UDC 636.4:619:578.835.2:615.371

doi: 10.15389/agrobiology.2014.4.70eng


N.I. Zakutskii, T.G. Shirokova, S.G. Yurkov, V.M. Balyshev

All-Russian Institute of Veterinary Virology and Microbiology, Russian Academy of Agricultural Sciences,
Pokrov, Petushinskii Region, Vladimir Province, 601120 Russia,
e-mail vniivvim@niiv.petush.elcom.ru

Received April 24, 2014

For many years, efforts of research scientists have been directed towards obtaining and testing of various attenuated variants of African swine fever (ASF) virus to develop specific methods for ASF prevention. Similar investigations were also carried out in the All-Russian Institute of Veterinary Virology and Microbiology which resulted in obtaining an attenuated ASF virus variant FC-135 through passaging a virulent strain France-32 of serotype 4 (V.A. Burlackov et al., 1979). The above strain grown in a porcine bone marrow (PBM) monolayer cell culture and taken at a 25-fold concentration provided 85 to 100 % protection from infection with a homologous virulent ASF virus. Because of higher manufacturability, performance and cost effectiveness of a suspension culture method as compared to those for a monolayer one, the investigation of antigenic and immunogenic activity of a virus-containing emulsified preparation produced in bioreactors from an ASF virus strain FC-135 grown in a PBM cell suspension was of some interest. The results of the reported research showed that when culturing the attenuated ASF virus strain FC-135 in PBM cell suspension in bioreactors, the highest virus accumulation levels of 7.50 to 8.00 lg HAU50/cm3 were observed after 4 day cultivation, and CF-antibody levels of 1:16 to 1:32 were indicated after 5 to 6 day cultivation. The titers of the virus grown in a PBM cell monolayer in mattresses were 0.25 to 0.50 lg HAU50/cm3 lower than those as seen in the previous case, although CF-antibody levels as observed within the same period were of 1:16. A concentrated emulsified preparation designed from the strain FC-135 grown in a PBM cell suspension using bioreactors provided protection of gilts against infection with a homologous virulent virus for 4 months (observation period). Also, we showed that the suspension method of the attenuated strain FC-135 culture in PBM cells using automated bioreactors, being more manufacturable, productive and cost-efficient as compared to the monolayer growth in mattresses, when applied under controlled conditions, provided production of homologous virus materials exhibiting high biological activity.

Keywords: African swine fever, ASF, ASF virus, porcine bone marrow cell culture, PBM cells, infectivity, haemadsorbing activity, complement-fixing (CF) activity, immunogenicity.


Full article (Rus)



1. Kovalenko YA.R., Sidorov M.A., Burba L.G. Afrikanskaya chuma svinei [African swine fever]. Moscow, 1972.
2. Hess W., Cox B., Heushele W., Stone S. Propagation and modification of African swine fever virus in cell cultures. Am. J. Vet. Res., 110, 1965: 141-146.
3. Burlakov V.A. Immunobiologicheskie svoistva virusa i problema razrabotki sredstv spetsificheskoi profilaktiki afrikanskoi chumy svinei. Doktorskaya dissertatsiya [Immonobiological propertied of African swine fever virus with respect to means for specific prevention. DSc Thesis]. Pokrov, 1979.
4. Vishnyakov I.F., Mitin N.I., Petrov Yu.I., Cheryatnikov L.L., Kiselev A.V., Burlakov V.A., Balyshev V.M., Fedorishchev I.V., Morgunov Yu.P. Materialy nauchno-prakticheskoi konferentsii  «Aktual'nye voprosy veterinarnoi virusologii» [Proc. Sci. Pract. Conf. «Current Problems of Veterinary Virology»]. Pokrov, 1995: 141-143.
5. Kiselev A.V., Rudobel'skii A.V., Zuev V.V. e.a. Materialy nauchnoi konferntsii VNIIVViM «Voprosy veterinarnoi virusologii, mikrobiologii i epizootologii» [Proc. Conf. «Veterinary Virology, Microbiology and Epizootology»]. Pokrov, 1992: 44-45.
6. Prudnikova E.I., Balyshev V.M., Belyanin S.A., Balysheva V.I. Materialy Mezhdunarodnoi nauchno-prakticheskoi konferentsii, posvyashchennoi 55-letiyu VNIIVViM [Proc. Int. Conf., devoted to 55th anniversary of VNIIVViM]. Pokrov, 2013: 54-57.
7. Kalantaenko Yu.F., Zhesterev V.I., Mishchanin V.A., Balyshev V.M. Attenuirovannyi shtamm virusa afrikanskoi chumy svinei 2-go serotipa dlya razrabotki diagnosticheskikh i vaktsinnykh preparatov. Patent RF ¹ 2452511. A61K39/187 S12N7/00. Zayavl. 16.11.2010. Opubl. 10.06.2012. Byul. ¹ 16 [Attenuated strain of African swine fever virus 2 serotype to develop diagnostic kits and vaccines. Patent RU № 2452511. A61K39/187 S12N7/00. Appl. 16.11.2010. Publ. 10.06.2012. Bul. № 16] .
8. Repin V.I., Cheryatnikov L.L., Petrov Yu.I., Kiselev A.V. Materialynauchno-prakticheskoikonferentsii «Aktual'nye voprosy veterinarnoi virusologii» [Proc. Sci. Pract. Conf. «Current Problems of Veterinary Virology»]. Pokrov, 1995: 140-141.
9. Ruban E.A., Solov'ev B.V., Gunin M.A. Materialy Vserossiiskoi nauchno - prakticheskoi konferentsii « Nauchnye osnovy proizvodstva veterinarnykh biologicheskikh preparatov» [Proc. All-Russia Conf. «Scientific Basis for Production of Veterinary Biopreparations»]. Shchelkovo, 2000: 290-293.
10. Slivko I.A. Immunobiologicheskie svoistva vaktsinnykh shtammov TS-80 i 7/1 BelNIEV - VGNKI virusa beshenstva. Avtoreferat kandidatskoi dissertatsii [Immunobiological features of TS-80 and 7/1 BelNiEV-VGNKI strains of the rabies virus. PhD Thesis]. Pokrov, 2003.