Аннотация

Plant polyphenols have recently been recognized as promising new-generation prebiotics. However, the concentrations and profiles of polyphenols vary significantly among different plants, so the effects of their extracts on probiotic strains are also highly variable. This study assessed the effects of ethanol and supercritical fluid extracts of black currant and sea buckthorn on the growth of bifidobacteria and their antagonism against intestinal pathogens.
Extracts of black currant and sea buckthorn were obtained using reflux ethanol extraction or supercritical fluid extraction. Total phenols in the extracts were determined by the Folin-Ciocalteu colorimetric method. Individual phenolic compounds were identified by UV-Vis spectroscopy. Effects of the extracts on bifidobacteria and the intestinal pathogens Bacillus cereus and Staphylococcus aureus were evaluated in monoculture. Antagonism of bifidobacteria against the pathogens was evaluated in coculture in the media with various ethanol extracts. Concentrations of organic acids were determined using HPLC.
Phenolic components of the berries had stimulated, neutral, or negative effects on probiotic growth and organic acid production, depending on the type of raw material and the extraction method. Bifidobacterium adolescentis and Bifidobacterium longum were the most sensitive to, and strongly inhibited by, the extracts, while no pronounced growth-stimulating effect was observed for any of the studied bifidobacteria strains. Additionally, the effect of total phenolic concentrations in the extracts on the strain growth was evaluated by diluting the native extracts. Minimum inhibitory concentrations in the ethanol extracts for bifidobacteria ranged from 30 to 140 μg GAE/mL, while those in the supercritical fluid extracts were lower (13–32 μg GAE/mL). The sea buckthorn supercritical fluid extract stimulated Bifidobacterium bifidum growth, while the ethanol extract of sea buckthorn and the supercritical fluid extract of black currant stimulated the growth of Bifidobacterium breve and B. longum subsp. infantis. B. cereus was not sensitive to changes in total phenolic concentrations. The minimum inhibitory concentration of total phenols for S. aureus was the lowest (7 μg GAE/mL) in the supercritical fluid extract of sea buckthorn. The extracts had mostly neutral or positive effects on lactic and acetic acids production by bifidobacteria. The greatest increase in acid concentrations was observed for B. breve in the medium with black currant ethanol extract. The greatest suppression of pathogen growth was achieved by a combination of B. breve with either of the ethanol extracts.
The results demonstrate that black currant and sea buckthorn extracts are highly selective prebiotics that exert different effects on the antagonism of bifidobacteria against pathogens. The study revealed the most effective combination of the bifidobacteria and berry extracts against the pathogens, as well as the most effective symbiotic composition.

Ключевые слова

Bifidobacteria, sea buckthorn, black currant, supercritical fluid extraction, ethanol extraction, flavonoids, antagonism, synbiotics

Список литературы

1. Stachelska MA, Karpiński P, Kruszewski B. A comprehensive review of biological properties of flavonoids and their role in the prevention of metabolic, cancer and neurodegenerative diseases. Applied Sciences. 2025;15(19):10840. https://doi.org/10.3390/app151910840
2. Williams RJ, Spencer JPE, Rice-Evans C. Flavonoids: Antioxidants or signalling molecules? Free Radical Biology and Medicine. 2004;36(7):838–849. https://doi.org/10.1016/j.freeradbiomed.2004.01.001
3. Winkel-Shirley B. Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiology. 2001;126(2):485–493. https://doi.org/10.1104/pp.126.2.485
4. Serafini M, Peluso I, Raguzzini A. Flavonoids as anti-inflammatory agents. Proceedings of the Nutrition Society. 2010;69(3):273–278. https://doi.org/10.1017/S002966511000162X
5. Kumar S, Pandey AK. Chemistry and biological activities of flavonoids: An overview. The Scientific World Journal. 2013;2013(1):162750. https://doi.org/10.1155/2013/162750
6. Manach C, Scalbert A, Morand C, Rémésy C, Jiménez L. Polyphenols: Food sources and bioavailability. The American Journal of Clinical Nutrition. 2004;79(5):727–747. https://doi.org/10.1093/ajcn/79.5.727
7. Kähkönen MP, Hopia AI, Heinonen M. Berry phenolics and their antioxidant activity. Journal of Agricultural and Food Chemistry. 2001;49(8):4076–4082. https://doi.org/10.1021/jf010152t
8. Guseva EV, Khromova NYu, Karetkin BA, Artemiev AI, Demkin KM, et al. Biological value of berry polyphenols and prospects for supercritical extraction application for their isolation: A review. Foods and Raw Materials. 2026;14(1):1–13. https://doi.org/10.21603/2308-4057-2026-1-653
9. Skrovankova S, Sumczynski D, Mlcek J, Jurikova T, Sochor J. Bioactive compounds and antioxidant activity in different types of berries. International Journal of Molecular Sciences. 2015;16(10):24673–24706. https://doi.org/10.3390/ijms161024673
10. Määttä K, Kamal-Eldin A, Törrönen R. Phenolic compounds in berries of black, red, green, and white currants (Ribes sp.). Antioxidants & Redox Signaling. 2001;3(6):981–993. https://doi.org/10.1089/152308601317203521
11. Tang J, Zhang H, Wu R, Liu H, He K, et al. Sea buckthorn: A potential prebiotic and promising functional ingredient for fermented dairy products. Journal of Future Foods. 2026;6(3):348–360. https://doi.org/10.1016/j.jfutfo.2025.04.006
12. Kodentsova VM, Risnik DV, Serba EM, Abramova IM, Sokolova EN, et al. Prospects for integrated processing of black currant. Food Processing: Techniques and Technology. 2024;54(3):621–632. (In Russ.) https://doi.org/10.21603/2074-9414-2024-3-2525
13. Pietta PG. Flavonoids as antioxidants. Journal of Natural Products. 2000;63(7):1035–1042. https://doi.org/10.1021/np9904509
14. García-Lafuente A, Guillamón E, Villares A, Rostagno MA, Martínez JA. Flavonoids as anti-inflammatory agents: Implications in cancer and cardiovascular disease. Inflammation Research. 2009;58:537–552. https://doi.org/10.1007/s00011-009-0037-3
15. Murota K, Nakamura Y, Uehara M. Flavonoid metabolism: The interaction of metabolites and gut microbiota. Bioscience, Biotechnology & Biochemistry. 2018;82(4):600–610. https://doi.org/10.1080/09168451.2018.1444467
16. Yadav MK, Kumari I, Singh B, Sharma KK, Tiwari SK. Probiotics, prebiotics and synbiotics: Safe options for next-generation therapeutics. Applied Microbiology and Biotechnology. 2022;106:505–521. https://doi.org/10.1007/s00253-021-11646-8
17. Vesnina A D, Frolova A S, Chekushkina DYu, Milentyeva IS, Luzyanin SL, et al. Gut microbiota and its role in development of chronic disease and aging. Foods and Raw Materials. 2026;14(1):174–197. https://doi.org/10.21603/2308-4057-2026-1-668
18. Levy M, Kolodziejczyk AA, Thaiss CA, Elinav E. Dysbiosis and the immune system. Nature Reviews Immunology. 2017;17:219–232. https://doi.org/10.1038/nri.2017.7
19. Ruiz de la Bastida A, Peirotén Á, Langa S, Álvarez I, Arqués JL, et al. Metabolism of flavonoids and lignans by lactobacilli and bifidobacteria strains improves the nutritional properties of flaxseed-enriched beverages. Food Research International. 2021;147:110488. https://doi.org/10.1016/j.foodres.2021.110488
20. Liang A, Leonard W, Beasley JT, Fang Z, Zhang P, et al. Anthocyanins-gut microbiota-health axis: A review. Critical Reviews in Food Science and Nutrition. 2024;64(21):7563–7588. https://doi.org/10.1080/10408398.2023.2187212
21. Louis P, Hold GL, Flint HJ. The gut microbiota, bacterial metabolites and colorectal cancer. Nature Reviews Microbiology. 2014;12:661–672. https://doi.org/10.1038/nrmicro3344
22. Kähkönen MP, Hopia AI, Heinonen M. Berry phenolics and their antioxidant activity. Journal of Agricultural and Food Chemistry. 2001;49(8):4076–4082. https://doi.org/10.1021/jf010152t
23. Heinonen M. Antioxidant activity and antimicrobial effect of berry phenolics – A Finnish perspective. Molecular Nutrition & Food Research. 2007;51:684–691. https://doi.org/10.1002/mnfr.200700006
24. Zeb A. Chemical and nutritional constituents of sea buckthorn juice. Pakistan Journal of Nutrition. 2004;3(2):99–106. https://doi.org/10.3923/pjn.2004.99.106
25. Gopalan A, Reuben SC, Ahmed S, Darvesh AS, Hohmann J, et al. The health benefits of blackcurrants. Food & Function. 2012;3(8):795–809. https://doi.org/10.1039/c2fo30058c
26. Donno D, Cerutti AK, Mellano MG, Prgomet Z, Beccaro GL. Serviceberry, a berry fruit with growing interest of industry: Physicochemical and quali-quantitative health-related compound characterisation. Journal of Functional Foods. 2016;26:157–166. https://doi.org/10.1016/j.jff.2016.07.014
27. Solverson PM, Rumpler WV, Leger JL, Redan BW, Ferruzzi MG, et al. Blackberry feeding increases fat oxidation and improves insulin sensitivity in overweight and obese males. Nutrients. 2018;10(8):1048. https://doi.org/10.3390/nu10081048
28. Wrona O, Rafińska K, Możeński C, Buszewski B. Supercritical fluid extraction of bioactive compounds from plant materials. Journal of AOAC International. 2017;100(6):1624–1635. https://doi.org/10.5740/jaoacint.17-0232
29. Williams JR, Clifford AA, Al-Saidi SH. Supercritical fluids and their applications in biotechnology and related areas. Molecular Biotechnology. 2002;22:263–286. https://doi.org/10.1385/MB:22:3:263
30. Singh S, Verma DK, Thakur M, Tripathy S, Patel AR, et al. Supercritical fluid extraction (SCFE) as green extraction technology for high-value metabolites of algae, its potential trends in food and human health. Food Research International. 2021;150(Part A):110746. https://doi.org/10.1016/j.foodres.2021.110746
31. Azmir J, Zaidul ISM, Rahman MM, Sharif KM, Mohamed A, et al. Techniques for extraction of bioactive compounds from plant materials: A review. Journal of Food Engineering. 2013;117(4):426–436. https://doi.org/10.1016/j.jfoodeng.2013.01.014
32. Frolova AS, Fokina AD, Milentyeva IS, Asyakina LK, Proskuryakova LA, et al. The biological active substances of Taraxacum officinale and Arctium lappa from the Siberian Federal District. International Journal of Molecular Sciences. 2024;25(6):3263. https://doi.org/10.3390/ijms25063263
33. Rossi M, Corradini C, Amaretti A, Nicolini M, Pompei A, et al. Fermentation of fructooligosaccharides and inulin by bifidobacteria: A comparative study of pure and fecal cultures. Applied and Environmental Microbiology. 2005;71(10):6150–6158. https://doi.org/10.1128/AEM.71.10.6150-6158.2005
34. Scherer R, Rybka ACP, Ballus CA, Meinhart AD, Filho JT, et al. Validation of a HPLC method for simultaneous determination of main organic acids in fruits and juices. Food Chemistry. 2012;135(3):150–154. https://doi.org/10.1016/j.foodchem.2012.03.111
35. Zaky AS, Pensupa N, Andrade-Eiroa Á, Tucker GA, Du C. A new HPLC method for simultaneously measuring chloride, sugars, organic acids and alcohols in food samples. Journal of Food Composition and Analysis. 2017;56:25–33. https://doi.org/10.1016/j.jfca.2016.12.010
36. Karetkin BA, Guseva EV, Evdokimova SA, Mishchenko AS, Khabibulina NV, et al. A quantitative model of Bacillus cereus ATCC 9634 growth inhibition by bifidobacteria for synbiotic effect evaluation. World Journal of Microbiology and Biotechnology. 2019;35:89. https://doi.org/10.1007/s11274-019-2665-2
37. Evdokimova S, Karetkin B, Nokhaeva V, Guseva E, Shakir I. Minimum inhibitory concentrations of organic acids against foodborne opportunistic microbial pathogens. In: Proceedings of the 21st Intern. Multidisciplinary Sci. GeoConf. SGEM. Albena, 2021:193–200. https://doi.org/10.5593/sgem2021/6.1/s25.25
38. Yilmaz Y, Toledo RT. Major flavonoids in grape seeds and skins: Antioxidant capacity of catechin, epicatechin, and gallic acid. Journal of Agricultural and Food Chemistry. 2004;52(2):255–260. https://doi.org/10.1021/jf030117h
39. Fu M, Zhang L, Killeen R, Onugwu KE, McCarrick RM, et al. Green tea polyphenol epigallocatechin gallate interactions with copper-serum albumin. Molecules. 2025;30(2):320. https://doi.org/10.3390/molecules30020320
40. Elgailani IEH, Ishak CY. Methods for extraction and characterization of tannins from some acacia species of Sudan. Pakistan Journal of Analytical & Environmental Chemistry. 2016;17(1):43–49. https://doi.org/10.21743/pjaec/2016.06.007
41. Pawlak-Lemanska K, Szymusiak H, Tyrakowska B, Zielinski R, Soffers AEMF, et al. The influence of pH on antioxidant properties and the mechanism of antioxidant action of hydroxyflavones. Free Radical Biology and Medicine. 2001;31(7):869–881. https://doi.org/10.1016/S0891-5849(01)00638-4
42. Giusti MM, Wrolstad RE. Characterization and measurement with UV-Visible spectroscopy. Current Protocols in Food Analytical Chemistry. Hoboken: John Wiley & Sons; 2001, pp. F1.2.1–F1.2.13. https://doi.org/10.1002/0471142913.faf0102s00
43. Frémont L. Biological effects of resveratrol. Life Sciences. 2000;66(8):663–673. https://doi.org/10.1016/S0024-3205(99)00410-5
44. Quideau S, Feldman KS. Ellagitannin chemistry. Chemical Reviews. 1996;96(1):475–504. https://doi.org/10.1021/cr940716a
45. Niemetz R, Gross GG. Enzymology of gallotannin and ellagitannin biosynthesis. Phytochemistry. 2005;66(17):2001–2011. https://doi.org/10.1016/j.phytochem.2005.01.009
46. Zhang Y, Yu J, Dong XD, Ji HY. Research on characteristics, antioxidant and antitumor activities of dihydroquercetin and its complexes. Molecules. 2018;23(1):20. https://doi.org/10.3390/molecules23010020
47. Hopia A, Heinonen M. Antioxidant activity of flavonol aglycones and their glycosides in methyl linoleate. Journal of the American Oil Chemists' Society. 1999;76:139–144. https://doi.org/10.1007/s11746-999-0060-0
48. Kahraman N, Karadag R. Characterization of sixteenth to nineteenth century ottoman silk brocades by scanning electron microscopy – energy dispersive X-ray spectroscopy and high-performance liquid chromatography. Analytical Letters. 2017;50(10):1553–1567. https://doi.org/10.1080/00032719.2016.1236264
49. De Souza LA, Soeiro MM, De Almeida WB. A DFT study of molecular structure and ¹H NMR, IR, and UV-Vis spectrum of Zn(II)-kaempferol complexes: A metal-flavonoid complex showing enhanced anticancer activity. International Journal of Quantum Chemistry. 2018;118:e25773. https://doi.org/10.1002/qua.25773
50. Robbins RJ. Phenolic acids in foods: An overview of analytical methodology. Journal of Agricultural and Food Chemistry. 2003;51(10):2866–2887. https://doi.org/10.1021/jf026182t
51. Gil-Izquierdo A, Gil MI, Ferreres F, Tomás-Barberán FA. In vitro availability of flavonoids and other phenols in orange juice. Journal of Agricultural and Food Chemistry. 2001;49(2):1035–1041. https://doi.org/10.1021/jf0000528
52. Gwiazdowska D, Juś K, Jasnowska-Małecka J, Kluczyńska K. The impact of polyphenols on Bifidobacterium growth. Acta Biochimica Polonica. 2015;62(4):895–901. https://doi.org/10.18388/abp.2015_1154
53. Téglás T, Mihok E, Cziáky Z, Oláh NK, Nyakas C, et al. The flavonoid rich black currant (Ribes nigrum) ethanolic gemmotherapy extract elicits neuroprotective effect by preventing microglial body swelling in hippocampus and reduces serum TNF-α level: Pilot study. Molecules. 2023;28(8):3571. https://doi.org/10.3390/molecules28083571
54. Pereira P, Cebola MJ, Oliveira MC, Bernardo-Gil MG. Supercritical fluid extraction vs conventional extraction of myrtle leaves and berries: Comparison of antioxidant activity and identification of bioactive compounds. The Journal of Supercritical Fluids. 2016;113:1–9. https://doi.org/10.1016/j.supflu.2015.09.006
55. Attri S, Sharma K, Raigond P, Goel G. Colonic fermentation of polyphenolics from Sea buckthorn (Hippophae rhamnoides) berries: Assessment of effects on microbial diversity by principal component analysis. Food Research International. 2018;105:324–332. https://doi.org/10.1016/j.foodres.2017.11.032
56. Kawabata K, Sugiyama Y, Sakano T, Ohigashi H. Flavonols enhanced production of anti-inflammatory substance(s) by Bifidobacterium adolescentis: Prebiotic actions of galangin, quercetin, and fisetin. Biofactors. 2013;39(4):422–429. https://doi.org/10.1002/biof.1081
57. Nina N, Bressa C, de Lucas B, Martin de la Torre I, Jiménez-Aspee F, et al. Polyphenol metabolism, short-chain fatty acids production, and microbiota changes during in vitro digestion and fermentation of Chilean beans (Phaseolus vulogaris L.). Food Chemistry. 2025;486:144669. https://doi.org/10.1016/j.foodchem.2025.144669
58. Yang K, Zhang L, Liao P, Xiao Z, Zhang F, et al. Impact of gallic acid on gut health: Focus on the gut microbiome, immune response, and mechanisms of action. Frontiers in Immunology. 2020;11:580208. https://doi.org/10.3389/fimmu.2020.580208
59. Li X, Xie E, Sun S, Shen J, Ding Y, et al. Flavonoids for gastrointestinal tract local and associated systemic effects: A review of clinical trials and future perspectives. Journal of Advanced Research. 2025;77:15–41. https://doi.org/10.1016/j.jare.2025.01.014
60. Makarewicz M, Drożdż I, Tarko T, Duda-Chodak A. The interactions between polyphenols and microorganisms, especially gut microbiota. Antioxidants. 2021;10(2):188. https://doi.org/10.3390/antiox10020188
61. Qabaha KI. Antimicrobial and free radical scavenging activities of five Palestinian medicinal plants. African Journal of Traditional, Complementary and Alternative Medicines. 2013;10(4):101–108. https://doi.org/10.4314/ajtcam.v10i4.17
62. Veličković DT, Randjelović NV, Ristić MS, Veličković AS, Šmelcerović AA. Chemical constituents and antimicrobial activity of the ethanol extracts obtained from the flower, leaf and stem of Salvia officinalis L. Journal of the Serbian Chemical Society. 2003;68(1):17–24. https://doi.org/10.2298/JSC0301017V
63. Bozyel ME, Şenturan M, Benek A, Bozyel EM, Canli K, et al. In vitro antimicrobial activity screening of Heliotropium europaeum against wide range of microorganisms and multi drug resistant (MDR) bacteria. European Journal of Biomedical and Pharmaceutical Sciences. 2019;6(3):113–117.
64. Al-Juraifani AA. Antimicrobial activity of some medicinal plants used in Saudi Arabia. Canadian Journal of Pure and Applied Sciences. 2011;5(2):1509–1512.
65. Rauha JP, Remes S, Heinonen M, Hopia A, Kähkönen M, et al. Antimicrobial effects of Finnish plant extracts containing flavonoids and other phenolic compounds. International Journal of Food Microbiology. 2000;56(1):3–12. https://doi.org/10.1016/S0168-1605(00)00218-X
66. Hirano R, Sakanaka M, Yoshimi K, Sugimoto N, Eguchi S, et al. Next-generation prebiotic promotes selective growth of bifidobacteria, suppressing Clostridioides difficile. Gut Microbes. 2021;13(1):1973835. https://doi.org/10.1080/19490976.2021.1973835
67. Alowo D, Olum S, Mukisa IM, Ongeng D. Prebiotic potential of oligosaccharides extracted from improved Ugandan varieties of millet, sesame, soybean, and sorghum: Enhancing probiotic growth and enteric pathogen inhibition. BMC Microbiology. 2025;25:307. https://doi.org/10.1186/s12866-025-04028-x
68. Higashi B, Mariano TB, de Abreu Filho BA, Gonçalves RAC, de Oliveira AJB. Effects of fructans and probiotics on the inhibition of Klebsiella oxytoca and the production of short-chain fatty acids assessed by NMR spectroscopy. Carbohydrate Polymers. 2020;248:116832. https://doi.org/10.1016/j.carbpol.2020.116832
69. Śliżewska K, Chlebicz-Wójcik A. The in vitro analysis of prebiotics to be used as a component of a synbiotic preparation. Nutrients. 2020;12(5):1272. https://doi.org/10.3390/nu12051272
70. Evdokimova SA, Nokhaeva VS, Karetkin BA, Guseva EV, Khabibulina NV, et al. A study on the synbiotic composition of Bifidobacterium bifidum and fructans from Arctium lappa roots and Helianthus tuberosus tubers against Staphylococcus aureus. Microorganisms. 2021;9(5):930. https://doi.org/10.3390/microorganisms9050930