In Vitro Evaluation of Antimicrobial and Biofilm-Modulating Effects of Commercial Cranberry (Vaccinium macrocarpon) Extracts Against Escherichia coli and Staphylococcus aureus

Anisa Veledar-Hamalukić; Monia Avdić; Amra Demirović

doi:10.5937/scriptamed56-57135

Anisa Veledar-Hamalukić International Burch University
Monia Avdić Faculty of Engineering, Natural and Medical Sciences, Department of Genetics and Bioengineering, International Burch University, Sarajevo, Bosnia and Herzegovina.
Amra Demirović Amsal Pharmaceuticals d.o.o. Sarajevo, Bosnia and Herzegovina.

DOI: https://doi.org/10.5937/scriptamed56-57135

Sažetak

Background/Aim: Cranberry (Vaccinium macrocarpon Ait) extracts are widely recognised for their antimicrobial properties, particularly in preventing urinary tract infections (UTIs). However, their effects on biofilm formation remain underexplored. This study aimed to authenticate commercially available cranberry extracts, quantify their proanthocyanidin (PAC) content and evaluate their antimicrobial and biofilm-modulating effects against Escherichia coli and Staphylococcus aureus.

Methods: Two commercially available cranberry extracts were analysed using high-performance liquid chromatography (HPLC) to confirm their authenticity based on anthocyanin profiles. PAC content was determined using a modified Bate-Smith method. Antimicrobial activity was assessed via the microbroth dilution method, determining the minimum inhibitory concentration (MIC) for each extract against two E coli and three S aureus strains. Biofilm formation was evaluated using crystal violet staining, with statistical analyses including paired sample t-tests, Friedman’s ANOVA and the Durbin-Conover test.

Results: HPLC analysis confirmed that both extracts were derived from the V macrocarpon. PAC content differed significantly between samples (p < 0.001), yet MIC values remained consistent at 125 µg/mL for E coli and 250 µg/mL for S aureus. Notably, at concentrations below the MIC, both extracts unexpectedly enhanced bacterial planktonic growth and biofilm formation, likely due to quorum sensing activation. Statistical analyses revealed significant differences in biofilm formation among S aureus strains, while E coli strains showed a more uniform response.

Conclusion: While cranberry extracts exhibit strong antibacterial properties at MIC levels, their ability to stimulate biofilm formation at sub-inhibitory concentrations raises concerns regarding their application. These findings highlight the need for further molecular studies to optimise cranberry-based interventions against biofilm-associated infections.

Reference

Di Martino P. Extracellular polymeric substances, a key element in understanding biofilm phenotype. AIMS Microbiol. 2018;4(2):274–88. doi: 10.3934/microbiol.2018.2.274.

Petrova OE, Sauer K. Sticky situations: key components that control bacterial surface attachment. J Bacteriol. 2012 May 15;194(10):2413–25. doi: 10.1128/JB.00003-12.

Zheng S, Bawazir M, Dhall A, Kim HE, He L, Heo J, et al. Implication of surface properties, bacterial motility, and hydrodynamic conditions on bacterial surface sensing and their initial adhesion. Front Bioeng Biotechnol. 2021 Feb 12;9:643722. doi: 10.3389/fbioe.2021.643722.

Sharma S, Mohler J, Mahajan SD, Schwartz SA, Bruggemann L, Aalinkeel R. Microbial biofilm: a review on formation, infection, antibiotic resistance, control measures, and innovative treatment. Microorganisms. 2023 Jun 19;11(6):1614. doi: 10.3390/microorganisms11061614.

Sauer K, Stoodley P, Goeres DM, Hall-Stoodley L, Burmølle M, Stewart PS, et al. The biofilm life cycle: expanding the conceptual model of biofilm formation. Nat Rev Microbiol. 2022 Oct;20(10):608–20. doi: 10.1038/s41579-022-00767-0.

Zhang L, Jin M, Sun M. Inhibition characteristics of biofilm structure of Staphylococcus aureus. Cell Mol Biol. 2020 May 16;66(2):204–11. doi: 10.14715/cmb/2020.66.2.32.

Detho H. Biofilm formation and antibiotic resistance in uropathogenic Escherichia coli: The quest for effective treatment of urinary tract infections. Pure Appl Biol. 2024 Mar 10;13(1). doi: 10.19045/bspab.2024.130004.

Sauer MM, Jakob RP, Eras J, Baday S, Eriş D, Navarra G, et al. Catch-bond mechanism of the bacterial adhesin FimH. Nat Commun. 2016 Mar 7;7(1):10738. doi: 10.1038/ncomms10738.

Kostakioti M, Hadjifrangiskou M, Hultgren SJ. Bacterial biofilms: development, dispersal, and therapeutic strategies in the dawn of the postantibiotic era. Cold Spring Harb Perspect Med. 2013 Apr 1;3(4):a010306–a010306. doi: 10.1101/cshperspect.a010306.

Ribić R, Meštrović T, Neuberg M, Kozina G. Effective anti-adhesives of uropathogenic Escherichia coli. Acta Pharm. 2018 Mar 1;68(1):1–18. doi: 10.2478/acph-2018-0004.

Vasudevan S, Thamil Selvan G, Bhaskaran S, Hari N, Solomon AP. Reciprocal cooperation of type A procyanidin and nitrofurantoin against multi-drug resistant (MDR) UPEC: A pH-dependent study. Front Cell Infect Microbiol. 2020 Aug 11;10:421. doi: 10.3389/fcimb.2020.00421.

Maximilian Matthias Sauer. Catch and release: mechanism and binding epitope of the bacterial adhesin fimh. Institute of Molecular Biology and Biophysics, ETH Zurich, Switzerland; 2017.

Brown PN, Shipley PR. Determination of anthocyanins in cranberry fruit and cranberry fruit products by high-performance liquid chromatography with ultraviolet detection: single-laboratory validation. J AOAC Int. 2011;94(2):459–66. PMID: 21563679.

Avdić M, Džuzić N, Hasanić O, Spahić A, Smajlović Skenderagić L, Badnjević A, et al. Development of a novel biofilm classification tool and comparative analysis of result interpretation methodologies for the evaluation of biofilm forming capacity of bacteria using tissue culture plate method. Med Glas. 2018 Aug 24;16(1):7–12. doi: 10.17392/997-19.

Gardana C, Scialpi A, Fachechi C, Simonetti P. Identification of markers for the authentication of cranberry extract and cranberry-based food supplements. Heliyon. 2020 Apr;6(4):e03863. doi: 10.1016/j.heliyon.2020.e03863.

Whelan S, Lucey B, Finn K. Uropathogenic Escherichia coli (UPEC)-Associated urinary tract infections: the molecular basis for challenges to effective treatment. Microorganisms. 2023 Aug 28;11(9):2169. doi: 10.3390/microorganisms11092169.

Harmsen M, Yang L, Pamp SJ, Tolker-Nielsen T. An update on Pseudomonas aeruginosa biofilm formation, tolerance, and dispersal. FEMS Immunol Med Microbiol. 2010 Aug;59(3):253–68. doi: 10.1111/j.1574-695X.2010.00690.x.

Parsek MR, Greenberg EP. Acyl-homoserine lactone quorum sensing in gram-negative bacteria: a signaling mechanism involved in associations with higher organisms. Proc Natl Acad Sci U S A. 2000 Aug 1;97(16):8789–93. doi: 10.1073/pnas.97.16.8789.

Prazdnova EV, Gorovtsov AV, Vasilchenko NG, Kulikov MP, Statsenko VN, Bogdanova AA, et al. Quorum-Sensing Inhibition by Gram-Positive Bacteria. Microorganisms. 2022 Feb 3;10(2):350. doi: 10.3390/microorganisms10020350.

Kungwani N, Shukla SK, Subba Rao T, Das S. Biofilm-mediated bioremediation of polycyclic aromatic hydrocarbons: current status and future perspectives. In: Microbial Biodegradation and Bioremediation. Amsterdam, NA: Elsevier; 2022. p. 547–70. doi: 10.1016/B978-0-323-85455-9.00021-7.

Zhou Y, Zhou Z, Zheng L, Gong Z, Li Y, Jin Y, et al. Urinary tract infections caused by uropathogenic Escherichia coli: mechanisms of infection and treatment options. Int J Mol Sci. 2023 Jun 23;24(13):10537. doi: 10.3390/ijms241310537.

Dodson KW, Pinkner JS, Rose T, Magnusson G, Hultgren SJ, Waksman G. Structural Basis of the interaction of the pyelonephritic E. coli adhesin to its human kidney receptor. Cell. 2001 Jun 15;105(6):733–43. doi: 10.1016/S0092-8674(01)00388-9

Hannon DB, Thompson JT, Khoo C, Juturu V, Vanden Heuvel JP. Effects of cranberry extracts on gene expression in THP ‐1 cells. Food Sci Nutr. 2017 Jan;5(1):148–59. doi: 10.1002/fsn3.374.

Xu K, Wang Y, Jian Y, Chen T, Liu Q, Wang H, et al. Staphylococcus aureus ST1 promotes persistent urinary tract infection by highly expressing the urease. Front Microbiol. 2023 Feb 22;14:1101754. doi: 10.3389/fmicb.2023.1101754.