doi: 10.15389/agrobiology.2016.6.853eng

UDC 619:578:57.083.2



E.A. Balashova, E.I. Baryshnickova, V.M. Lyska, O.L. Kolbasova,
A.V. Lunitsin, D.V. Kolbasov

All-Russian Institute of Veterinary Virology and Microbiology, Federal Agency of Scientific Organizations, 1, ul. Akademika Bakuleva, pos. Vol’ginskii, Petushinskii Region, Vladimir Province, 601125 Russia, e-mail

Received July 31, 2016


Presently, due to the variety and diversity of economic and tourist ties of Russia, episodes of accidental or maybe purposeful (i.e., biological terrorism) entry of exotic infectious pathogens including Akabane disease to the Russian Federation should not be excluded. Akabane disease is a viral transmissible infection. Its recurrent outbreaks are characterized by abortions, still or premature births, or malformations (e.g., arthrogryposis and/or hydrocephaly) for calves, lambs and kids. Akabane disease virus can persist both in animal body and in vitro (e.g., in continuous cell lines). A study of the sensitivity of African green monkey kidney cell line to Akabane virus carried out earlier showed that Akabane virus caused definite cytopathic changes resulting in cell rounding followed by cytolysis and detachment of the cell monolayer within 48 hours post infection. In this paper we first reported on the cytomorphological changes caused by Akabane virus in the primary lamb synovial membrane diploid cell culture (LSMCC) prepared according to an earlier developed procedure, and on a suitability of this culture for the virus accumulation in titers sufficient for study and making diagnosis. It has been formerly determined that the lamb synovial membrane cell culture is sensitive to small ruminant lentiviruses like caprine arthritis encephalitis virus or Visna-Maedi virus in sheep. LSMCC was prepared using a method for tissue explant culture. On day 4 post inoculation of the cell monolayer with Akabane virus the cytopathic effect appeared which manifested as formation of symplasts that grew larger due to their fusion on day 5 to 6. The Akabane virus activity was 6.0±0.05 lg TCID50/cm3for strain V8935, and 5.0±0.05 lg TCID50/cm3 for strain P. As many as three passages and also the primary cell culture (after freezing) kept the virus-producing activity, and the Akabane virus retained its infective properties. The lamb synovial membrane cells can be re-cultured, and excessive diploid culture can be frozen to preserve and thawed as required. It is expedient to use a strain of diploid lamb synovial membrane cells deposited and patented earlier. One more advantage of the primary LSMCC as compared to monkey cell lines is that the latter ones may be a source of simian herpes B virus.

Keywords: lamb synovial membrane cell culture, Akabane disease, Orthobunyavirus.


Full article (Rus)

Full text (Eng)



  1. Omori T., Inaba Y., Kurogi H., Miura Y., Nobuto K., Motumoto M. Viral abortion arthrogryposis-hydranencephaly syndrome in cattle in Japan 1972-1974. Bull. Off. Int. Epiz., 1974, 81: 447-458.
  2. Kono R., Hirata M., Kaji M., Goto Y., Ikeda S., Yanase T., Kato T., Tanaka S., Tsutsui T., Imada T., Yamakawa M. Bovine epizootic encephalomyelitis caused by Akabane virus in southern Japan. Vet Res., 2008, 13: 4-20.
  3. Oem J.K., Yoon H.J., Kim H.R., Roh I.S., Lee K.H., Lee O.S., Bae Y.C. Genetic and pathogenic characterization of Akabane viruses isolated from cattle with encephalomyelitis in Korea. Vet. Microbiol., 2012, 158(3-4): 259-266 CrossRef
  4. Della-Porta A., Murray M., Cybinski D. Congenital bovine epizootic artrogryposis and hydranencephaly in Australia. Distribution of antibodies to Akabane virus in Australian cattle after the 1974 epixootic. Aust. Vet. J., 1976, 52: 496-501 CrossRef
  5. Kalmar E., Peleg B., Savir D. Arthogryposis-hydranencephaly syndrome in newborn cattle, sheep and goats. Serological survey for antibodies against the Akabane virus. Ref. Vet., 1975, 32: 47-54.
  6. Al-Busaidy S.M., Hamblin C., Taylor V. Neutralising antibodies to Akabane virus in free-living wild animals in Africa. Tropical Animal Health and Production, 1987, 19: 197-202.
  7. Al-Busaidy S.M., Mellor P.S., Taylor W.P. Prevalence of neutralizing antibodies to Akabane virus in Arabian Peninsula. Vet. Microbiol., 1988, 17(2): 141-149.
  8. Oya A. Annual reports of WHO Regional Reference Center for Arbovirus. National Institute of Health, Tokyo, 1971.
  9. Nakajima Y., Takahashi E., Konno S. Encephalomyelitis in mice experimentally infected with Akabane virus. Natl. Inst. Anim. Health Q. (Japan), 1979, 19: 47-52.
  10. Miah A., Spradbrow P. The growth of Akabane virus in chicken embryos. Research in Veterinary Science, 1978, 25: 253-254.
  11. Nakamura T., Matsuyama T., Okuno T., Oya A. Arbovirus antibody survey. Igaku no Ayumi (in Japanese), 1967, 60: 72-73.
  12. Nikitina E.G., Sal'nikov N.I., Balashova E.A., Tsybanov S.Zh., Kolbasov D.V. Akabane and Schmallenberg diseases: similarities and differences. Agricultural Biology, 2013, 4: 48-52 CrossRef (in Engl.).
  13. Matumoto M., Inaba Y. Akabane disease and Akabane virus. Kitasato Arch. Exp. Med., 1980, 53(1-2): 1-21.
  14. Kato T., Shirafuji H., Tanaka S., Sato M., Yamakawa M., Tsuda T., Yanase T. Bovine arboviruses in Culicoides biting midges and sentinel cattle in Southern Japan from 2003 to 2013. Transbound. Emerg. Dis., 2015, 63(6): 160-172.
  15. Motumoto M., Oya A., Ogata T., Kobayashi I., Nakamure T., Taka-
    hashi H., Kitacka M. Isolation of arbor viruses from mosquitoes collected at live-stook pens in Gumma Prefecture in 1959. Jap. J. Med. Sci. Biol., 1960, 13: 191-198.
  16. Brinner J., Tsuda T., Yadin H., Chai D., Stram Y., Kato T. Serological and clinical evidence of a teratogenic Simbu serogroup virus infection of cattle in Israel 2001-2003. Veterinaria Italiana, 2004, 40(3): 119-123.
  17. Yang D.K., Kim B.H., Kweon C.H., Nah J.J., Kim H.J., Lee K.W., Yang Y.J., Mun K.W. Serosurveillance for Japanese encephalitis, Akabane, and Aino viruses for Thoroughbred horses in Korea. J. Vet. Sci., 2008, 9(4): 381-385.
  18. Oem J.K., Kim Y.H., Kim S.H., Lee M.H., Lee K.K. Serological characteristics of affected cattle during an outbreak of bovine enzootic encephalomyelitis caused by Akabane virus. Trop. Anim. Health Prod., 2014, 46: 261-263.
  19. Oem J.K., Lee K.H., Kim H.R., Bae Y.C., Chung J.Y., Lee O.S., Roh I.S. Bovin epizootic encephalomyelitis caused by Akabane virus infection in Korea. J. Comp. Pathol., 2012, 147(2-3): 101-105 CrossRef
  20. Murray M.D. Akabane epizoonotis in New South Wales: evidence for the long-distance dispersal of the biting midge Culicoides brevitarsis. Australian Veterinary Journal, 1987, 64(10): 305-308.
  21. Yilmaz H., Hoffmann B., Turan N., Cizmecigil U.Y., Richt J.A., Van der Poel W.H. Detection and partial sequencing of Schmallenberg virus in cattle and sheep in Turkey. Vector-Borne and Zoonotic Diseases, 2014, 14: 223-225.
  22. Sellers R., Herniman K. Neutralising antibodies to Akabane virus in ruminants in Ciprus (transmitted by Culicoides midges). Trop. Anim. Health Prod., 1981, 13(1): 57-60.
  23. Sellers R., Pedgley D. Possible windborne spread to western Turkey of bluetongue virus in 1977 and of Akabane virus in 1979. Journal of Hygiene, 1995, 1: 149-158.
  24. Elhassan A.M., Mansour M.E., Shamon A.A., EI Hussein A.M. A serological survey of Akabane virus infection in cattle in Sudan. ISRN Veterinary Science, 2014: Article ID 123904 CrossRef
  25. Metselaar D., Robin Y. Akabane virus isolated in Kenya. Vet. Rec., 1976, 99(5): 86 CrossRef
  26. Anderson A.A., Campbell C.H. Experimental placental transfer of Akabane of virus in the hamster. Am. J. Vet. Res., 1978, 39: 301-304.
  27. Kurogi H., Inaba Y., Takahashi H., Sato K., Akashi H., Satoda K., Omori T. An attenuated strain of Akabane virus: a candidate for live virus vaccine. Natl. Inst. Anim. Health Q (Tokyo), 1979, 19(1-2): 12-22.
  28. Kurogi H., Inaba Y., Takahashi H., Sato K., Omori T., Miura Y., Goto Y.M., Fujiwara Y., Hatano Y., Kodama K., Fukuyama S., Sasaki N., Matumoto M.  Epizootik congenital artrogryposis-hydranencephaly syndrom in cattle of Akabane virus from affected fetuses. Arch. Virol., 1976, 51: 67-74.
  29. Malakhova M.S., Balashova E.A. Tezisy konferentsii «Voprosy veterinarnoi virusologii, mikrobiologii i epizootologii» [Proc. Conf. «Veterinary virology, microbiology and epizootology»]. Pokrov, 1990: 83-84 (in Russ.).
  30. Nikitina E.G., Sal’nikov N.I., Katorkin S.A., Balashova E.A., Tsybanov S.Zh., Kolbasov D.V., Lunitsin A.V. Detection of Akabane virus genome in organs and blood of experimentally infected cavies. Agricultural Biology, 2014, 6: 67-72 CrossRef (in Engl.).
  31. Kotova O.Yu., Khan E.O., Kushnir S.D., Ponomarev V.N., Yurkov S.D., Balashova E.A. NauchnyizhurnalKubGau (Krasnodar), 2012, 83(09). Available No date (in Russ.).
  32. Kotova O.Yu., Suvorova Yu.A., Kushnir S.D., Yurkov S.G., Balashova E.A. Veterinariya, 2015, 3: 57-59 (in Russ.).
  33. Baryshnikova E.I., Kolbasova O.L. Shtamm diploidnykh kletok sinovial'noi membrany yagnenka Ovis aries, ispol'zuemyi dlya virusologicheskikh issledovanii. Pat. RF 2507255. GNU NIIVViM Rossel'khozakademii. Zayavl. 23.07.2012. Opubl. 20.02.2014 [A strain of diploid cells of lamb Ovis aries synovial membrane for virological research. Patent RF 2507255. GNU NIIVViM RAS. Appl. July 23, 2012. Publ. February 20, 2014] (in Russ.).
  34. Sidel'nikov  G.D., Kolbasova O.L., Zhigaleva O.N., Tsybanov S.Zh., Kolbasov D.V. Veterinariya, 2009, 4: 52-55 (in Russ.).