Study of the Wobenzym Enzyme Preparation Effect on the Formation of Bacterial Biofilms

Background. According to the International Center for Disease Control (CDC), 65–80% of all bacterial infections recorded in countries around the world are associated with the ability of their pathogens to form biofilms. To eliminate biofilms, various methods are being developed, including those involving the use of enzymes, proteins, plant extracts, and composite antibacterial coatings. Both antibiotic-resistant and sensitive strains have the ability to form biofilms, which confirms the relevance of the problem of biofilm formation by bacterial cells and the necessity of finding a solution to the treatment of infections caused by film-forming isolates.

The aim was to study the influence of the Wobenzym enzyme preparation on the formation of biofilms of clinical bacterial isolates and to determine the presence of a potentiating effect on the action of antibiotics.

Material and methods. A bacteriological method was used in the study. In this work, 20 strains that differed in their ability to form films were studied.

Results. The most pronounced film-forming ability was exhibited by Escherichia coli (OD=1.0) and Enterococcus faecalis (OD=0.649), isolated from the discharge of the cervical canal, as well as Enterobacter aerogenes (OD=0.406), isolated from the discharge of the pharynx of a newborn child. Cultivation of all studied strains in the presence of Wobenzym significantly reduces their film-forming ability (OD without the addition of enzyme — 0.255±0.005; with enzyme — 0.084±0.006, P=0.0009). Potentiation of the antibiotics ampicillin and amikacin by the Wobenzym enzyme preparation was confirmed by a more than two-fold decrease in CFU/µl.

Conclusion. Cultivation of Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, and Enterobacter aerogenes strains in the presence of Wobenzym significantly reduces their ability to form biofilms, which can be used to prevent biofilm formation and eradicate strains of opportunistic microorganisms that cause infectious and inflammatory processes in normally non-sterile mucous membrane loci of the human body. The combined use of Wobenzym with antibacterial therapy has a direct potentiating effect of antibacterial drugs, as well as an indirect effect that increases the clinical effectiveness of antibacterial therapy by reducing the film-forming ability.

1. Matosova E.V., Besednova N.N., Kusaikin M.I., Andryukov B.G., Makarenkova I.D., Shchelkanov M.Yu., Lyapun I.N., Bynina M.P., Ermakova S.P., Zvyagintseva T.N. Antibiofilm activity of fukoidans isolated from brown algae. Antibiot Khimioter = Antibiotics and Chemotherapy. 2023; 68 (9–10): 5–11. doi: https://doi.org/10.37489/0235-2990-2023-68-9-10-5-11. (in Russian)]

2. Khryanin A.A. Microbial biofilms: modern concepts. Antibiotics and Chemotherapy. 2020; 65 (5–6): 70–77. doi: https://doi.org/10.37489/0235-2990-2020-65-5-6-70-77. (in Russian)]

3. Tolker-Nielsen T. Biofilm development. Microbiol Spectr. 2015 Apr; 3 (2): MB-0001-2014. doi: 10.1128/microbiolspec.MB-0001-2014. PMID: 26104692.

4. Mirzaei R., Mohammadzadeh R., Alikhani M.Y., Shokri Moghadam M., Karampoor S., Kazemi S., Barfipoursalar A., Yousefimashouf R. The biofilm-associated bacterial infections unrelated to indwelling devices. IUBMB Life. 2020 Jul; 72 (7): 1271–1285. doi: 10.1002/iub.2266. Epub 2020 Mar 9. PMID: 32150327.

5. Nikolenko M.V., Baryshnikova N.V., Malishevskaya O.I., Enoktaeva O.V., Vaseva E.M. A 24-hour Сandida sp. biofilm formation dynamically assesed with modified macrometric method. Russian Journal of Infection and Immunity = Infektsiya i immunitet. 2022; 12 (6): 1129–1135. doi: https://doi.org/10.15789/2220-7619-AHC-1929. (in Russian)]

6. Ciofu O., Tolker-Nielsen T. Antibiotic tolerance and resistance in biofilms. In: T. Bjarnsholt, P. Jensen, C. Moser, N. Høiby (eds.). Biofilm Infections. Springer, New York, NY. 2010. doi: 10.1007/978-1-4419-6084-9_13.

7. Polygach O.A., Dabizheva A.N., Voroshilova N.N. Effect of the composition of lytic bacteriophages of P. aeruginosa formation and destruction of bacterial biofilms. Epidemiology and Vaccinal Prevention. 2018; 17 (4): 20–25. doi: https://doi.org/10.31631/2073-3046-2018-17-4-20-25. (in Russian)]

8. Song Y.J., Yu H.H., Kim Y.J., Lee N.K., Paik H.D. Anti-Biofilm Activity of Grapefruit Seed Extract against Staphylococcus aureus and Escherichia coli. J Microbiol Biotechnol. 2019 Aug 28; 29 (8): 1177–1183. doi: 10.1041/jmb.1905.05022. PMID: 31370119.

9. Lagha R., Ben Abdallah F., Al-Sarhan B.O., Al-Sodany Y. Antibacterial and biofilm inhibitory activity of medicinal plant essential oils against Escherichia coli isolated from UTI patients. Molecules. 2019 Mar 23; 24 (6): 1161. doi: 10.3390/molecules24061161. PMID: 30909573; PMCID: PMC6471185.

10. Giedraitiene A., Pereckaite L., Bredelyte-Gruodiene E., Virgailis M., Ciapiene I., Tatarunas V. CTX-M-producing Escherichia coli strains: resistance to temocillin, fosfomycin, nitrofurantoin and biofilm formation. Future Microbiol. 2022 Jul; 17: 789–802. doi: 10.2217/fmb-2021-0202. Epub 2022 May 13. PMID: 35549350.