Evaluation ofthe additive effect ofthe interaction of phytobiotics with zinc on Quorum Sensing P. aeruginosa in an in vitro mode

Relevance. The development of the aquaculture sector is one of the promising developing areas that contribute to ensuring the food security of mankind in the world. However, all aquatic animals are susceptible to the impact of negative factors leading to a decrease in growth rates, a decrease in the quality of finished products, etc.
Methods. The paper presents experimental data on the study of the additive effect of a combination of various commercial phytobiotic drugs with zinc on the sense of quorum and inhibitory characteristics on a model of a polyresistant strain of P. aeruginosa. The choice of the strain is due to its high resistance characteristics, ability to biofilm formation, as well as the ability to visually assess the impact of the tested compounds on the Quorum Sensing (QS) system by suppressing the formation of the pyocyanin pigment, which provides virulence factors and biofilm growth. Butitan, Probiocid®-Phyto, and Intebio preparations were used as factors regulating the growth of the tested strain; ZnSO₄ was used as a source of zinc cations. The use of the diffusion method of agar wells allowed us to evaluate not only the level of the inhibitory effect of the studied compounds, but also the presence of their influence (QS) of the system.
Results. The experimental data obtained indicate a pronounced effect of zinc cations on the production of the pyocyanin pigment (0.25 mM/ml), as well as the tested preparations from the group of fodder phytobiotics at concentrations of 100 mg/ml. The presence of significantly significant differences (р ≤ 0.001) in the impact on (QS) in combinations of zinc with phytobiotics at concentrations of 0.13 mM/ml ZnSO₄ and extracts of drugs 50 mg/ml was established, with the highest rates in “Probiocid®-Phyto”. Thus, the data obtained allow us to conclude that the use of the studied phytobiotics in combination with essential elements is promising as an alternative to feed antibiotics in fish feeding, for the prevention of infectious diseases.

1. Dawood M.A.O. et al. Exploring the Roles of Dietary Herbal Essential Oils in Aquaculture: A Review. Animals. 2022; 12(7): 823. https://doi.org/10.3390/ani12070823

2. Bertocci F., Mannino G. Can Agri-Food Waste Be a Sustainable Alternative in Aquaculture? A Bibliometric and Meta-Analytic Study on Growth Performance, Innate Immune System, and Antioxidant Defenses. Foods. 2022; 11(13): 1861. https://doi.org/10.3390/foods11131861

3. Thi M.T.T., Wibowo D., Rehm B.H.A. Pseudomonas aeruginosa Biofilms. International Journal of Molecular Sciences. 2020; 21(22): 8671. https://doi.org/10.3390/ijms21228671

4. O’Loughlin C.T., Miller L.C., Siryaporn A., Drescher K., Semmelhack M.F., Bassler B.L. A quorum-sensing inhibitor blocks Pseudomonas aeruginosa virulence and biofilm formation. Proceedings of the National Academy of Sciences. 2013; 110(44): 17981–17986. https://doi.org/10.1073/pnas.1316981110

5. Lee J., Zhang L. The hierarchy quorum sensing network in Pseudomonas aeruginosa. Protein & Cell. 2015; 6(1): 26–41. https://doi.org/10.1007/s13238-014-0100-x

6. Rosier A., Bishnoi U., Lakshmanan V., Sherrier D.J., Bais H.P. A perspective on inter-kingdom signaling in plant-beneficial microbe interactions. Plant Molecular Biology. 2016; 90(6): 537–548. https://doi.org/10.1007/s11103-016-0433-3

7. Paczkowski J.E. et al. Flavonoids Suppress Pseudomonas aeruginosa Virulence through Allosteric Inhibition of Quorum-sensing Receptors. Journal of Biological Chemistry. 2017; 292(10): 4064–4076. https://doi.org/10.1074/jbc.M116.770552

8. Anju V.T. et al. Sesamin and sesamolin rescues Caenorhabditis elegans from Pseudomonas aeruginosa infection through the attenuation of quorum sensing regulated virulence factors. Microbial Pathogenesis. 2021; 155: 104912. https://doi.org/10.1016/j.micpath.2021.104912

9. Luciardi M.C., Blázquez M.A., Alberto M.R., Cartagena E., Arena M.E. Grapefruit essential oils inhibit quorum sensing of Pseudomonas aeruginosa. Food Science and Technology International. 2020; 26(3): 231–241. https://doi.org/10.1177/1082013219883465

10. Gholami M., Zeighami H., Bikas R., Heidari A., Rafiee F., Haghi F. Inhibitory activity of metal-curcumin complexes on quorum sensing related virulence factors of Pseudomonas aeruginosa PAO1. AMB Express. 2020; 10: 111. https://doi.org/10.1186/s13568-020-01045-z

11. Seryakova A.A., Panov V.P., Prosekova E.A., Komarchev A.S., Voronin K.O., Tsvetkova V.A. Тhe effect of the feed additive “Butitan” (“Farmatan BCO”) on the histophysiological state of the intestinal tube and the productive qualities of broiler chickens. Agrarian science. 2021; 4S: 60–65 (in Russian). https://doi.org/10.32634/0869-8155-2021-347-4-60-65

12. Manukyan V.A., Kharitonova D.I., Baykovskaya E.Yu. The Effects of an Acidifier with Essential Oil of Lemongrass on Non-Specific Immunity, Intestinal Microbiota, and Productive Performance in Broilers. Ptitsevodstvo. 2021; 7–8: 34–37 (in Russian). https://doi.org/10.33845/0033-3239-2021-70-7-8-34-37

13. Egorov I.A. et al. Poultry diets without antibiotics. II. Intestinal microbiota and performance of broiler (Gallus gallus L.) breeders fed diets with a phytobiotic. Agricultural Biology. 2019; 54(4): 798–809. https://doi.org/10.15389/agrobiology.2019.4.798eng

14. Gündel S.d.S. et al. Nanoemulsions containing Cymbopogon flexuosus essential oil: Development, characterization, stability study and evaluation of antimicrobial and antibiofilm activities. Microbial Pathogenesis. 2018; 118: 268–276. https://doi.org/10.1016/j.micpath.2018.03.043

15. Gao S. et al. Antimicrobial Activity of Lemongrass Essential Oil (Cymbopogon flexuosus) and Its Active Component Citral Against Dual-Species Biofilms of Staphylococcus aureus and Candida Species. Frontiers in Cellular and Infection Microbiology. 2020; 10: 603858. https://doi.org/10.3389/fcimb.2020.603858

16. Rossi G.G. et al. Antibiofilm activity of nanoemulsions of Cymbopogon flexuosus against rapidly growing mycobacteria. Microbial Pathogenesis. 2017; 113: 335–341. https://doi.org/10.1016/j.micpath.2017.11.002

17. Duplantier M, Lohou E, Sonnet P. Quorum Sensing Inhibitors to Quench P. aeruginosa Pathogenicity. Pharmaceuticals. 2021; 14(12): 1262. https://doi.org/10.3390/ph14121262