Effect of ozonation on resistance of ovine and human erythrocytes to hypothermic storage

  • K. M. Holovina Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine
  • O. M. Bobrova Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine
  • S. Y. Kovalenko Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine
  • Y. S. Hovorova Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine
  • O. A. Nardid Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine
Keywords: red blood cells; Alsever’s solution; mannitol; preservation medium; osmotic fragility; sphericity index; rosette formation.


Long-term hypothermic storage of animal blood can lead to the loss of its quality and can cause complications in recipient animals after transfusion, so the search for new methods of increasing the preservation of erythrocytes after hypothermic storage continues. The article presents the data of the ozonation effect on the preservation rate of ovine and human erythrocytes during hypothermic storage with Alsever’s solution and mannitol medium. Hemolysis, osmotic fragility and distribution density of ovine and human erythrocytes by the sphericity index were determined at different stages of hypothermic storage. The ovine erythrocytes in the control had a lower osmotic resistance compared to human erythrocytes; however, their osmotic fragility did not change significantly after hypothermic storage for 8 weeks, and for human erythrocytes, it significantly increased. Storage of ovine and human erythrocytes longer than 8 weeks led to a sharp hemolysis, while the level of hemolysis of ovine erythrocytes was lower than that of human erythrocytes. Preservation of ozonated erythrocytes is higher than non-ozonated ones during prolonged hypothermic storage. The effect of ozonation on the preservation rate of red blood cells depended on the composition of the preservation media. Hypothermal storage of human erythrocytes in Alsever’s solution for up to 8 weeks led to a shift in the density of distribution according to the sphericity index towards spheroidization of cells; in a medium with mannitol, the number of flattened cell forms increased. After 8 weeks of hypothermic storage of ovine erythrocytes, most of the cells had high sphericity indices. Pretreatment of human and sheep erythrocytes with ozone allows the distribution of cells to be kept closer to the control during long-term storage, which is especially pronounced in mannitol medium. Ovine erythrocytes retained the ability to form rosettes with human T-lymphocytes after hypothermic storage for up to 12 weeks. Thus, ozone pretreatment and the use of mannitol as part of hypothermic storage medium could be an approach to improve the quality of preserved ovine erythrocytes.


Adili, N., & Melizi, M. (2014). Preliminary study of the influence of red blood cells morphometry on the species determinism of domestic animals. Veterinary World, 7(4), 219–223.

Baskurt, O. K., Hardeman, M. R., & Rampling, M. W. (2007). Handbook of hemorheology and hemodynamics. IOS press, Amsterdam.

Bell, G. (2006). Blood transfusions in cattle. Livestock, 11(3), 39–43.

Chandler, K., Fitzpatrick, J., Mellor, D., Milne, M., & Fishwick, G. (1998). Intraperitoneal administration of whole blood as a treatment for anaemia in lambs. The Veterinary Record, 142(7), 175–176.

Davidow, B. (2013). Transfusion medicine in small animals. Small Animal Practice, 43(4), 735–756.

Durham, A. E. (1996). Blood and plasma transfusion in the horse. Equine Veterinary Education, 8(1), 8–12.

Fung, Y. L., Simonova, G., & Tung, J. P. (2016). Lessons from sheep models of transfusion. ISBT Science Series, 11(2), 73–78.

Gordiyenko, O. I., Gordiyenko, Y. E., & Makedonska, V. O. (2004). Estimation of erythrocyte population state by the spherical index distribution. Bioelectrochemistry, 62, 119–122.

Jikuya, T., Tsutsui, T., Shigeta, O., Sankai, Y., & Mitsui, T. (1998). Species differences in erythrocyte mechanical fragility: Comparison of human, bovine, and ovine cells. American Society for Artificial Internal Organs Journal, 44(5), 452–455.

Kisielewicz, C., & Self, I. A. (2014). Canine and feline blood transfusions: Controversies and recent advances in administration practices. Veterinary Anaesthesia and Analgesia, 41(3), 233–242.

Klaus, J. (1990). Lymphocytes: Methods. Mir, Moskow (in Russian).

Kumar, R. (2017). Blood transfusion in veterinary medicine. Hematology and Transfusion International Journal, 4(4), 116–122.

Ladics, G. S. (2007). Primary immune response to sheep red blood cells (SRBC) as the conventional T-cell dependent antibody response (TDAR) test. Journal of Immunotoxicology, 4(2), 149–152.

Le Gal, A., Thomas, E. K., & Humm, K. R. (2020). Xenotransfusion of canine blood to cats: A review of 49 cases and their outcome. Journal of Small Animal Practice, 61(3), 145–208.

Lomako, V. V., Kovalenko, I. F., & Shilo, A. V. (2012). Peripheral blood erythrocytes at various types of hypothermia of homoiothermal organism problems of cryobiology, 22 (4), 389–397.

Martinez-Sogues, L, Blois, S. L., Manzanilla, E. G., Abrams-Ogg, A. O., & Cosentino, P. (2020). Exploration of risk factors for non-survival and for transfusion-associated complications in cats receiving red cell transfusions: 450 cases (2009 to 2017). Journal of Small Animal Practice, 61(3), 177–184.

Matei, H., Frentescu, L., & Benga, G. (2000). Comparative studies of the protein composition of red blood cell membranes from eight mammalian species. Journal of Cellular and Molecular Medicine, 4(4), 270–276.

Matsuzawa, T., & Ikarashi, Y. (1979). Haemolysis of various mammalian erythrocytes in sodium chloride, glucose and phosphate-buffer solutions. Laboratory Animals, 13(4), 329–331.

McAllister, E. J., Apgar, J. R., Leung, C. R., Rickert, R. C., & Jellusova, J. (2017). New methods to analyze B cell immune responses to thymus-dependent antigen sheep red blood cells. The Journal of Immunology, 199(8), 2998–3003.

Ostras, D. A., Lutsenko, D. G., Lomako, V. V., Kovalenko, I. F., & Shylo, O. V. (2019). Changes in erythrocytes osmotic fragility and sphericity index in rat’s of different ages under cold exposures. Problems of Cryobiology and Cryomedicine, 29(2), 172.

Park, H. J., Lee, S. Y., Ji, M., Kim, K., Son, Y. H., Jang, S., & Park, Y. K. (2016). Measuring cell surface area and deformability of individual human red blood cells over blood storage using quantitative phase imaging. Scientific Reports, 6, 34257.

Ryskina, E. A., & Gilmiyarova, F. N. (2015). Group antigens of various animals. RUDN Journal of Agronomy and Animal Industries, 1, 25–34 (in Russian).

Saganuwan, S. A. (2019). Effects of therapeutic and toxic agents on erythrocytes of different species of animals. In: Tombak, A. (Ed.). Erythrocyte. IntechOpen, London.

Saritha, G., Haritha, G. S., Kumari, K. N., & Sundar, N. S. (2016). Blood transfusion in a calf with life-threatening anemia. IOSR Journal of Agriculture and Veterinary Science, 9(5), 69–70.

Shinbrot, T. (2019). Biomedical fluid dynamics: Flow and form. Oxford University Press, Oxford.

Simonova, G., Tung, J. P., Fraser, J. F., Do, H. L., Staib, A., Chew, M. S., Dunster, K. R., Glenister, K. M., Jackson, D. E., & Fung, Y. L. (2014). A comprehensive ovine model of blood transfusion. Vox Sanguinis, 106(2), 153–160.

Sousa, R. S., Minervino, A. H. H., Araújo, C. A. S. C., Rodrigues, F. A. M. L., Oliveira, F. L. C., Mori, C. S., Zaminhan, J. L. R., Moreira, T. R., Sousa, I. K. F., Ortolani, E. L., & Barrêto Júnior, R. A. (2014). Clinical response and transfusion reactions of sheep subjected to single homologous blood transfusion. The Scientific World Journal, 2014, 734397.

Wain, E. B., & Redpath, J. A. (1985). Blood transfusion as a treatment of anaemia in lambs. The Veterinary Record, 116(19), 527–527.

Wardrop, K. J., Reine, N., Birkenheuer, A., Hale, A., Hohenhaus, A., Crawford, C., & Lappin, M. R. (2005). Canine and feline blood donor screening for infectious disease. Journal of Veterinary Internal Medicine, 19(1), 135–142.

Webb, G. (2019). Canine and feline blood transfusions. The Veterinary Nurse, 10(3), 139–145.

Windberger, U., Pöschl, C., Peters, S., Huber, J., & Van den Hoven, R. (2017). Whole blood of mammalian species in the oscillating shear field: Influence of erythrocyte aggregation. In: IOP Conference Series: Materials Science and Engineering, 175(1), 012005.

Wybran, J., Carr, M. C., & Fudenberg, H. H. (1972). The human rosette-forming cell as a marker of a population of thymus-derived cells. The Journal of Clinical Investigation, 51(10), 2537–2543.

Yagi, K., & Holowaychuk, M. (2016). Manual of veterinary transfusion medicine and blood banking. Wiley Blackwell.

How to Cite
Holovina, K. M., Bobrova, O. M., Kovalenko, S. Y., Hovorova, Y. S., & Nardid, O. A. (2021). Effect of ozonation on resistance of ovine and human erythrocytes to hypothermic storage . Regulatory Mechanisms in Biosystems, 12(1), 116-120. https://doi.org/10.15421/022118