Food Processing: Techniques and Technology Food Processing: Techniques and Technology Техника и технология пищевых производств 2074-9414 2313-1748 72956 10.21603/2074-9414-2023-4-2472 ОРИГИНАЛЬНАЯ СТАТЬЯ ORIGINAL ARTICLE ОРИГИНАЛЬНАЯ СТАТЬЯ Antibiotic Resistance of Enterobacteriaceae in Microbiomes Associated with Poultry Farming Антибиотикорезистентность Enterobacteriaceae в микробиомах цыплят-бройлеров https://orcid.org/0000-0003-1918-3030 Кривоногова Анна Сергеевна Krivonogova Anna S. https://orcid.org/0000-0001-8349-3004 Донник Ирина Михайловна Donnik Irina M.

доктор биологических наук;

doctor of sciences in biology;

 https://orcid.org/0000-0001-8395-1247 Исаева Альбина Геннадьевна Isaeva Albina G. isaeva.05@bk.ru https://orcid.org/0009-0007-0810-8365 Логинов Егор Александрович Loginov Egor Aleksandrovich https://orcid.org/0000-0002-9892-6092 Петропавловский Максим Валерьевич Petropavlovskiy Maxim V. https://orcid.org/0000-0003-4347-6554 Беспамятных Елисей Николаевич Bespamyatnykh Elisey N. Уральский государственный аграрный университет Екатеринбург Россия Ural State Agrarian University Yekaterinburg Russian Federation Уральский государственный аграрный университет Екатеринбург Россия Ural State Agrarian University Yekaterinburg Russian Federation Уральский государственный аграрный университет Екатеринбург Россия Ural State Agrarian University Yekaterinburg Russian Federation Уральский государственный аграрный университет Екатеринбург Россия Ural State Agrarian University Yekaterinburg Russian Federation Уральский государственный аграрный университет Екатеринбург Россия Ural State Agrarian University Yekaterinburg Russian Federation Уральский государственный аграрный университет Екатеринбург Россия Ural State Agrarian University Yekaterinburg Russian Federation 30 12 2023 30 12 2023 53 4 710 717 17 05 2023 04 07 2023 https://fptt.ru/en/issues/22269/22222/

Постоянное применение антибиотиков в нетерапевтических целях привело к тому, что представители родов Escherichia, Enterobacter, Klebsiella и Proteus являются резервуарами для генетических детерминант резистентности и способствуют распространению резистентности и контаминации сырья и продукции. Сегодня учеными ведется поиск альтернативных антибиотикам средств сохранения здоровья и поддержания высокой продуктивности птицы – про- и фитобиотиков, органических кислот и др. Целью работы являлось изучение влияния применения антибиотика и фитобиотика на энтеропатогенные бактерии в микробиомах цыплят-бройлеров. Мы провели опыт in vitro c культивированием стандартного штамма Escherichia coli на питательных средах, содержавших антибиотики в подпороговых концентрациях в течение 37 суток. Оценили влияние низких доз антибиотиков на чувствительность изолятов. Исследовали микробиоценозы цыплят-бройлеров, получавших в качестве добавки к рациону антибиотик авиламицин А и фитобиотик на основе Brassica juncea, Linum usitatissimum и Nigella sativa L. Изучили видовой состав условно-патогенных Enterobacteriaceae, фенотипическую чувствительность к антибиотикам и генетические детерминанты резистентности. Провели анализ антимикробного потенциала фитобиотиков. Культивирование in vitro стандартных штаммов E. coli в присутствии антибиотиков в дозах ниже минимальных показало, что на протяжении 37 суток резистентность к антибиотикам не развивалась. В смывах и образцах подстилки всех групп цыплят-бройлеров преобладали условно-патогенные грамотрицательные бактерии порядка Enterobacterales. На долю E. coli, Klebsiella pneumoniae и Proteus mirabilis приходилось более 30 % от всех выделенных штаммов. Введение в рацион бройлеров авиламицина А и фитобиотика повлияло на кокковую микрофлору и не сказалось на динамике родовидового состава Enterobacteriaceae. Изоляты K. pneumoniae имели высокий уровень фенотипической устойчивости к ципрофлоксацину и содержали гены blaDHA. Микробиота подстилки имеет значение в сохранении и циркуляции агентов резистентности при содержании бройлеров. Наименьший уровень устойчивости к ципрофлоксацину мы выявили в группе цыплят, которые получали только фитобиотик, наибольший – в группе с авиламицином А. На коротких временных отрезках, сопоставимых со сроком выращивания бройлеров до убоя, изолированные in vitro бактерии, которые не имеют в геноме активных детерминант резистентности, сохраняют чувствительность к антибиотикам даже при постоянном контакте с ними. Применение фитобиотиков перспективно в технологии выращивания бройлеров в качестве дополнения к существующим рационам для снижения (отказа) количества применяемых кормовых антибиотиков.

Antibiotics have long been overused for non-therapeutic purposes. As a result, Escherichia, Enterobacter, Klebsiella, and Proteus in avian microbiomes have become reservoirs for genetic determinants of resistance, thus spreading resistance to antibiotics and contaminating raw materials and finished products. The food industry is looking for alternative means to preserve health and maintain high productivity of commercial poultry, e.g., probiotics, phytobiotics, organic acids, etc. The research featured the effect of antibiotics and phytobiotics on enteropathogenic bacteria in the microbiomes of broiler chicken. Escherichia coli bacteria were cultivated in vitro in subthreshold concentrations on nutrient media with antibiotics for 37 days to study the effect of low doses of antibiotics on the sensitivity of isolates. The study involved microbiocenoses of broiler chicken that received avilamycin A and a phytobiotic based on Brassica juncea, Linum usitatissimum, and Nigella sativa L. A set of experiments covered the species composition of opportunistic Enterobacteriaceae, the phenotypic sensitivity to antibiotics, and the genetic determinants of resistance, as well as the antimicrobial potential of phytobiotics. E. coli developed no resistance for 37 days when the antibiotic dose remained below minimal inhibitory. Opportunistic gram-negative Enterobacterales predominated in all litter samples. E. coli, Klebsiella pneumoniae, and Proteus mirabilis accounted for more than 30% of all isolated strains. Avilamycin A and the phytobiotic affected the coccal microflora but had no effect on the genus-species composition of Enterobacteriaceae. Litter samples from both experimental and control groups demonstrated K. pneumoniae with severe phenotypic resistance to ciprofloxacin, as well as blaDHA genes. In broiler farming, maintenance and circulation of resistance agents depends on litter microbiota. In this research, the chicken that received the phytobiotic showed the lowest level of resistance to ciprofloxacin while the groups that received avilamycin A had the highest resistance results. During a broiler’s life span, bacteria with no active resistance determinants in their genome remained sensitive to antibiotics, even though the contact with the latter was constant. Phytobiotics showed good prospects for broiler farming as food additive that could reduce and eventually eliminate the intake of antibiotics.

 Антибиотикорезистентность энтеробактерии антибиотик фитобиотик микробиом бройлеры Antibiotic resistance enterobacteria antibiotic phytobiotic microbiome broilers Исследование выполнено при финансовой поддержке Российского научного фонда (РНФ) , грант № 18-16-00040. The research was supported by the Russian Science Foundation ( RSF), Project No. 18-16-00040.

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