According to the main results obtained in the course of the performed studies for 2017–2020, a review of the methods and protocols for studying differential expression by full transcriptomic sequencing (RNASeq) of genes associated with traits of economic importance was carried out, and the corresponding technology was developed.
The resulting list of 34 selected genes involved in the formation of productivity traits and immunity of laying hens, and the developed method for determining the expression of genes associated with productive traits and resistance indicators in laying hens, using RT-PCR, made it possible to assess the effect of feed additives of various types on the microflora intestines, productivity and resistance in the egg type poultry.
Results of studying the development of gastrointestinal tract microbiocenosis in embryos of industrial crosses during embryonic development and at different periods of oviposition, as well as the results of T-RFLP analyzes of the contents of the intestines of chickens and the results of using the developed system for monitoring pathogenic bacteria at various stages of the technological process of growing laying hens allow for a targeted correction of the microbiocenosis of laying hens using feed factors to increase the productivity in the egg type poultry.
In the course of studying the effect of the phytobiotic Intebio® and the probiotic Liquipro® on the microflora of the digestive tract of poultry, the microbiocenoses of the digestive tract of chicks and adult chickens were tested. Protocols for studying the microflora of poultry intestinal contents by T-RFLP and NGS sequencing and protocols for testing the expression of genes associated with poultry productivity and immunity (two interleukin genes, three gallinacin genes, caspase gene, etc.) in response to salmonella infection were obtained.
The study of the relationship between the use of feed additives, gastrointestinal microflora, productivity and expression levels of productivity genes was carried out. The effects of the used feed additives (prebiotics and phytobiotics) on the productivity in hens of an egg layer cross and on the expression of genes associated with productivity and immunity have been described.
Mathematical models of the dependence of the productivity and expression of associated genes of poultry on the composition of the diet and intestinal microbiocenosis have been developed.
Based on the results of the development of a system for monitoring and preventing pathogenic bacteria by correcting the diet of laying hens with the help of antimicrobial additives (probiotics, phytobiotics), the appropriate guidelines were published with a description of the developed system.
In 2020-2021, the scientific research is aimed at conducting industrial experience to study the effect of a feed additive on gene expression (related to productivity and resistance) in laying hens and setting up a feeding experiment in industrial crosses of laying hens exposed to stress, when using an antimicrobial feed additive.
In the process of carrying out research within the framework of the mega-grant, scientific approaches and practical methods of managing the intestinal microbiota were developed. The developed technology was tested in the conditions of industrial egg production, and its adaptation to the conditions of modern egg poultry farms in Russia was conducted. At the same time, special attention is paid to the organic integration of the developed microbiota management technology into the already existing technological approaches and techniques of egg poultry farming. Particular attention is paid to the synergistic interaction of various feed additives and their effect on the intestinal microbiota in order to maximize the genetic potential of modern egg poultry crosses.
Implemented results of research:
During the project implementation period, the following were developed and applied in practice:
– "The Method for Determining the Expression of Genes Associated with Productive Traits in Laying Hens" using RT-PCR was introduced at the International Laboratory of Molecular Genetics and Genomics of Poultry using equipment: analyzer for real-time PCR analysis LightCycler 96 Instrument, mini-homogenizer Minilys, laboratory centrifuge Eppendorf 5424R, as well as in the educational process at the Department of Zoohygiene and Poultry Farming named after A. K. Danilova in the areas of training "Veterinary Medicine", "Zootechnics", and "Biotechnology" (Extract from Minutes No. 10 dated December 14, 2017 of the meeting of the Educational and Methodological Commission).
– "Methodological Recommendations for the Implementation of the Developed System for the Prevention of Pathogenic Bacteria by Correcting the Diet of Laying Hens and the Use of Antimicrobial Additives (Probiotics, Phytobiotics)" were introduced in the International Laboratory of Molecular Genetics and Genomics of Poultry and in the educational process at the Department of Zoohygiene and Poultry Farming named after A. K. Danilova in the area of training "Veterinary Medicine" (Extract from Minutes No. 11 dated December 24, 2018 of the meeting of the Educational and Methodological Commission; Implementation Act No. 2 dated December 27, 2018).
– "Methodical Recommendations on the Application of the Foundations of the Technology of Feeding Laying Hens, Providing a High Percentage of the Realization of the Genetic Potential of Productivity" were published and tested in the International Laboratory of Molecular Genetics and Genomics of Poultry and introduced into the educational process at the Department of Zoohygiene and Poultry Farming named after A.K. Danilova in the areas of training "Veterinary" and "Zootechnics" (Extract from Minutes No. 13 dated November 3, 2019 of the meeting of the Educational and Methodological Commission of the Faculty of Zootechnology and Agribusiness FSBEI HE MGAVMiB – MVA named after K.I. Skryabin; Implementation Act No. 2 dated December 5, 2019).
– "Practical Recommendations for the Use of Feed Additives to Improve the Productivity and Stress Resistance of Egg-type Poultry" were published and tested in the International Laboratory of Molecular Genetics and Poultry Genomics and introduced into the educational process at the Department of Zoohygiene and Poultry Farming named after A.K. Danilova in the areas of training "Veterinary" and "Zootechnics" (Extract from Minutes No. 13 dated November 3, 2019 of the meeting of the Educational and Methodological Commission of the Faculty of Zootechnology and Agribusiness FSBEI HE MGAVMiB – MVA named after K.I. Skryabin; Implementation Act No. 3 of December 31, 2019; Review of practical recommendations).
– "Methodological Recommendations on the Use of Modern Biotechnology Created to Assess the Expression of Genes Associated with the Productivity and Resistance of Poultry to Adverse Factors" were published and tested in the International Laboratory of Molecular Genetics and Genomics of Poultry and introduced into the educational process at the Department of Zoohygiene and Poultry Farming named after A.K. Danilova in the areas of training "Veterinary" and "Zootechnics" (Extract from Minutes No. 13 dated November 3, 2019 of the meeting of the Educational and Methodological Commission of the Faculty of Zootechnology and Agribusiness FSBEI HE MGAVMiB – MVA named after K.I. Skryabin; Implementation Act No. 2 dated December 10, 2019).
– "Methodological recommendations for the use of a feed additive that has been tested in industrial conditions on highly productive industrial laying hens" were tested in the International Laboratory of Molecular Genetics and Genomics of Poultry and introduced into the educational process at the Department of Zoohygiene and Poultry Farming named after A. K. Danilova in the areas of training "Veterinary" and "Zootechnics" (Implementation Act No. 1 dated December 25, 2020).
Education and career development:
Supervision of students and graduate students, which is carried out by the leading scientist and laboratory staff:
As part of the implementation of the project stages, members of the research team, I.I.Kochish and V.I. Smolensky supervised the following post-graduate students: I.G. Grebenchikov, R.R. Kolesnikova, E.U. Shikhbabaev, D.L. Bogdanova, E.D. Suprunov, E.E. Zimin, and G.R. Sharafetdinov.
During the implementation of the project in 2018, the thesis was defended for the degree of Candidate of Biological Sciences in the direction of scientific research on the topic "The Use of Mixed Feeds of Various Compositions and Their Effect on Broiler Intestinal Microbiota" by the member of the research team I.N. Nikonov (Order of the Higher Attestation Commission No. 426/nk dated May 7, 2019).
The final qualifying work of the mega-grant participant V.A. Dogadaev was defended on the topic "Evaluation of the Meat Qualities of Poultry" by qualification: Master in the Area of Training 04/36/02 – Zootechnics, under the guidance of the key member of the research team, Academician of the Russian Academy of Sciences I.I. Kochish (Master's Degree with Honors No. 107705 0051133 dated June 25, 2019; Minutes No. 1 dated June 7, 2019).
The final qualifying work of the mega-grant participant K.E. Bernikova was defended on the topic "Comparison of the Effect of Feeding the Phytobiotic "Intebio" and the Antibiotic "Stafak 110" on the Expression of the Productivity Gene Ovocalyxin-32 (OCX-32) in Laying Hens of the Lohmann White LSL Cross" by qualification: Bachelor in the Area of Training 04.04.02 – Animal Science, under the guidance of the member of the research team, Associate Professor O.V. Myasnikova.
The final qualification work was defended in the area of training 06.06.01 – Veterinary and Animal Science by the mega-grant participant, post-graduate student R.R. Kolesnikova on the topic "Influence of Feed Factors and Intestinal Microbiocenosis on the Expression of Genes That Control the Productivity and Viability of Chickens", and the qualification "Researcher; Research Teacher" was awarded; the scientific adviser being Doctor of Agricultural Sciences, Academician of the Russian Academy of Sciences, key executor of the mega-grant I.I. Kochish (Diploma No. 1077050011039 dated June 29, 2020).
The final qualification work was defended in the area of training 06.06.01 – Veterinary and Animal Science by the mega-grant participant, post-graduate E.U. Shikhbabaev on the topic "Influence of Feed Factors and Intestinal Microbiocenosis on the Expression of Genes That Control the Natural Resistance of Poultry", and the qualification "Researcher; Research teacher" was awarded; the scientific adviser being Doctor of Biological Sciences, Professor, executor of the mega-grant V.I. Smolenskiy (Diploma No. 1077050013137 dated June 29, 2020).
The final qualification work was defended for the qualification of a Bachelor in the Area of Training 03/36/02 – Zootechnics by the megagrant participant T.K. Kuvanov on the topic "Changes in the Microbial Profile in the Intestines of Chickens with the Use of Various Feed Additives" under the guidance of Associate Professor, executor of the mega-grant O.V. Myasnikova. (Bachelor's Degree with Honors No. 107705 0112844 dated July 2, 2020; Minutes No. 19 dated June 19, 2020).
Laboratory staff admitted to postgraduate and doctoral studies in the declared direction of scientific research in 2020:
- Sharafetdinov Grigory Rustamovich, date of birth on February 06, 1998, was admitted to postgraduate studies in the area of training 06.06.01 – Veterinary and Animal Science; Department of Animal Hygiene and Poultry Farming named after A.K. Danilova, FSBEI HE MGAVMiB – MVA named after K.I. Skryabin (Order No. 637i dated August 31, 2020), on the topic "The Use of Biologically Active Substances to Increase the Resistance and Productivity of Poultry"; scientific supervisor, Academician of the Russian Academy of Sciences, Professor, Doctor of Agricultural Sciences I.I. Kochish.
- Zimin Evgeniy Evgenievich, date of birth on March 20, 1997, was admitted to graduate school in the area of training 06.06.01 – Veterinary and Animal Science; Department of Animal Hygiene and Poultry Farming named after A.K. Danilova, FSBEI HE MGAVMiB – MVA named after K.I. Skryabin (Order No. 637i dated August 31, 2020), on the topic "The Effect of Dietary Supplements from Algae on Intestinal Microbiota and Expression of Genes for Productivity and Resistance of Chickens"; scientific adviser, Academician of the Russian Academy of Sciences, Professor, Doctor of Agricultural Sciences I.I. Kochish.
Teaching educational courses for undergraduate and graduate students of Russian organizations in the direction of scientific research:
During the implementation of the project, the leading scientist M.N. Romanov developed and delivered educational courses for undergraduate and graduate students:
- for 3rd and 4th year students of the direction of training "Zootechnics" (bachelor), in the form of lectures on the following topics:
- Genomics of poultry.
- Molecular markers and approaches for genotyping poultry populations.
- The structure of genes in different breeds of chickens.
- Mapping and sequencing of avian genomes.
- Results of recent studies using genomic technologies.
- for 5th year students of the specialty "Veterinary Medicine", in the form of lectures on the following topics:
- Effects of various types of feed additives on intestinal microflora and poultry productivity.
- Molecular markers and approaches for genotyping in poultry farming on the example of research at the University of Kent, UK.
- Gene structure in different chicken breeds and molecular markers and approaches for genotyping in poultry.
- for postgraduates of 1st year of study of the specialty 06.02.05 – Veterinary Sanitation, Ecology,Zzoohygiene and Veterinary and Sanitary Examination in the form of lectures:
- Gene expression in poultry; Study of genes responsible for resistance in birds using feeding factors.
- Determination of microbiocenoses in the intestines of chickens during ontogenesis using the T-RFLP method.
- Microsatellites and molecular genetic markers used in the study of avian genomes and genotyping of populations.
Preparation of textbooks, websites and other educational materials on the subject of the project:
For students of higher educational institutions in the specialty "Veterinary Medicine" and the areas of training "Animal Science" and "Veterinary and Sanitary Expertise", as well as graduate students, researchers, students of institutes and faculties of advanced training, and specialists of poultry enterprises, the following textbooks were prepared and published on the research topic:
- "Biology and Pathology of Poultry" / Kochish I.I., Smolensky V.I., Shcherbatov V.I. Moscow, Agricultural Technologies Publishing House, 2018;
- "Biology and Pathology of Poultry" / Kochish I.I., Smolensky V.I., Shcherbatov V.I. 2nd edition, added. Moscow, Agricultural Technologies Publishing House, 2019;
as well as monographs:
- "Forage and Veterinary Aspects of the State of the Intestinal Microbiota of Laying Hens" / Kochish I.I., Laptev G.Yu., Nikonov I.N., Smolenskiy V.I. Moscow, Agricultural Technologies Publishing House, 2017, 182 p. – Gold medal of the competition "Golden Autumn-2018";
- "Molecular Mechanisms of Maintaining the Health of the Intestines of Poultry: the Role of Microbiota" / Surai P.F., Kochish I.I. with coauthors. Moscow, Agricultural Technologies Publishing House, 2018, 344 p.;
- "Prevention of Free Radical Anomalies in Chickens in Early Ontogenesis" / Kochish I.I. Moscow, Agricultural Technologies Publishing House, 2019. ISBN 978-5-604-2808-5-0;
- "Non-infectious Diseases and Prevention of Feed Disturbances in Modern Poultry Farming" / Kochish I.I., Podobed L.I., Smolenskiy V.I., Nikonov I.N. Moscow, Agricultural Technologies Publishing House, 2021. ISBN 978-5-6045650-8-7.
Monographs on the research topic have been published in international publishing houses:
- Pigments from Microalgae Handbook / Surai P.F., Kochish I.I. (2020) Carotenoids in Aviculture / Jacob-Lopes E., Queiroz M., Zepka L. (Eds.). Springer, Cham. P. 515–540. doi: 10.1007/978-3-030-50971-2_20. ISBN 978-3-030-50970-5. https://link.springer.com/chapter/10.1007%2F978-3-030-50971-2_20;
- Chapter in monograph «Heat Shock Proteins in Veterinary Medicine and Sciences» / Surai P.F., Kochish I.I. (2019) / Part II Poultry Animals 5 Antioxidant Systems and Vitagenes in Poultry Biology: Heat Shock Proteins. Springer, Cham. Р. 123–177. https://doi.org/10.1007/978-3-319-73377-7.
- Chapter in monograph «Recent Advances in Trace Elements» / Chapter 20 «Dietary Food and Feed Supplements with Trace Elements» / Athanasios C. Pappas, Katarzyna Godlewska, and Peter F. Surai, 2018. John Wiley & Sons Ltd, 2019. P. 421–443.
Collaborations:
- Federal State Budgetary Scientific Institution Federal Research Center "All-Russian Poultry Research and Technological Institute" of the Russian Academy of Sciences, Sergiev Posad, Moscow Region;
- Molecular Genetic Laboratory of LLC "Biotrof +", St. Petersburg;
- Saint Petersburg State University of Veterinary Medicine.
The format of cooperation is joint research and joint scientific events.
