{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T01:41:26Z","timestamp":1792806086814,"version":"4.5.0"},"reference-count":58,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2024,12,26]],"date-time":"2024-12-26T00:00:00Z","timestamp":1735171200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Research Council of Lithuania","award":["09.3.3-LMT-K-712-10-0232"],"award-info":[{"award-number":["09.3.3-LMT-K-712-10-0232"]}],"id":[{"id":"https:\/\/ror.org\/02gs16m83","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Research Council of Lithuania","award":["09.3.3-LMT-K-712-16-0030"],"award-info":[{"award-number":["09.3.3-LMT-K-712-16-0030"]}],"id":[{"id":"https:\/\/ror.org\/02gs16m83","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Instrumental Analysis Open Access Centre of Vytautas Magnus University","award":["09.3.3-LMT-K-712-10-0232"],"award-info":[{"award-number":["09.3.3-LMT-K-712-10-0232"]}],"id":[{"id":"https:\/\/ror.org\/04y7eh037","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Instrumental Analysis Open Access Centre of Vytautas Magnus University","award":["09.3.3-LMT-K-712-16-0030"],"award-info":[{"award-number":["09.3.3-LMT-K-712-16-0030"]}],"id":[{"id":"https:\/\/ror.org\/04y7eh037","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background\/Objectives: A variety of phytochemicals from different plants are collected by bees into bee pollen granules. This research focused on evaluating the effects of lactic acid fermentation and enzymatic hydrolysis on the antibacterial activity of bee pollen and its interaction with antibiotics. There is limited knowledge regarding the interactions between treated bee pollen extracts and antibiotics, and this study contributes to the field by providing new insights into the antibacterial activity of pollen subjected to eight distinct treatment methods. Methods: Bee pollen\u2019s bacterial fermentation using a Lacticaseibacillus rhamnosus culture and spontaneous fermentation were performed. Bee pollen hydrolysis was performed using commercial enzymes, including enzyme mixtures as well as pure enzymes. The agar well diffusion assay was employed to assess the antibacterial activity against Staphylococcus aureus, Listeria monocytogenes, and Salmonella enterica serovars Enteritidis and Typhimurium, as well as their interaction with antibiotics (ceftazidime, ciprofloxacin, oxytetracycline dihydrate, and erythromycin). Results: This study showed an enhancement in bee pollen\u2019s antibacterial activity after both fermentation and enzymatic hydrolysis. The increase varied with the pollen\u2019s origin, treatment type, and culture used for antimicrobial tests. More than 77% of bee pollen extracts demonstrated a synergistic effect with antibiotics across all tested bacterial strains, while antagonistic interactions were comparatively rare. Conclusions: The applied treatment methods can improve the antibacterial properties of bee pollen. Bee pollen extracts, in combination with antibiotics, can enhance their effectiveness. These findings provide new insights into the potential use of bee pollen in combating bacterial infections.<\/jats:p>","DOI":"10.3390\/ph18010015","type":"journal-article","created":{"date-parts":[[2024,12,26]],"date-time":"2024-12-26T19:33:07Z","timestamp":1735241587000},"page":"15","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Improved Antibacterial Properties of Fermented and Enzymatically Hydrolyzed Bee Pollen and Its Combined Effect with Antibiotics"],"prefix":"10.3390","volume":"18","author":[{"given":"Vaida","family":"Damulien\u0117","sequence":"first","affiliation":[{"name":"Instrumental Analysis Open Access Centre, Vytautas Magnus University, LT-44404 Kaunas, Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1939-9408","authenticated-orcid":false,"given":"Vilma","family":"Ka\u0161konien\u0117","sequence":"additional","affiliation":[{"name":"Instrumental Analysis Open Access Centre, Vytautas Magnus University, LT-44404 Kaunas, Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paulius","family":"Ka\u0161konas","sequence":"additional","affiliation":[{"name":"Institute of Metrology, Kaunas University of Technology, LT-51368 Kaunas, Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6798-3708","authenticated-orcid":false,"given":"R\u016bta","family":"Mickien\u0117","sequence":"additional","affiliation":[{"name":"Instrumental Analysis Open Access Centre, Vytautas Magnus University, LT-44404 Kaunas, Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Audrius","family":"Maru\u0161ka","sequence":"additional","affiliation":[{"name":"Instrumental Analysis Open Access Centre, Vytautas Magnus University, LT-44404 Kaunas, Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,12,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1582","DOI":"10.1016\/j.biotechadv.2015.08.001","article-title":"Discovery and resupply of pharmacologically active plant-derived natural products: A review","volume":"33","author":"Atanasov","year":"2015","journal-title":"Biotechnol. Adv."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1038\/s41573-020-00114-z","article-title":"Natural products in drug discovery: Advances and opportunities","volume":"20","author":"Atanasov","year":"2021","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2973","DOI":"10.1016\/j.phytochem.2007.09.004","article-title":"Phytochemicals: The good, the bad and the ugly?","volume":"68","author":"Molyneux","year":"2007","journal-title":"Phytochemistry"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Yuan, H., Ma, Q., Ye, L., and Piao, G. (2016). The traditional medicine and modern medicine from natural products. Molecules, 21.","DOI":"10.3390\/molecules21050559"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/j.ijantimicag.2005.09.002","article-title":"Antimicrobial activity of flavonoids","volume":"26","author":"Cushnie","year":"2005","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_6","first-page":"684","article-title":"Evaluation of antifungal activity and phytochemical screening of Solanum surattense seeds","volume":"3","author":"David","year":"2010","journal-title":"J. Pharm. Res."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Egamberdieva, D., Wirth, S., Behrendt, U., Parvaiz, A., and Berg, G. (2017). Antimicrobial activity of medicinal plants correlates with the proportion of antagonistic endophytes. Front. Microbiol., 8.","DOI":"10.3389\/fmicb.2017.00199"},{"key":"ref_8","first-page":"697","article-title":"Antimicrobial activity of some plant extracts against bacterial pathogens isolated from feces of red deer (Cervus elaphus)","volume":"20","author":"Gnat","year":"2017","journal-title":"Pol. J. Vet. Sci."},{"key":"ref_9","first-page":"2145","article-title":"Antibacterial, antifungal and antioxidant activities of some medicinal plants","volume":"6","author":"Wazir","year":"2014","journal-title":"Pak. J. Pharm. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"535","DOI":"10.3390\/futurepharmacol3030034","article-title":"Use of plant extracts, bee-derived products, and probiotic-related applications to fight multidrug-resistant pathogens in the post-antibiotic era","volume":"3","author":"Machado","year":"2023","journal-title":"Future Pharmacol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e20180117","DOI":"10.1590\/0001-3765201920180117","article-title":"Plant extracts display synergism with different classes of antibiotics","volume":"91","author":"Silva","year":"2019","journal-title":"An. Acad. Bras. Cienc."},{"key":"ref_12","first-page":"59","article-title":"The antibacterial effect of some medicinal plant extracts and their synergistic effect with antibiotics","volume":"46","author":"Jouda","year":"2016","journal-title":"World J. Pharm. Pharm. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Algethami, J.S., El-Wahed, A.A.A., Elashal, M.H., Ahmed, H.R., Elshafiey, E.H., Omar, E.M., Naggar, Y.A., Algethami, A.F., Shou, Q., and Alsharif, S.M. (2022). Bee pollen: Clinical trials and patent applications. Nutrients, 14.","DOI":"10.3390\/nu14142858"},{"key":"ref_14","first-page":"S483","article-title":"Bee pollen: A superfood with health benefits and digestibility challenges","volume":"9","author":"Agrawal","year":"2023","journal-title":"J. Food Chem. Nanotechnol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4303","DOI":"10.1002\/jsfa.7729","article-title":"Biological and therapeutic properties of bee pollen: A review","volume":"96","author":"Denisow","year":"2016","journal-title":"J. Sci. Food Agric."},{"key":"ref_16","first-page":"297425","article-title":"Bee pollen: Chemical composition and therapeutic application","volume":"2015","author":"Olczyk","year":"2015","journal-title":"eCAM"},{"key":"ref_17","first-page":"6","article-title":"Antioxidant properties of pollen","volume":"3","year":"2014","journal-title":"Hrana Zdr. Boles. Znan. Stru\u010dni \u010casopis Za Nutr. Dijetetiku"},{"key":"ref_18","first-page":"306","article-title":"Relation between composition, antioxidant and antibacterial activities, and botanical origin of multi-floral bee pollen","volume":"90","author":"Retamal","year":"2017","journal-title":"J. Appl. Bot. Food Qual."},{"key":"ref_19","first-page":"678","article-title":"Bacteriocins and lactic acid bacteria\u2014A minireview","volume":"5","author":"Savadogo","year":"2006","journal-title":"Afr. J. Biotechnol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Chen, J., Pang, H., Wang, L., Ma, C., Wu, G., Liu, Y., Guan, Y., Zhang, M., Qin, G., and Tan, Z. (2022). Bacteriocin-producing lactic acid bacteria strains with antimicrobial activity screened from Bamei pig feces. Foods, 11.","DOI":"10.3390\/foods11050709"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1323","DOI":"10.1007\/s00253-017-8088-9","article-title":"Current state of purification, isolation, and analysis of bacteriocins produced by lactic acid bacteria","volume":"101","author":"Malakauskas","year":"2017","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_22","first-page":"119","article-title":"Digestibility and availability of nutrients in bee pollen applying different pretreatments","volume":"19","author":"Quicazan","year":"2017","journal-title":"Ing. Compet."},{"key":"ref_23","first-page":"79","article-title":"Bee-pollen structure modification by physical and biotechnological processing: Influence on the availability of nutrients and bioactive compounds","volume":"43","author":"Zuluaga","year":"2015","journal-title":"Chem. Eng. Trans."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"21732","DOI":"10.3390\/molecules201219800","article-title":"Polyphenols from bee pollen: Structure, absorption, metabolism and biological activity","volume":"20","author":"Stojko","year":"2015","journal-title":"Molecules"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2744","DOI":"10.1002\/jsfa.10902","article-title":"In vitro and in vivo digestion comparison of bee pollen with or without wall-disruption","volume":"101","author":"Wu","year":"2021","journal-title":"J. Sci. Food Agric."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.foodres.2015.05.039","article-title":"Cell wall disruption of rape bee pollen treated with combination of protamex hydrolysis and ultrasonication","volume":"75","author":"Dong","year":"2015","journal-title":"Food Res. Int."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.ifset.2008.08.004","article-title":"Breaking the cells of rape bee pollen and consecutive extraction of functional oil with supercritical carbon dioxide","volume":"10","author":"Xiang","year":"2009","journal-title":"Innov. Food Sci. Emerg. Technol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Ada\u0161kevi\u010di\u016bt\u0117, V., Ka\u0161konien\u0117, V., Bar\u010dauskait\u0117, K., Ka\u0161konas, P., and Maru\u0161ka, A. (2022). The impact of fermentation on bee pollen polyphenolic compounds composition. Antioxidants, 11.","DOI":"10.3390\/antiox11040645"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2115","DOI":"10.1007\/s11696-018-0417-7","article-title":"The impact of solid state fermentation on bee pollen phenolic compounds and radical scavenging capacity","volume":"72","year":"2018","journal-title":"Chem. Pap."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"100532","DOI":"10.1016\/j.fbio.2020.100532","article-title":"Antimicrobial and antioxidant activities of natural and fermented bee pollen","volume":"34","year":"2020","journal-title":"Food Biosci."},{"key":"ref_31","first-page":"e13983","article-title":"Effect of enzymatic hydrolysis on structural characteristics and bioactive composition of bee-pollen","volume":"43","year":"2019","journal-title":"J. Food Process."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Damulien\u0117, V., Ka\u0161konien\u0117, V., Ka\u0161konas, P., and Maru\u0161ka, A. (2023). The influence of enzymatic hydrolysis on bee pollen antioxidant and antibacterial activities. Foods, 12.","DOI":"10.3390\/foods12193582"},{"key":"ref_33","first-page":"131","article-title":"The role of bee products in the control of antimicrobial resistance and biofilm formation","volume":"30","author":"Acaroz","year":"2024","journal-title":"Kafkas Univ. Vet. Fak. Derg."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Al-Ani, I., Zimmermann, S., Reichling, J., and Wink, M. (2018). Antimicrobial activities of European propolis collected from various geographic origins alone and in combination with antibiotics. Medicines, 5.","DOI":"10.3390\/medicines5010002"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Hanafy, N.A., Eltonouby, E.A.B., Salim, E.I., Mahfouz, M.E., Leporatti, S., and Hafez, E.H. (2023). Simultaneous administration of bevacizumab with bee-pollen extract-loaded hybrid protein hydrogel NPs is a promising targeted strategy against cancer cells. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24043548"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1","DOI":"10.18103\/mra.v12i9.5924","article-title":"Evaluation of the potential synergistic effect of Tetragonisca angustula pot-pollen with amikacin and meropenem against extensively drug-resistant bacteria of clinical origin","volume":"12","author":"Araque","year":"2024","journal-title":"Med. Res. Arch."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Ilie, C.I., Oprea, E., Geana, E.I., Spoiala, A., Buleandra, M., Gradisteanu Pircalabioru, G., Badea, I.A., Ficai, D., Andronescu, E., and Ficai, A. (2022). Bee pollen extracts: Chemical composition, antioxidant properties, and effect on the growth of selected probiotic and pathogenic bacteria. Antioxidants, 11.","DOI":"10.3390\/antiox11050959"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1016\/j.tifs.2021.06.016","article-title":"Food safety hazards of bee pollen\u2014A review","volume":"114","author":"Sipos","year":"2021","journal-title":"Trends Food Sci. Technol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"117935","DOI":"10.1016\/j.trac.2024.117935","article-title":"Unveiling bee pollen\u2019s contamination with pesticides and mycotoxins: Current analytical procedures, results and regulation","volume":"180","author":"Carrera","year":"2024","journal-title":"TrAC Trends Anal. Chem."},{"key":"ref_40","unstructured":"(2024, October 14). Regulation (EU) 2023\/915 of 25 April 2023 on Maximum Levels for Certain Contaminants in Food and Repealing Regulation (EC) No 1881\/2006. Available online: http:\/\/data.europa.eu\/eli\/reg\/2023\/915\/2024-07-22."},{"key":"ref_41","unstructured":"(2024, October 14). Regulation (EC) No 396\/2005 of the European Parliament and of the Council of 23 February 2005 on Maximum Residue Levels of Pesticides in or on Food and Feed of Plant and Animal Origin and Amending Council Directive 91\/414\/EEC (Text with EEA relevance). Available online: http:\/\/data.europa.eu\/eli\/reg\/2005\/396\/2024-11-06."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1080\/00173130260045468","article-title":"Immunochemical and cytochemical detection of wall components of germinated pollen of gymnosperms","volume":"41","author":"Yatomi","year":"2002","journal-title":"Grana"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Zhang, D., and Li, H. (2014). Exine export in pollen. Plant ABC Transporters, Springer International Publishing.","DOI":"10.1007\/978-3-319-06511-3_4"},{"key":"ref_44","first-page":"71","article-title":"Effectiveness of ethanol and methanol alcohols on different isolates of Staphylococcus species","volume":"7","author":"Elzain","year":"2019","journal-title":"J. Bacteriol. Mycol. Open Access"},{"key":"ref_45","unstructured":"Rutala, W.A., Weber, D.J., and Guideline for Disinfection and Sterilization in Healthcare Facilities, 2008 (2024, December 17). Update: May 2019, Available online: https:\/\/www.cdc.gov\/infection-control\/hcp\/disinfection-sterilization\/chemical-disinfectants.html."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"110338","DOI":"10.1016\/j.lwt.2020.110338","article-title":"Purification and antimicrobial mechanism of a novel bacteriocin produced by Lactobacillus rhamnosus 1.0320","volume":"137","author":"Xu","year":"2021","journal-title":"LWT"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"927","DOI":"10.2298\/ABS1203927K","article-title":"The antimicrobial activity of honey, bee pollen loads, and beeswax from Slovakia","volume":"64","author":"Kacaniova","year":"2012","journal-title":"Arch. Biol. Sci."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1080\/03601234.2013.727664","article-title":"Antioxidant and antimicrobial properties of monofloral bee pollen","volume":"48","author":"Kacaniova","year":"2013","journal-title":"J. Environ. Sci. Health"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1111\/jfbc.12160","article-title":"Phenolic extract from propolis and bee pollen: Composition, antioxidant and antibacterial activities","volume":"39","author":"Mohdaly","year":"2015","journal-title":"J. Food Biochem."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Stefanovi\u0107, O.D. (2018). Synergistic activity of antibiotics and bioactive plant extracts: A study against Gram-positive and Gram-negative bacteria. Bacterial Pathogenesis and Antibacterial Control, IntechOpen.","DOI":"10.5772\/intechopen.72026"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"482","DOI":"10.3934\/microbiol.2018.3.482","article-title":"An overview of the antimicrobial resistance mechanisms of bacteria","volume":"4","author":"Reygaert","year":"2018","journal-title":"AIMS Microbiol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/S0074-7696(08)61093-1","article-title":"Pollen wall and sporopollenin","volume":"140","author":"Wiermann","year":"1992","journal-title":"Int. Rev. Cytol."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Amin, M.U., Khurram, M., Khattak, B., and Khan, J. (2015). Antibiotic additive and synergistic action of rutin, morin and quercetin against methicillin resistant Staphylococcus aureus. BMC Complement. Altern. Med., 15.","DOI":"10.1186\/s12906-015-0580-0"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1089\/mdr.2012.0244","article-title":"Antibacterial activity and mode of action of ferulic and gallic acids against pathogenic bacteria","volume":"19","author":"Borges","year":"2013","journal-title":"Microb. Drug Resist."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1016\/j.micpath.2018.11.033","article-title":"Ferulic acid potentiates the antibacterial activity of quinolone-based antibiotics against Acinetobacter baumannii","volume":"126","author":"Ibitoye","year":"2019","journal-title":"Microb. Pathog."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Yougbare, S., Chang, T.K., Tan, S.H., Kuo, J.C., Hsu, P.H., Su, C.Y., and Kuo, T.R. (2019). Antimicrobial gold nanoclusters: Recent developments and future perspectives. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20122924"},{"key":"ref_57","unstructured":"(2023). Bee Pollen\u2014Specifications (Standard No. ISO 24382:2023)."},{"key":"ref_58","unstructured":"Ada\u0161kevi\u010di\u016bt\u0117, V., Ka\u0161konien\u0117, V., and Maru\u0161ka, A. (2021, January 24\u201328). Optimization of conditions for bee pollen lactic acid fermentation. Proceedings of the CYSENI 2021: The 17th International Conference of Young Scientists on Energy Issues, Kaunas, Lithuania."}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/18\/1\/15\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T17:00:39Z","timestamp":1760115639000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/18\/1\/15"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,26]]},"references-count":58,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,1]]}},"alternative-id":["ph18010015"],"URL":"https:\/\/doi.org\/10.3390\/ph18010015","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,12,26]]}}}