The effect of cryofraction of the bovine spleen and placenta on the frequency of dominant lethal mutations in mice under genotoxic conditions

The aim of the work was to study the effect of HCBS, LCBS and LCBSP on the frequency of dominant lethal mutations (DLM) in mice under the genotoxic effect of the experimental mutagen - cyclophosphamide (CP). Eleven groups of outbred mice were formed for the work, 6 males and 36 females in each group. HCBS, LCBS and LCBSP were administered intramuscularly to males once at a dose of 0.5 ml/kg separately or together with intraperitoneal administration of CP at a dose of 60 mg/kg. The studied pharmaceutical substances were also administered three times with an interval of 24 hours before the administration of CP in doses similar to a single administration. To study DLM, female mice were implanted with males three times with an interval of one week. On the 15-17th day after implantation with males, pregnant females were removed from the experiment, autopsied and the frequency of postimplantation mortality (PIM) of embryos was determined, which correlates with damage to spermatozoa, mature and immature spermatids in males. The value of PIM in the experimental groups was compared with that of the negative and positive control groups. Single and course administration of HCBS to male mice with CPinduced cytogenetic instability resulted in decreased clastogenic damage to spermatozoa and late spermatids by CP. Course administration of LCBS and LCBSP to male mice before CP injection induced a significant decrease in postimplantation mortality of mouse embryos, indicating a decrease in the mutagenic effect of CP on mature spermatozoa. Thus, HCBS, LCBS, and LCBSP exerted an antimutagenic effect on germ cells of male mice under conditions of CPinduced cytogenetic instability.

1. Kumaresan, A. Sperm DNA Integrity and Male Fertility in Farm Animals: A Review./ A., Kumaresan, M., Das Gupta, T.K., Datta, J.M., Morrell, // Frontiers in veterinary science. 2020: 7: 321. DOI 10.3389/fvets.2020.00321. URL: https://pubmed.ncbi.nlm.nih.gov/32637425/.

2. Raudsepp. T., Chowdhary BP. Chromosome Aberrations and Fertility Disorders in Domestic Animals / T., Raudsepp, B.P., Chowdhary // Annual review of animal biosciences. 2016: 4: 15-43. DOI 10.1146/annurev-animal-021815-111239. URL: https://pubmed.ncbi.nlm.nih.gov/26884101/.

3. Beketova, D.D. Analysis of mutagenic activity. Test for accounting of dominant lethal mutations in mammals / D. D. Beketova, K. L. Kryshen, A. E. Kastornova, A. A. Atsapkina, M. N. Makarova, V. G. Makarov // International Bulletin of Veterinary Science. 2014: 3: 95-104. URL: https://elibrary.ru/item.asp?id=22307726.

4. Yılmaz, Ö.F. Some reproductive defects in farm animals / Ö.F., Yılmaz, M.A., Çam // Black Sea Journal of Agriculture. 2025: 8 (1): 118-123. URL: https://dergipark.org.tr/en/pub/bsagriculture/issue/88938/1593697.

5. Andrade, M.F. Embryonic and Fetal Mortality in Dairy Cows: Incidence, Relevance, and Diagnosis Approach in Field Conditions/ M.F., Andrade, J., Simões, // Dairy. 2024: 5: 526-541. DOI 10.3390/dairy5030040. URL: https://doi.org/10.3390/dairy5030040.

6. Vostroilova, G.A. Study of the antimutagenic effect of hydrophilic cryofraction of cattle spleen / G. A. Vostroilova, S. V. Shabunin, D. I. Shabanov, Khokhlova N. A., A. A. Korchagina, M. Yu. Syromyatnikov, A. V. Nekrasov, M. A. Selyutina, D. D. Morozova // Agricultural Biology. 2024: 59(6): 1192-1203. DOI 10.15389/agrobiology.2024.6.1192rus. URL: https://agrobiology.ru/6-2024vostroilova.html.

7. Vostroilova, G.A., Study of the effect of lipophilic cryofractions of bovine spleen and placenta on mice under the genotoxic effect of mitomycin / G. A. Vostroilova, D. I. Shabanov, N. A. Khokhlova, A. A. Korchagina, A. V. Nekrasov, M. Yu. Syromyatnikov, N. A. Strelnikov // Agrarian science. 2025: 398 (09): 22–29. DOI 10.32634/0869-8155-2025-398-09-22-29. URL: https://www.vetpress.ru/jour/issue/current.

8. Mironov, A. N. Guidelines for conducting preclinical studies of drugs. Part one / Ed. A.N. Mironova. - Moscow, Publisher: "Grif i K", 2012. - 944 p. URL: https:// rsmu.ru/fileadmin/templates/DOC/Zakon_RF/Mironov_Rukovodstvo_po_provedeniju_doklinicheskikh_issledovanii_lekarstvennykh_sredstv.pdf.

9. Lapin, K. N. The effect of cyclophosphamide on the reproductive system of male rats / K. N. Lapin, I. A. Ryzhkov, N. M. Zakharova // Experimental and clinical pharmacology. 2020: 83(11): 16-19. DOI 10.30906/0869-2092-2020-83-11-16-19. URL: https://elibrary.ru/item.asp?id=44315493.

10. Voelcker G. The Mechanism of Action of Cyclophosphamide and Its Consequences for the Development of a New Generation of Oxazaphosphorine Cytostatics / G. Voelcker // Scientia Pharmaceutica. 2020: 88 (4): 42. DOI 10.3390/scipharm88040042. URL: https://www.mdpi.com/2218-0532/88/4/42.

11. Spears N. Ovarian damage from chemotherapy and current approaches to its protection / N., Spears, F., Lopes, A., Stefansdottir, V., Rossi, M., De Felici, R.A., Anderson, F.G., Klinger, // Human reproduction update. 2019: 25(6): 673-693. DOI 10.1093/humupd/dmz027. URL: https://pubmed.ncbi.nlm.nih.gov/31600388/.

12. Olowe, T.G. Cytotoxic Properties of Cyclophosphamide: A Focus on Its Mechanistic Impacts on Male Gonadal Functions / T.G., Olowe, M.O., Oyovwi, K.E., Nwangwa, E.P., Ohwin, O.B. Oghenetega, // J Explor Res Pharmacol. 2024: 9(2): 106–115. DOI 10.14218/JERP.2023.00025S. URL: https://www.xiahepublishing.com/2572-5505/JERP-2023-00025S

13. Ebokaiwe A.P. Cyclophosphamideinduced testicular oxidative-inflammatory injury is accompanied by altered immunosuppressive indoleamine 2, 3-dioxygenase in Wister rats: Influence of dietary quercetin / A.P., Ebokaiwe, D.O., Obasi, R.C., Njoku, S. Osawe, // Andrologia. 2022: 54(3): e14341. DOI 10.1111/and.14341. URL: https://onlinelibrary.wiley.com/doi/10.1111/and.14341.

14. Shabunin S.V., Belyaev V.I., Vostroilova G.A., Kabitsky S.N. Organopreparations (medicines from animal organs and tissues). Voronezh, 2013.; URL: https://elibrary.ru/item.asp?id=26618629&ysclid=mfmhl8wtge466492286.

15. Kawakatsu M., Placental extract protects bone marrow-derived stem/progenitor cells against radiation injury through antiinflammatory activity. / M., Kawakatsu, Y., Urata, S., Goto, Y., Ono, T. S. Li, // Journal of Radiation Research. 2013: 54(2): 268-276. DOI 10.1093/jrr/rrs105. URL: https://pubmed.ncbi.nlm.nih.gov/23154884/.

16. Shabanov, D. I. Study of cytogenetic stability of mouse bone marrow using hydrophilic cryofraction of cattle spleen / D. I. Shabanov // Veterinary Pharmacological Bulletin. 2024: 4(29): 36-43. DOI 10.17238/issn2541-8203.2024.4.36. URL: http://en.vfvrn.ru/netcat_files/6/2/VFV_2024_4_29_.pdf.

17. Shabunin S.V. The influence of aminoseleton on the state of the prooxidant and

18. antioxidant blood systems in sows / S. V. Shabunin, A. G. Shakhov, G. A. Vostroilova, P. A. Parshin, T. G. Ermolova, N. A. Khokhlova, G. N. Bliznetsova // Achievements of science and technology of the agroindustrial complex. 2019: 33 (7): 71-74. DOI 10.24411/0235-2451-2019-10716. URL: https://elibrary.ru/item.asp?id=39205932.