect, 18, 268-81.
Makabenta, J. M. V., Nabawy, A., Li, C. H., Schmidt-Malan, S., Patel, R. & Rotello, V. M. 2021. Nanomaterial-based therapeutics for antibiotic-resistant bacterial infections. Nature Reviews Microbiology, 19, 23-36.
Mikhailova, E. O. 2025. Green Silver Nanoparticles: An Antibacterial Mechanism. Antibiotics, 14, 5.
Momenah, A. M., Bakri, R. A., Jalal, N. A., Ashgar, S. S., Felemban, R. F., Bantun, F., Hariri, S. H., Barhameen, A. A., Faidah, H. & Al-Said, H. M. 2023. Antimicrobial Resistance Pattern of Pseudomonas aeruginosa: An 11-Year Experience in a Tertiary Care Hospital in Makkah, Saudi Arabia. Infect Drug Resist, 16, 4113-4122.
Pastorino, G., Cornara, L., Soares, S., Rodrigues, F. & Oliveira, M. 2018. Liquorice (Glycyrrhiza glabra): A phytochemical and pharmacological review. Phytother Res, 32, 2323-2339.
Rajivgandhi, G. N., Ramachandran, G., Maruthupandy, M., Manoharan, N., Alharbi, N. S., Kadaikunnan, S., Khaled, J. M., Almanaa, T. N. & Li, W.-J. 2020. Anti-oxidant, anti-bacterial and anti-biofilm activity of biosynthesized silver nanoparticles using Gracilaria corticata against biofilm producing K. pneumoniae. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 600, 124830.
Rani, K., Devi, N., Banakar, P., Kharb, P. & Kaushik, P. 2022. Nematicidal Potential of Green Silver Nanoparticles Synthesized Using Aqueous Root Extract of Glycyrrhiza glabra. Nanomaterials, 12, 2966.
Singh, P., Pandit, S., Garnæs, J., Tunjic, S., Mokkapati, V. R. S. S., Sultan, A., Thygesen, A., Mackevica, A., Mateiu, R. V., Daugaard, A. E., Baun, A. & Mijakovic, I. 2018. Green synthesis of gold and silver nanoparticles from Cannabis sativa (industrial hemp) and their capacity for biofilm inhibition. International Journal of Nanomedicine, 13, 3571-3591.
Skóra, B., Krajewska, U., Nowak, A., Dziedzic, A., Barylyak, A. & Kus-Liśkiewicz, M. 2021. Noncytotoxic silver nanoparticles as a new antimicrobial strategy. Scientific Reports, 11.
Sondi, I. & Salopek-Sondi, B. 2004. Silver nanoparticles as antimicrobial agent: a case study on E. coli as a model for Gram-negative bacteria. Journal of Colloid and Interface Science, 275, 177-182.
Thi, M. T. T., Wibowo, D. & Rehm, B. H. A. 2020. Pseudomonas aeruginosa Biofilms. Int J Mol Sci, 21.
Tuon, F. F., Dantas, L. R., Suss, P. H. & Tasca Ribeiro, V. S. 2022. Pathogenesis of the Pseudomonas aeruginosa Biofilm: A Review. Pathogens, 11.
Wu, L., Ma, T., Zang, C., Xu, Z., Sun, W., Luo, H., Yang, M., Song, J., Chen, S. & Yao, H. 2024. Glycyrrhiza, a commonly used medicinal herb: Review of species classification, pharmacology, active ingredient biosynthesis, and synthetic biology. Journal of Advanced Research.
Yang, A. F., Huang, V., Samaroo-Campbell, J. & Augenbraun, M. 2023. Multi-drug resistant Pseudomonas aeruginosa: a 2019–2020 single center retrospective case control study. Infection Prevention in Practice, 5, 100296.
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