mesЭкстремальная биомедицинаExtreme Medicine2713-27572713-2765Centre for Strategic Planning of the Federal Medical and Biological Agency10.47183/mes.2024-26-4-58-65mes-239Research ArticleМИКРОБИОЛОГИЯMICROBIOLOGYВлияние комбинаций антибиотиков, фагов и деполимеразы на биопленки лекарственно-устойчивого штамма Klebsiella pneumoniaeEffect of combinations of antibiotics, phages, and depolymerase on biofilms of the drug-resistant Klebsiella pneumoniae strainhttps://orcid.org/0009-0007-7117-1519КривуляА. О.KrivuliaA. O.

Кривуля Анастасия Олеговна

Москва

Moscow

kimnimka@gmail.comhttps://orcid.org/0000-0002-1685-1307ГородничевР. Б.GorodnichevR. B.

Городничев Роман Борисович

Москва

Moscow

gorodnichev.r.b@gmail.comhttps://orcid.org/0000-0002-1966-8452КорниенкоМ. А.KornienkoM. A.

Корниенко Мария Андреевна, канд. биол. наук

Москва

Moscow

kornienkomariya@gmail.comhttps://orcid.org/0000-0002-8187-0272АбдраймоваН. К.AbdraimovaN. K.

Абдраймова Нарина Казбековна

Москва

Moscow

abdraimovanarina@gmail.comhttps://orcid.org/0000-0001-7510-163XМалаховаМ. В.MalakhovaM. V.

Малахова Майя Владимировна, канд. биол. наук

Москва

Moscow

maja_m@mail.ruhttps://orcid.org/0000-0002-5016-2089ЗайчиковаМ. В.ZaychikovaM. V.

Зайчикова Марина Викторовна, канд. биол. наук

Москва

Moscow

marinaz15@yandex.ruhttps://orcid.org/0000-0002-4865-6004ШитиковЕ. А.ShitikovE. A.

Шитиков Егор Александрович, д-р биол. наук

Москва

Moscow

eshitikov@mail.ruФедеральный научно-клинический центр физико-химической медицины им. академика Ю.М. Лопухина ФМБА РоссииРоссияLopukhin Federal Research and Clinical Center of Physical-Chemical MedicineRussian Federation2024241120242645865Copyright © Кривуля А.О., Городничев Р.Б., Корниенко М.А., Абдраймова Н.К., Малахова М.В., Зайчикова М.В., Шитиков Е.А., 20242024Кривуля А.О., Городничев Р.Б., Корниенко М.А., Абдраймова Н.К., Малахова М.В., Зайчикова М.В., Шитиков Е.А.Krivulia A.O., Gorodnichev R.B., Kornienko M.A., Abdraimova N.K., Malakhova M.V., Zaychikova M.V., Shitikov E.A.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.extrememedicine.ru/jour/article/view/239Введение

Введение. Klebsiella pneumoniae представляет серьезную угрозу глобальному здравоохранению из-за высокой доли изолятов с множественной лекарственной устойчивостью. Более того, формирование бактерией биопленок значительно усложняет лечение инфекций.

Цель

Цель. Оценка эффективности индивидуального и комбинированного действия антибиотиков и бактериофагов или полисахарид-деполимеразы на биопленки клинически значимого штамма K. pneumoniae.

Материалы и методы

Материалы и методы. В работе использовали штамм K. pneumoniae с множественной лекарственной устойчивостью 9faiz, 4 антибиотика различных классов (гентамицин, левофлоксацин, меропенем и хлорамфеникол), 3 бактериофага различных родов (Dlv622, Seu621 и FRZ284) и 1 полисахарид-деполимеразу (Dep622). Эксперименты проводили на сформированных биопленках путем обработки 24-часовых пленок K. pneumoniae антимикробными агентами индивидуально или в комбинациях. Способность штамма образовывать биопленки оценивали окрашиванием кристаллическим фиолетовым. Сравнение между средними значениями оптической плотности проводилось с помощью t-теста и считалось значимым при p ≤ 0,05.

Результаты

Результаты. Индивидуальное применение антибиотиков в пиковых концентрациях (Cmax) или деполимеразы в концентрации 100 МДК (минимальная действующая концентрация) не приводило к значимому снижению биомассы биопленки, тогда как бактериофаги в титре 5×109 БОЕ/мл статистически значимо снижали ее биомассу на 27–31% (p ≤ 0,05). Большинство комбинаций фагов и антибиотиков не приводило к значимому повышению эффективности разрушения биопленок; лишь сочетание фага FRZ284 с гентамицином статистически значимо показало дополнительное снижение биомассы биопленки на 27% (p ≤ 0,05). Комбинации деполимеразы со всеми антибиотиками, кроме меропенема, приводили к значимому увеличению биомассы биопленки на 27–39% (p ≤ 0,05).

Выводы

Выводы. Результаты показывают необходимость индивидуального подбора антимикробных комбинаций для борьбы с биопленками K. pneumoniae из-за возможного влияния эффектов синергии и антагонизма на исход терапии.

Introduction

Introduction. Klebsiella pneumoniae poses a serious threat to global healthcare due to the high proportion of multidrug-resistant isolates. Moreover, the formation of biofilms by bacteria significantly complicates the treatment of infections.

Objective

Objective. To evaluate the effectiveness of the individual and combined action of antibiotics and bacteriophages or polysaccharide depolymerase on biofilms of a clinically significant strain K. pneumoniae.

Materials and methods

Materials and methods. The work used the K. pneumoniae strain with multidrug resistance (9faiz), 4 antibiotics of various classes (gentamicin, levofloxacin, meropenem and chloramphenicol), 3 bacteriophages of various genera (Dlv622, Seu621 and FRZ284), and 1 polysaccharide depolymerase (Dep622). Experiments were carried out on the formed biofilms by treating 24-hour K. pneumoniae films with antimicrobial agents individually or in combinations. The ability of the strain to form biofilms was evaluated by staining with crystalline violet. The comparison between the average optical density values was carried out using a t-test and was considered significant at p ≤ 0.05.

Results

Results. The individual use of antibiotics peak concentrations (Cmax) or depolymerase concentration of 100 MED (minimum effective dose — MED) did not lead to a significant decrease in biofilm biomass, whereas bacteriophages in a titer of 5×109  PFU/mL (plaque-forming unit per mL) statistically significantly reduced its biomass by 27–31% (p < 0.05) Most combinations of phages and antibiotics did not lead to a significant increase in the efficiency of biofilm destruction. Only the combination of phage FRZ284 with gentamicin statistically significantly showed an additional decrease in biofilm biomass by 27% (p < 0.05).

Conclusions

Conclusions. The results show the need for individual selection of antimicrobial combinations to combat K. pneumoniae biofilms due to the possible effect of synergy and antagonism effects on the outcome of therapy.

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The authors declare that there are no conflicts of interest present.