DOI: https://doi.org/10.22263/2312-4156.2021.1.46
Artsiukh T.V., Sokolova T.N., Astrowskaja A.B.
The resistance peculiarities of E.coli and C.albicans clinical isolates forming a biofilm
Grodno State Medical University, Grodno, Republic of Belarus
Vestnik VGMU. 2021;20(1):46-54.
Abstract.
Objectives. To model biofilms of mixed paired cultures of C.albicans and E.coli in vitro and study the characteristics of their sensitivity to antimicrobial drugs (doxycycline, amoxiclav, ofloxacin, fluconazole, clotrimazole) in planktonic form, as well as in the composition of mono- and mixed biofilms.
Material and methods. In the course of the study, the properties and ability to film formation of C.albicans 2924 and E.coli 2646 isolated from mucous membranes in patients with clinical signs of vaginitis were investigated. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of antibiotics were determined for planktonic forms of microorganisms: bacteria and candida in the composition of a mono-biofilm, as well as in the composition of a mixed biofilm.
Results. It has been revealed that the studied microorganisms are capable of forming a biofilm. On studying the sensitivity of microorganisms in biofilms (BF) it has been found that the MIC of preparations for mono-biofilms increases 2-4 times, for mixed biofilms 2.5-8.5 times compared to the MIC of preparations for planktonic forms of E.coli and C.albicans. Bactericidal concentrations of amoxiclav, ofloxacin, doxycycline, fluconazole, clotrimazole for planktonic forms are not effective for biofilm forms of the same microorganisms.
Conclusions. To determine the sensitivity of microorganisms in the composition of biofilms to antibacterial drugs is necessary for the administration of rational antibiotic therapy and the development of empirical antibiotic therapy protocols. It is important to study the mechanisms of bacterial resistance in the composition of biofilms, develop new pharmaceuticals with a high potential for action on biofilms, since standard therapy with high doses of antibiotics (AB) is toxic for the macroorganism, which dictates the need to test new agents.
Key words: resistance, microbial films, antibiotics, antimycotic drugs, minimum inhibitory concentration.
Acknowledgement: The authors express their gratitude to the senior laboratory assistant of the Chair of Microbiology, Virology and Immunology named after S.I. Gelberg of Grodno State Medical University Domostoy E.A. for the help provided while conducting investigations.
References
1. VOZ, Evrop Region biuro. Antimicrobial resistance surveillance in Central Asia and Europe [Elektronnyi resurs]: CAESAR 2019 g: prakt posobie. Rezhim dostupa: https://www.euro.who.int/__data/assets/pdf_file/0013/430132/WHO-CAESAR-manual-2019-RUS.pdf. Data dostupa: 26.01.2021. (In Russ.)
2. VOZ, Evrop Region komitet. European strategic action plan on antibiotic resistance [Elektronnyi resurs]: 61-ia ses, Baku, Azerbaidzhan, 2011 g. Rezhim dostupa: https://www.euro.who.int/__data/assets/pdf_file/0015/150612/RC61_Res_r06.pdf. Data dostupa: 26.01.2021. (In Russ.)
3. Gavrilova IA, Titov LP. Resistance of hospital isolates of staphylococci and Pseudomonas aeruginosa to disinfectants. Zdravookhranenie. 2011;(11):18-20. (In Russ.)
4. Afinogenova AG, Darovskaia EN. Microbial biofilms of wounds: state of the art. Travmatologiia Ortopediia Rossii. 2011;(3):119-25. (In Russ.)
5. Oliveira NM, Martinez-Garcia E, Xavier J, Durham WM, Kolter R, Kim W, et al. Biofilm formation as a response to ecological competition. PLoS Biol. 2015 Jul 9;13(7):e1002191. doi: http://dx.doi.org/10.1371/journal.pbio.1002191
6. Shlepotina NM, Peshikova MV, Kolesnikov OL, Shishkova IuS. Modern concepts of the mechanisms of interaction between biofilm and cellular immunity factors. Zhurn Mikrobiologii Epidemiologii Immunologii. 2020;97(1):83-90. (In Russ.)
7. Bukharin OV, Gintcburg AL, Romanova IuM, El-Registan GI. Bacterial survival mechanisms. Moscow, RF: Meditsina; 2005. 367 р. (In Russ.)
8. Biradar B, Devi P. Quorum sensing in plaque biofilms: challenges and future prospects. J Contemp Dent Pract. 2011 Nov;12(6):479-85. doi: http://dx.doi.org/10.5005/jp-journals-10024-1080
9. Simonova IR, Golovin SN, Verkina LM, Berezniak EA, Titova SV. Cultivation and study methods of bacterial biofilms. Izv Vyssh Ucheb Zavedenii Severo-Kavkaz Region Estestv Nauki. 2017;(1):75-81. (In Russ.)
10. Hooshangi S, Bentley WE. From unicellular properties to multicellular behavior: bacteria quorum sensing circuitry and applications. Curr Opin Biotechnol. 2008 Dec;19(6):550-5. doi: http://dx.doi.org/10.1016/j.copbio.2008.10.007
11. Høiby N, Bjarnsholt T, Givskov M, Molin S, Ciofu O. Antibiotic resistance of bacterial biofilms. Int J Antimicrob Agents. 2010 Apr;35(4):322-32. doi: http://dx.doi.org/10.1016/j.ijantimicag.2009.12.011
12. Petrova OE, Sauer K. Escaping the biofilm in more than one way: desorption, detachment or dispersion. Curr Opin Microbiol. 2016 Apr;30:67-78. doi: http://dx.doi.org/10.1016/j.mib.2016.01.004
13. Gopal R, Kim YG, Lee JH, Lee SK, Chae JD, Son BK, et al. Synergistic effects and antibiofilm properties of chimeric peptides against multidrug-resistant Acinetobacter baumannii strains. Antimicrob Agents Chemother. 2014;58(3):1622-9. doi: http://dx.doi.org/10.1128/AAC.02473-13
14. Cherniavskii VI. Bacterial biofilms and infections. Ann Mechnikov Inst. 2013;(1):86-90. (In Russ.)
15. Mardanova AM, Kabanov DA, Rudakova NL, Sharipova MR. Biofilms: basic principles of organization and research methods: ucheb-metod posobie. Kazan, RF: Tsentr pechati Link; 2016. 48 р. (In Russ.)
16. Liuis K. Persistent cells and the mystery of biofilm survival. Biokhimiia. 2005;70(2):327-36. (In Russ.)
17. Lafleur MD, Qi Q, Lewis K. Patients with long-term oral carriage harbor high-persister mutants of Candida albicans. Antimicrob Agents Chemother. 2010 Jan;54(1):39-44. doi: http://dx.doi.org/10.1128/AAC.00860-09
18. Jorge P, Lourenco A, Pereira MO. New trends in peptide-based anti-biofilm strategies: a review of recent achievements and bioinformatic approaches. Biofouling. 2012;28(10):1033-61. doi: http://dx.doi.org/10.1080/08927014.2012.728210
19. Volosach OS, Petrova SE, Markovich NS. Leading causative agents of urinary tract infections in patients of Grodno region in 2018 [Elektronnyi resurs]. V: Aktual'nye voprosy mikrobiologii, infektologii i immunologii: sb materialov mezhvuz konf, posviashch pamiati prof SI Gel'berga, Grodno, 29 apr 2019 g. Grodno, RB; 2019. P. 30-3. 1 elektron opt disk. (In Russ.)
20. Pozdeev OK, Fedorov RV. Enterobacteriaceae: ruk dlia vrachei. Moscow, RF: GEOTAR-Media; 2007. 720 р. (In Russ.)
21. Donlan RM, Costerton JW. Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev. 2002 Apr;15(2):167-93. doi: http://dx.doi.org/10.1128/cmr.15.2.167-193.2002
22. Delaloye J, Calandra T. Invasive candidiasis as a cause of sepsis in the critically ill patient. Virulence. 2014 Jan;5(1):161-9. doi: http://dx.doi.org/10.4161/viru.26187
23. Mills BB. Vaginitis: beyond the basics. Obstet Gynecol Clin North Am. 2017 Jun;44(2):159-177. doi: http://dx.doi.org/10.1016/j.ogc.2017.02.010
24. Sherry L, Kean R, McKloud E, O'Donnell LE, Metcalfe R, Jones BL, et al. Biofilms formed by isolates from recurrent vulvovaginal candidiasis patients are heterogeneous and insensitive to fluconazole. Antimicrob Agents Chemother. 2017 Aug 24;61(9):e01065-17. doi: http://dx.doi.org/10.1128/AAC.01065-17
25. Bairamova GR, Baranov II, Priputnevich TV, Chernova VF. Vulvovaginal candidiasis: clinical and therapeutic aspects in the practice of an obstetrician-gynecologist. Akusherstvo i ginekologiia: novosti, mneniia, obucheniia. 2017;(4):63-9. (In Russ.)
26. Kontcevaia II. Microbiology: Cultivation and Growth of Bacteria: prakt ruk dlia studentov biol spets vuzovia. Chernihiv, Ukraine: Desna poligraf; 2017. 44 р. (In Russ.)
27. Tapalskii DV, Bilskii IA. Determination of antibiotic susceptibility by broth microdilution: a modification available to all: modifikatsiia, dostupnaia dlia vsekh. Klin Mikrobiologiia Antimikrob Khimioterapiia. 2018;20(1):62-7. (In Russ.)
28. Biosan SIA. DEN-1 DEN-1В. Densitometer. Suspension turbidity detector [Elektronnyi resurs]: instruktsiia pol'zovatelia. Rezhim dostupa: https://biosan.lv/media/products/files/den-1-den-1b-ru204-0319_D7XD9aA.pdf. Data dostupa: 26.01.2021. (In Russ.)
29. Ramage G, Walle KV, Wickes BL, Lo´pez-Ribot JL. Standardized method for in vitro antifungal susceptibility testing of Candida albicans biofilms. Antimicrob Agents Chemother. 2001 Sep;45(9):2475-9. doi: http://dx.doi.org/10.1128/aac.45.9.2475-2479.2001
30. Lysak VV, Zheldakova RA, Fomina OV. Microbiology: praktikum. Minsk, RB: BGU; 2015. 115 р. (In Russ.)
31. Sokolova TN. Microbial biofilms and methods for their detection. Zhurn GrGMU. 2014;(4):12-5. (In Russ.)
32. Pierce CG, Uppuluri P, Tristan AR, Wormley FL, Mowat E, Ramage G, et al. A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing. Nat Protoc. 2008;3(9):1494-500. doi: http://dx.doi.org/10.1038/nport.2008.141
33. Zinchenko AI. Biofilms of microorganisms and methods of combating them. V: Kolomiets EI, Lobanok AG, Zinchenko AI, Galushko VM, Ivshina IB, Mikhailova RV, i dr, redkol. Mikrobnye biotekhnologii: fundamental'nye i prikladnye aspekty: sb st. Minsk, RB; 2016. T 8. Р. 334-52. (In Russ.)
Information about authors:
Artsiukh T.V. – lecturer of the Chair of Microbiology, Virology and Immunology named after S.I. Gelberg, Grodno State Medical University,
ORCID: https://orcid.org/0000-0001-7368-0623
Sokolova T.N. – Candidate of Medical Sciences, associate professor of the Chair of Microbiology, Virology and Immunology named after S.I. Gelberg, Grodno State Medical University,
ORCID: https://orcid.org/0000-0002-4075-4515
Astrowskaja A.B. – Candidate of Medical Sciences, associate professor of the Chair of Microbiology, Virology and Immunology named after S.I. Gelberg, senior researcher of morphology group with electron microscopy of the Research Laboratory, Grodno State Medical University,
ORCID: https://orcid.org/0000-0003-3513-2014
Correspondence address: Republic of Belarus, 230009, Grodno, 80 Gorky str., Grodno State Medical University, Chair of Microbiology, Virology and Immunology named after S.I. Gelberg. E-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра. –Tatiana V. Artsiukh.
