IRN AP23484620

The relevance of the project: 

Production of high-quality and safe milk is a key priority for farmers and dairy processing plants. Low milk productivity, despite the large livestock population, is caused by several factors, including extensive livestock production practices, insufficient realization of the animals’ genetic potential, and the prevalence of diseases transmitted among domestic livestock. Among infectious diseases in dairy farming, mastitis is widespread and causes substantial economic losses. Mastitis in dairy cows deteriorates milk quality and thereby constrains the development of the dairy industry. Mastitis develops when pathogenic bacteria enter the mammary gland through the teat canals during machine milking. Mechanical impact, pathogenic microbial infection, and chemical injury are the three main categories into which the causes of this disease may be divided. An enteromammary route of infection also cannot be excluded. In cases of gastrointestinal microbiota dysbiosis, microorganisms may spread to the mammary gland through the lymphatic and circulatory systems, resulting in the development of mastitis in cows. Implementation of this project will help address this problem and improve milk quality indicators, because the need to administer antibiotics to animals will be reduced. Introduction of a domestically produced Kazakhstani product to the market will reduce the risk of dependence on imports.

Current mastitis prevention strategies are based on culling infected cows or administering antibiotics to eliminate the disease.

The advantages of medicinal plants include their lower toxicity and the possibility of long-term use without the development of adverse effects. Plant families will be selected according to their chemical composition. Particular attention will be paid to plants characterized by a high content of essential oils, bioflavonoids, gums, tannins, and other compounds capable of exerting regenerative, fungicidal, anesthetic, and film-forming effects.

The concept and novelty of this project consist in the use of newly developed therapeutic and preventive phytogenic and phytobiotic products made from components of medicinal plants growing in the Republic of Kazakhstan. The phytogenic products are presented as medicated extruded feed granules, whereas the phytobiotic products are presented as a teat-treatment product.

The final project outcome is a prototype in the form of biological products ready for commercialization, distinguished from products currently available on the Kazakhstani market by their safety and efficacy and enabling the production of high-quality and safe dairy foods, including environmentally friendly products. At present, Kazakhstan needs to develop domestic production of veterinary preparations, which is the main objective of the project.

The purpose of the project: To develop and test phytogenics and phytobiotics in production conditions for the prevention and treatment of animal mastitis with an assessment of the quality and safety of milk.

Achieving the goal of increasing the quantity, quality and safety of milk produced on farms

Expected and achieved results: 

Patent and information search

A patent and information search was conducted in accordance with the established WIPO rules on the “PCT Minimum Documentation,” including searches in international databases such as Web of Science, Scopus, SpringerLink, Elsevier, and others.

Microbiological studies

Swab samples from the teats of cows and goats were examined, and microorganisms were isolated. The main mastitis pathogens detected on the farms were bacteria of the genera Staphylococcus, including S. aureus, Bacillus, Escherichia coli, Candida spp., and Aerococcus viridans.

Development of biological products

More than 10 formulations of therapeutic and preventive biological products were developed and manufactured in the form of phytobiotic udder-treatment products and phytogenic products consisting of extruded feed granules. The effects of the developed formulations on the isolated microbiota were studied, and their antimicrobial activity was determined.

Chemical composition and antioxidant activity

The chemical composition of the main biologically active components of the developed composite formulations was determined, including total phenols, phenolic acids, flavonoids, and catechins, and their antioxidant activity was assessed. The obtained data indicate a high concentration of phenolic compounds in poplar buds, amounting to 42.86 mg/mL. Poplar buds also demonstrated the highest content of flavonoids, 34.064 mg/mL, and phenolic acids, 12.058 mg/mL. Assessment of antioxidant activity based on inhibition of the DPPH radical and expressed as IC₅₀ showed that poplar bud extract had the strongest activity, at 1.89 mg/g, followed by clove, 2.9 mg/g; wormwood, 7.5 mg/g; greater plantain, 9.4 mg/g; and yarrow, 10.2 mg/g. Weaker activity was demonstrated by hop, 32.7 mg/g, and coriander, 41.8 mg/g, whereas milk thistle and tinder fungus extracts were inactive under the experimental conditions.

In vitro antimicrobial activity

In vitro testing showed that the plant extracts demonstrated moderate to high antimicrobial activity. Populus balsamifera produced inhibition zones of 15.1 mm against the field Candida strain and 19.1 mm against S. aureus, while Humulus lupulus produced inhibition zones of 18.2 mm against the field Candida strain and 18.4 mm against S. aureus; these extracts showed the largest zones of inhibition against key mastitis pathogens. Syzygium aromaticum demonstrated excellent activity against Candida ATCC, 14.5 mm, and E. coli ATCC, 17.5 mm. Plantago major showed its strongest activity against the field Candida strain, 8.1 mm, but produced inconsistent results against different strains. Humulus lupulus inhibited a broad spectrum of most target microorganisms, with inhibition zones ranging from 9.0 to 18.4 mm. The formulations and commercial products differed in efficacy: CP2 reached its maximum activity against Candida ATCC, 28.8 mm; F2 demonstrated broad activity ranging from 12.6 to 17.2 mm; whereas F1 had limited activity against field isolates, with inhibition zones of 3.2–7.6 mm. Reference strains were generally more susceptible than field isolates, reflecting the higher resistance of clinical mastitis pathogens.

Results of commercial farm trials

Commercial farm trials were conducted on two farms in Akmola Region. The results obtained from the combined use of a phytogenic product, namely an extruded feed additive, and a phytobiotic product, namely an udder-treatment preparation, showed the following:

• the proportion of healthy animals in the experimental group was 29.8%, compared with 18.4% in the control group, representing an increase of 11.4 percentage points;
• the overall incidence of mastitis was reduced by 11.4%;
• hematological indicators improved: leukocyte counts decreased by 43%, lymphocyte counts by 41%, and granulocyte counts by 75%;
• milk productivity increased by 3.85%, reaching 40.4 kg compared with 38.9 kg, and the increase per cow reached 6.2 kg/day, or 18.1%;
• feed intake increased by 19.7%, and feed efficiency increased by 3.5%;
• in the experimental group, milk fat content increased by 29.3%, protein content increased by 14.9%, and the somatic cell count decreased by 39.3%, from 1,400 to 850 thousand cells/mL;
• clinical recovery was recorded in 35 of 118 diseased animals, or 29.7%.

Thus, the integrated use of both types of biological products produced a pronounced positive effect on animal health, productivity, and milk quality.

Documentation and information materials

Regulatory and technical documentation has been developed, including instructions for the application and production of biopreparations, as well as an organizational standard. Production trial and implementation reports have been obtained. Packaging label designs have been prepared. Packaging label designs were also prepared. Practical production recommendations entitled “Application of the 7-Phyt Phytogenic Extruded Feed Additive and the PhytoDip Phytobiotic Product for the Prevention and Control of Mastitis in Cows” were prepared. The monograph “Modern Aspects of Food Quality and Safety Control” was published, together with brochures and leaflets for livestock and veterinary specialists.

Information dissemination and promotion

To promote the project results and increase the likelihood of their implementation and commercialization, an additional webpage was created on the website https://biofeed.kz/scientific-project/nauchnyj-proekt-1, along with a “Patents” page at https://biofeed.kz/patents. These pages are actively used and regularly updated.

Information on the developed biological products was published on social media platforms, including Instagram, TikTok, Facebook, and Telegram:

1. https://www.instagram.com/reel/DQZ1f4_iEqY/?igsh=MXE3YjI2N3F2cjNxeQ==

2. https://www.instagram.com/p/DUdh1c-CLkX/?igsh=cnRtOTBjazVpYXNy

3. https://www.instagram.com/p/DVfjEAdiKoO/?igsh=MTJoMnkxNGk1N3J1ag==

4. https://biofeed.kz/scientific-project/nauchnyj-proekt-1

5. https://vt.tiktok.com/ZSQMm9Bhp/

6. https://www.instagram.com/reel/DZzcqlaoRZq/?igsh=dDB4bmxjZDEwODRi

Project exhibits were demonstrated at 18 international and national exhibitions, including the KazAgro/KazFarm 2025 International Exhibition, the RES-2026 Expo International Exhibition of Green and Sustainable Technologies, the opening of the KATIU Agrotechnology Park, and the KATIU Exhibition of Scientific Achievements attended by deputies of the Mazhilis of the Parliament of the Republic of Kazakhstan, among others.

Editing of a video training course for livestock farmers, veterinary specialists, and students is currently underway.

The development was implemented at the farm of Individual Entrepreneur “Semenova” in Akmola Region, as documented by the Production Implementation Act dated July 10, 2026.

Human resource development

Within the framework of the project, 2 PhD candidates have been prepared for dissertation defense, 3 Master of Veterinary Science graduates have completed their studies, and 5 veterinary specialists have graduated:

1. Abakanova G.N. PhD dissertation: "The Influence of Multifunctional Feed Bioadditives on Milk Quality and Safety."
2. Alyonova M.E. PhD dissertation: "The Effect of Phytobiotic Products and Phytogenic Feed Additives on Milk Quality and Safety in Animals with Mastitis."
3. 2025 – Tolendiyev Zh.Ye. Master's thesis: "Assessment of the Quality and Safety of Extruded Feeds Produced from Non-Grade Potatoes and Spent Mushroom Substrate." Participated in the development and production of phytobiotics.
4. 2026 – Gaidukevich V.V. Master's thesis: "Development of Phytobiotic Products for Teat Treatment and Evaluation of Their Effects on the Milk Quality of Cows and Goats."
5. 2026 – Gorchkhanova Kh.A. Master's thesis: "Development of a Phytogenic Feed Additive for the Prevention of Mastitis in Dairy Cows with Evaluation of Milk Quality."
6. 2025 – Zhitina A.A. Bachelor's thesis: "Development of a Phytobiotic Composition with Antimicrobial Properties for the Prevention of Mastitis in Dairy Cows."
7. 2025 – Moskalenko V.A. Bachelor's thesis: "Evaluation of the Effects of Phytobiotics with Sedative Properties on the Productivity of Dairy Cows and Milk Quality."
8. 2026 – Kurmanbayeva A.R. Bachelor's thesis: "Effect of Extruded Phytobiotic Feed Additives on Milk Production and Milk Quality in Dairy Cows."
9. 2026 – Sairan A.T. Bachelor's thesis: "Development and Evaluation of a Phytobiotic Product for the Prevention of Mastitis in Animals."
10. 2026 – Shakishev A.N. Bachelor's thesis: "Comparison of the Effectiveness of Plant Extracts Obtained by Different Extraction Methods against Mastitis Pathogens in Animals."

Research team

The project team includes specialists with experience in food safety, the development and manufacture of therapeutic and preventive biological products, phytogenic products, and phytobiotic products. These specialists have previously completed research projects successfully, including commercialization projects.

The project team comprises professors, associate professors, candidates of sciences, PhD holders, and master’s degree holders. The project consultants include three international scientists and one national scientist:

1) Galia Zamaratskaia – Associate Professor at the Swedish University of Agricultural Sciences, Uppsala, Sweden. h-index – 37; Web of Science ResearcherID: I-6155-2016; ORCID: 0000-0003-0926-4849; Scopus Author ID: 57299699500.

2) Martin Knicky – Senior Researcher at RISE Research Institutes of Sweden, Uppsala, Sweden. h-index – 6; Web of Science ResearcherID: GYD-8437-2022; ORCID: 0000-0001-5121-1957; Scopus Author ID: 8254820200.

3) Aleko Kalandia – Doctor of Chemical Sciences, Professor, Head of the Western Georgian Chromatography Center, Batumi, Georgia. h-index – 6; Web of Science ResearcherID: AAH-4645-2019; ORCID: 0000-0001-9570-9263; Scopus Author IDs: 60015410100, 56580232900, and 57204110464.

4) Vladilen Vasilievich Polyakov – Doctor of Chemical Sciences, Professor at M. Kozybayev North Kazakhstan State University, Petropavlovsk, Republic of Kazakhstan. h-index – 4; Web of Science ResearcherID: ABG-5962-2020; ORCID: 0000-0003-4700-7560; Scopus Author ID: 57224838039.

The research results of the consultants have been published in more than 150 high-ranking journals indexed in Scopus, Web of Science, and other databases. The scientific consultants select and validate ingredients for therapeutic and preventive products, advise on experimental design and the manufacture of extruded granules, perform statistical analysis of the results, and support publication in peer-reviewed journals.

Production specialists from livestock farms also participate in the project.

The research group includes university students and trainees, namely two doctoral students, three master’s students, and two undergraduate students. This participation will enable young specialists to improve their professional competencies and develop as competitive professionals. Data obtained during the experiments were incorporated into dissertations and graduation theses.

The Project Principal Investigator is Yuriy Alexandrovich Balji, Candidate of Veterinary Sciences and Professor, a specialist in food quality and safety, the production of phytobiotic products, and extruded compound feeds. He was twice awarded a scholarship in the competition for talented young scientists of the Republic of Kazakhstan, in 2010 and 2014. He was twice awarded the title “Best University Teacher of the Republic of Kazakhstan,” in 2011 and 2022. He received a Certificate of Honor from the Ministry of Agriculture of the Republic of Kazakhstan in 2015 and the honorary title “Honored Inventor of the Republic of Kazakhstan” in 2019. On the basis of previously obtained patents and registered results of scientific and technical activities, he established a startup company and organized the production and sale of feed bioadditives for animals.

The present project is a logical continuation of his scientific work in food safety and animal production. He has published more than 270 scientific and methodological works and obtained 65 patents. He has completed professional development at more than 20 universities and production organizations in Europe, the United States, and Kazakhstan and has received more than 30 certificates, 17 of which were issued by international organizations. He also completed training as an expert in organic products under CCAA.

Composition of the research group conducting the scientific studies, including international scientists and young researchers, such as postdoctoral researchers and doctoral, master’s, and undergraduate students, for 2024–2026

Published scientific works as of July 14, 2026

Two articles were published in journals indexed in Scopus, Q1 and Q2, and Web of Science:

1) Balji Y, Sultanayeva L, Mustafina R, Alyonova M, Kalandia A, Djafaridze I, Zamaratskaia G. In Vitro Antimicrobial Potential of Medicinal Plant Extracts and Their Combinations Against Mastitis-Causing Bacteria in Dairy Cows. Molecules. 2026;31(7):1089. https://doi.org/10.3390/molecules31071089. Scopus Q1, percentile – 89; Web of Science – Q2.

2) Balji, Yu., Abakanova, G., Sultanayeva, L., Mayer, Y., Kravchenko, O., Zamaratskaia, G. (2026). Optimization of extruded enriched feed additives production process and their effect on blood parameters of Holstein cows. Agriculture and Forestry, 72(2):123–135. https://doi.org/10.17707/AgricultForest.72.2.09; https://agricultforest.ac.me/data/20260616-09%20Balji%20et%20al.pdf. Scopus Q2, percentile – 50.

Two articles were published in journals recommended by the Committee for Quality Assurance in Science and Higher Education:

1) Abakanova G.N., Balji Yu.A., Sultanayeva L.Z., Zamaratskaia G. Assessment of the milk produced by mastitis affected cows using an extruded feed supplement containing phytogenics. Herald of Science of S. Seifullin Kazakh Agrotechnical Research University: Veterinary Sciences. Astana: S. Seifullin Kazakh Agrotechnical Research University, 2025. No. 1 (009). P. 85–91. ISSN 2958-5430, ISSN 2958-5449. https://doi.org/10.51452/kazatuvc.2025.5(009).1881.

2) Alyonova M.E., Mustafina R.Kh., Baboshina L.D., Balji Yu.A. Application of a phytobiotic product in “DIP” form for the prevention of mastitis in animals. 3i: Intellect, Idea, Innovation. No. 2, Part 1, 2026. P. 5–13. https://ojs.ksu.edu.kz/index.php/3i/article/view/1746.

Seven conference abstracts were published in the proceedings of international conferences, including conferences held in Sweden, Ukraine, Lithuania, and Serbia:

1) Gaidukevich V.V., Balji Yu.A. Methods for treating mastitis with phytobiotics. Proceedings of the International Scientific and Practical Conference “120 Years of Kazakh Veterinary Science: Achievements and New Challenges in Ensuring Biological Safety,” dedicated to the 120th anniversary of the Kazakh Scientific Research Veterinary Institute. Almaty: KazSRVI LLP, 2025. P. 161–165.

2) Balji Yu.A., Polyakov V.V., Mustafina R.Kh., Sultanayeva L.Z., Abakanova G.N., Musagiyeva D.K. Prospects for the use of phytobiotics and phytogenics in veterinary medicine. Proceedings of the 13th International Scientific and Practical Conference “Current Problems of Natural Sciences,” dedicated to the 75th anniversary of Doctor of Chemical Sciences, Professor B.E. Begenova, April 11, 2025.

3) Alyonova M.E., Mustafina R.Kh., Sultanayeva L.Z., Gaidukevich V.V., Zamaratskaia G., Balji Yu.A. Phytobiotic product for the prevention and treatment of mastitis in dairy animals. Proceedings “One Health in Practice: Animal Welfare, Management, and Food Safety,” Animal Science. Kyiv: NUBiP of Ukraine, 2026. P. 39.

4) Meruyert Alyonova, Raikhan Mustafina, Leila Sultanayeva, Galia Zamaratskaia, Yuriy Balji. Dysbiosis of the udder microbiota as a risk factor for mastitis in cows. IDF Mastitis and Milking Technology Joint Conference 2026 Proceedings, March 10–13, 2026. P. 131–134.

5) Meruyert Alyonova, Y. Balji, R. Mustafina, K. Gorchkhanova, Galia Zamaratskaia. Assessment of Radiation Safety and Phytochemical Composition of Selected Medicinal Plants from Kazakhstan for Veterinary Use. Second International Veterinary Students Conference held on April 24, 2026, at the Lithuanian University of Health Sciences, Veterinary Academy, Lithuania. P. 20.

6) Alyonova M.E., Sultanayeva L.Z., Gaidukevich V.V., Baboshina L.D., Gorchkhanova Kh.A., Mayer Y.G., Zhitina A.A., Mustafina R.Kh., Balji Yu.A., Zamaratskaia G. Antimicrobial potential of phytobiotics against causative agents of bovine mastitis. Proceedings of the International Scientific and Practical Conference “Seifullin Readings – 22: Agricultural Systems of the Future: Sustainable Development, Innovation, and Low-Carbon Solutions.” Astana, April 10, 2026. P. 182–186.

7) Yuriy Balji, Viktoriya Gaidukevich, Leila Sultanayeva, Meruyert Alyonova, Gulim Abakanova, Galia Zamaratskaia. Dietary plant-based bioactive compounds for sustainable mastitis control in livestock. Proceedings of the 15th International Symposium “Modern Trends in Livestock Production.” October 29–31, 2025, Belgrade, Serbia. P. 32–45.

A monograph incorporating the results obtained during the implementation of the project was published: “Modern Aspects of Food Quality and Safety Control” (Astana, 2025, 280 pages).

Intellectual property protection documents

1. Patent of the Republic of Kazakhstan for Utility Model No. 9502 was granted on the basis of Application No. 2023/1194.2 dated December 7, 2023, Bulletin No. 50 dated December 13, 2024: “Feed Additive for the Prevention and Treatment of Mastitis in Animals and a Method for Its Production.”

2. Patent of the Republic of Kazakhstan for Invention No. 37796, “Product for the Prevention and Treatment of Mastitis in Farm Animals,” was granted on the basis of Application No. 2024/0961.1, Bulletin No. 5 dated February 6, 2026.

3. Application No. 2025/1024.1 dated October 18, 2025, was filed for a patent for an invention of the Republic of Kazakhstan entitled “Method for Manufacturing a Phytobiotic Product for Treating the Teats of Animal Udders.”

4. Application No. 2026106356 dated March 10, 2026, was filed for a patent for an invention of the Russian Federation entitled “Method for Manufacturing a Phytobiotic Product for Treating the Teats of Animal Udders.”