AP09258412 Development of a domestic probiotic drug that increases the resistance and productivity of valuable fish species in aquaculture
Publication date: 09.09.2021 13:43
The AP09258412 project "Development of a domestic probiotic drug that increases the resistance and productivity of valuable fish species in aquaculture" is carried out under the contract for the implementation of scientific, scientific and technical projects for grant financing dated 07.04.2021 No.148/36-21-23 concluded between LLP "NPC of Microbiology and Virology" and the State Institution "Committee of Science of the Ministry of Education and Science of the Republic of Kazakhstan".
Applied research. Scope of application: fisheries, aquaculture, industrial biotechnology.

Relevance. The important tasks facing the Ministry of Ecology, Geology and Natural Resources of the Republic of Kazakhstan are the multiplication of fish resources by implementing measures to create conditions for natural reproduction, the implementation of artificial reproduction by the factory method and increasing the export of fish products. The increase in the level of organic pollution, the number of conditionally pathogenic microorganisms in the aquatic environment of fish farms and global contamination of feed raw materials and feed with mycotoxigenic fungi are serious problems of industrial cultivation of valuable fish species. At the same time, there is a weakening of the general condition of fish, the occurrence of various diseases complicated by drug resistance, the accumulation of antibiotics, chemotherapy drugs, toxins in tissues and immunosuppression. One of the biological methods of maintaining and restoring the normal physiological state of fish, increasing their resistance and productivity is the use of probiotic drugs. In many countries of the world, including Kazakhstan, ongoing research is underway to find new probiotic strains and study their antagonistic properties, develop new probiotic drugs for farm animals, birds and fish based on them. The disadvantages of domestic developments are the low fungicidal activity of probiotic drugs. In addition, despite the demand, none of the domestic developments were brought to the market of Kazakhstan in the format of an established industrial production of domestic probiotics for fish, not to mention exports. Kazakhstan's probiotic market is represented by imported probiotic drugs, often of questionable quality and not adapted to our raw materials.

The scientific novelty of the proposed research is associated with the search for new domestic strains of lactic acid and propionic acid bacteria with highly enzymatic, antagonistic and fungicidal activity; a comprehensive study of the molecular and biochemical nature of their antagonism, followed by the development on their basis of a highly active stable probiotic preparation for feeding fish in aquaculture, capable of combating pathogenic and mycotoxigenic microflora of feeds and reservoirs, increasing resistance and the productivity of fish. Compounds synthesized by lactic acid and propionic acid bacteria with fungicidal action are of particular scientific and practical value.

The aim of the project is to increase the resistance and productivity of valuable fish species in aquaculture, through the development of a new domestic probiotic drug for therapeutic and prophylactic feeds based on highly active cultures of lactic acid and propionic acid bacteria with increased antibacterial, fungicidal, proteolytic properties.

Expected results:
- for 2021: New strains of lactic acid and propionic acid bacteria will be isolated from the raw components of fish feeds; screening of new and collectible strains of lactic acid and propionic acid bacteria with high activity will be carried out; associations of lactic acid and propionic acid bacteria will be compiled and an antagonistically active consortium selected. An article (review) will be published in the journal recommended by the CCSON RK.
- for 2022: The identification of the selected crops will be carried out, the hazard class will be determined and an application for an invention will be filed. The spectrum of volatile compounds will be investigated and the primary chemical nature of antagonistically active metabolites of selected microorganisms/consortium will be determined. The nutrient medium will be selected, the cultivation modes of the consortium, the methods of stabilization of the liquid preparation and the drying modes of the preparation will be worked out. An application for the invention of the Republic of Kazakhstan will be submitted; participation in a scientific conference will be taken
- for 2023: The technology for obtaining the drug in laboratory conditions will be developed; the physico-chemical, biochemical and microbiological properties of the resulting drug will be investigated and the shelf life of the drug will be determined. Pilot-industrial approbation of the technology for obtaining the drug will be carried out and a pilot batch will be developed; the physicochemical, biochemical and microbiological properties of the resulting drug will be investigated and its shelf life will be determined. The effectiveness of the use of the developed probiotic preparation will be determined when feeding with full-fledged compound feeds and/or as part of production compound feeds for valuable fish species in aquaculture conditions. NTDs and guidelines for the use of probiotic drugs will be developed. An application for the invention of the Republic of Kazakhstan will be submitted; participation in a scientific conference will be taken.
According to the research results, for the entire period of the project implementation, at least 2 articles and (or) reviews will be published in peer-reviewed scientific publications indexed in the SCIE database of Web of Science and (or) having a CiteScore percentile in the Scopus database of at least 35; or at least 1 article or review in a peer-reviewed scientific publication indexed in the SCIE of the Web of Science database and (or) having a CiteScore percentile in the Scopus database of at least 35, and 1 patent included in the Derwent Innovations Index database; or at least 1 article or review in a peer-reviewed scientific publication indexed in the SCIE of the Web of Science database and (or) having a CiteScore percentile in the Scopus database of at least 35, and an act of introduction of scientific and technical products prepared as a result of the project implementation or a license agreement for it; as well as at least 1 article or review in a peer-reviewed foreign or domestic publication recommended by COXON; or at least 1 article or review in a peer-reviewed scientific publication included in the 1st or 2nd quartile in the Web of Science database and (or) having a CiteScore percentile in the Scopus database of at least 65.
The developed domestic probiotic preparation for feeding valuable fish species in aquaculture conditions based on safe highly active cultures of microorganisms will ensure the production of healthy, biologically valuable, high-quality and safe fish products, which will directly affect the health and working capacity of people. The results of the project are relevant, economically viable, meet modern production requirements and priority tasks of national socio-economic development – import substitution, food security, renewal of fish resources, environmental friendliness.

Achieved results (for the first half of 2021):
- New 15 isolates of lactic acid bacteria and 2 isolates of propionic acid bacteria, previously assigned to 5 genera: R. Lactococcus, R. Lactobacillus, R. Pediococcus, R. Leuconoctoc and R. Propionibacterium, were isolated from raw components of plant and animal origin of various regions of Kazakhstan included in the formulations of fish feeds.
- As a result of screening of the physiological and biochemical properties of 17 newly isolated bacterial strains, as well as 2 strains of lactic acid bacteria and 1 strain of propionic acid bacteria, 6 strains with increased enzymatic, antibacterial and fungicidal properties with high resistance to adverse environmental factors and promising for the creation of probiotic were selected from the collection of the Laboratory of Food Microbiology of the NPC of Microbiology and Virology the drug.
- Research has begun on the creation of a new consortium of lactic acid and propionic acid bacteria based on selected strains with increased proteolytic, antibacterial and fungicidal properties for its use in a feed probiotic preparation for fish.

Publications:
- 2 reports have been published in collections of materials of international conferences:
1. Chizhaeva A.V., Amangeldi A.A., Alybayeva A.Zh., Sadanov A.K. Prospects of using lactic acid bacteria to create a probiotic preparation for valuable fish species in aquaculture// Actual directions of scientific research: technologies, quality and safety: collection of materials of the II National (All-Russian) Conference of Scientists / under the general editorship of A. Y. Prosekov; Kemerovo State University. – Kemerovo, 2021. – from 287-289.
The article presents the results of a study of the acid-forming and antagonistic activity of new strains of lactic acid bacteria isolated from raw components of plant and animal origin included in fish feed formulations. The prospects of their use for the creation of a probiotic preparation for valuable fish species in aquaculture are substantiated.
2. A.V. Chizhaeva, E.A. Oleinikova, A.A. Amangeldi, A.Zh. Alybaeva, V.I. Sidorova. Production of probiotics for aquaculture in Kazakhstan// Materials of the II International Scientific and Practical Conference "Modern problems of biotechnology: from laboratory research to production" dedicated to the 80th anniversary of a major microbiologist, academician of the Kazakhstan National Academy of Natural Sciences, Doctor of Biological Sciences, Professor Zhubanova Azhar Akhmetovna, June 4-5, 2021 - Almaty: Kazakh University. - pp. 38-41.
The article substantiates the use of probiotic drugs as one of the effective biological methods of maintaining and restoring the normal physiological state of fish in aquaculture, increasing their productivity. An overview of domestic scientific developments and the market of probiotic preparations for fish in Kazakhstan, with their disadvantages and prospects for use in aquaculture, is given.
- 1 article has been prepared for publication in a domestic publication recommended by COXON – Bulletin of KazNU, Biological series.


Members of the research group:
Project Manager - A.V. Chizhaeva, Candidate of Biological Sciences, specialty 03.00.07 Microbiology, Professor of RAE; technical expert of the KazInSt Technical Committee; leading researcher of the Scientific and Production Center of Microbiology and Virology LLP, 26 years of scientific experience, author of more than 120 scientific papers, including 13 patents, h-index-1.
Author Identifier:
https://orcid.org/0000-0001-6081-0273; Scopus ID 57219126169, ResearcherID Web of Science (Chizhayeva Anna); Scopus ID 57219963267, ResearcherID Web of Science (Chizhaeva Anna).

Main performers:
Alybayeva A.Zh., PhD in biotechnology, SNC "NPC of Microbiology and Virology", 8 years of scientific experience, author of more than 20 scientific papers, h-index-1. Author Identifier: https://orcid.org/0000-0002-0166-1666
Amangeldi A.A., Master of Technical Sciences, biotechnologist, NS "SPC of Microbiology and Virology", 8 years of scientific experience, more than 20 scientific papers, 3 patents, h-index-1. Author Identifier: https://orcid.org/0000-0003-2208-9184

Information for potential users:
Scientific and Production Center of Microbiology and Virology LLP
Republic of Kazakhstan, 050010, Almaty, Bogenbai batyr str., 105
+7(727) 291-84-97, ext. 113; Mobile: +77772440224
E-mail: anna_chizhaeva@mail.ru
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