Veterynarna biotehnologija – Veterinary biotechnology, 2016, 29, 92-99 [in Ukrainian].

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

National University of Life and Environmental Sciences of Ukraine

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

Ltd. "BioTestLab"

SELECTION OF ADJUVANTS FOR THE CONSTRUCTION OF INACTIVATED VACCINE AGAINST INFECTIOUS BOVINE RHINOTRACHEITIS

Introduction. An important step to create an inactivated vaccine and make it effective along with the virus highly immunogenic strain is the selection of adjuvant that characterized with high sorbing abilities and potency. Therefore, during development of any vaccine the selection of the most effective immunostimulatory adjuvant is a necessary step.

The goal of the work. To test three samples of adjuvants for the vaccine composition to increase their potency. To conduct a comparative assessment and identify the most effective design of inactivated vaccines against infectious bovine rhinotracheitis.

Materials and methods. “BM” virus strain of infectious bovine rhinotracheitis was cultivated in continuous bovine kidney cells culture MDBK. Infectious activity of infectious bovine rhinotracheitis virus was 7.0 lg TCID50/cm³. For virus inactivation we used formaldehyde in final concentration 0.05 %. As adjuvants were used 3% aluminum hydroxide gel with glycerin, silicon emulsion and highly purified mineral oil mixed with aerosil. Samples of the vaccine were prepared mixing 40 % virus antigen and 60 % adjuvant. All vaccine samples undergone bacterial and fungal contamination control. Safety of produced vaccine samples was tested on white mice. To study vaccine samples we formed 3 test and 1 control groups of 2–3 months old rabbits (m=2–2.5 kg). Vaccine samples were injected intramuscularly at a dose of 1 cm³ twice with 14 days interval. Blood sampling was performed before the vaccine administration, in 14 days after the first injection, and in 7 and 14 days after the second injection. Presence of virus specific antibodies was detected in the virus neutralization test (VN).

Results of research and discussion. After immunization there were not registered any complications or negative reactions in rabbits of all groups. The level of antibodies in animals of the first group, where aluminum hydroxide with glycerin adjuvant was used and second group, where a silicon emulsion adjuvant was used, were almost equal, and by 14 days after revaccination were 6.87±0.18 lоg₂ and 7.02±0.15 lоg₂ respectively. The highest level of antibodies was in animals of the third group vaccinated with vaccine included mixture of highly purified mineral oil and aerosol. In 14 days after revaccination it was 9.0±0.13 log₂.

Conclusions and prospects for further research. Inactivated vaccine against infectious bovine rhinotracheitis designed using strain “BM” with infectious activity 7.0 lg TCID50/cm³ before inactivation, produced on MDBK cell culture and inactivated with formaldehyde included highly purified mineral oil mixed with aerosil has shown the best potency index. Perspectives for further research are to study the immunogenic response of vaccine samples on farm animals, calves and pregnant cows.

Keywords: adjuvant, infectious bovine rhinotracheitis, inactivated vaccine, immunogenicity of vaccine.

REFERENCES

1. Mashero, V.A. (2006). Infekcionnye bolezni teljat [Infectious diseases of calves]. Vitebsk: UO VGAVM [in Russian].
2. Straub, O.C. (1990). Bovine Rhinotracheitis Virus. Virus Infection of Ruminants. Amsterdam ets. Elsevier. sci. publ., 9, 71-108.
3. OIE Terrestrial Manual (2016). Chapter 2.4.12. — Infectious bovine rhinotracheitis / infectious pustular vulvovaginitis. oie.int. Retrieved from http://www.oie.int/international-standard-setting/terrestrial-manual/access-online/.
4. Hulyanych, M.M., Nedosekov, V.V., & Sobko, Y.A. (2016). Cultural Properties of Infectious Bovine Rhinotracheitis virus. Proceedings from The Conference “Non sibi, sed omnibus – not for themselves, but for everybody”: 2nd International Scientific Conference of Veterinary Medicine Students (15 May 2016). (p. 76). Warsaw: Warsaw University of Life Sciences.
5. Hulyanych, M.M., & Nedosekov, V.V. (2016). Porivniannia chutlyvosti pereshchepliuvanykh kultur klityn do virusu infektsiinoho rynotrakheitu velykoi rohatoi khudoby [Сomparison of sensitivity continuous cell cultures to the infectious bovine rhinotracheitis virus]. Naukovyi zhurnal «Molodyi vchenyi» – Scientific journal “Young Scientist”, 7 (34), 296-298 [in Ukraine].
6. Hulyanych, M.M., Nedosekov, V.V., & Kleimanov, I.S. (2015). Tekhnolohichni aspekty vyhotovlennia inaktyvovanykh vaktsyn proty infektsiinoho rynotrakheitu velykoi rohatoi khudoby [Тechnology aspects for production of inactivated vaccines against infectious bovine rhinotracheitis]. Naukovo-tekhnichnyi biuleten Naukovo-doslidnyi tsentr biobezpeky ta ekolohichnoho kontroliu resursiv APK – Scientific and technical bulletin Research Center biosafety and environmental control resources AIC, 3, 3, 58-63 [in Ukraine].
7. Medunicyn, N.V. (2004). Vakcinologija [Vaccinology]. M.: Triada-H [in Russian].
8. Reed, L.J., & Muench, H. (1938). A simple method of estimating fifty percent endpoints. Am. J. Hygiene, 27, 493-497.

Download full text in PDF