The relevance of this study is conditioned upon epidemic growth of nosocomial infections, which include Escherichia Coli (E. coli). One of the factors of pathogenicity of such microorganisms is the ability to form a biofilm – a complex community, within which bacteria acquire increased resistance to environmental factors, primarily to antibacterial drugs, which considerably complicates the course of the infectious process. In this regard, the purpose of this study was to determine the features of the formation and dependence of the density of the formed biofilm on the antibiotic resistance of pathogenic and commensal E. coli strains isolated from dogs and cats. The resistance of E. coli isolates to antibacterial drugs was established according to the disk diffusion method, according to EUCAST recommendations. The ability of microorganisms to form biofilms and determine their density was investigated in sterile plastic 96-well plates. The ability to form biofilms was assessed visually and microscopically, the density of biofilms was determined in units, spectrophotometrically, by the optical density of the washing solution. The paper presents the results of a study of 63 samples of pathological (wound infections) and biological material. From them, 10 E. coli isolates were obtained (6 from dogs and 4 from cats), which were selected for further research. It was established that all E. coli isolates had the ability to form phenotypic biofilm. The study investigated the interdependence of antibiotic resistance of E. coli isolates and their ability to form biofilms. Thus, isolates that were parted from pathological material and had a positive reaction on the CHROMagar™ ESBL medium for the determination of extended-spectrum beta-lactamases had greater resistance to various groups of antibacterial drugs and formed high- and medium-density biofilms, while E. coli isolates parted from pathological and biological materials with a negative reaction on CHROMagar™ ESBL medium formed a low-density biofilm and had less resistance to different groups of antibacterial drugs. The results obtained allow searching for innovative, sometimes alternative, methods of treatment and prevention of pathologies caused by them
microorganism, biofilm, antibiotic resistance, antibacterial drugs, wound infection
[1] Ushkalov, V., Salmanov, A., Kalachniuk, L., Vishovan, Y., Boianovskiy, S., Ushkalov, A., Granate, A., Huwiage, G.M., & Kepple, O. (2020). The influence of cultivation temperature on some phenotypic traits of Yersinia pseudotuberculosis. One Health & Risk Management, 1(2), 34-40. doi: 10.38045/ohrm.2020.1.14.
[2] Tallawi, M., Opitz, M., & Lieleg, O. (2017). Modulation of the mechanical properties of bacterial biofilms in response to environmental challenges. Biomaterials Science, 5, 887-900. doi: 10.1039/C6BM00832A.
[3] Akter, S., Chowdhury, A., & Mina, S.A. (2021). Antibiotic resistance and plasmid profiling of Escherichia coli isolated from human sewage samples. Microbiology Insights, 14. doi: 10.1177/11786361211016808.
[4] Albert, M.J., Bulach, D., Alfouzan, W., Izumiya, H., Carter, G., Alobaid, K., Alatar, F., Sheikh, A.R., & Poirel, L. (2019). Non-typhoidal Salmonella blood stream infection in Kuwait: Clinical and microbiological characteristics. Plos Neglected Tropical Diseases, 13(4). doi: 10.1371/journal.pntd.0007293.
[5] Mahmud, Z.H., Shirazi, F.F., Hossainey, M., Islam, M.I., Ahmed, M.A., Nafiz, T.N., Imran, K.M., Sultana, J., Islam, M.S., Islam, M.A., & Islam, M.S. (2019). Presence of virulence factors and antibiotic resistance among Escherichia coli strains isolated from human pit sludge. Journal of Infection in Developing Countries, 13(3), 195-203. doi: 10.3855/jidc.10768.
[6] Gonzales-Rodriguez, A.O., Infante Varillas, S.F., Barrón Pastor, H.J., Llimpe Mitma, Y., Huerta Canales, D., Wong Chero, P.A., Gutierrez, C., & Suarez Cunza, S. (2020). Immunological and biochemical response of older adults with urinary tract infection to uropathogenic Esherichia coli virulence factors. Rev Peru Med Exp Salud Publica, 37(3), 527-531. doi: 10.17843/rpmesp.2020.373.4918.
[7] Hall, C.W., & Mah, T.F. (2017). Molecular mechanisms of biofilm-based antibiotic resistance and tolerance in pathogenic bacteria. FEMS Microbiology Reviews, 41(3), 276-301. doi: 10.1093/femsre/fux010.
[8] Del Pozo, J.L. (2018). Biofilm-related disease. Expert Review of Anti-Infective Therapy, 16(1), 51-65. doi: 10.1080/14787210.2018.1417036.
[9] Huang, J., Liu, S., Zhang, C., Wang, X., Pu, J., Ba, F., Xue, S., Ye, H., Zhao, T., Li, K., Wang, Y., Zhang, J., Wang, L., Fan, C., Lu, T.K., & Zhong, C. (2019). Programmable and printable Bacillus subtilis biofilms as engineered living materials. Nature Chemical Biology, 15(1), 34-41. doi: 10.1038/s41589-018-0169-2.
[10] Vishovan, Y., Ushkalov, V., Vygovska, L., Ishchenko, L., Salmanov, A., Bilan, A., Kalakailo, L., Hranat, A., & Boianovskiy, S. (2021). Biofilm formation and antibiotic resistance in Staphylococcus isolated from different objects. EUREKA: Life Sciences, (4), 58-65. doi: 10.21303/2504-5695.2021.001925.
[11] Miquel, S., Lagrafeuille, R., Souweine, B., & Forestier, C. (2016). Anti-biofilm activity as a health issue. Frontiers in Microbiology, 7, 592. doi: 10.3389/fmicb.2016.00592.
[12] Puligundla, P., & Mok, C. (2017). Potential applications of nonthermal plasmas against biofilm-associated micro-organisms in vitro. Journal of Applied Microbiology, 122(5), 1134-1148.
doi: 10.1111/jam.13404. [13] Baker, K.S. (2015). Demystifying Escherichia coli pathovars. Nature Reviews Microbiology, 13(1), 5. doi: 10.1038/nrmicro3411.
[14] Rohatgi, A., & Gupta, P. (2021). Natural and synthetic plant compounds as anti-biofilm agents against Escherichia coli O157:H7 biofilm. Infection, Genetics and Evolution: Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases, 95, article number 105055. doi: 10.1016/j.meegid.2021.105055.
[15] Kot, B. (2019). Antibiotic resistance among uropathogenic Escherichia coli. Polish Journal of Microbiology, 68(4), 403-415. doi: 10.33073/pjm-2019-048.
[16] Poolman, J.T., & Wacker, M. (2016). Extraintestinal pathogenic Escherichia coli, a common human pathogen: Challenges for vaccine development and progress in the field. The Journal of Infectious Diseases, 213(1), 6-13. doi: 10.1093/infdis/jiv429.
[17] Raeispour, M., & Ranjbar, R. (2018). Antibiotic resistance, virulence factors and genotyping of uropathogenic Escherichia coli strains. Antimicrobial Resistance and Infection Control, 7, 118. doi: 10.1186/s13756-018-0411-4.
[18] Roy, R., Tiwari, M., Donelli, G., & Tiwari, V. (2018). Strategies for combating bacterial biofilms: A focus on anti-biofilm agents and their mechanisms of action. Virulence, 9(1), 522-554. doi: 10.1080/21505594.2017.1313372.
[19] Potochylova, V., Rudnyeva, K., Pokas, O., & Vyshnyakova, H. (2020). Sensitivity to antibacterial drugs and phenotypic determination of resistance factors in microorganisms of the family Enterobacteriaceae – pathogens of wound infections. Bulletin of Problems of Biology and Medicine, 4(158), 259-263. doi: 10.29254/2077-4214- 2020-4-158-259-263.
[20] Kukhtin, M.D., & Krushelnytska, N.V. (2014). Formation of biofilms by microorganisms isolated from milking equipment. Animal Biology, 16(1), 95-103. doi: 10.15587/1729-4061.2017.110488.
[21] Mishyna, M.M., Makieieva, N.I., Marchenko, I.A., Golovachova, V.A., & Osolodchenko, T.P. (2020). Biofilms formation by pyelonephritis causative agents in infants as a mechanism of resistance to antimicrobial agents. Ukrainian Journal of Medicine, Biology and Sport, 2(24), 104-111. doi: 10.26693/jmbs05.02.104.
[22] Eucast. The European committee on antimicrobial susceptibility testing. Retrieved from https://www.eucast.org/.
[23] Sergeant, ESG. Epitools Epidemiological Calculators. Ausvet. (2018). Retrieved from https://epitools.ausvet.com.au.