{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T20:26:23Z","timestamp":1791491183527,"version":"4.3.4"},"reference-count":54,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2024,6,28]],"date-time":"2024-06-28T00:00:00Z","timestamp":1719532800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Estonian Center of Analytical Chemistry (ECAC)","award":["SS22004"],"award-info":[{"award-number":["SS22004"]}]},{"name":"Estonian Research Council (TT4)","award":["SS22004"],"award-info":[{"award-number":["SS22004"]}]},{"DOI":"10.13039\/501100005789","name":"Tallinn University of Technology","doi-asserted-by":"publisher","award":["SS22004"],"award-info":[{"award-number":["SS22004"]}],"id":[{"id":"10.13039\/501100005789","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Lyme disease, caused by Borrelia burgdorferi sensu lato infection, is the most widespread vector-borne illness in the Northern Hemisphere. Unfortunately, using targeted antibiotic therapy is often an ineffective cure. The antibiotic resistance and recurring symptoms of Lyme disease are associated with the formation of biofilm-like aggregates of B. burgdorferi. Plant extracts could provide an effective alternative solution as many of them exhibit antibacterial or biofilm inhibiting activities. This study demonstrates the therapeutic potential of Plantago major and Plantago lanceolata as B. burgdorferi inhibitors. Hydroalcoholic extracts from three different samples of each plant were first characterised based on their total concentrations of polyphenolics, flavonoids, iridoids, and antioxidant capacity. Both plants contained substantial amounts of named phytochemicals and showed considerable antioxidant properties. The major non-volatile constituents were then quantified using HPLC-DAD-MS analyses, and volatile constituents were quantified using HS-SPME-GC-MS. The most prevalent non-volatiles were found to be plantamajoside and acteoside, and the most prevalent volatiles were \u03b2-caryophyllene, D-limonene, and \u03b1-caryophyllene. The B. burgdorferi inhibiting activity of the extracts was tested on stationary-phase B. burgdorferi culture and its biofilm fraction. All extracts showed antibacterial activity, with the most effective lowering the residual bacterial viability down to 15%. Moreover, the extracts prepared from the leaves of each plant additionally demonstrated biofilm inhibiting properties, reducing its formation by 30%.<\/jats:p>","DOI":"10.3390\/ijms25137112","type":"journal-article","created":{"date-parts":[[2024,6,28]],"date-time":"2024-06-28T06:51:53Z","timestamp":1719557513000},"page":"7112","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Plantago major and Plantago lanceolata Exhibit Antioxidant and Borrelia burgdorferi Inhibiting Activities"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2276-2925","authenticated-orcid":false,"given":"Pille-Riin","family":"Laanet","sequence":"first","affiliation":[{"name":"Department of Chemistry and Biotechnology, Tallinn University of Technology, Akadeemia tee 15, 12618 Tallinn, Estonia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olga","family":"Bragina","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biotechnology, Tallinn University of Technology, Akadeemia tee 15, 12618 Tallinn, Estonia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8373-9895","authenticated-orcid":false,"given":"Piia","family":"J\u00f5ul","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biotechnology, Tallinn University of Technology, Akadeemia tee 15, 12618 Tallinn, Estonia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3818-1906","authenticated-orcid":false,"given":"Merike","family":"Vaher","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biotechnology, Tallinn University of Technology, Akadeemia tee 15, 12618 Tallinn, Estonia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,6,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Radolf, J.D., Strle, K., Lemieux, J.E., and Strle, F. (2021). Lyme Disease in Humans. Curr. Issues Mol. Biol., 42.","DOI":"10.21775\/9781913652616.24"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Meril\u00e4inen, L., Herranen, A., Schwarzbach, A., and Gilbert, L. (2015). Morphological and Biochemical Features of Borrelia Burgdorferi Pleomorphic Forms. Microbiology, 161.","DOI":"10.1099\/mic.0.000027"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1001\/jama.2022.0081","article-title":"Diagnosis of Lyme Disease","volume":"327","author":"Pitrak","year":"2022","journal-title":"J. Am. Med. Assoc."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"16090","DOI":"10.1038\/nrdp.2016.90","article-title":"Lyme Borreliosis","volume":"2","author":"Steere","year":"2016","journal-title":"Nat. Rev. Dis. Prim."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1277","DOI":"10.1056\/NEJMe1502350","article-title":"Time for a Different Approach to Lyme Disease and Long-Term Symptoms","volume":"374","author":"Melia","year":"2016","journal-title":"N. Engl. J. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"598","DOI":"10.1111\/j.1469-0691.2004.00895.x","article-title":"Lyme Borreliosis: From Infection to Autoimmunity","volume":"10","author":"Singh","year":"2004","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Di Domenico, E.G., Cavallo, I., Bordignon, V., D\u2019Agosto, G., Pontone, M., Trento, E., Gallo, M.T., Prignano, G., Pimpinelli, F., and Toma, L. (2018). The Emerging Role of Microbial Biofilm in Lyme Neuroborreliosis. Front. Neurol., 9.","DOI":"10.3389\/fneur.2018.01048"},{"key":"ref_8","first-page":"S5","article-title":"Metamorphosis of Borrelia Burgdorferi Organisms\u2014RNA, Lipid and Protein Composition in Context with the Spirochetes\u2019 Shape","volume":"50","author":"Dihazi","year":"2010","journal-title":"J. Basic Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1127","DOI":"10.1128\/AAC.39.5.1127","article-title":"Effects of Penicillin, Ceftriaxone, and Doxycycline on Morphology of Borrelia Burgdorferi","volume":"39","author":"Kersten","year":"1995","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Aberer, E., and Duray, P.H. (1991). Morphology of Borrelia Burgdorferi: Structural Patterns of Cultured Borreliae in Relation to Staining Methods. J. Clin. Microbiol., 29.","DOI":"10.1128\/jcm.29.4.764-772.1991"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Sapi, E., Bastian, S.L., Mpoy, C.M., Scott, S., Rattelle, A., Pabbati, N., Poruri, A., Burugu, D., Theophilus, P.A.S., and Pham, T.V. (2012). Characterization of Biofilm Formation by Borrelia Burgdorferi In Vitro. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0048277"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1038\/nrmicro1820","article-title":"Morphological Plasticity as a Bacterial Survival Strategy","volume":"6","author":"Justice","year":"2008","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4","DOI":"10.3390\/ijpb13010003","article-title":"Bioactive Secondary Metabolites from Plant Sources: Types, Synthesis, and Their Therapeutic Uses","volume":"13","author":"Twaij","year":"2022","journal-title":"Int. J. Plant Biol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Saar-Reismaa, P., Bragina, O., Kuhtinskaja, M., Reile, I., Laanet, P.-R., Kulp, M., and Vaher, M. (2022). Extraction and Fractionation of Bioactives from Dipsacus Fullonum L. Leaves and Evaluation of Their Anti-Borrelia Activity. Pharmaceuticals, 15.","DOI":"10.3390\/ph15010087"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.indcrop.2015.12.038","article-title":"The Medicinal Potential of Plants from the Genus Plantago (Plantaginaceae)","volume":"83","author":"Romano","year":"2016","journal-title":"Ind. Crops Prod."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0378-8741(00)00212-9","article-title":"The Traditional Uses, Chemical Constituents and Biological Activities of Plantago Major L. A Review","volume":"71","author":"Samuelsen","year":"2000","journal-title":"J. Ethnopharmacol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1016\/S0031-9422(98)00210-6","article-title":"Effects of Cinnamic Acid on Polyphenol Production in Plantago lanceolata","volume":"49","author":"Fons","year":"1998","journal-title":"Phytochemistry"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/j.jep.2008.11.010","article-title":"Medicinal Herbs for the Treatment of Rheumatic Disorders\u2014A Survey of European Herbals from the 16th and 17th Century","volume":"121","author":"Adams","year":"2009","journal-title":"J. Ethnopharmacol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1016\/j.jevs.2011.03.014","article-title":"The Healing Potential of Plantago lanceolata Ointment on Collagenase-Induced Tendinitis in Burros (Equus asinus)","volume":"31","author":"Oloumi","year":"2011","journal-title":"J. Equine Vet. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1016\/S0031-9422(00)00306-X","article-title":"Chemotaxonomy of Plantago. Iridoid Glucosides and Caffeoyl Phenylethanoid Glycosides","volume":"55","author":"Franzyk","year":"2000","journal-title":"Phytochemistry"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"65","DOI":"10.2174\/157015909787602823","article-title":"Oxidative Stress and Neurodegenerative Diseases: A Review of Upstream and Downstream Antioxidant Therapeutic Options","volume":"7","author":"Uttara","year":"2009","journal-title":"Curr. Neuropharmacol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"750","DOI":"10.1016\/j.jep.2013.06.007","article-title":"Ethnopharmacological in Vitro Studies on Austria\u2019s Folk Medicine\u2014An Unexplored Lore in Vitro Anti-Inflammatory Activities of 71 Austrian Traditional Herbal Drugs","volume":"149","author":"Vogl","year":"2013","journal-title":"J. Ethnopharmacol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1002\/1099-1573(200012)14:8<617::AID-PTR674>3.0.CO;2-N","article-title":"Immunoenhancing Properties of Plantago Major Leaf Extract","volume":"14","author":"Calderon","year":"2000","journal-title":"Phyther. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1016\/j.jpba.2010.02.014","article-title":"Liquid Chromatography\/Tandem Mass Spectrometry Study of Anti-Inflammatory Activity of Plantain (Plantago L.) Species","volume":"52","author":"Beara","year":"2010","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1211\/0022357055605","article-title":"In-Vitro Anti-Inflammatory Activity of Pinus Sylvestris and Plantago lanceolata Extracts: Effect on Inducible NOS, COX-1, COX-2 and Their Products in J774A.1 Murine Macrophages","volume":"57","author":"Vigo","year":"2005","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1080\/09712119.2010.9707093","article-title":"Effect of Plantago Major on Burn Wound Healing in Rat","volume":"37","author":"Amini","year":"2010","journal-title":"J. Appl. Anim. Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1134\/S0003683811020086","article-title":"Study of a New Group of Bioregulators Isolated from the Greater Plantain (Plantago Major L.)","volume":"47","author":"Krasnov","year":"2011","journal-title":"Appl. Biochem. Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1379","DOI":"10.1258\/ebm.2012.012139","article-title":"Evaluation of Healing Wound and Genotoxicity Potentials from Extracts Hydroalcoholic of Plantago Major and Siparuna Guianensis","volume":"237","author":"Pereira","year":"2012","journal-title":"Exp. Biol. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1111\/jphp.12316","article-title":"Plantago lanceolata L. Water Extract Induces Transition of Fibroblasts into Myofibroblasts and Increases Tensile Strength of Healing Skin Wounds","volume":"67","author":"Sabol","year":"2014","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2487","DOI":"10.1080\/00032711003725581","article-title":"Quantitative Determination of Aucubin in Seven Plantago Species Using HPLC, HPTLC, and LC-ESI-MS Methods","volume":"43","author":"Beara","year":"2010","journal-title":"Anal. Lett."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1305","DOI":"10.1002\/cbdv.201200210","article-title":"Simultaneous RP-HPLC-DAD Separation, and Determination of Flavonoids and Phenolic Acids in Plantago L. Species","volume":"10","author":"Kremer","year":"2013","journal-title":"Chem. Biodivers."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1470","DOI":"10.1002\/jssc.201200010","article-title":"A Novel Strategy for Target Profiling Analysis of Bioactive Phenylethanoid Glycosides in Plantago Medicinal Plants Using Ultra-Performance Liquid Chromatography Coupled with Tandem Quadrupole Mass Spectrometry","volume":"35","author":"Qi","year":"2012","journal-title":"J. Sep. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2577","DOI":"10.1002\/elps.201300121","article-title":"Determination of Phenylethanoid Glycosides and Iridoid Glycosides from Therapeutically Used Plantago Species by CE-MEKC","volume":"34","author":"Gonda","year":"2013","journal-title":"Electrophoresis"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/j.jpba.2013.11.014","article-title":"Pharmacokinetics of Plantamajoside and Acteoside from Plantago asiatica in Rats by Liquid Chromatography\u2013Mass Spectrometry","volume":"89","author":"Li","year":"2014","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.3897\/pharmacia.69.e96526","article-title":"Study of Phytochemical Compounds of Plantago Major Leaves Grown in Kazakhstan","volume":"69","author":"Turgumbayeva","year":"2022","journal-title":"Pharmacia"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"183","DOI":"10.32598\/PBR.9.3.1061.4","article-title":"Antibacterial Activities and Chemical Compounds of Plantago lanceolata (Ribwort Plantain) and Plantago Major (Broadleaf Plantain) Leaf Extracts","volume":"9","author":"Bagheri","year":"2023","journal-title":"Pharm. Biomed. Res."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Laanet, P.-R., Saar-Reismaa, P., J\u00f5ul, P., Bragina, O., and Vaher, M. (2023). Phytochemical Screening and Antioxidant Activity of Selected Estonian Galium Species. Molecules, 28.","DOI":"10.3390\/molecules28062867"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Naqvi, S.A.R., Nadeem, S., Komal, S., Naqvi, S.A.A., Mubarik, M.S., Qureshi, S.Y., Ahmad, S., Abbas, A., Zahid, M., and Khan, N.-U.-H. (2019). Antioxidants: Natural Antibiotics. Antioxidants, IntechOpen.","DOI":"10.5772\/intechopen.84864"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1039\/D2NP00053A","article-title":"The Action of Phytochemicals in Biofilm Control","volume":"40","author":"Borges","year":"2023","journal-title":"Nat. Prod. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1768","DOI":"10.1021\/jf803011r","article-title":"Comparative Study of Antioxidant Properties and Total Phenolic Content of 30 Plant Extracts of Industrial Interest Using DPPH, ABTS, FRAP, SOD, and ORAC Assays","volume":"57","author":"Vitrac","year":"2009","journal-title":"J. Agric. Food Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.hermed.2012.03.001","article-title":"Antioxidant Capacity and Phenolic Constituents of Malva Neglecta Wallr. and Plantago lanceolata L. from Eastern Anatolia Region of Turkey","volume":"2","author":"Dalar","year":"2012","journal-title":"J. Herb. Med."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"5459","DOI":"10.3390\/molecules17055459","article-title":"Isolation and Characterization of Phenolic Antioxidants from Plantago Herb","volume":"17","author":"Amakura","year":"2012","journal-title":"Molecules"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Xiao, Y., Ren, Q., and Wu, L. (2022). The Pharmacokinetic Property and Pharmacological Activity of Acteoside: A Review. Biomed. Pharmacother., 153.","DOI":"10.1016\/j.biopha.2022.113296"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1576","DOI":"10.1007\/s13197-015-2083-x","article-title":"Chemical Composition of Essential Oils from Plantago lanceolata L. Leaves Extracted by Hydrodistillation","volume":"53","author":"Bajer","year":"2016","journal-title":"J. Food Sci. Technol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Hashiesh, H.M., Sharma, C., Goyal, S.N., Sadek, B., Jha, N.K., Al Kaabi, J., and Ojha, S. (2021). A Focused Review on CB2 Receptor-Selective Pharmacological Properties and Therapeutic Potential of \u03b2-Caryophyllene, a Dietary Cannabinoid. Biomed. Pharmacother., 140.","DOI":"10.1016\/j.biopha.2021.111639"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Anandakumar, P., Kamaraj, S., and Vanitha, M.K. (2021). D-Limonene: A Multifunctional Compound with Potent Therapeutic Effects. J. Food Biochem., 45.","DOI":"10.1111\/jfbc.13566"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"162","DOI":"10.3329\/ajmbr.v5i3.43584","article-title":"Chinese Star Anise and Anise, Magic Herbs in Traditional Chinese Medicine and Modern Pharmaceutical Science","volume":"5","author":"Shahrajabian","year":"2019","journal-title":"Asian J. Med. Biol. Res."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Di Somma, A., Moretta, A., Can\u00e8, C., Cirillo, A., and Duilio, A. (2020). Inhibition of Bacterial Biofilm Formation. Bacterial Biofilms, IntechOpen.","DOI":"10.5772\/intechopen.90614"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1016\/j.jaap.2013.06.011","article-title":"Quantification of Total Phenols in Bio-Oil Using the Folin-Ciocalteu Method","volume":"104","author":"Rover","year":"2013","journal-title":"J. Anal. Appl. Pyrolysis"},{"key":"ref_50","unstructured":"Badal, S., and Delgoda, R. (2017). Pharmacognosy: Fundamentals, Applications and Strategies, Elsevier, Inc."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.bse.2012.02.013","article-title":"Comparative Study of Some Polyphenols in Plantago Species","volume":"42","author":"Beara","year":"2012","journal-title":"Biochem. Syst. Ecol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1006\/abio.2000.4691","article-title":"A Fluorometric Method for Measurement of Oxygen Radical-Scavenging Activity of Water-Soluble Antioxidants","volume":"284","author":"Naguib","year":"2000","journal-title":"Anal. Biochem."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Feng, J., Wang, T., Zhang, S., Shi, W., and Zhang, Y. (2014). An Optimized SYBR Green I\/PI Assay for Rapid Viability Assessment and Antibiotic Susceptibility Testing for Borrelia Burgdorferi. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0111809"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"114203","DOI":"10.1016\/j.ejmech.2022.114203","article-title":"Development of Small Molecules That Work Cooperatively with Ciprofloxacin to Clear Salmonella Biofilms in a Chronic Gallbladder Carriage Model","volume":"232","author":"Woolard","year":"2022","journal-title":"Eur. J. Med. Chem."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/13\/7112\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:06:53Z","timestamp":1760108813000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/13\/7112"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,6,28]]},"references-count":54,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2024,7]]}},"alternative-id":["ijms25137112"],"URL":"https:\/\/doi.org\/10.3390\/ijms25137112","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,6,28]]}}}