Antibiofilm activity and phytochemical compounds of Selaginella plana extract against human pathogenic microorganisms

Juen Carla Warella; Khairunnida Rahma; Dominggus Rumahlatu; Johan Bruyif Bension

doi:10.55779/nsb18212853

Authors

Juen Carla Warella Universitas Pattimura, Faculty of Medicine, Department of Microbiology, Ir. M. Putuhena, Poka, 97233, Ambon (ID) https://orcid.org/0000-0003-2341-521X
Khairunnida Rahma Universitas Mulawarman, Faculty of Medicine, Department of Parasitology, Kuaro, 75119, Samarinda (ID) https://orcid.org/0000-0002-6089-3619
Dominggus Rumahlatu Universitas Pattimura, Faculty of Teacher Training and Education, Study Program of Biology Education, Ir. M. Putuhena, Poka, 97233, Ambon (ID) https://orcid.org/0000-0002-4466-5528
Johan Bruyif Bension Universitas Pattimura, Faculty of Medicine, Department of Medical Education, Ir. M. Putuhena, Poka, 97233, Ambon (ID) https://orcid.org/0009-0005-6215-443X

DOI:

https://doi.org/10.55779/nsb18212853

Keywords:

biofilm inhibition, secondary metabolites, medical plants, pathogenic microbes, ethanol extract, bioprospecting

Abstract

Selaginella plana (Desv.) Heiron, traditionally used in Indonesia, is known for its efficacy in treating infections in open wounds. Untreated wounds can lead to infections, cellulitis, sepsis, and biofilm formation, all of which may delay the wound-healing process. This study aimed to investigate the secondary metabolite content and antibiofilm activity of S. plana extract in preventing, inhibiting, and degrading biofilm formation by Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), Streptococcus pyogenes, and Candida albicans using the 96-well microtiter plate method. Three 96-well plates were inoculated with microbial suspensions, S. plana extract, and trypticase soy broth with 2% glucose, then incubated for 24–72 hours. Biofilms were stained with 1% crystal violet, and absorbance was measured at 595 nm. The results showed that S. plana extract contains secondary metabolites such as flavonoids, terpenoids, steroids, and saponins. The antibiofilm assay showed the highest biofilm prevention in S. aureus (49.2%), C. albicans (47%), and S. pyogenes (46%), with the lowest in MRSA (8.5%). Biofilm inhibition was greatest in S. aureus (42%), S. pyogenes (40%), and C. albicans (35.2%), while the highest degradation occurred in S. pyogenes (52%), C. albicans (48%), and S. aureus (42%). The study concluded that S. plana exhibits notable antibiofilm activity against S. aureus, S. pyogenes, and C. albicans, though it demonstrates limited efficacy against MRSA. These findings suggest that S. plana leaf extract holds potential for antibacterial applications and may serve as a promising candidate for bioprospecting in drug development.

Metrics

Metrics Loading ...

References

Adnan M, Siddiqui AJ, Hamadou WS, Patel M, Ashraf SA, Jamal A, et al. (2021). Phytochemistry, bioactivities, pharmacokinetics and toxicity prediction of Selaginella repanda with its anticancer potential against human lung, breast and colorectal carcinoma cell lines. Molecules, 26(3). https://doi.org/10.3390/molecules26030768

Alelah SMA, Abdulrazzaq AI, Hussein MO, Nasser SJ, Mssdf AK (2026). Overview of extraction methods for phytochemicals: principles, applications, and future directions. Results in Chemistry, 24: 103221. https://doi.org/10.1016/j.rechem.2026.103221

Alsolami A, Dirar AI, Konozy EHE, Osman ME-FM, Ibrahim MA, Alshammari KF, et al. (2023). Genome-wide mining of Selaginella moellendorffii for hevein-like lectins and their potential molecular mimicry with SARS-CoV-2 spike glycoprotein. Current Issues in Molecular Biology, 45(7): 5879–5901. https://doi.org/10.3390/cimb45070372

Antony A, Farid M (2022). Effect of temperatures on polyphenols during extraction. Applied Sciences, 12(4): 2107. https://doi.org/10.3390/app12042107

Bailly C (2021). The traditional and modern uses of Selaginella tamariscina (P.beauv.) spring, in medicine and cosmetic: applications and bioactive ingredients. Journal of Ethnopharmacology, 280: 114444. https://doi.org/10.1016/j.jep.2021.114444

Batiha GE-S, Alqahtani A, Ojo OA, Shaheen HM, Wasef L, Elzeiny M, et al. (2020). Biological properties, bioactive constituents, and pharmacokinetics of some Capsicum spp. and capsaicinoids. International Journal of Molecular Sciences, 21(15): 5179. https://doi.org/10.3390/ijms21155179

Bhadange YA, Carpenter J, Saharan VK (2024). A Comprehensive review on advanced extraction techniques for retrieving bioactive components from natural sources. ACS Omega, 9(29): 31274–31297. https://doi.org/10.1021/acsomega.4c02718

Bitwell C, Indra SS, Luke C, Kakoma MK (2023). A review of modern and conventional extraction techniques and their applications for extracting phytochemicals from plants. Scientific African, 19: e01585. https://doi.org/10.1016/j.sciaf.2023.e01585

Chen X, Yang Y, He X (2025). Ethnomedicinal uses, phytochemistry and pharmacological properties of Selaginella uncinata (Desv.) spring: A review. Fitoterapia, 184: 106634. https://doi.org/10.1016/j.fitote.2025.106634

Cho W-K, Choi H-J, Ma JY (2024). Selaginella tamariscina ethanol extract attenuates influenza a virus infection by inhibiting hemagglutinin and neuraminidase. Nutrients, 16(14): 2377. https://doi.org/10.3390/nu16142377

Elboughdiri N (2018). Effect of time, solvent-solid ratio, ethanol concentration and temperature on extraction yield of phenolic compounds from olive leaves. Engineering, Technology & Applied Science Research, 8(2): 2805–2808. https://doi.org/10.48084/etasr.1983

Gautam A, Pal LC, Rao C V., Kumar V (2023). The role of indian magical herb Selaginella bryopteris L. (selaginaceae) in pharmacotherapeutic perspective: an overview. Pharmacognosy Journal, 15(1): 14–20. https://doi.org/10.5530/pj.2023.15.3

Gemmell CT, Parreira VR, Farber JM (2022). Controlling Listeria monocytogenes growth and biofilm formation using flavonoids. Journal of Food Protection, 85(4): 639–646. https://doi.org/10.4315/JFP-21-135

Gu X, Hao D, Xiao P (2022). Research progress of chinese herbal medicine compounds and their bioactivities: fruitful 2020. Chinese Herbal Medicines, 14(2): 171–186. https://doi.org/10.1016/j.chmed.2022.03.004

Gulati M, Lohse MB, Ennis CL, Gonzalez RE, Perry AM, Bapat P, et al. In vitro culturing and screening of Candida albicans biofilms. Current Protocols Microbiology, 50(1): e60. https://doi.org/10.1002/cpmc.60

Hatcher CR, Ryves DB, Millett J (2020). The function of secondary metabolites in plant carnivory. Annals of Botany, 125(3): 399–411. https://doi.org/10.1093/aob/mcz191

Hernández-Cuellar E, Tsuchiya K, Valle-Ríos R, Medina-Contreras O (2023). Differences in biofilm formation by methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains. Diseases, 11(4): 160. https://doi.org/10.3390/diseases11040160

Huang Y, Xie F-J, Cao X, Li M-Y (2021). Research progress in biosynthesis and regulation of plant terpenoids. Biotechnology & Biotechnological Equipment, 35(1): 1799–1808. https://doi.org/10.1080/13102818.2021.2020162

Kamal RM, Abo-Elghiet F, Hal DM, Hammam WE, El-Dessouki AM, Mahrous MH, et al. (2026). From conventional to green extraction of saponins: A review on plant matrices and byproducts. Journal of Agriculture and Food Research, 25: 102607. https://doi.org/10.1016/j.jafr.2025.102607

Klahs PC, McMurchie EK, Nikkel JJ, Clark LG (2023). A maceration technique for soft plant tissue without hazardous chemicals. Applications in Plant Sciences, 11(5): e11543. https://doi.org/10.1002/aps3.11543

Kovács R, Majoros L (2020). Fungal Quorum-Sensing Molecules: A review of their antifungal effect against Candida biofilms. Journal of Fungi, 6(3): 99. https://doi.org/10.3390/jof6030099

Křížkovská B, Kumar R, Řehořová K, Sýkora D, Dobiasová S, Kučerová D, et al. (2021). Comparison of chemical composition and biological activities of eight Selaginella species. Pharmaceuticals, 14(1): 16. https://doi.org/10.3390/ph14010016

Kumar A, Nirmal P, Kumar M, Jose A, Tomer V, Oz E, et al. (2023). Major phytochemicals: recent advances in health benefits and extraction method. Molecules, 28(2): 887. https://doi.org/10.3390/molecules28020887

Lee J-H, Kim Y-G, Park I, Lee J (2024). Antifungal and antibiofilm activities of flavonoids against Candida albicans: Focus on 3,2′-dihydroxyflavone as a potential therapeutic agent. Biofilm, 8: 100218. https://doi.org/10.1016/j.bioflm.2024.100218

Liu X, Yao H, Zhao X, Ge C (2023). Biofilm formation and control of foodborne pathogenic bacteria. Molecules, 28(6): 2432. https://doi.org/10.3390/molecules28062432

Lu X, Li W, Wang Q, Wang J, Qin S (2023). Progress on the extraction, separation, biological activity, and delivery of natural plant pigments. Molecules, 28(14): 5364. https://doi.org/10.3390/molecules28145364

Macé S, Hansen LT, Rupasinghe HPV (2017). Anti-bacterial activity of phenolic compounds against Streptococcus pyogenes. Medicines, 4(2): 25. https://doi.org/10.3390/medicines4020025

Mirzaei A, Karamolah KS, Mahnaie MP, Mousavi F, Moghadam PM, Mahmoudi H (2020). Antibacterial activity of honey against methicillin-resistant and sensitive Staphylococcus aureus isolated from patients with diabetic foot ulcer. The Open Microbiology Journal, 14(1): 260–265. https://doi.org/10.2174/1874434602014010260

Muema FW, Liu Y, Zhang Y, Chen G, Guo M (2022). Flavonoids from Selaginella doederleinii hieron and their antioxidant and antiproliferative activities. Antioxidants, 11(6): 1189. https://doi.org/10.3390/antiox11061189

Neenu RS, Prakash GW, Praveen Dhar T, Biju C, Brijithlal ND (2024). Antioxidant, antimicrobial and phytochemical analysis of four species of Selaginella P. Beauv. International Journal of Plant and Environment, 10(03): 102–109. https://doi.org/10.18811/ijpen.v10i03.11

Nguyen HTT, Nguyen TH, Otto M (2020). The staphylococcal exopolysaccharide PIA – biosynthesis and role in biofilm formation, colonization, and infection. Computational and Structural Biotechnology Journal, 18: 3324–3334. https://doi.org/10.1016/j.csbj.2020.10.027

Olawuwo OS, Famuyide IM, McGaw LJ (2022). Antibacterial and antibiofilm activity of selected medicinal plant leaf extracts against pathogens implicated in poultry diseases. Frontiers in Veterinary Science, 9. https://doi.org/10.3389/fvets.2022.820304

Pang Z, Chen J, Wang T, Gao C, Li Z, Guo L, et al. (2021). Linking plant secondary metabolites and plant microbiomes: a review. Frontiers in Plant Science, 12. https://doi.org/10.3389/fpls.2021.621276

Pengpid S, Peltzer K (2018). Utilization of traditional and complementary medicine in Indonesia: Results of a national survey in 2014–15. Complementary Therapies in Clinical Practice, 33: 156–163. https://doi.org/10.1016/j.ctcp.2018.10.006

Qiu D, Wen Y, Xie Y, Lin Q (2020). The complete chloroplast genome sequence of medicinal plant, Selaginella involvens. Mitochondrial DNA Part B, 5(3): 2605–2606. https://doi.org/10.1080/23802359.2020.1781574

Rodrigues CF, Černáková L (2020). Farnesol and tyrosol: secondary metabolites with a crucial quorum-sensing role in Candida biofilm development. Genes, 11(4): 444. https://doi.org/10.3390/genes11040444

Shariati A, Didehdar M, Razavi S, Heidary M, Soroush F, Chegini Z (2022). Natural compounds: a hopeful promise as an antibiofilm agent against Candida species. Frontiers in Pharmacology, 13. https://doi.org/10.3389/fphar.2022.917787

Sharma S, Mohler J, Mahajan SD, Schwartz SA, Bruggemann L, Aalinkeel R (2023). Microbial biofilm: a review on formation, infection, antibiotic resistance, control measures, and innovative treatment. Microorganisms, 11(6): 1614. https://doi.org/10.3390/microorganisms11061614

Shree P, Singh CK, Sodhi KK, Surya JN, Singh DK (2023). Biofilms: understanding the structure and contribution towards bacterial resistance in antibiotics. Medicine in Microecology, 16: 100084. https://doi.org/10.1016/j.medmic.2023.100084

Sivaraj D, Shanmugam S, Rajan M, Sasidharan SP, Sathyanarayanan S, Muniyandi K, et al. (2018). Evaluation of Aristolochia indica L. and Piper nigrum L. methanol extract against centipede Scolopendra moristans L. using wistar albino rats and screening of bioactive compounds by high pressure liquid chromatography: a polyherbal formulation. Biomedicine & Pharmacotherapy, 97: 1603–1612. https://doi.org/10.1016/j.biopha.2017.11.114

Solis-Velazquez OA, Gutiérrez-Lomelí M, Guerreo-Medina PJ, Rosas-García M de L, Iñiguez-Moreno M, Avila-Novoa MG (2021). Nosocomial pathogen biofilms on biomaterials: different growth medium conditions and components of biofilms produced in vitro. Journal of Microbiology, Immunology and Infection, 54(6): 1038–1047. https://doi.org/10.1016/j.jmii.2020.07.002

Sulaiman M, Jannat K, Nissapatorn V, Rahmatullah M, Paul AK, de Lourdes Pereira M, et al. (2022). Antibacterial and antifungal alkaloids from Asian angiosperms: distribution, mechanisms of action, structure-activity, and clinical potentials. Antibiotics, 11(9): 1146. https://doi.org/10.3390/antibiotics11091146

Vaou N, Stavropoulou E, Voidarou C, Tsigalou C, Bezirtzoglou E (2021). Towards advances in medicinal plant antimicrobial activity: a review study on challenges and future perspectives. Microorganisms, 9(10): 2041. https://doi.org/10.3390/microorganisms9102041

Warella JC, Rahma K, Widodo ADW, Setiabudi RJ (2023). Antifungal activity of Selaginella plana (Desv. ex Poir.) Hieron extract against Candida albicans in vitro. Folia Medica Indonesiana, 59(3): 295–301. https://doi.org/10.20473/fmi.v59i3.44165

Wen Q-H, Wang R, Zhao S-Q, Chen B-R, Zeng X-A (2021). Inhibition of biofilm formation of foodborne Staphylococcus aureus by the citrus flavonoid naringenin. Foods, 10(11): 2614. https://doi.org/10.3390/foods10112614

Wijesundara NM, Lee SF, Rupasinghe HPV (2022). Carvacrol inhibits Streptococcus pyogenes biofilms by suppressing the expression of genes associated with quorum-sensing and reducing cell surface hydrophobicity. Microbial Pathogenesis, 169: 105684. https://doi.org/10.1016/j.micpath.2022.105684

World Health Organization (2022). Global strategies and plans of action that are scheduled to expire within one year - WHO traditional medicine strategy: 2014–2023. World Health Organization (WHO). Available at: https://www.who.int/publications/i/item/9789241506096

Wu X, Wang H, Xiong J, Yang G-X, Hu J-F, Zhu Q, et al. (2024). Staphylococcus aureus biofilm: formulation, regulatory, and emerging natural products-derived therapeutics. Biofilm, 7: 100175. https://doi.org/10.1016/j.bioflm.2023.100175

Yeh T, Sung W, Shih W (2024). Antibacterial and antibiofilm effects of Camellia oleifera seed dreg extract and its application in cosmetics. Journal of Cosmetic Dermatology, 23(3): 1055–1065. https://doi.org/10.1111/jocd.16068

Zafer MM, Mohamed GA, Ibrahim SRM, Ghosh S, Bornman C, Elfaky MA (2024). Biofilm-mediated infections by multidrug-resistant microbes: a comprehensive exploration and forward perspectives. Archives of Microbiology, 206(3): 101. https://doi.org/10.1007/s00203-023-03826-z

Zhang L, Liang E, Cheng Y, Mahmood T, Ge F, Zhou K, et al. (2020). Is combined medication with natural medicine a promising therapy for bacterial biofilm infection? Biomedicine & Pharmacotherapy, 128: 110184. https://doi.org/10.1016/j.biopha.2020.110184

Zhang X, Liu Y, Deng G, Huang B, Kai G, Chen K, et al. (2021). A purified biflavonoid extract from Selaginella moellendorffii alleviates gout arthritis via NLRP3/ASC/Caspase-1 axis suppression. Frontiers in Pharmacology, 12: 676297. https://doi.org/10.3389/fphar.2021.676297

Zhao A, Sun J, Liu Y (2023). Understanding bacterial biofilms: from definition to treatment strategies. Frontiers in Cellular and Infection Microbiology, 13. https://doi.org/10.3389/fcimb.2023.1137947

Zhydzetski A, Głowacka-Grzyb Z, Bukowski M, Żądło T, Bonar E, Władyka B (2024). Agents targeting the bacterial cell wall as tools to combat gram-positive pathogens. Molecules, 29(17): 4065. https://doi.org/10.3390/molecules29174065