veterinaryВетеринария сегодняVeterinary Science Today2304-196X2658-6959"Veinard"10.29326/2304-196X-2025-14-3-302-309veterinary-946Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИ | ВЕТЕРИНАРНАЯ МИКРОБИОЛОГИЯORIGINAL ARTICLES | VETERINARY MICROBIOLOGYИдентификация и некоторые биологические свойства изолятов бактерий рода Bacillus, выделенных из толстого отдела кишечника птицыIdentification and some biological characteristics of Bacillus strains isolated from poultry large intestinehttps://orcid.org/0000-0002-4695-1077МаликН. И.MalikN. I.

Малик Нина Ивановна - д-р биол. наук, профессор, главный научный сотрудник отдела научного планирования и НИР ФГБУ «ВГНКИ».

Звенигородское шоссе, 5, Москва, 123022

Nina I. Malik - Dr. Sci. (Biology), Professor, Chief Researcher, Department of Scientific Planning and Research, The Russian State Centеr for Quality and Standardization of Veterinary Drugs and Feed.

5 Zvenigorodskoe shosse, Moscow 123022

nimalik@vgnki.ruhttps://orcid.org/0009-0000-8358-0618ЧупахинаН. А.ChupahinaN. A.

Чупахина Наталия Александровна - канд. биол. наук, ведущий научный сотрудник отдела бактериологии ФГБУ «ВГНКИ».

Звенигородское шоссе, 5, Москва, 123022

Nataliya A. Chupahina - Cand. Sci. (Biology), Leading Researcher, Department of Bacteriology, The Russian State Centеr for Quality and Standardization of Veterinary Drugs and Feed.

5 Zvenigorodskoe shosse, Moscow 123022

n.chupahina@vgnki.ruhttps://orcid.org/0009-0008-3388-2990РусановИ. А.RusanovI. A.

Русанов Иван Анатольевич - старший научный сотрудник научно-технологической лаборатории ФГБУ «ВГНКИ».

Звенигородское шоссе, 5, Москва, 123022

Ivan A. Rusanov - Senior Researcher, Scientific and Technological Laboratory, The Russian State Centеr for Quality and Standardization of Veterinary Drugs and Feed.

5 Zvenigorodskoe shosse, Moscow 123022

rusanov@vgnki.ruhttps://orcid.org/0000-0001-7836-3010МаликЕ. В.MalikE. V.

Малик Евгений Васильевич - канд. вет. наук, ведущий научный сотрудник отдела научного планирования и НИР ФГБУ «ВГНКИ».

Звенигородское шоссе, 5, Москва, 123022

Evgeniy V. Malik - Cand. Sci. (Veterinary Medicine), Leading Researcher, Department of Scientific Planning and Research, The Russian State Centеr for Quality and Standardization of Veterinary Drugs and Feed.

5 Zvenigorodskoe shosse, Moscow 123022

evmalik@vgnki.ruhttps://orcid.org/0000-0003-3149-8625МаленковаЛ. А.MalenkovaL. A.

Маленкова Лия Андреевна - старший научный сотрудник отдела бактериологии ФГБУ «ВГНКИ».

Звенигородское шоссе, 5, Москва, 123022

Liya A. Malenkova - Senior Researcher, Department of Bacteriology, The Russian State Centеr for Quality and Standardization of Veterinary Drugs and Feed.

5 Zvenigorodskoe shosse, Moscow 123022

l.malenkova@vgnki.ruhttps://orcid.org/0009-0002-3519-7838СамохваловаН. С.SamokhvalovaN. S.

Самохвалова Надежда Сергеевна - научный сотрудник отдела бактериологии ФГБУ «ВГНКИ».

Звенигородское шоссе, 5, Москва, 123022

Nadezhda S. Samokhvalova - Researcher, Department of Bacteriology, The Russian State Centеr for Quality and Standardization of Veterinary Drugs and Feed.

5 Zvenigorodskoe shosse, Moscow 123022

n.samokhvalova@vgnki.ruhttps://orcid.org/0009-0003-1870-4292СурогинМ. В.SuroginM. V.

Сурогин Михаил Витальевич - канд. вет. наук, заведующий отделом бактериологии, ФГБУ «ВГНКИ».

Звенигородское шоссе, 5, Москва, 123022

Mikhail V. Surogin - Cand. Sci. (Veterinary Medicine), Head of Department of Bacteriology, The Russian State Centеr for Quality and Standardization of Veterinary Drugs and Feed.

5 Zvenigorodskoe shosse, Moscow 123022

surogin@vgnki.ruФГБУ «Всероссийский государственный Центр качества и стандартизации лекарственных средств для животных и кормов» (ФГБУ «ВГНКИ»)РоссияThe Russian State Centеr for Quality and Standardization of Veterinary Drugs and FeedRussian Federation202517092025143302309Copyright © Малик Н.И., Чупахина Н.А., Русанов И.А., Малик Е.В., Маленкова Л.А., Самохвалова Н.С., Сурогин М.В., 20252025Малик Н.И., Чупахина Н.А., Русанов И.А., Малик Е.В., Маленкова Л.А., Самохвалова Н.С., Сурогин М.В.Malik N.I., Chupahina N.A., Rusanov I.A., Malik E.V., Malenkova L.A., Samokhvalova N.S., Surogin M.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://veterinary.arriah.ru/jour/article/view/946Введение

Введение. Проведенная за последнее десятилетие перепись филогенетического разнообразия бактерий, колонизирующих кишечный тракт клинически здоровой птицы, показывает, что до 60% родóв, присутствующих в микробиоме кишечника, содержат спорообразующие бактерии, и эти роды составляют 30% от общей кишечной микробиоты. Были зарегистрированы преимущества, связанные с использованием пробиотиков, содержащих спорообразующие бактерии рода Bacillus. Анализ широты распространения гемолитической и потенциальной биопленкообразующей активности, а также антибиотикорезистентности у кишечной популяции споробиоты птицы необходим для понимания истинной роли аэробных спорообразователей рода Bacillus в экологии кишечного микробиома птицы.

Цель работы

Цель работы. Идентификация и исследование биологических характеристик (гемолитическая активность, потенциальная способность к биопленкообразованию и антибиотикорезистентность) изолятов бактерий рода Bacillus, выделенных из толстого отдела кишечника птицы.

Материалы и методы

Материалы и методы. Выделение спорообразующих бактерий из содержимого слепых отростков толстого кишечника птицы проводили путем прогревания образцов. Фенотипическую идентификацию изолятов осуществляли с использованием биохимических тест-панелей API 50CHB (bioMérieux, Франция). Гемолитические свойства определяли на колумбийском агаре (HiMedia Laboratories Pvt Ltd., Индия) с добавлением 5% стерильной дефибринированной крови барана; каталазную активность – в тесте с 10%-й перекисью водорода по ОФС.1.7.2.0012.15; чувствительность к антибиотикам – диско-диффузионным методомсостандартными дисками, импрегнированными антибиотиками вконцентрациях от5до 30 μg/disk. Скрининг спорообразующих бактерий – продуцентов биопленки проводили качественным методом на сердечно-мозговом агаре (HiMedia Laboratories Pvt Ltd., Индия) с добавлением индикатора конго красного и 5% сахарозы.

Результаты

Результаты. Установлено, что кишечная популяция аэробной споробиоты слепых отростков толстого кишечника птицы представлена видами B. licheniformis, B. subtilis/amyloliquefaciens, B. mycoides, B. megaterium и B. сereus. Все изученные изоляты были каталазоположительными, не обладали α-гемолитической активностью. У части изолятов отмечена β-гемолитическая активность. Подавляющее большинство изолятов относились к биопленкообразующим фенотипам и проявляли чувствительность к тестируемым антибиотикам.

Заключение

Заключение. Вегетативные формы спорообразующих бактерий рода Bacillus потенциально могут сохраняться в сложной экосистеме кишечника или временно ассоциироваться с ней. Гемолитически активные кишечные изоляты спорообразующих бактерий не могут считаться безопасными до выяснения действия этого фактора вирулентности на организм животных. Результаты исследований могут быть использованы при отборе кандидатных штаммов бактерий рода Bacillus, выбранных в качестве пробиотиков.

Introduction

Introduction. Censuses of phylogenetic diversity of bacteria colonizing the intestinal tract of clinically healthy poultry conducted over the past decade indicate that up to 60% of genera present in the gut microbiome contain spore-forming bacteria, accounting for 30% of total gut microbiota. Benefits associated with using probiotics containing Bacillus spore-forming bacteria have been documented. Analysis of the prevalence of hemolytic and potential biofilm-forming activity, as well as antibiotic resistance in poultry gut spore-forming microbiota is essential for understanding the true role of aerobic spore-formers of the Bacillus genus in avian gut microbiome ecology.

Objective

Objective. Identification and investigation of biological characteristics (hemolytic activity, potential biofilm-forming capacity and antibiotic resistance) of Bacillus bacterial isolates obtained from the large intestine of poultry.

Materials and methods

Materials and methods. Spore-forming bacteria were isolated from cecal content through sample heat treatment. Phenotypic identification was performed using API 50CHB biochemical test panels (bioMérieux, France). Hemolytic properties were assessed using Columbia agar (HiMedia Laboratories Pvt Ltd., India) supplemented with 5% sterile defibrinated sheep blood; catalase activity was assessed using 10% hydrogen peroxide according to General Pharmacopoeia Monograph GPM.1.7.2.0012.15; antibiotic sensitivity was assessed with disk diffusion test involving standard antibiotic-impregnated disks (5–30 μg/disk). Biofilm-producing, spore-forming bacteria were tested qualitatively using brain-heart infusion agar (BHI; HiMedia Laboratories Pvt Ltd., India) supplemented with Congo red and 5% sucrose.

Results

Results. It was established that the cecal aerobic spore-forming microbiota in poultry comprised B. licheniformis, B. subtilis/amyloliquefaciens, B. mycoides, B. megaterium and B. cereus. All tested isolates were catalase-positive and lacked α-hemolytic activity. Some isolates demonstrated β-hemolytic activity. The overwhelming majority exhibited biofilm-forming phenotypes and showed susceptibility to tested antibiotics.

Conclusion

Conclusion. Vegetative forms of Bacillus spore-forming bacteria may potentially persist in or temporarily associate with the complex gut ecosystem. Hemolytically active intestinal isolates cannot be considered safe until the effects of this virulence factor on animals are clarified. These findings provide a basis for selecting candidate Bacillus strains for probiotic development.

споробиотагемолизбиопленкаантибиотикорезистентностьsporobiotahemolysisbiofilmantibiotic resistanceИсследование финансировалось Федеральной службой по ветеринарному и фитосанитарному надзору, научно-исследовательский проект АААА-А20-120012790050-7 по теме «Диагностика состояния нормальной микробиоты желудочно-кишечного тракта сельскохозяйственной птицы под воздействием антимикробных и пробиотических препаратов для разработки и осуществления мер по ее сохранению или восстановлению».The study was funded by the Federal Service for Veterinary and Phytosanitary Surveillance (Rosselkhoznadzor) within Research Project No. AAAA-A20-120012790050-7“Diagnostic monitoring of poultry gut microbiota responses to antimicrobial and probiotic agents for developing and implementing measures to ensure microbiome preservation and restoration”.ReferencesVon Martels J. Z., Sadabad M. S., Bourgonje A. R., Blokzijl T., Dijkstra G., Faber K. N., Harmsen H. J. M. 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The authors declare that there are no conflicts of interest present.