FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / J Virol / A live attenuated equine infectious anemia virus proviral va...
Journal of virology2003; 77(13); 7244-7253; doi: 10.1128/jvi.77.13.7244-7253.2003

A live attenuated equine infectious anemia virus proviral vaccine with a modified S2 gene provides protection from detectable infection by intravenous virulent virus challenge of experimentally inoculated horses.

Abstract: Previous evaluations of inactivated whole-virus and envelope subunit vaccines to equine infectious anemia virus (EIAV) have revealed a broad spectrum of efficacy ranging from highly type-specific protection to severe enhancement of viral replication and disease in experimentally immunized equids. Among experimental animal lentivirus vaccines, immunizations with live attenuated viral strains have proven most effective, but the vaccine efficacy has been shown to be highly dependent on the nature and severity of the vaccine virus attenuation. We describe here for the first time the characterization of an experimental attenuated proviral vaccine, EIAV(UK)deltaS2, based on inactivation of the S2 accessory gene to down regulate in vivo replication without affecting in vitro growth properties. The results of these studies demonstrated that immunization with EIAV(UK)deltaS2 elicited mature virus-specific immune responses by 6 months and that this vaccine immunity provided protection from disease and detectable infection by intravenous challenge with a reference virulent biological clone, EIAV(PV). This level of protection was observed in each of the six experimental horses challenged with the reference virulent EIAV(PV) by using a low-dose multiple-exposure protocol (three administrations of 10 median horse infectious doses [HID(50)], intravenous) designed to mimic field exposures and in all three experimentally immunized ponies challenged intravenously with a single inoculation of 3,000 HID(50). In contrast, naïve equids subjected to the low- or high-dose challenge develop a detectable infection of challenge virus and acute disease within several weeks. Thus, these data demonstrate that the EIAV S2 gene provides an optimal site for modification to achieve the necessary balance between attenuation to suppress virulence and replication potential to sufficiently drive host immune responses to produce vaccine immunity to viral exposure.
Get Full Text
Publication Date: 2003-06-14 PubMed ID: 12805423PubMed Central: PMC164776DOI: 10.1128/jvi.77.13.7244-7253.2003Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research presents a study in which an experimental vaccine for equine infectious anemia virus (EIAV) is developed, resulting in protection against the disease. The vaccine, named EIAV(UK)deltaS2, modifies the S2 accessory gene to limit in vivo replication without affecting in vitro growth properties, and its efficacy was tested in horses and ponies.

Background

  • Equine infectious anemia virus (EIAV) affects horses and other members of the equid family.
  • Previous vaccine studies against EIAV were mentioned and their limitations explained: whole-virus and envelope subunit vaccines have shown inconsistent results, ranging from highly type-specific protection to severe enhancement of viral replication and disease.
  • Vaccines using live attenuated viral strains have proven effective in some animal trials, but their efficacy depends on the nature and severity of the vaccine virus attenuation.

Creation of the EIAV(UK)deltaS2 Vaccine

  • The researchers created EIAV(UK)deltaS2, an attenuated proviral vaccine that inactivates the S2 accessory gene.
  • The purpose of modifying the S2 gene was to control the in vivo replication of the virus without affecting its in vitro growth properties.

Testing the EIAV(UK)deltaS2 Vaccine

  • The vaccine was tested on horses, and the results indicated that it sparked mature virus-specific immune responses by 6 months.
  • The immunized horses were then exposed to a virulent biological clone of EIAV, called EIAV(PV), via intravenous challenge. Regardless of the dosing protocol applied, all experimental animals showed protection from the disease and detectable infection.

Comparing the Results with Control Group

  • The researchers compared the results to naïve (unvaccinated) horses exposed to the same EIAV(PV) challenge. The unvaccinated horses developed a detectable infection and disease symptoms within several weeks.

Conclusions

  • The research showed the potential of the EIAV S2 gene as a site for modification to balance between attenuation to suppress virulence and replication potential to stimulate host immune responses. This creation resulted in a vaccine that can immunize against viral exposure.

It is clear that the research proposes a potential solution to effectively vaccinate horses and related species against EIAV.

Cite This Article

APA
Li F, Craigo JK, Howe L, Steckbeck JD, Cook S, Issel C, Montelaro RC. (2003). A live attenuated equine infectious anemia virus proviral vaccine with a modified S2 gene provides protection from detectable infection by intravenous virulent virus challenge of experimentally inoculated horses. J Virol, 77(13), 7244-7253. https://doi.org/10.1128/jvi.77.13.7244-7253.2003

Publication

Journal of virology
ISSN: 0022-538X
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 77
Issue: 13
Pages: 7244-7253

Researcher Affiliations

Li, Feng
  • Department of Molecular Genetics and Biochemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.
Craigo, Jodi K
    Howe, Laryssa
      Steckbeck, Jonathan D
        Cook, Sheila
          Issel, Charles
            Montelaro, Ronald C

              MeSH Terms

              • Animals
              • Antibody Formation
              • Equine Infectious Anemia / immunology
              • Equine Infectious Anemia / prevention & control
              • Horses
              • Immunity, Cellular
              • Infectious Anemia Virus, Equine / pathogenicity
              • Viral Proteins / genetics
              • Viral Vaccines / administration & dosage
              • Viral Vaccines / immunology
              • Virulence

              Grant Funding

              • R01 AI025850 / NIAID NIH HHS
              • R01 AI25850 / NIAID NIH HHS

              References

              This article includes 29 references
              1. Bogers WM, Cheng-Mayer C, Montelaro RC. Developments in preclinical AIDS vaccine efficacy models.. AIDS 2000;14 Suppl 3:S141-51.
                pubmed: 11086857
              2. Clements JE, Montelaro RC, Zink MC, Amedee AM, Miller S, Trichel AM, Jagerski B, Hauer D, Martin LN, Bohm RP. Cross-protective immune responses induced in rhesus macaques by immunization with attenuated macrophage-tropic simian immunodeficiency virus.. J Virol 1995 May;69(5):2737-44.
                pmc: PMC188966pubmed: 7707496doi: 10.1128/jvi.69.5.2737-2744.1995google scholar: lookup
              3. Coggins L, Norcross NL. Immunodiffusion reaction in equine infectious anemia.. Cornell Vet 1970 Apr;60(2):330-5.
                pubmed: 4986043
              4. Cole KS, Murphey-Corb M, Narayan O, Joag SV, Shaw GM, Montelaro RC. Common themes of antibody maturation to simian immunodeficiency virus, simian-human immunodeficiency virus, and human immunodeficiency virus type 1 infections.. J Virol 1998 Oct;72(10):7852-9.
                pmc: PMC110105pubmed: 9733822doi: 10.1128/jvi.72.10.7852-7859.1998google scholar: lookup
              5. Cole KS, Rowles JL, Jagerski BA, Murphey-Corb M, Unangst T, Clements JE, Robinson J, Wyand MS, Desrosiers RC, Montelaro RC. Evolution of envelope-specific antibody responses in monkeys experimentally infected or immunized with simian immunodeficiency virus and its association with the development of protective immunity.. J Virol 1997 Jul;71(7):5069-79.
                pmc: PMC191740pubmed: 9188572doi: 10.1128/jvi.71.7.5069-5079.1997google scholar: lookup
              6. Cole KS, Rowles JL, Murphey-Corb M, Clements JE, Robinson J, Montelaro RC. A model for the maturation of protective antibody responses to SIV envelope proteins in experimentally immunized monkeys.. J Med Primatol 1997 Feb-Apr;26(1-2):51-8.
                pubmed: 9271189doi: 10.1111/j.1600-0684.1997.tb00319.xgoogle scholar: lookup
              7. Cook RF, Leroux C, Cook SJ, Berger SL, Lichtenstein DL, Ghabrial NN, Montelaro RC, Issel CJ. Development and characterization of an in vivo pathogenic molecular clone of equine infectious anemia virus.. J Virol 1998 Feb;72(2):1383-93.
                pmc: PMC124617pubmed: 9445039doi: 10.1128/jvi.72.2.1383-1393.1998google scholar: lookup
              8. Hammond SA, Cook SJ, Falo LD Jr, Issel CJ, Montelaro RC. A particulate viral protein vaccine reduces viral load and delays progression to disease in immunized ponies challenged with equine infectious anemia virus.. Virology 1999 Feb 1;254(1):37-49.
                pubmed: 9927572doi: 10.1006/viro.1998.9550google scholar: lookup
              9. Hammond SA, Cook SJ, Lichtenstein DL, Issel CJ, Montelaro RC. Maturation of the cellular and humoral immune responses to persistent infection in horses by equine infectious anemia virus is a complex and lengthy process.. J Virol 1997 May;71(5):3840-52.
                pmc: PMC191535pubmed: 9094660doi: 10.1128/jvi.71.5.3840-3852.1997google scholar: lookup
              10. Hammond SA, Li F, McKeon BM Sr, Cook SJ, Issel CJ, Montelaro RC. Immune responses and viral replication in long-term inapparent carrier ponies inoculated with equine infectious anemia virus.. J Virol 2000 Jul;74(13):5968-81.
                pmc: PMC112093pubmed: 10846078doi: 10.1128/jvi.74.13.5968-5981.2000google scholar: lookup
              11. Hammond SA, Raabe ML, Issel CJ, Montelaro RC. Evaluation of antibody parameters as potential correlates of protection or enhancement by experimental vaccines to equine infectious anemia virus.. Virology 1999 Sep 30;262(2):416-30.
                pubmed: 10502520doi: 10.1006/viro.1999.9939google scholar: lookup
              12. Harrold SM, Cook SJ, Cook RF, Rushlow KE, Issel CJ, Montelaro RC. Tissue sites of persistent infection and active replication of equine infectious anemia virus during acute disease and asymptomatic infection in experimentally infected equids.. J Virol 2000 Apr;74(7):3112-21.
                pmc: PMC111810pubmed: 10708426doi: 10.1128/jvi.74.7.3112-3121.2000google scholar: lookup
              13. Issel CJ, Foil LD. Studies on equine infectious anemia virus transmission by insects.. J Am Vet Med Assoc 1984 Feb 1;184(3):293-7.
                pubmed: 6321420
              14. Issel CJ, Horohov DW, Lea DF, Adams WV Jr, Hagius SD, McManus JM, Allison AC, Montelaro RC. Efficacy of inactivated whole-virus and subunit vaccines in preventing infection and disease caused by equine infectious anemia virus.. J Virol 1992 Jun;66(6):3398-408.
                pmc: PMC241120pubmed: 1316455doi: 10.1128/jvi.66.6.3398-3408.1992google scholar: lookup
              15. Johnson RP, Lifson JD, Czajak SC, Cole KS, Manson KH, Glickman R, Yang J, Montefiori DC, Montelaro R, Wyand MS, Desrosiers RC. Highly attenuated vaccine strains of simian immunodeficiency virus protect against vaginal challenge: inverse relationship of degree of protection with level of attenuation.. J Virol 1999 Jun;73(6):4952-61.
                pmc: PMC112539pubmed: 10233957doi: 10.1128/jvi.73.6.4952-4961.1999google scholar: lookup
              16. Leroux C, Craigo JK, Issel CJ, Montelaro RC. Equine infectious anemia virus genomic evolution in progressor and nonprogressor ponies.. J Virol 2001 May;75(10):4570-83.
                pmc: PMC114210pubmed: 11312327doi: 10.1128/jvi.75.10.4570-4583.2001google scholar: lookup
              17. Leroux C, Issel CJ, Montelaro RC. Novel and dynamic evolution of equine infectious anemia virus genomic quasispecies associated with sequential disease cycles in an experimentally infected pony.. J Virol 1997 Dec;71(12):9627-39.
                pmc: PMC230271pubmed: 9371627doi: 10.1128/jvi.71.12.9627-9639.1997google scholar: lookup
              18. Li F, Leroux C, Craigo JK, Cook SJ, Issel CJ, Montelaro RC. The S2 gene of equine infectious anemia virus is a highly conserved determinant of viral replication and virulence properties in experimentally infected ponies.. J Virol 2000 Jan;74(1):573-9.
                pmc: PMC111574pubmed: 10590152doi: 10.1128/jvi.74.1.573-579.2000google scholar: lookup
              19. Li F, Puffer BA, Montelaro RC. The S2 gene of equine infectious anemia virus is dispensable for viral replication in vitro.. J Virol 1998 Oct;72(10):8344-8.
                pmc: PMC110207pubmed: 9733881doi: 10.1128/jvi.72.10.8344-8348.1998google scholar: lookup
              20. Lichtenstein DL, Rushlow KE, Cook RF, Raabe ML, Swardson CJ, Kociba GJ, Issel CJ, Montelaro RC. Replication in vitro and in vivo of an equine infectious anemia virus mutant deficient in dUTPase activity.. J Virol 1995 May;69(5):2881-8.
                pmc: PMC188985pubmed: 7707512doi: 10.1128/jvi.69.5.2881-2888.1995google scholar: lookup
              21. Montelaro R, Ball JM, Rushlow K. Equine retroviruses. p. 257-360. In J. A. Levy (ed.), Τhe Retroviridae. Plenum Press, New York, N.Y..
              22. Montelaro RC, Cole KS, Hammond SA. Maturation of immune responses to lentivirus infection: implications for AIDS vaccine development.. AIDS Res Hum Retroviruses 1998 Oct;14 Suppl 3:S255-9.
                pubmed: 9814952
              23. Montelaro RC, Grund C, Raabe M, Woodson B, Cook RF, Cook S, Issel CJ. Characterization of protective and enhancing immune responses to equine infectious anemia virus resulting from experimental vaccines.. AIDS Res Hum Retroviruses 1996 Mar 20;12(5):413-5.
                pubmed: 8882322doi: 10.1089/aid.1996.12.413google scholar: lookup
              24. Nilsson C, Mäkitalo B, Thorstensson R, Norley S, Binninger-Schinzel D, Cranage M, Rud E, Biberfeld G, Putkonen P. Live attenuated simian immunodeficiency virus (SIV)mac in macaques can induce protection against mucosal infection with SIVsm.. AIDS 1998 Dec 3;12(17):2261-70.
                pubmed: 9863867doi: 10.1097/00002030-199817000-00006google scholar: lookup
              25. Raabe ML, Issel CJ, Cook SJ, Cook RF, Woodson B, Montelaro RC. Immunization with a recombinant envelope protein (rgp90) of EIAV produces a spectrum of vaccine efficacy ranging from lack of clinical disease to severe enhancement.. Virology 1998 May 25;245(1):151-62.
                pubmed: 9614876doi: 10.1006/viro.1998.9142google scholar: lookup
              26. Rwambo PM, Issel CJ, Adams WV Jr, Hussain KA, Miller M, Montelaro RC. Equine infectious anemia virus (EIAV) humoral responses of recipient ponies and antigenic variation during persistent infection.. Arch Virol 1990;111(3-4):199-212.
                pubmed: 2162160doi: 10.1007/bf01311054google scholar: lookup
              27. Shen R-X, Wang Z. Development and use of an equine infectious anemia donkey leucocyte attenuated vaccine. EIAV: a national review of policies, programs, and future objectives. American Quarter Horse Association, Amarillo, Tex..
              28. Ui M, Kuwata T, Igarashi T, Ibuki K, Miyazaki Y, Kozyrev IL, Enose Y, Shimada T, Uesaka H, Yamamoto H, Miura T, Hayami M. Protection of macaques against a SHIV with a homologous HIV-1 Env and a pathogenic SHIV-89.6P with a heterologous Env by vaccination with multiple gene-deleted SHIVs.. Virology 1999 Dec 20;265(2):252-63.
                pubmed: 10600597doi: 10.1006/viro.1999.0049google scholar: lookup
              29. Warren J. Preclinical AIDS vaccine research: survey of SIV, SHIV, and HIV challenge studies in vaccinated nonhuman primates.. J Med Primatol 2002 Aug;31(4-5):237-56.
                pubmed: 12390546doi: 10.1034/j.1600-0684.2002.02010.xgoogle scholar: lookup

              Citations

              This article has been cited 23 times.
              1. Hull-Nye D, Meadows T, Smith SR, Schwartz EJ. Key Factors and Parameter Ranges for Immune Control of Equine Infectious Anemia Virus Infection.. Viruses 2023 Mar 6;15(3).
                doi: 10.3390/v15030691pubmed: 36992401google scholar: lookup
              2. Uversky VN. On the irrationality of rational design of an HIV vaccine in light of protein intrinsic disorder.. Arch Virol 2021 May;166(5):1283-1296.
                doi: 10.1007/s00705-021-04984-5pubmed: 33606110google scholar: lookup
              3. Ahmad I, Li S, Li R, Chai Q, Zhang L, Wang B, Yu C, Zheng YH. The retroviral accessory proteins S2, Nef, and glycoMA use similar mechanisms for antagonizing the host restriction factor SERINC5.. J Biol Chem 2019 Apr 26;294(17):7013-7024.
                doi: 10.1074/jbc.RA119.007662pubmed: 30862674google scholar: lookup
              4. de Pablo-Maiso L, Doménech A, Echeverría I, Gómez-Arrebola C, de Andrés D, Rosati S, Gómez-Lucia E, Reina R. Prospects in Innate Immune Responses as Potential Control Strategies against Non-Primate Lentiviruses.. Viruses 2018 Aug 17;10(8).
                doi: 10.3390/v10080435pubmed: 30126090google scholar: lookup
              5. Wang HN, Rao D, Fu XQ, Hu MM, Dong JG. Equine infectious anemia virus in China.. Oncotarget 2018 Jan 2;9(1):1356-1364.
                doi: 10.18632/oncotarget.20381pubmed: 29416700google scholar: lookup
              6. Wang XF, Lin YZ, Li Q, Liu Q, Zhao WW, Du C, Chen J, Wang X, Zhou JH. Genetic Evolution during the development of an attenuated EIAV vaccine.. Retrovirology 2016 Feb 3;13:9.
                doi: 10.1186/s12977-016-0240-6pubmed: 26842878google scholar: lookup
              7. Craigo JK, Ezzelarab C, Cook SJ, Liu C, Horohov D, Issel CJ, Montelaro RC. Protective efficacy of centralized and polyvalent envelope immunogens in an attenuated equine lentivirus vaccine.. PLoS Pathog 2015 Jan;11(1):e1004610.
                doi: 10.1371/journal.ppat.1004610pubmed: 25569288google scholar: lookup
              8. Craigo JK, Montelaro RC. Lessons in AIDS vaccine development learned from studies of equine infectious, anemia virus infection and immunity.. Viruses 2013 Dec 2;5(12):2963-76.
                doi: 10.3390/v5122963pubmed: 24316675google scholar: lookup
              9. Craigo JK, Ezzelarab C, Cook SJ, Chong L, Horohov D, Issel CJ, Montelaro RC. Envelope determinants of equine lentiviral vaccine protection.. PLoS One 2013;8(6):e66093.
                doi: 10.1371/journal.pone.0066093pubmed: 23785473google scholar: lookup
              10. Meng Q, Lin Y, Ma J, Ma Y, Zhao L, Li S, Yang K, Zhou J, Shen R, Zhang X, Shao Y. A pilot study comparing the development of EIAV Env-specific antibodies induced by DNA/recombinant vaccinia-vectored vaccines and an attenuated Chinese EIAV vaccine.. Viral Immunol 2012 Dec;25(6):477-84.
                doi: 10.1089/vim.2012.0014pubmed: 23171359google scholar: lookup
              11. Issel CJ, Scicluna MT, Cook SJ, Cook RF, Caprioli A, Ricci I, Rosone F, Craigo JK, Montelaro RC, Autorino GL. Challenges and proposed solutions for more accurate serological diagnosis of equine infectious anaemia.. Vet Rec 2013 Feb 23;172(8):210.
                doi: 10.1136/vr-2012-100735pubmed: 23161812google scholar: lookup
              12. Craigo JK, Barnes S, Cook SJ, Issel CJ, Montelaro RC. Divergence, not diversity of an attenuated equine lentivirus vaccine strain correlates with protection from disease.. Vaccine 2010 Nov 29;28(51):8095-104.
                doi: 10.1016/j.vaccine.2010.10.003pubmed: 20955830google scholar: lookup
              13. Liu L, Wan Y, Wu L, Sun J, Li H, Li H, Ma L, Shao Y. Broader HIV-1 neutralizing antibody responses induced by envelope glycoprotein mutants based on the EIAV attenuated vaccine.. Retrovirology 2010 Sep 1;7:71.
                doi: 10.1186/1742-4690-7-71pubmed: 20807451google scholar: lookup
              14. Qi X, Wang X, Wang S, Lin Y, Jiang C, Ma J, Zhao L, Lv X, Shen R, Wang F, Kong X, Su Z, Zhou J. Genomic analysis of an effective lentiviral vaccine-attenuated equine infectious anemia virus vaccine EIAV FDDV13.. Virus Genes 2010 Aug;41(1):86-98.
                doi: 10.1007/s11262-010-0491-6pubmed: 20526660google scholar: lookup
              15. Covaleda L, Fuller FJ, Payne SL. EIAV S2 enhances pro-inflammatory cytokine and chemokine response in infected macrophages.. Virology 2010 Feb 5;397(1):217-23.
                doi: 10.1016/j.virol.2009.11.005pubmed: 19945727google scholar: lookup
              16. Craigo JK, Barnes S, Zhang B, Cook SJ, Howe L, Issel CJ, Montelaro RC. An EIAV field isolate reveals much higher levels of subtype variability than currently reported for the equine lentivirus family.. Retrovirology 2009 Oct 20;6:95.
                doi: 10.1186/1742-4690-6-95pubmed: 19843328google scholar: lookup
              17. Mealey RH, Leib SR, Littke MH, Wagner B, Horohov DW, McGuire TC. Viral load and clinical disease enhancement associated with a lentivirus cytotoxic T lymphocyte vaccine regimen.. Vaccine 2009 Apr 21;27(18):2453-68.
                doi: 10.1016/j.vaccine.2009.02.048pubmed: 19368787google scholar: lookup
              18. Tagmyer TL, Craigo JK, Cook SJ, Even DL, Issel CJ, Montelaro RC. Envelope determinants of equine infectious anemia virus vaccine protection and the effects of sequence variation on immune recognition.. J Virol 2008 Apr;82(8):4052-63.
                doi: 10.1128/JVI.02028-07pubmed: 18234792google scholar: lookup
              19. Craigo JK, Zhang B, Barnes S, Tagmyer TL, Cook SJ, Issel CJ, Montelaro RC. Envelope variation as a primary determinant of lentiviral vaccine efficacy.. Proc Natl Acad Sci U S A 2007 Sep 18;104(38):15105-10.
                doi: 10.1073/pnas.0706449104pubmed: 17846425google scholar: lookup
              20. Craigo JK, Durkin S, Sturgeon TJ, Tagmyer T, Cook SJ, Issel CJ, Montelaro RC. Immune suppression of challenged vaccinates as a rigorous assessment of sterile protection by lentiviral vaccines.. Vaccine 2007 Jan 15;25(5):834-45.
                doi: 10.1016/j.vaccine.2006.09.040pubmed: 17023099google scholar: lookup
              21. Steckbeck JD, Grieser HJ, Sturgeon T, Taber R, Chow A, Bruno J, Murphy-Corb M, Montelaro RC, Cole KS. Dynamic evolution of antibody populations in a rhesus macaque infected with attenuated simian immunodeficiency virus identified by surface plasmon resonance.. J Med Primatol 2006 Aug;35(4-5):248-60.
                doi: 10.1111/j.1600-0684.2006.00173.xpubmed: 16872288google scholar: lookup
              22. Craigo JK, Li F, Steckbeck JD, Durkin S, Howe L, Cook SJ, Issel C, Montelaro RC. Discerning an effective balance between equine infectious anemia virus attenuation and vaccine efficacy.. J Virol 2005 Mar;79(5):2666-77.
                doi: 10.1128/JVI.79.5.2666-2677.2005pubmed: 15708986google scholar: lookup
              23. Jin S, Issel CJ, Montelaro RC. Serological method using recombinant S2 protein to differentiate equine infectious anemia virus (EIAV)-infected and EIAV-vaccinated horses.. Clin Diagn Lab Immunol 2004 Nov;11(6):1120-9.
                doi: 10.1128/CDLI.11.6.1120-1129.2004pubmed: 15539516google scholar: lookup
              Subscribe to our newsletter
              Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.