Phytochemical analysis and antimicrobial activity of blackberry (Rubus fruticosus) fruit extract against Gram-negative multidrug-resistant bacteria isolated from clinical samples
DOI:
https://doi.org/10.12923/cipms-2025-0041Keywords:
blackberry fruit, anthocyanins, multidrug-resistant strains, molecular dockingAbstract
While antibiotics remain foundational in treating microbial infections, their overuse has accelerated the emergence and global spread of multidrug-resistant (MDR) bacterial strains. This study aimed to characterize the phytochemical composition of a concentrated blackberry (Rubus fruticosus) extract and evaluate its antimicrobial potential against clinical MDR strains of Acinetobacter baumannii, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Enterobacter cloacae. Phytochemical quantification was performed using spectrophotometric, titrimetric, and HPLC analyses, while antimicrobial activity was determined via the well diffusion method and minimum inhibitory concentration (MIC) assays. The blackberry extract contained 0.54% total phenolic compounds and 4.60% organic acids; in comparison, a green tea leaf extract exhibited higher phenolic content (10.10%) but lower organic acids (1.60%). The total anthocyanin content of the blackberry extract was 159.81 mg/100 g, with cyanidin-3-O-glucoside (134.56 ± 0.10 mg/100 g) identified as the predominant constituent.
In silico molecular docking studies indicated that neither individual antibiotics nor isolated anthocyanins could effectively inhibit the full array of resistance mechanisms in Gram-negative bacteria. In contrast, in vitro assays demonstrated that the whole blackberry extract significantly inhibited the growth of all tested MDR strains. These integrated findings suggest that combating complex MDR pathogens requires a multi-target strategy. The broad-spectrum efficacy of the crude extract, likely arising from synergistic interactions among its constituents, underscores the potential of complex phytochemical preparations. Such natural formulations represent a promising avenue for the development of novel antimicrobial agents and may offer a viable strategy to restore the efficacy of existing antibiotics through adjunctive therapy.
References
1. Park H, Cho D, Huang E, Seo JY, Kim WG, Todorov SD, et al. Amelioration of alcohol induced gastric ulcers through administration of Lactobacillus plantarum APSulloc 331261 isolated from green tea. Front Microbiol. 2020;11:420. doi:10.3389/fmicb.2020.00420
2. Kadasah S, Al Eid AS, Alawad SS, Al Shahrani AS, Alruwaihi AS, Elfaki I, et al. Gastroprotective influence of topiramate in ethanol produced gastric ulcers in rats. Toxicol Rep. 2021;8:1031–1039. doi:10.1016/j.toxrep.2021.05.004
3. Sanpinit S, Chonsut P, Punsawad C, Wetchakul P. Gastroprotective and antioxidative effects of the traditional Thai polyherbal formula Phy Blica D against ethanol induced gastric ulcers in rats. Nutrients. 2022;14(1):172. doi:10.3390/nu14010172
4. Fernandes HB, Silva FV, Passos FFB, Bezerra RDS, Chaves MH, Oliveira FA, et al. Gastroprotective effect of the ethanolic extract of Parkia platycephala Benth. leaves against acute gastric lesion models in rodents. Biol Res. 2010;43:451–457.
5. Yoo JH, Lee JS, Lee YS, Kim S, Lee HJ. Protective effect of bovine milk against HCl/ethanol induced gastric ulcer in mice. J Dairy Sci. 2018;101:3758–3770. doi:10.3168/jds.2017-13872
6. El Meligy RM, Awaad AS, Soliman GA, Kenawy SA, Alqasoumi SI. Prophylactic and curative anti ulcerogenic activity and possible mechanisms of action of some desert plants. Saudi Pharm J. 2017;25:387–396.
7. Zeghad N, Ejaz A, Belkhiri A, Vander Heyden Y, Demeyer K. Antioxidant activity of Vitis vinifera, Punica granatum, Citrus aurantium and Opuntia ficus indica fruits cultivated in Algeria. Heliyon. 2019;5:e01575.
8. Sorrenti V, Randazzo CL, Caggia C, Ballistreri G, Romeo FV, Fabroni S, et al. Beneficial effects of pomegranate peel extract and probiotics on preadipocyte differentiation. Front Microbiol. 2019;10:1–11
9. Zeghad N, Madi A, Helmi S, Belkhiri A. In vivo analgesic activity and safety assessment of Vitis vinifera L. and Punica granatum L. fruit extracts. Trop J Pharm Res. 2016;15(9):1319–1326.
10. Zeghad N, Ahmed E, Khan MZ, Belkhiri A. Exploring the potential use of pomegranate (Punica granatum L.) and prickly pear (Opuntia ficus indica L.) peels as sources of cosmeceutical sunscreen agents. Pharm Sci Asia. 2023;50(4):273–280.
11. Moghaddam G, Sharifzadeh M, Hassanzadeh G, Khanavi M, Hajimahmoodi M. Anti ulcerogenic activity of pomegranate peel (Punica granatum) methanol extract. Food Nutr Sci. 2013;4:43–48.
12. Shah Ayub MA, Garg SK, Garg KM. Subacute toxicity studies on pendimethalin in rats. Indian J Pharmacol. 1997;29(5):322–324.
13. Bürger C, Fischer DR, Cordenunzzi DA, Batschauer APB, Cechinel Filho V, Soares ARS. Acute and subacute toxicity of the hydroalcoholic extract from Wedelia paludosa (Acmela brasiliensis) in mice. J Pharm Sci. 2005;8(2):370–373.
14. Zeghad N, Ejaz A, Madi A, Helmi S. Acute toxicity and analgesic activity of the aerial parts of Ajuga iva (L.) Schreb. grown in eastern Algeria. FABAD J Pharm Sci. 2020;45(1):1–7.
15. Sargul HS, Sheila MN, Zahra AA, Al Bustany HA, Nadir MQ. Gastroprotective activity of Hypericum perforatum extract in ethanol induced gastric mucosal injury in Wistar rats. Heliyon. 2020;6(7):e05249. doi:10.1016/j.heliyon.2020.e05249
16. Hamedi S, Arian AA, Farzaei MH. Gastroprotective effect of aqueous stem bark extract of Ziziphus jujuba L. against HCl/ethanol induced gastric mucosal injury in rats. J Tradit Chin Med. 2015;35(6):666–670.
17. Abdel Aziz Ibrahim I, Abdulla MA, Hajrezaie M, Bader A, Shahzad N, Al Ghamdi S, et al. Gastroprotective effects of hydroalcoholic extract of Monolluma quadrangula against ethanol induced gastric mucosal injuries in Sprague Dawley rats. Drug Des Devel Ther. 2016;10:93–105. doi:10.2147/DDDT.S99887
18. Sobreira F, Hernandes LS, Vetore Neto A, Diaz IEC, Santana FC, Filho JM, et al. Gastroprotective activity of hydroethanolic extract and ethyl acetate fraction from Kalanchoe pinnata (Lam.) Pers. Braz J Pharm Sci. 2017;53(1):e16027.
19. Abdulla MA, Al Bayaty FH, Younis LT, Abu Hassan MI. Anti ulcer activity of Centella asiatica leaf extract against ethanol induced gastric mucosal injury in rats. J Med Plants Res. 2010;4(13):1253–1259.
20. Boligon AA, Freitas RB, Brum TF, Waczuk EP, Klimaczewski CV, Avila DS, et al. Antiulcerogenic activity of Scutia buxifolia on ethanol induced gastric ulcers in rats. Acta Pharm Sin B. 2014;4(5):358–367. doi:10.1016/j.apsb.2014.07.005
21. Kennedy GL, Ferenz RL, Burgess BA. Estimation of acute oral toxicity in rats by determination of the approximate lethal dose rather than the LD50. J Appl Toxicol. 1986;6:145–148.
22. Oluwabunmi IJ, Abiola T. Gastroprotective effect of methanolic extract of Gomphrena celosioides on indomethacin induced gastric ulcer in Wistar rats. Int J Appl Basic Med Res. 2015;5(1):41–45.
23. Sumbul S, Ahmad MA, Asif M, Akhtar M. Role of phenolic compounds in peptic ulcer: an overview. J Pharm Bioallied Sci. 2011;3:361–367. doi:10.4103/0975-7406.84433
24. Serafim C, Araruna ME, Júnior EA, Diniz M, Hiruma Lima C, Batista LM. Role of flavonoids in peptic ulcer disease: a review (2010–2020). Molecules. 2020;25(22):5431. doi:10.3390/molecules25225431
25. de Oliveira AC, Miyagawa LM, Monteiro KM, Dias ALS, Longato BL, Spindola H, et al. Phenolic composition, antiproliferative and antiulcerogenic activities of a polyphenol rich extract from açaí (Euterpe oleracea) fruits. Int J Food Sci. 2021;2021:1–9.
26. Zeghad N, Ejaz A, Belkhiri A, Demeyer K, Vander Heyden Y. Phenolic compound profiling of Algerian pomegranate (Punica granatum L.) fruit extract by UPLC DAD ESI MS. Chem Afr. 2022;5:1295–1303. doi:10.1007/s42250-022-00408-6
27. Zhang W, Lian Y, Li Q, Sun L, Chen R, Lai X, et al. Preventive and therapeutic potential of flavonoids in peptic ulcers. Molecules. 2020;25(1):1–31. doi:10.3390/molecules25010001
28. Demarque DP, Callejon DR, de Oliveira GG, Silva DB, Carollo CA, Lopes NP. The role of tannins as antiulcer agents: a fluorescence imaging based study. Rev Bras Farmacogn. 2017;28(4):1–8.
29. Abdel Rady N, Dahpy MA, Ahmed A, Elgalal DA, Hadiya S, Ahmed MAM, et al. Interplay of biochemical, genetic and immunohistochemical factors in gastric ulcer pathogenesis: effect of pomegranate loaded nanoparticles versus pomegranate peel extract. Front Physiol. 2021;12:1–20. doi:10.3389/fphys.2021.680943
30. Ahmed TJ, Alibraheem S, Taresh FJ, Mhalhal SL. Evaluation of anti ulcer activity of pomegranate peel powder (Punica granatum L.) in rabbits with aspirin induced peptic ulcer. Int J Pharm Res. 2020;12(3):2980–2987.
31. Moghaddam G, Sharifzadeh M, Hassanzadeh G, Khanavi M, Hajimahmoodi M. Anti ulcerative potential of Punica granatum L. hydroalcoholic fruit peel extract. Trop J Pharm Res. 2014;13(7):1093–1097.
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