Bulletin "Veterinary biotechnology"

Veterynarna biotehnologija – Veterinary biotechnology, 2018, 32(1), 121-126 [in Ukrainian]. https://doi.org/10.31073/vet_biotech32(1)-14

KOVALENKO V.L., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.ZOTSENKO I.А.

State Scientific Control Institute of Biotechnology and strains (DNKIBSHM)

GARKAVENKO V.M., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

State Scientific and Research Institute of Laboratory Diagnostics and Veterinary and Sanitary Expertise

PONOMARENKO G.V., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.IHNATIEVA T.M., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

Kharkiv State Zooveterinary Academy

BALATSKYI Y.О., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

Bila Tserkva National Agrarian University

RESEARCH OF ANTIVIRUS AND TOXIC INFLUENCE DISINFECTANT ORGASPET IN CELLS CULTURE PKEV, CTP

Introduction. Toxicity research methods with the use of laboratory animals are inhumane, cost-intensive and inconsistent with the international standards of biotic evaluation of experiments. Therefore, a cell culture-based toxicity research method are more efficient in the nowadays research scientific practice than conducting experiments on animals.

The goal of the work. Examine of disinfectant Orgasept toxicity and antiviral actions on virus strain “Klon-B” of Auezky disease having been tested and adapted to continues culture cells of PKEV and CTP.

Materials and methods. A disinfection agent Orgasept (15 % lactic acid, 5,0 % benzylalkonium chloride, argentum nanoparticles) was the object of the research. The process and framework of the research were laid out and carried out according to recommendations of the usage. Its cache concentrations were action on during 60 min. In experiments used: continuous versenised culture cells of pig embryo ridney (PKEV) and continuous cell cultures of pig testicles (CTP) growed as monolayer on bottom microplate. For growing of continuous cell lines used of RPMI-1640, DMEM, GLA medius, bload serum of cattle.

Results of research and discussion. Orgasept in 1,0 % concentration on cell cultures of PKEV and CTP caused of cytotoxic effect that is why this concentration did not evaluate regarding disinfectant influence on virus of Auezky disease. In microplate wells with cell cultures in which added the means in less concentrations as toxicity threshold the cell monolayer remained unhurt and visually did not different from control wells with culture. As a result of our studies to determine the toxic effects of product after cell culture passaging by exposure of 0.1 % solution of Orhasept was found that the drug concentration did not affect the change in culture monolayer compared to control. Subsequent studies found that 0,1 – 0,5 % – Orhasept solutions showed of inactivating effect on Aujeszky disease virus. Therefore, the drug in these concentrations can be recommended for disinfection to prevent of viral diseases. For statistical analysis of the research results, in addition to visual evaluation of cell culture monolayer, the method of cells counting on each carrier with cell culture, taken in the experiment separately. Influenced of 0,1 % Orhasept concentration for 4 days on PKEV number of cells was 14730,0 ± 21,1, for 6 days 15910,0 ± 34,3. In the control experiment observed 16202,0 ± 24,3 PKEV cells. The same results with PTP cell culture where influenced of Orhasept in 0.1% concentration for 4 day PTP number of cells was 17520,0 ± 24,9, for 6 days 18350,0 ± 24,3. In the control experiment observed 19050,0 ± 34,4 cells of PTP. In these cases, the observed untoxic effect of this disinfectant.

Conclusions. The disinfecting agent Orgasept with 0,1 ‒ 0,5% concentration exposes no toxicity on cell cultures and an inactivating affect on Suid herpesvirus. Therefore, it can be applied for a preventive disinfection on veterinary medicine objects with animals in recommended doses and expositions.

Keywords: disinfectants, culture cells, toxicity, Aujeszky disease virus (ADV).

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