Modelling effects of phytobiotic administration on coherent responses to Salmonella infection in laying hens

Practice of layer poultry farming and commercial egg production relies on the optimal use and improvement of the welfare and genetically determined functional abilities of laying hens, their efficient intake of feed and its components, adaptation to housing conditions and resistance to infectious diseases including salmonellosis. Previous studies were focussed on relationships of chicken performance and resistance with the expression profiles of individual genes involved in metabolic processes and immune system, or with genetic markers that can be closely associated with these processes in chickens. In this study, mathematical models of coherent changes in laying hens were developed for the expression of eight genes involved in immunity and metabolism, on the one hand, and biochemical and immunological blood parameters, on the other hand, in response to Salmonella infection and administration of a phytobiotic Intebio. The proposed modelling approach can be a further basis for an in-depth research of the relationship between the gene expression, functional state and welfare of poultry, impact of pathogenic microorganisms and use of immunomodulatory drugs

[Mathematical assessment of BAC-based interspecies hybridization data in the process of genomic mapping in the white-throated sparrow as an avian behavioral model] Математическая оценка данных межвидовой БАК-гибридизации в процессе геномного картирования у белошейной зонотрихии как модели поведения птиц

The white-throated sparrow (Zonotrichia albicollis) known for its morphological, behavioral and chromosomal polymorphisms represents a quite new model system to study genomic mechanisms underlying variable behavioral repertoire interwoven with population biology, reproduction and adaptation in this species. It was previously shown that these polymorphisms could be due to chromosomal rearrangements (inversions) on sparrow chromosome 2 (ZAL2) that is characterized by a heterogeneity in two distinct morphs, tan (ZAL2/ZAL2) and white (ZAL2/ZAL2m). To construct a comparative genomic map of ZAL2 and other chromosomes, we used a sparrow genomic BAC library, CHORI-264. Following a cross-species overgo hybridization approach, we screened the library and developed a first-generation BAC-based comparative physical map using the chicken and zebra finch reference genomes. The map includes 640 BAC-gene assignments for 77 loci and serves for further refining the genomic regions and identifying candidate genes that are affected by rearrangements and contribute to the observed behavioral polymorphisms. Mathematical assessment of the BAC-based hybridization data was undertaken to show evolutionary relationships of avian genomes. В настоящей работе представлена математическая оценка межвидовой БАК-гибридизации в процессе геномного картирования у белошейной зонотрихии – вида воробьев, рассматриваемого в качестве удобной модели поведения птиц

Examination of the expression of immunity genes and bacterial profiles in the caecum of growing chickens infected with Salmonella Enteritidis and fed a phytobiotic

This study was performed to investigate the differential expression of eight immunity genes and the bacterial profiles in the caecum of growing chickens challenged with Salmonella enterica serovar Enteritidis (SE) at 1 and 23 days post inoculation (dpi) in response to SE infection at 19 days of age and administration of the phytobiotic Intebio. Following infection, the genes CASP6 and IRF7 were upregulated by greater than twofold. Chicks fed Intebio showed at 1 dpi upregulation of AvBD10, IL6, IL8L2, CASP6 and IRF7. At 23 dpi, expression of AvBD11, IL6, IL8L2, CASP6 and IRF7 lowered in the experiment subgroups as compared with the control. Examination of the caecal contents at 1 dpi demonstrated a significant decrease in the microbial biodiversity in the infected subgroup fed normal diet. Bacterial content of Lactobacillus and Bacillus declined, while that of Enterobacteriaceae rose. In the infected subgroup fed Intebio, a pronounced change in composition of the microflora was not observed. In the early infection stages, the phytobiotic seemed to promote response to infection. Subsequently, an earlier suppression of the inflammatory reaction took place in chickens fed Intebio. Thus, use of Intebio as a drug with phytobiotic activity in chickens, including those infected with Salmonella, proved to be promising

Effects of essential oils-based supplement and Salmonella infection on gene expression, blood parameters, cecal microbiome and egg production in laying hens

Simple Summary Salmonellosis is one of the most severe zoonotic diseases transmitted to humans through animal products (especially poultry meat and eggs). Essential oils (EOs)-based feed additives in poultry nutrition are a possible alternative replacement of antimicrobials to fight this infection. In the present study on laying hens, we tested a phytobiotic, Intebio®, and elucidated formation of immune response and changes in cecal microbiocenosis and biochemical/immunological variables in blood caused by Salmonella. Changes in differential gene expression were observed at both one and seven days post-inoculation in the hens’ intestines, revealing similarities with known mammalian/human tissue-specific expression. The results of this study suggest that the challenge of birds with Salmonella had a negative effect, while phytobiotic intake had a positive effect on the status of their gastrointestinal microbiome, their level of metabolism, and their performance. Abstract One of the main roles in poultry resistance to infections caused by Salmonella is attributed to host immunity and intestinal microbiota. We conducted an experiment that involved challenging Lohmann White laying hens with Salmonella Enteritidis (SE), feeding them a diet supplemented with an EOs-based phytobiotic Intebio®. At 1 and 7 days post-inoculation, the expression profiles of eight genes related to immunity, transport of nutrients in the intestine, and metabolism were examined. Cecal microbiome composition and blood biochemical/immunological indices were also explored and egg production traits recorded. As a result, the SE challenge of laying hens and Intebio® administration had either a suppressive or activating effect on the expression level of the studied genes (e.g., IL6 and BPIFB3), the latter echoing mammalian/human tissue-specific expression. There were also effects of the pathogen challenge and phytobiotic intake on the cecal microbiome profiles and blood biochemical/immunological parameters, including those reflecting the activity of the birds’ immune systems (e.g., serum bactericidal activity, β-lysine content, and immunoglobulin levels). Significant differences between control and experimental subgroups in egg performance traits (i.e., egg weight/number/mass) were also found. The phytobiotic administration suggested a positive effect on the welfare and productivity of poultry

[Determination of intestinal microbiocenoses of chickens of the Hisex breed by the T-RFLP method in ontogenesis] Определение микробиоценозов кишечника кур породы «Хайсекс» методом T-RFLP в онтогенезе

Организаторы: Сибирское отделение Российской академии наук; Институт химической биологии и фундаментальной медицины Лаптев Г.Ю., Ильина Л.А., Никонов И.Н., Кочиш И.И., Романов М.Н., Смоленский В.И., Панин А.Н., Йылдырым Е.А., Новикова Н.И., Филиппова В.А., Дубровин А.В. Цель работы: выявление структуры и таксономического состава микроорганизмов слепых отростков ЖКТ кур породы «Хайсекс» в онтогенезе с применением комплекса молекулярно-генетических методов. Исследования состава бактериального сообщества слепых отростков кишечника 40-, 155- и 315-суточных кур-несушек (по 3 из каждой группы) проводили молекулярно-генетическими методами (Т-RFLP и ПЦР в реальном времени). У исследуемой птицы в онтогенезе происходило развитие микробного сообщества ЖКТ, изменение содержания и появление новых микроорганизмов. Отмечено, что спектр выявляемых бактерий был выше у 40- и 155-суточной птицы (221 11 и 258 9 филотипов соответственно) по сравнению с 315-суточными курами-несушками (178 8 филотипов). Также у 315-суточной птицы выявлено наименьшее содержание неидентифицированных филотипов. В слепых отростках ЖКТ взрослой птицы выявлено изменение доминирующих таксономических групп микроорганизмов - более высокая доля бактерий кислот-утилизирующих бактерий класса Negativicutes, целлюлозолитиков класса Clostridia. Обратная тенденция наблюдалась в отношении бактерий классов Bifidobacteriales, Bacillales. Большее содержание лактобактерий порядка Lactobacillales наблюдалась у 315-суточных кур-несушек (33,15 1,05%) по сравнению с 40- (5,13 0,23%) и 155-суточной (24,58 0,86%) птицей. Разнообразие и количество бактерий в слепых отростков ЖКТ, которых традиционно относят к возбудителям различных заболеваний птицы, из родов Enterobacter, Pantoea, Listeria, Acinetobacter, Mycoplasma, семейств Campylobacteraceae, Pasteurellaceae, филума Fusobacteria увеличивается с возрастом птицы. Таким образом, в ходе молекулярно-генетических исследований был определен видовой состав микробиоценозов слепых отростков ЖКТ кур яичной породы «Хайсекс» в онтогенезе. Исследования выполнены при поддержке Министерства образования и науки Российской Федерации, Договор № 14.W03.31.0013 от 20.02.2017 г

Comparison of gut microbiota in hens of the crosses Hisex Brown and Lohmann Brown

Changes in the composition of the intestinal microbiota in the cecum of poultry could have a direct impact on the quality and safety of poultry products. This study presents the results of comparative molecular genetic analysis of the cecal microbiocoenoses in laying hens of two commercial crosses, Hisex Brown and Lohmann Brown, during ontogeny. According to the analysis of overall taxonomic representation, more than 70% phylotypes determined can be attributed to three: phyla, Firmicutes, Bacteroidetes, and Proteobacteria. Less represented were Actinobacteria, Tenericutes and Fusobacteria, and the presence of significant amounts of unidentified bacteria was also revealed. During ontogenesis, birds exhibited marked changes in the ratio of the number of phylotypes and taxonomic groups of the intestinal microbiota. Chickens of both crosses went through several stages in the development of microbial communities, including a stabilization period at the age of 20 to 40 weeks, as evidenced by the biodiversity assessment using ecological indexes. The stabilization period was characterised with a significant increase in representatives of class Clostridia involved in the metabolism of carbohydrates, and in bacteria with high antagonistic properties (genera Lactobacillus and Bacillus). There was also a significant reduction of number of opportunistic and pathogenic taxa, such as families Campylobacteraceae and Enterobacteriaceae, order Pseudomonadales, and phylum Tenericutes. Despite the similar conditions of housing and feeding, the Lohmann Brown hens had a maximum level of representatives of the normal flora observed by 40 weeks of age. This probably determines a smaller number of pathogens like Staphylococcus, family Campylobacteraceae, and phyla Tenericutes and Fusobacteria found by 40 to 60 weeks of age and greater stability of intestinal microbiocoenosis in the Lohmann Brown birds as compared with the Hisex Brown chickens

Determination of microbiocoenosis in the intestine of the Hisex Brown hens in ontogenesis using T-RFLP method

Microbiocoenosis in the gastrointestinal tract, especially in the ceca, play an important part in life processes of poultry. Identification of the structure and taxonomic composition of microorganisms in the cecum using molecular genetic methods serves as a crucial approach in understanding how a cecal microbiota interplays with the chicken organism during ontogenesis. For this purpose, we studied an intestinal bacterial community composition in the ceca of the Hisex Brown laying hens at age of 40, 155 and 315 days using T-RFLP and RTPCR. In the chickens studied, development of the cecal microbial communities, changes in their content, and appearance of new microorganisms occurred in the ontogeny. A broader spectrum of bacteria was found in 40- and 155-day-old birds (221 ± 11 and 258 ± 9 phylotypes, respectively) as compared with 315-day-old laying hens (178 ± 8 phylotypes). Also, 315-day-old birds showed the least content of unidentified phylotypes. In the ceca of adult hens, there was a change in the dominant microbial taxonomic groups including a higher proportion of acidutilising bacteria of the class Negativicutes and cellulolytic bacteria of the class Clostridia, with a lower content of the classes Bifidobacteriales and Bacillales. Lactobacteria (order Lactobacillales) showed a greater content in 315-day-old laying hens (33.15 ± 1.05%) as compared with 40- and 155-day-old birds (5.13 ± 0.23% and 24.58 ± 0.86%, respectively). The variety and number of bacteria in the ceca conventionally attributed to various pathogens of poultry diseases, including the genera Enterobacter, Pantoea, Listeria, Acinetobacter and Mycoplasma, families Campylobacteraceae and Pasteurellaceae, and phylum Fusobacteria, increased with the age of birds. Thus, during molecular genetic studies, the species composition and dynamics of the microbiocoenosis in the cecum of the Hisex Brown laying hens was determined as related to their ontogeny

[Development of modern biotechnologies for evaluating gene expression in connection with disease resistance and productivity in poultry] Разработка современных биотехнологий для оценки экспрессии генов в связи с устойчивостью к болезням и продуктивностью у домашней птицы

The aim of this study was to examine the expression of immunity genes as well as the cecal bacterial profiles of chicks when infected with a strain of Salmonella enterica serovar Enteritidis (SE). The effectiveness of the phytobiotic Intebio based on essential oils was also investigated. In general, Salmonella infection led to an increase in the expression level of most of the studied immunity genes. The addition of Intebio into the feed contributed to an increase in the expression of interleukin genes as compared with the control in birds both challenged with SE and non-challenged. In 3 weeks after infection, a decrease in the expression level of these genes or the same expression level as compared with the control was shown. A study of the cecal microbiological profiles of birds using T-RFLP in one day after infection revealed that infection of birds with SE led to a significant reduction in the biodiversity of microorganisms in the group without additives. At the same time, the content of bacteria of the genera Lactobacillus and Bacillus fell sharply, while the microorganisms of the Enterobacteriaceae family increased. In the group fed the phytobiotic and challenged with SE, a pronounced change in the composition of the microflora was not observed. The study of microbiocenosis in the cecum of birds in three weeks after infection revealed a general tendency to increase bacterial diversity with age, regardless of the treatments. The data obtained indicate that infection with SE leads to activation of the expression of certain immunity genes, and also causes dysbiotic disturbances in the composition of the intestinal microflora. In the early stages of infection, the introduction of the phytobiotic into the diet contributed to the enhancement of the immune response in birds due to SE. Subsequently, in birds fed Intebio, an earlier suppression of the inflammatory reaction occurred. Thus, the experiment of using the Intebio feed additive as an immunomodulator and a drug with phytobiotic activity, when artificially challenging birds with a dangerous pathogen, was successful. Одна из наиболее распространённых инфекционных болезней в птицеводстве в Российской Федерации — это сальмонеллез, вызываемый Salmonella enterica serovar Enteritidis (SE). Патологический процесс, вызванный сальмонеллой, инициирует у птиц экспрессию генов, в том числе интерлейкинов (цитокинов) и β-дефензинов (галлинацинов). Еще одна группа защитных факторов макроорганизма включает симбиотические микроорганизмы кишечника. Целью данного исследования было изучение экспрессии генов иммунитета, а также бактериальных профилей слепых отростков кишечника цыплят при инфицировании штаммом бактерии SE. Исследовали также эффективность действия фитобиотика Интебио на основе эфирных масел. Эксперимент проводили на цыплятах с 1-суточного до 43-суточного возраста. Как показали результаты исследования методом количественной ПЦР, в целом заражение сальмонеллой привело к увеличению уровня экспрессии большинства исследованных генов иммунитета в 2 и более раз. Введение в корм Интебио способствовало увеличению экспрессии генов интерлейкинов по сравнению с контролем как на фоне заражения сальмонеллой (экспрессия выросла в 4–6 раз), так и без него (экспрессия выросла в 3,5–5 раз). Через 3 недели после заражения показано снижение уровня экспрессии данных генов или тот же уровень экспрессии по сравнению с контролем. Изучение микробиологического профиля слепых отростков кишечника птиц методом T-RFLP через сутки после инфицирования выявило, что заражение птиц SE привело к значительному снижению биоразнообразия микроорганизмов в варианте без добавок. При этом содержание бактерий родов Lactobacillus и Bacillus резко падало, а микроорганизмов семейства Enterobacteriaceae увеличивалось. В группе с введением фитобиотика на фоне заражения выраженного изменения состава микрофлоры не наблюдалось. Исследование микробиоценоза в слепых отростках кишечника птиц через три недели после инфицирования выявило общую тенденцию увеличения бактериального разнообразия с возрастом вне зависимости от вариантов. Полученные данные свидетельствуют о том, что инфицирование SE приводит к активации экспрессии некоторых генов иммунитета, а также вызывает дисбиотические нарушения состава микрофлоры кишечника. На ранних стадиях заражения введение в рацион фитобиотика способствовало усилению иммунного ответа у птиц в ответ на заражение сальмонеллой. Впоследствии в вариантах с применением Интебио происходило более раннее подавление воспалительной реакции. Таким образом, опыт применения кормовой добавки Интебио в качестве иммуномодулятора и препарата с фитобиотической активностью, в том числе на фоне искусственного заражения птицы опасным патогеном, оказался весьма успешным

[Methodical recommendations on the application of the foundations of the technology of feeding egg-type chickens, providing a high percentage of realization of their genetic potential for productivity] Методические рекомендации по применению основ технологии кормления яичных кур, обеспечивающей высокий процент реализации их генетического потенциала продуктивности

The recommendations are intended for specialists and managers of poultry farms, farmers, researchers, teachers and students of agricultural universities as well as students of the advanced training system. Рекомендации предназначены для специалистов и руководителей птицеводческих хозяйств, фермеров, научных работников, преподавателей и студентов сельскохозяйственных вузов, слушателей системы повышения квалификации

[Methodical recommendations on the implementation of the developed system for the prevention of pathogen bacteria by correcting diets in laying hens and using antimicrobial additives] Методические рекомендации по внедрению разработанной системы профилактики бактерий-патогенов путем коррекции рационов питания у кур несушек и применения антимикробных добавок

The recommendations are intended for specialists and managers of poultry farms, farmers, researchers, teachers and students of agricultural universities as well as students of the advanced training system. Рекомендации предназначены для специалистов и руководителей птицеводческих хозяйств, фермеров, научных работников, преподавателей и студентов сельскохозяйственных вузов, слушателей системы повышения квалификации