vuzbiochemiИзвестия вузов. Прикладная химия и биотехнологияProceedings of Universities. Applied Chemistry and Biotechnology2227-29252500-1558ИРНИТУ10.21285/achb.912PCUTZFvuzbiochemi-1234Research ArticleФИЗИКО-ХИМИЧЕСКАЯ БИОЛОГИЯPHYSICOCHEMICAL BIOLOGYИзучение способности бактерий активного ила к образованию биопленок in vitroStudy on the ability of activated sludge bacteria to form biofilms in vitrohttps://orcid.org/0009-0004-3730-4080ХасановаА. А.KhasanovaA. A.

Айгуль Айратовна Хасанова, аспирант

420015; ул. Карла Маркса, 68; Казань

Aigul A. Khasanova, Postgraduate Student

420015; 68, Karl Marx St.; Kazan

hasanovaaigyl@mail.ruhttps://orcid.org/0000-0002-4480-9907СироткинА. С.SirotkinA. S.

Александр Семенович Сироткин, д. т. н., профессор, заведующий кафедрой

420015; ул. Карла Маркса, 68; Казань

Aleksandr S. Sirotkin, Dr. Sci. (Engineering), Professor, Head of the Department

420015; 68, Karl Marx St.; Kazan

asirotkin66@gmail.comhttps://orcid.org/0000-0002-2631-4724ПерушкинаЕ. В.PerushkinaE. V.

Елена Вячеславовна Перушкина, к. т. н., доцент

420015; ул. Карла Маркса, 68; Казань

Elena V. Perushkina, Cand. Sci. (Engineering), Associate Professor

420015; 68, Karl Marx St.; Kazan

perushkina_elena@mail.ruКазанский национальный исследовательский технологический университетРоссияKazan National Research Technological UniversityRussian Federation202407072024142207214Copyright © Хасанова А.А., Сироткин А.С., Перушкина Е.В., 20242024Хасанова А.А., Сироткин А.С., Перушкина Е.В.Khasanova A.A., Sirotkin A.S., Perushkina E.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vuzbiochemi.elpub.ru/jour/article/view/1234

   Целью работы являлась сравнительная характеристика биопленкообразования в условиях in vitro бактериальных культур, выделенных из активного ила, а также музейных культур, способных к биодеструкции ксенобиотиков: Alcaligenes faecalis 2, Acinetobacter guillouiae 11h, Rhodococcus erythropolis ИЛБИО, Achromobacter pulmonis ПНОС.

   Согласно результатам анализа нуклеотидной последовательности гена 16S рРНК идентифицированы штаммы, выделенные из активного ила: Paenibacillus odorifer, Bacillus subtilis, Micrococcus yunnanensis и Bacillus proteolyticus. Исследовано формирование биопленок микроорганизмами на среде LB и синтетической питательной среде (источник углерода – ацетат натрия). При росте клеток на среде LB биомасса биопленки увеличивается у бактерий Paenibacillus odorifer, Bacillus subtilis, Alcaligenes faecalis 2, Achromobacter pulmonis ПНОС. Продолжительность стадии культивирования 72 и 144 часа и дополнительное дозирование субстратов оказали влияние на процесс биопленкообразования: к 144 часам культивирования показатели биомассы составили 0,6–1,3 опт. ед. Отмечено, что для клеток Bacillus subtilis и Paenibacillus odorifer наблюдается увеличение биомассы биопленок в среднем на 63–77 % по сравнению с 72-часовым процессом. На заключительном этапе культивирования (144 часа) содержание экзополисахаридов в матриксе для микроорганизмов Bacillus subtilis и Paenibacillus odorifer составило более 0,02 опт. ед. Метаболическая активность бактерий активного ила, формирующих биопленку, достигла 628–3609 Фл./ОП540. Таким образом, показано, что в процессе роста микроорганизмы активного ила в составе биопленки сохраняют жизнеспособность и метаболическую активность в условиях in vitro.

   The study aims to comparatively characterize in vitro biofilm formation in bacterial cultures isolated from activated sludge, as well as archival cultures capable of xenobiotics biodegradation: Alcaligenes faecalis 2, Acinetobacter guillouiae 11h, Rhodococcus erythropolis ILBIO, and Achromobacter pulmonis PNOS. An analysis of the 16S rRNA nucleotide sequence identified strains isolated from activated sludge: Paenibacillus odorifer, Bacillus subtilis, Micrococcus yunnanensis, and Bacillus proteolyticus.

   The formation of biofilms by microorganisms was studied on LB medium and synthetic culture medium (with sodium acetate as a carbon source). With cell growth on LB medium, an increase in biofilm biomass was observed in Paenibacillus odorifer, Bacillus subtilis, Alcaligenes faecalis 2, and Achromobacter pulmonis PNOS. The cultivation stage duration (72 and 144 h), as well as the additional dosing of substrates, had an effect on the biofilm formation process: by 144 h of cultivation, the biomass values amounted to 0.6–1.3 optical units. An average 63–77% increase in biofilm biomass was noted for Bacillus subtilis and Paenibacillus odorifer cells as compared to the 72-hour process. At the final stage of cultivation (144 h), the values of exopolysaccharides in the matrix amounted to over 0.02 optical units for Bacillus subtilis and Paenibacillus odorifer. The metabolic activity of activated sludge bacteria forming the biofilm reached 628–3609 Fl./OD540. Thus, activated sludge microorganisms forming the biofilm were shown to retain viability and metabolic activity during growth under in vitro conditions.

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