Bulletin "Veterinary biotechnology"

Veterynarna biotehnologija – Veterinary biotechnology, 2016, 28, 114-125 [in Ukrainian].

KRYVOSHIYA P.YU., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it., KOT L.B., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it., ROMANKO M.V., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

Experimental Station of Epizootology of the Institute of Veterinary Medicine of NAAS


Introduction. According to FAO Equine Infectious Anemia is registered in 28 countries. It is registered in the United States, Canada, countries of Latin America, Great Britain, Italy, Australia, and in some parts of Venezuela, Norway, France, almost on all territory of Mongolia. Sporadic cases are observed in Libya, Mexico and others.

The goal of the work was to analyze and summarize the regulatory base of developed countries, research sources and submit the information about current approaches to the prevention and elimination of Equine Infectious Anemia (EIA).

Materials and methods. During the research we used and processed available sources: national legislation of a number of countries and directives of supranational organizations, articles in scientific journals.

Results of research and discussion. By the standards of OIE for the prevention and elimination of Equine Infectious Anemia Coggins test and ELISA are accurate and reliable serological methods of diagnosis of EIA in horses. Positive ELISA results must be confirmed by Coggins test. Virus isolation and identification are performed in order to identify infected animals. The virus can be isolated in a white blood cells culture of healthy horses and identified in ELISA, RIF, by molecular genetics methods or bioassay on susceptible horses. Intravenous injection of 1.25 ml of whole blood usually pretty for infection. Clinical inspection carried out the experimentally infected animals within 45 days. Polymerase chain reaction (PCR) – a sensitive method used to identify the viral genome. The advantage of the method of RT-PCR is that the virus can be detected already on the third day after infection. Main methods of virus detection are serological. The conventional method for diagnosis is serologic test in the agar gel immunodiffusion – Coggins test. Control of horses on EIA in Coggins test – a mandatory requirement for international traffic. Additional methods of laboratory diagnosis are PCR, immunoblotting, IFA, RIF and others. The first two methods are used in the system of diagnostic tests for preventive and health measures in the US and Australia. Positive ELISA results should be confirmed in RDP or immunoblotting. Immunoblotting used as an additional method of diagnosis of EIA in case of obtaining questionable results in Coggins test or ELISA. This property is used for early laboratory diagnosis In the United States in 2006 approved the «Equine Infectious Anemia – the only diagnostic methods and measures for prevention and liquidation». Scheduled research of horses carried out least once a year. Coggins test and ELISA are the official diagnostic laboratory methods. Immunoblotting is used to clarify the questionable results. In Australia, the Department of Agriculture and Water Resources has recognized EIA as exotic disease. Australia State structures adhere to the requirements of «Terrestrial Animal Health Code» and «Manual of Diagnostic Tests and Vaccines for Terrestrial Animals» developed by OIE. On the territory of the EU member states are allowed to import horses from other countries safe from EIA. Infected animals are sent to slaughter, other re-examine, establish quarantine. If the results of two consecutive studies of all animals with an interval of three months remain negative, the quarantine can be canceled. Veterinary certificate or submitted a declaration of health must be received for all of horses. A regulatory document «Protection against Equine Infectious Anemia» was elaborated in Germany. Serologic, clinical, haematological and pathological studies of horses for EIA are conducted. Patients and, in some cases, suspicious about the disease horses are sent for slaughter and disposal. In France, prevention and elimination of diseases regulated by a number of documents (JORF 11/08/90, JORF 26/09/92, JORF 29/12/94, JORF 07/08/03). Horses from outside the EU and to sale are exploring in Coggins test. Infected animals slaughtered for 15 days, and for the abolition of restrictions two negative results in Coggins test with an interval of three months for all horses are required. In Italy, the Ministry of Health issued a decree on December 4, 1976 «Prevention of Equine Infectious Anemia». Animal suspicious about the disease and all the animals of quarantine zone, check in Coggins test. Horse farm can be considered safe about the EIA within three months no animals have symptoms, all animals over six months with negative results of two studies with an interval of 40 days in Coggins test, annual inspections did not reveal positively reacting animals, and all the horses have the certificates of health.

Conclusions and prospects for further research. The main method of diagnosis EIA is serologic test in the agar gel immunodiffusion (Coggins test), which in some cases (with dubious results of the first test examination, especially valuable horses) supplemented by alternative methods: ELISA, immunoblotting and RT-PCR.

Keywords: Coggins test, Equine Infectious Anemia, immunoblotting.


  1. Coggins L., & Norcross N.L. (1970). Immunodiffusion reaction in equine infectious anemia. Cornell Vet., 60, 330-335.
  2. Coggins L., Norcross N.L., & Nusbaum S.R. (1972). Diagnosis of equine infectious anemia by immunodiffusion test. Am. J. Vet. Res., 33, 11-18.
  3. Coggins L. (1975). Mechanism of viral persistence in in equine infectious anemia. Cornell Vet., 65, N 2-3, 151.
  4. Archambault D., Wang A.M., & Lacal J.C. (1989). Development of an enzyme-linked immunosorbent assay for equine [Review article: Advances in diagnosis of EIA Nat. Inst. Anim. Health]. J. Clin. Microbiol., 27, 1167-1173.
  5. Kono Y., Kobayashi K., & Fukunaga Y. (1973). Antigenic drift of equine infectious anemia virus in chronically infected horses. Arch. Virol., 41, 1-10.
  6. McConnell S., & Katada M. (1981). Transmission of equine infectious anemia virus from a horse negative to agar gel immunodiffusion test. EqVet., 13, 123-126.
  7. Valpotic T., Kostelan M., & Rudolf M. (1989). T- and B-lymphocytes of horses persistently infected with equine infectious anemia virus. Veterinary Research Communication, 13, 56-65.
  8. Evans K., Carpenter S., & Sevoin M. (1984). Detection of equine infectious anemia virus in horse leukocyte cultures taken from horses in various stages of equine infectious anemia viral infection. Am. J. Vet. Res., 45, 20-25.
  9. McGuire T.C., Crawford Т.В., & Hensen J.B. (1971). Immunofluorescent localization of equine infectious anemia virus in tissue. Am. J. Path., 62, 283-292.
  10. Rwambo P.M., Issel C.J., & Hussain K.A. (1990). In vitro isolation of neutralization escape mutant of equine infectious anemia virus (EIAV). Arch. Virol., 111, 275-280.
  11. Campbell C.L. (1977) Report of the Committee on Infectious Diseases of Horses. Proc. US Animal Health Association, 81, 312- 313.
  12. Issel C.J., Adams W.V., & Foil L.D. (1985). Prospective study of progeny of inapparent equine carriers of equine infectious anemia virus. Am. J. Vet. Res., 5, 1114-1116.
  13. O'Rourke I.U., Besola M.L., & McGuire T.C. (1991). Proviral sequences detected by polymerase chain reaction in peripheral blood cells of horses with equine infectious anemia lentivirus. Arch. Virol., 117, 109-119.
  14. Montelaro R.C., Parer H., & Orrega A. (1984). Antigenic variation during persistent infection by equine infectious anemia virus, a retrovirus. J. Biol. Chem., 259, 10539-10544.
  15. Kono Y., Sentsui H., & Murakami Y. (1976). Development of specific in vitro lymphocyte stimulation responses in horses infected with equine infectious anemia virus. Veterinary Microbiology, 1, 31-34.
  16. Nishimura M., & Narajima H. (1984). Structural proteins of equine infectious anemia virus and their antigenic activity. Amer. J. Vet. Res., 45, 5-10.
  17. Chong Y., Payne S.L., & Issel C.J. (1991). Characterization of the antigenic domains of the major core protein (p26) of equine infectious anemia virus. J. Virol., 65, 1007-1012.
  18. O'Rourke I.U., Perryman L.E., & McGuire T.C. (1988). Antiviral, anti-glycoprotein and neutralizing of antibodies of horses with equine infectious anemia virus. J. gemr. Virol., 3, 667-674.
  19. Boulanger P., Bamieter G., & Carrier S. (1972). Equine infectious anemia: Preparation of antigen extract liquid for the Agargel Immunodiffusion and Complement-fixation tests. Canadian Journal of comparative Med., 36, 116-122.
  20. Pearson J.E., & Gipson C.A. (1987). Standartization of equine infectious anemia immunodiffusion and ELISA tests and their application to control of the disease in the United States. Abstracts. World veterinary congr. Montreal, 23, 233-320.

Download full text in PDF