ANALYSIS OF STATIONARY CULTIVATION OF THE PASTEUR STRAIN OF RABIES VIRUS

511 Volume 5, Issue 10: Special Issue (EJAR) ISSN: 2181-2020 MPHAPP THE 6TH INTERNATIONAL SCIENTIFIC AND PRACTICAL CONFERENCE “MODERN PHARMACEUTICS: ACTUAL PROBLEMS AND PROSPECTS” TASHKENT, OCTOBER 17, 2025 in-academy.uz ANALYSIS OF STATIONARY CULTIVATION OF THE PASTEUR STRAIN OF RABIES VIRUS Nurmukhambetova A.B. Ustenova G.O. Krykbayev E.A. Akhmetsadykov N.N. JSC "Kazakh National Medical University named after S.D. Asfendiyarov", Almaty, Republic of Kazakhstan e-mail: [email protected], tel . +7(701)724-77-45 https://doi.org/10.5281/zenodo.17343650 Relevance Modern production of immunobiological preparations and diagnostic test systems primarily relies on economic efficiency. Traditionally, a stationary cultivation method is used to accumulate virus-containing material, which has a number of advantages associated primarily with the lack of need for expensive equipment , consumables , and greater opportunities for monitoring fungal and bacterial contamination, with low personnel qualifications. The aim of the research was to analyze stationary cultivation to determine the most effective ways of accumulation of viral antigen and control the kinetics of accumulation. Materials and methods Pasteur strain of rabies virus was obtained from the OIE (OIE) – the reference laboratory for rabies (Nancy, France). The strain is adapted to continuous cell cultures. The continuous cell culture BHK-21/C-13, presented by the laboratory "cell culture" of TOO Scientific and Production Enterprise "Antigen" (Kazakhstan, Almaty), was originally obtained from ATCC ® (CCL-10). Stationary cultivation of the Pasteur strain was carried out in ventilated culture mattresses on a monolayer of 23 day old BHK-21 cell culture, with different infection doses. Samples were kept for up to 96 hours in a thermostat. Daily microscopy was carried out with control of the level of cytopathic action . Control of the biological activity of the Pasteur strain of the rabies virus in animals was carried out by the method of sequential titration on the BHK-21 cell culture according to the Reed and Mench method. Results As a result of the study, it was revealed that the stationary growth phase is very long, and lasts from 24 to 30 hours, before the first signs of cytopathogenic action appear, this is due to the time of rabies virus replication in cells, followed by infection of neighboring cells of the monolayer. The exponential growth phase is presented from 24 to 72 hours, since the virus begins to massively infect the monolayer. The stationary growth phase is presented from 72 to 96 hours, and is distinguished by a slowdown in the accumulation of virus-containing material, since all cells on the monolayer are already infected, and further accumulation is limited. The slow growth phase is presented from 96 to 120 hours, and is distinguished by a decrease in the concentration of viable viruses, since the viruses that replicated in phase I are already beginning to die off. Table 1 - Result of stationary cultivation No. Name Growth phase/hour Concentration I II III IV 1 Ratio 1:10 Pasteur 24-30 24-72 72-96 96-120 6.00 lg TCD 50 /cm 3 512 Volume 5, Issue 10: Special Issue (EJAR) ISSN: 2181-2020 MPHAPP THE 6TH INTERNATIONAL SCIENTIFIC AND PRACTICAL CONFERENCE “MODERN PHARMACEUTICS: ACTUAL PROBLEMS AND PROSPECTS” TASHKENT, OCTOBER 17, 2025 in-academy.uz Conclusions. The conducted analysis of stationary cultivation will allow optimizing the conditions of stationary cultivation, with the introduction of antirabies immunobiological preparations and diagnostic test systems into technological processes. In particular, the biological activity of the Pasteur strain has been increased with 6.00 lg TCID 50 /cm 3 , which, when recalculated, is 1 million viral particles in 1 cm 3.