Probiotic properties of lactic acid bacteria isolated from the reproductive system of cows on eco‑farms in the North-West of Russia
Abstract
Background. The application of probiotics capable of restoring the normal microbiota of the cow’s reproductive tract and enhancing resistance to infections represents a promising direction.
Purpose. The aim of this study was to isolate, identify, and characterize promising lactic acid bacteria strains from the reproductive tract of healthy cows for the development of a novel biopreparation aimed at the prevention of endometritis.
Materials and methods. Biological material (vaginal wall swabs) for the isolation of prospective Lactobacilli strains was collected from ten clinically healthy dairy cows on three eco-farms (located in the North-Western region of the Russian Federation and the Republic of Karelia). Isolation of pure cultures was performed by plating serial dilutions onto a selective agar medium. Identification was carried out using standard microbiological methods, including PCR analysis. Antagonistic activity against ten pathogenic and opportunistic microorganisms was assessed in vitro using the agar diffusion method.
Results. A total of 95 Lactobacilli isolates were obtained. Screening of 42 lactobacilli strains revealed 19 active and moderately active variants exhibiting antimicrobial activity against pathogens. Strain E-49 (Lactobacillus sp.) demonstrated the highest antagonistic activity against Clostridium perfringens (38 ± 1,0 mm) and Enterococcus faecalis (32 ± 1,9 mm). Strain E-52 (Lactobacillus sp.) actively inhibited the growth of Penicillium sp. (37 ± 1,5 mm) and Streptococcus agalactiae (21 ± 0,9 mm). None of the tested strains exhibited hemolytic activity. Several active strains, such as E-49, E-52, and E-10, demonstrated stable titers of viable cells over 30 days of storage (2,4×10⁹ ± 1,5×10⁸ and 1,4×10⁹ ± 7,0×10⁷ CFU/mL for E-52 at 10 and 30 days, and 7,3×10⁹ ± 6,2×10⁸ CFU/mL for E-10 at 30 days) and optimal pH values (3,8-4,2).
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References
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Prabhurajeshwar, C., & Chandrakanth, R. K. (2017). Probiotic potential of Lactobacilli with antagonistic activity against pathogenic strains: An in vitro validation for the production of inhibitory substances. Biomedical journal, 40(5), 270–283. https://doi.org/10.1016/j.bj.2017.06.008
Saini, P., Singh, M., & Kumar, P. (2019). Fungal endometritis in bovines. Open veterinary journal, 9(1), 94–98. https://doi.org/10.4314/ovj.v9i1.16
Shokryazdan, P., Sieo, C. C., Kalavathy, R., Liang, J. B., Alitheen, N. B., Faseleh Jahromi, M., & Ho, Y. W. (2014). Probiotic potential of Lactobacillus strains with antimicrobial activity against some human pathogenic strains. BioMed research international, 2014, 927268. https://doi.org/10.1155/2014/927268
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Singh, R. P., Shadan, A., & Ma, Y. (2022). Biotechnological Applications of Probiotics: A Multifarious Weapon to Disease and Metabolic Abnormality. Probiotics and antimicrobial proteins, 14(6), 1184–1210. https://doi.org/10.1007/s12602-022-09992-8. EDN: https://elibrary.ru/MGSNWT
Van Hul, M., Cani, P. D., Petitfils, C., De Vos, W. M., Tilg, H., & El-Omar, E. M. (2024). What defines a healthy gut microbiome?. Gut, 73(11), 1893–1908. https://doi.org/10.1136/gutjnl-2024-333378. EDN: https://elibrary.ru/GIQFKH
Várhidi, Z., Csikó, G., Bajcsy, Á. C., & Jurkovich, V. (2024). Uterine Disease in Dairy Cows: A Comprehensive Review Highlighting New Research Areas. Veterinary sciences, 11(2), 66. https://doi.org/10.3390/vetsci11020066. EDN: https://elibrary.ru/HEABXN
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Markowiak-Kopeć, P., & Śliżewska, K. (2020). The Effect of Probiotics on the Production of Short-Chain Fatty Acids by Human Intestinal Microbiome. Nutrients, 12(4), 1107. https://doi.org/10.3390/nu12041107. EDN: https://elibrary.ru/DFSSQL
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Meziti, A., Tsementzi, D., Rodriguez-R, L. M., Hatt, J. K., Karayanni, H., Kormas, K. A., & Konstantinidis, K. T. (2019). Quantifying the changes in genetic diversity within sequence-discrete bacterial populations across a spatial and temporal riverine gradient. The ISME Journal, 13(3), 767–779. https://doi.org/10.1038/s41396-018-0307-6. EDN: https://elibrary.ru/ARKMLC
Miranda, C., Contente, D., Igrejas, G., Câmara, S. P. A., Dapkevicius, M. L. E., & Poeta, P. (2021). Role of Exposure to Lactic Acid Bacteria from Foods of Animal Origin in Human Health. Foods, 10(9), 2092. https://doi.org/10.3390/foods10092092. EDN: https://elibrary.ru/VIIFBK
Monteagudo-Mera, A., Rastall, R. A., Gibson, G. R., Charalampopoulos, D., & Chatzifragkou, A. (2019). Adhesion mechanisms mediated by probiotics and prebiotics and their potential impact on human health. Applied Microbiology and Biotechnology, 103(16), 6463–6472. https://doi.org/10.1007/s00253-019-09978-7. EDN: https://elibrary.ru/VMEXGW
Mor, A., Driggers, P. H., & Segars, J. H. (2015). Molecular characterization of the human microbiome from a reproductive perspective. Fertility and Sterility, 104(6), 1344–1350. https://doi.org/10.1016/j.fertnstert.2015.10.008. EDN: https://elibrary.ru/WVKXWB
Papadimitriou, K., Zoumpopoulou, G., Foligné, B., Alexandraki, V., Kazou, M., Pot, B., & Tsakalidou, E. (2015). Discovering probiotic microorganisms: In vitro, in vivo, genetic and omics approaches. Frontiers in Microbiology, 6, 58. https://doi.org/10.3389/fmicb.2015.00058. EDN: https://elibrary.ru/XONSIX
Prabhurajeshwar, C., & Chandrakanth, R. K. (2017). Probiotic potential of Lactobacilli with antagonistic activity against pathogenic strains: An in vitro validation for the production of inhibitory substances. Biomedical Journal, 40(5), 270–283. https://doi.org/10.1016/j.bj.2017.06.008
Saini, P., Singh, M., & Kumar, P. (2019). Fungal endometritis in bovines. Open Veterinary Journal, 9(1), 94–98. https://doi.org/10.4314/ovj.v9i1.16
Shokryazdan, P., Sieo, C. C., Kalavathy, R., Liang, J. B., Alitheen, N. B., Faseleh Jahromi, M., & Ho, Y. W. (2014). Probiotic potential of Lactobacillus strains with antimicrobial activity against some human pathogenic strains. BioMed Research International, 2014, 927268. https://doi.org/10.1155/2014/927268
Singh, N., & Sethi, A. (2022). Endometritis — Diagnosis, treatment and its impact on fertility — A scoping review. JBRA Assisted Reproduction, 26(3), 538–546. https://doi.org/10.5935/1518-0557.20220015. EDN: https://elibrary.ru/FLRPCJ
Singh, R. P., Shadan, A., & Ma, Y. (2022). Biotechnological Applications of Probiotics: A Multifarious Weapon to Disease and Metabolic Abnormality. Probiotics and Antimicrobial Proteins, 14(6), 1184–1210. https://doi.org/10.1007/s12602-022-09992-8. EDN: https://elibrary.ru/MGSNWT
Van Hul, M., Cani, P. D., Petitfils, C., De Vos, W. M., Tilg, H., & El-Omar, E. M. (2024). What defines a healthy gut microbiome? Gut, 73(11), 1893–1908. https://doi.org/10.1136/gutjnl-2024-333378. EDN: https://elibrary.ru/GIQFKH
Várhidi, Z., Csikó, G., Bajcsy, Á. C., & Jurkovich, V. (2024). Uterine Disease in Dairy Cows: A Comprehensive Review Highlighting New Research Areas. Veterinary Sciences, 11(2), 66. https://doi.org/10.3390/vetsci11020066. EDN: https://elibrary.ru/HEABXN
Zangirolamo, A. F., Souza, A. K., Yokomizo, D. N., Miguel, A. K. A., Costa, M. C. D., Alfieri, A. A., & Seneda, M. M. (2024). Updates and Current Challenges in Reproductive Microbiome: A Comparative Analysis between Cows and Women. Animals, 14(13), 1971. https://doi.org/10.3390/ani14131971. EDN: https://elibrary.ru/QLHPJU
Copyright (c) 2026 Elena A. Yildirim, Valentina A. Filippova, Irina A. Klyuchnikova, Ksenia A. Sokolova, Larisa A. Ilina, Alesya A. Savicheva, Elena A. Korochkina
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