Along with the relevance of preventing the formation of biofilms, their damage, antimicrobial action on fungi and bacteria, especially antibiotic-resistant, it is important to find substances that can inhibit the adhesion process, since attachment of microbial cells of the pathogen to the biological object is the beginning of the development infectious process and primary stage of formation of biofilm of pathogens, which are much more complex to struggle than planktonic forms. The purpose of the work is to study in vitro the ability of metabolites complexes of Lactobacillus rhamnosus GG and Saccharomyces boulardii, obtained by the author's method, to influence the adhesion of microbial cells to substantiate the prospects of their use in the development of new multifunctional preparations. Material and methods. The in vitro study of the adhesion of bacterial cells of Staphylococcus aurues 209-P was carried out after their treatment or treatment of erythrocytes with disintegrates (obtained by ultrasonic irradiation of cells of Lactobacillus rhamnosus GG or Saccharomyces boulardii) and metabolites (obtained by culturing lactobacteria and saccharomycetes in their own structural components). Results and discussion. The highest statistically significant inhibition of adhesion of the treated S. aurues cells occurred with L. rhamnosus metabolites obtained by culturing producers in their own structural components (17.19%, P=0.0005) and S. boulardii metabolites obtained by culturing producents in structural components L.°rhamnosus (11.03%, P=0.006). The maximum suppression of adhesion of S. aurues during the processing of erythrocytes was also carried out by metabolites of L. rhamnosus (by 14.9%, P=0.005) and metabolites of S. boulardii (by 12.98%, P=0.02). Regardless of the treatment method (erythrocytes or staphylococcus cells) less anti-adhesive properties had the disintegrate of lactobacteria and the combination of metabolites L. °rhamnosus and S. boulardii (by 4.12% - 5.63%). The absence of suppression of the adhesion of pathogen cells to erythrocytes was noted during the treatment of staphylococcus with metabolites of S. boulardii, obtained by culturing the producents in their own structural components, and during the processing of erythrocytes also by the disintegrate of S. boulardii. Conclusion. The presented results can be useful when creating antimicrobial preparations new generation based on the metabolite complexes Lactobacillus and Saccharomyces.

Keywords: adhesion, metabolites, anti-adhesive properties, Lactobacillus rhamnosus GG, Saccharomyces boulardii

Full text: PDF (Ukr) 277K

1. Nougayrède JP, Homburg S, Taieb F, Boury M, Brzuszkiewicz E, Gottschalk G, et al. Escherichia coli Induces DNA Double-Strand Breaks in Eukaryotic Cells. Science. 2006; 313(5788): 848-51. https://www.ncbi.nlm.nih.gov/pubmed/16902142. https://doi.org/10.1126/science.1127059
2. Cuevas-Ramos G, Isberg RR. Escherichia coli induces DNA damage in vivo and triggers genomic instability in mammalian cells. Proc Natl Acad Sci USA. 2010; 107(25): 11537-42. https://www.ncbi.nlm.nih.gov/pubmed/20534522. https://www.ncbi.nlm.nih.gov/pmc/articles/2895108. https://doi.org/10.1073/pnas.1001261107
3. Million M, Raoult D. Publication biases in probiotics. Eur J Epidemiol. 2012; 27(11): 885-6. https://www.ncbi.nlm.nih.gov/pubmed/23086285. https://doi.org/10.1007/s10654-012-9740-4
4. Isayenko OY, Knysh OV, Babych YM, Ryzhkova TN, Dyukareva GI. Effect of disintegrates and metabolites of Lactobacillus rhamnosus and Saccharomyces boulardii on biofilms of antibiotic resistant conditionally pathogenic and pathogenic bacteria. Regul Mech Biosyst. 2019; 10(1): 3-8. [Ukrainian]. https://doi.org/10.15421/021901
5. Isayenko OYu. Anti-diphtheria properties of structural-metabolites complexes of Lactobacteria and Saccharomyces probiotic strains. [Protydyfteriyni vlastyvosti strukturno-metabolitnykh kompleksiv probiotychnykh shtamiv laktobakteriy i sakharomitsetiv v testakh in vitro ta in vivo]. Fiziol Zh. 2019; 65(6): 51-60. [Ukrainian]. https://doi.org/10.15407/fz65.06.051
6. Isayenko OY. Synergistic activity of filtrates Lactobacillus rhamnosus GG and Saccharomyces boulardii and antibacterial preparations against Corynebacterium. Regul Mech Biosyst. 2019; 10(4): 445-53. [Ukrainian]. https://doi.org/10.15421/021968
7. Patent 123122 Ukraine, МПК: C12R 1/25, A61K 35/74, C12N 1/20, A61K 35/741. Method of producing metabolites of probiotic bacterial strains [Sposib oderzhannia metabolitov probioticheskikh shtammov bakterii] / Horbach TV, Isayenko OYu, Babych YeM, Kivva FV, Knysh OV, Balak OK. (UA); zayavnik i vlasnik patentu DU«IMI NAMN» (UA). № 123122; zayavl 04.09.17 ; opubl 12.02.18. Byul № 3. [Ukrainian]
8. Patent 126603 Ukraine. МПК: C12N 1/00, A61K 36/06, A61K 35/66. A method of obtaining a combination of metabolites of probiotic strains of fungi and bacteria. [Sposib oderzhannia kombіnatcії metabolіtіv probіotichnikh shtamіv gribіv і bakterіi] / Isayenko OYu, Knysh OV, Babych YeM, Vashchenko V, Zachepylo SV, Polyanska VP, Kovalenko OI, Balak OK. (UA); zayavnik i vlasnik patentu DU«IMI NAMN» (UA). № 126603; zayavl 05.02.18; opubl 25.06.18. Byul № 12. [Ukrainian]
9. Ivonin AG, Oborin VA. Development and prospects for application of the photometric method for determining the bacteriofixing activity of erythrocytes in veterinarium. Letters of the orenburg state agricultural university. 2008; 3(19): 80-82. [Russian]
10. Pirog Т, Grytsenko N, Konon А, Shevchuk Т, Іutynska G. Antiadhesive potencial of Rhodococcus erythropolis IMВ Aс-5017 biosurfactants. Microbiol Zh. 2014; 6(76): 9-16.
11. Iglesias MB, Viñas I, Colás-Medà P, Collazo С, Serrano JCE. Adhesion and invasion of Listeria monocytogenes and interaction with Lactobacillus rhamnosus GG after habituation on fresh-cut pear. J Funct Foods. 2017; 34: 453-60. https://doi.org/10.1016/j.jff.2017.05.011
12. Nantavisai К, Puttikamonkul S, Chotelersak К, Taweechotipatr М. In vitro adhesion property and competition against enteropathogens of Lactobacillus strains isolated from Thai infants. Songklanakarin J Sci Technol. 2018; 40 (1): 69-74. https://doi.org/10.14456/sjst-psu.2018.14.
13. Zeraik AE, Nitschke M. Biosurfactants as agents to reduce adhesion of pathogenic bacteria to polystyrene surfaces: effect of temperature and hydrophobicity. Curr Microbiol. 2010; 61(6): 554 -59. https://www.ncbi.nlm.nih.gov/pubmed/20422191. https://doi.org/10.1007/s00284-010-9652-z
14. Mataraci E, Dosler S. In vitro activities of antibiotics and antimicrobial cationic peptides alone and in combination against methicillin-resistant Staphylococcus aureus biofilms. Antimicrob Agents Chemother. 2012; 56(12): 6366-71. https://www.ncbi.nlm.nih.gov/pubmed/23070152. https://www.ncbi.nlm.nih.gov/pmc/articles/3497160. https://doi.org/10.1128/AAC.01180-12
15. Dunne C, Kelly P, O'Halloran S, Soden D, Bennett M, von Wright A, et al. Mechanisms of adherence of a probiotic Lactobacillus strain during and after in vivo assessment in ulcerative colitis patients. Microbial Ecology in Health and Disease. 2004; 16: 96-104.https://doi.org/10.1080/08910600410032295
16. Nitschke M, Araujo LV, Costa SG, Pires RC, Zeraik AE, Fernandes AC, et al. Surfactin reduces the adhesion of food-borne pathogenic bacteria to solid surfaces. Lett Appl Microbiol. 2009; 49(2): 241-47. https://www.ncbi.nlm.nih.gov/pubmed/19486287. https://doi.org/10.1111/j.1472-765X.2009.02646.x
17. Kalyani R. Bishwambhar M, Suneetha V. Recent potential usage of surfactant from microbial origin in pharmaceutical and biomedical arena: a perspective. Int Res J Pharm. 2011; 2(8): 11-5.
18. Lebeer S, Vanderleyden J, De Keersmaecker SC. Genes and molecules of lactobacilli sup-porting probiotic action. MMBR. 2008; 72(4): 728-64. https://www.ncbi.nlm.nih.gov/pubmed/19052326. https://www.ncbi.nlm.nih.gov/pmc/articles/2593565. https://doi.org/10.1128/MMBR.00017-08
19. He X, Zeng Q, Puthiyakunnon S, Zeng Z, Yang W, Qiu J, et al. Lactobacillus rhamnosus GG supernatant enhance neonatal resistance to systemic Escherichia coli K1 infection by accelerating development of intestinal defense. Sci Rep. 2017; 7: 43305. https://www.ncbi.nlm.nih.gov/pubmed/28262688. https://www.ncbi.nlm.nih.gov/pmc/articles/5338013. https://doi.org/10.1038/srep43305
20. Polishchuk LZ, Ryabtseva OD, Lukyanova NYu, Chekhun VF. Adhesion molecules and their importance in the development of malignant tumors. Oncology. 2011; 13(1): 4-11.