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Clinical and vaccine immunology : CVI2013; 20(11); 1752-1757; doi: 10.1128/CVI.00479-13

Serum antibodies from a subset of horses positive for Babesia caballi by competitive enzyme-linked immunosorbent assay demonstrate a protein recognition pattern that is not consistent with infection.

Abstract: Tick-borne pathogens that cause persistent infection are of major concern to the livestock industry because of transmission risk from persistently infected animals and the potential economic losses they pose. The recent reemergence of Theileria equi in the United States prompted a widespread national survey resulting in identification of limited distribution of equine piroplasmosis (EP) in the U.S. horse population. This program identified Babesia caballi-seropositive horses using rhoptry-associated protein 1 (RAP-1)-competitive enzyme-linked immunosorbent assay (cELISA), despite B. caballi being considered nonendemic on the U.S. mainland. The purpose of the present study was to evaluate the suitability of RAP-1-cELISA as a single serological test to determine the infection status of B. caballi in U.S. horses. Immunoblotting indicated that sera from U.S. horses reacted with B. caballi lysate and purified B. caballi RAP-1 protein. Antibody reactivity to B. caballi lysate was exclusively directed against a single ∼50-kDa band corresponding to a native B. caballi RAP-1 protein. In contrast, sera from experimentally and naturally infected horses from regions where B. caballi is endemic bound multiple proteins ranging from 30 to 50 kDa. Dilutions of sera from U.S. horses positive by cELISA revealed low levels of antibodies, while sera from horses experimentally infected with B. caballi and from areas where B. caballi is endemic had comparatively high antibody levels. Finally, blood transfer from seropositive U.S. horses into naive horses demonstrated no evidence of B. caballi transmission, confirming that antibody reactivity in cELISA-positive U.S. horses was not consistent with infection. Therefore, we conclude that a combination of cELISA and immunoblotting is required for the accurate serodiagnosis of B. caballi.
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Publication Date: 2013-09-18 PubMed ID: 24049108PubMed Central: PMC3837787DOI: 10.1128/CVI.00479-13Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • U.S. Gov't
  • Non-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 study reveals that tick-born Babesia caballi, typically nonendemic in the U.S., has shown up in some horses as per a nationwide survey. The study analyses the validity of using RAP-1-cELISA as an exclusive method for determining infection of the pathogen in U.S. horses. The findings suggest that cELISA positive horses in the U.S. displayed antibody reactivity not consistent with infection, indicating the need for an integrated approach involving cELISA and immunoblotting for accurate serodiagnosis.

Background

  • The research revolves around tick-borne pathogens that pose a huge risk to the livestock industry due to their potential transmission from persistently infected animals leading to significant economic loss.
  • A nationwide survey in the U.S. unveiled a surprising reemergence of Theileria equi and some degree of equine piroplasmosis in the horse population.
  • The survey also discovered horses testing positive for Babesia caballi using a test method called RAP-1-competitive enzyme-linked immunosorbent assay (cELISA), which is peculiar since B. caballi is generally considered nonendemic on the U.S. mainland.

Purpose of the Study

  • The study sought to determine the effectiveness of the RAP-1-cELISA as a single test in detecting B. caballi infection in U.S horses.

Research Methodology and Findings

  • Immunoblotting indicated that serum from U.S. horses reacted with B. caballi lysate and purified B. caballi RAP-1 protein.
  • However, the antibody reaction against the B. caballi lysate was exclusively directed against a single protein band approximately measuring 50-kDa, related to the native B. caballi RAP-1 protein.
  • In contrast, serum from horses in regions where B. caballi is endemic showed bound multiple proteins ranging from 30 to 50 kDa.
  • Dilution of serum from cELISA positive U.S. horses revealed low levels of antibodies as compared to serum obtained from horses in regions where B. caballi is endemic.
  • Blood transfer experiments from seropositive U.S. horses into naive horses showed no evidence of B. caballi transmission, confirming that the antibody reactivity in cELISA positive U.S. horses was not in line with the infection.

Conclusion

  • The results drawn suggest that merely relying on cELISA might not suffice in accurately diagnosing B. caballi in horses.
  • Therefore, the researchers concluded that a combined test methodology involving both cELISA and immunoblotting is required for precise serodiagnosis of B. caballi.

Cite This Article

APA
Awinda PO, Mealey RH, Williams LB, Conrad PA, Packham AE, Reif KE, Grause JF, Pelzel-McCluskey AM, Chung C, Bastos RG, Kappmeyer LS, Howe DK, Ness SL, Knowles DP, Ueti MW. (2013). Serum antibodies from a subset of horses positive for Babesia caballi by competitive enzyme-linked immunosorbent assay demonstrate a protein recognition pattern that is not consistent with infection. Clin Vaccine Immunol, 20(11), 1752-1757. https://doi.org/10.1128/CVI.00479-13

Publication

Clinical and vaccine immunology : CVI
ISSN: 1556-679X
NlmUniqueID: 101252125
Country: United States
Language: English
Volume: 20
Issue: 11
Pages: 1752-1757

Researcher Affiliations

Awinda, Peter O
  • Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, Washington, USA.
Mealey, Robert H
    Williams, Laura B A
      Conrad, Patricia A
        Packham, Andrea E
          Reif, Kathryn E
            Grause, Juanita F
              Pelzel-McCluskey, Angela M
                Chung, Chungwon
                  Bastos, Reginaldo G
                    Kappmeyer, Lowell S
                      Howe, Daniel K
                        Ness, SallyAnne L
                          Knowles, Donald P
                            Ueti, Massaro W

                              MeSH Terms

                              • Animals
                              • Antibodies, Protozoan / blood
                              • Antigens, Protozoan / immunology
                              • Babesia / immunology
                              • Babesiosis / diagnosis
                              • Babesiosis / immunology
                              • Babesiosis / veterinary
                              • Enzyme-Linked Immunosorbent Assay
                              • Horse Diseases / diagnosis
                              • Horse Diseases / immunology
                              • Horses
                              • Protozoan Proteins / immunology
                              • Serum / chemistry
                              • United States

                              References

                              This article includes 37 references
                              1. Waters WR, Buddle BM, Vordermeier HM, Gormley E, Palmer MV, Thacker TC, Bannantine JP, Stabel JR, Linscott R, Martel E, Milian F, Foshaug W, Lawrence JC. Development and evaluation of an enzyme-linked immunosorbent assay for use in the detection of bovine tuberculosis in cattle.. Clin Vaccine Immunol 2011 Nov;18(11):1882-8.
                                pmc: PMC3209037pubmed: 21918115doi: 10.1128/CVI.05343-11google scholar: lookup
                              2. Short MA, Clark CK, Harvey JW, Wenzlow N, Hawkins IK, Allred DR, Knowles DP, Corn JL, Grause JF, Hennager SG, Kitchen DL, Traub-Dargatz JL. Outbreak of equine piroplasmosis in Florida.. J Am Vet Med Assoc 2012 Mar 1;240(5):588-95.
                                pubmed: 22332629doi: 10.2460/javma.240.5.588google scholar: lookup
                              3. Rodning SP, Givens MD, Marley MS, Zhang Y, Riddell KP, Galik PK, Hathcock TL, Gard JA, Prevatt JW, Owsley WF. Reproductive and economic impact following controlled introduction of cattle persistently infected with bovine viral diarrhea virus into a naive group of heifers.. Theriogenology 2012 Oct 15;78(7):1508-16.
                                pubmed: 22980084doi: 10.1016/j.theriogenology.2012.05.031google scholar: lookup
                              4. Knowles DP Jr. Control of Babesia equi parasitemia.. Parasitol Today 1996 May;12(5):195-8.
                                pubmed: 15275213doi: 10.1016/0169-4758(96)10007-7google scholar: lookup
                              5. Grause JF, Ueti MW, Nelson JT, Knowles DP, Kappmeyer LS, Bunn TO. Efficacy of imidocarb dipropionate in eliminating Theileria equi from experimentally infected horses.. Vet J 2013 Jun;196(3):541-6.
                                pubmed: 23199699doi: 10.1016/j.tvjl.2012.10.025google scholar: lookup
                              6. Schwint ON, Ueti MW, Palmer GH, Kappmeyer LS, Hines MT, Cordes RT, Knowles DP, Scoles GA. Imidocarb dipropionate clears persistent Babesia caballi infection with elimination of transmission potential.. Antimicrob Agents Chemother 2009 Oct;53(10):4327-32.
                                pmc: PMC2764191pubmed: 19620328doi: 10.1128/AAC.00404-09google scholar: lookup
                              7. Eriks IS, Stiller D, Palmer GH. Impact of persistent Anaplasma marginale rickettsemia on tick infection and transmission.. J Clin Microbiol 1993 Aug;31(8):2091-6.
                                pmc: PMC265702pubmed: 8370734doi: 10.1128/jcm.31.8.2091-2096.1993google scholar: lookup
                              8. Ueti MW, Mealey RH, Kappmeyer LS, White SN, Kumpula-McWhirter N, Pelzel AM, Grause JF, Bunn TO, Schwartz A, Traub-Dargatz JL, Hendrickson A, Espy B, Guthrie AJ, Fowler WK, Knowles DP. Re-emergence of the apicomplexan Theileria equi in the United States: elimination of persistent infection and transmission risk.. PLoS One 2012;7(9):e44713.
                                doi: 10.1371/journal.pone.0044713pmc: PMC3435266pubmed: 22970295google scholar: lookup
                              9. de Waal DT. Equine piroplasmosis: a review.. Br Vet J 1992 Jan-Feb;148(1):6-14.
                                pubmed: 1551016doi: 10.1016/0007-1935(92)90061-5google scholar: lookup
                              10. Scoles GA, Ueti MW, Noh SM, Knowles DP, Palmer GH. Conservation of transmission phenotype of Anaplasma marginale (Rickettsiales: Anaplasmataceae) strains among Dermacentor and Rhipicephalus ticks (Acari: Ixodidae).. J Med Entomol 2007 May;44(3):484-91.
                                pubmed: 17547235doi: 10.1603/0022-2585(2007)44[484:cotpoa]2.0.co;2google scholar: lookup
                              11. Ueti MW, Palmer GH, Kappmeyer LS, Statdfield M, Scoles GA, Knowles DP. Ability of the vector tick Boophilus microplus to acquire and transmit Babesia equi following feeding on chronically infected horses with low-level parasitemia.. J Clin Microbiol 2005 Aug;43(8):3755-9.
                                pmc: PMC1233951pubmed: 16081906doi: 10.1128/JCM.43.8.3755-3759.2005google scholar: lookup
                              12. Scoles GA, Hutcheson HJ, Schlater JL, Hennager SG, Pelzel AM, Knowles DP. Equine piroplasmosis associated with Amblyomma cajennense Ticks, Texas, USA.. Emerg Infect Dis 2011 Oct;17(10):1903-5.
                                pmc: PMC3310643pubmed: 22000367doi: 10.3201/eid1710.101182google scholar: lookup
                              13. Knowles DP Jr, Gorham JR. Advances in the diagnosis of some parasitic diseases by monoclonal antibody-based enzyme-linked immunosorbent assays.. Rev Sci Tech 1993 Jun;12(2):425-33.
                                pubmed: 8400383doi: 10.20506/rst.12.2.692google scholar: lookup
                              14. McGuire TC, Van Hoosier GL Jr, Henson JB. The complement-fixation reaction in eguine infectious anemia: demonstration of inhibition by IgG (T).. J Immunol 1971 Dec;107(6):1738-44.
                                pubmed: 5000991
                              15. Wagner B, Miller DC, Lear TL, Antczak DF. The complete map of the Ig heavy chain constant gene region reveals evidence for seven IgG isotypes and for IgD in the horse.. J Immunol 2004 Sep 1;173(5):3230-42.
                                pubmed: 15322185doi: 10.4049/jimmunol.173.5.3230google scholar: lookup
                              16. Schwint ON, Knowles DP, Ueti MW, Kappmeyer LS, Scoles GA. Transmission of Babesia caballi by Dermacentor nitens (Acari: Ixodidae) is restricted to one generation in the absence of alimentary reinfection on a susceptible equine host.. J Med Entomol 2008 Nov;45(6):1152-5.
                                pubmed: 19058641doi: 10.1603/0022-2585(2008)45[1152:tobcbd]2.0.co;2google scholar: lookup
                              17. Kappmeyer LS, Perryman LE, Hines SA, Baszler TV, Katz JB, Hennager SG, Knowles DP. Detection of equine antibodies to babesia caballi by recombinant B. caballi rhoptry-associated protein 1 in a competitive-inhibition enzyme-linked immunosorbent assay.. J Clin Microbiol 1999 Jul;37(7):2285-90.
                                pmc: PMC85139pubmed: 10364599doi: 10.1128/JCM.37.7.2285-2290.1999google scholar: lookup
                              18. Packham AE, Conrad PA, Wilson WD, Jeanes LV, Sverlow KW, Gardner IA, Daft BM, Marsh AE, Blagburn BL, Ferraro GL, Barr BC. Qualitative evaluation of selective tests for detection of Neospora hughesi antibodies in serum and cerebrospinal fluid of experimentally infected horses.. J Parasitol 2002 Dec;88(6):1239-46.
                                pubmed: 12537119doi: 10.1645/0022-3395(2002)088[1239:QEOSTF]2.0.CO;2google scholar: lookup
                              19. Duarte PC, Ebel ED, Traub-Dargatz J, Wilson WD, Conrad PA, Gardner IA. Indirect fluorescent antibody testing of cerebrospinal fluid for diagnosis of equine protozoal myeloencephalitis.. Am J Vet Res 2006 May;67(5):869-76.
                                pubmed: 16649923doi: 10.2460/ajvr.67.5.869google scholar: lookup
                              20. Ness SL, Peters-Kennedy J, Schares G, Dubey JP, Mittel LD, Mohammed HO, Bowman DD, Felippe MJ, Wade SE, Shultz N, Divers TJ. Investigation of an outbreak of besnoitiosis in donkeys in northeastern Pennsylvania.. J Am Vet Med Assoc 2012 Jun 1;240(11):1329-37.
                                pubmed: 22607601doi: 10.2460/javma.240.11.1329google scholar: lookup
                              21. Friedhoff KT. [Piroplasmas of horses--impact on the international horse trade].. Berl Munch Tierarztl Wochenschr 1982 Oct 1;95(19):368-74.
                                pubmed: 6756387
                              22. Friedhoff KT, Tenter AM, Müller I. Haemoparasites of equines: impact on international trade of horses.. Rev Sci Tech 1990 Dec;9(4):1187-94.
                                pubmed: 2132711
                              23. Bhoora R, Quan M, Zweygarth E, Guthrie AJ, Prinsloo SA, Collins NE. Sequence heterogeneity in the gene encoding the rhoptry-associated protein-1 (RAP-1) of Babesia caballi isolates from South Africa.. Vet Parasitol 2010 May 11;169(3-4):279-88.
                                pubmed: 20138703doi: 10.1016/j.vetpar.2010.01.009google scholar: lookup
                              24. Böse R, Daemen K. Demonstration of the humoral immune response of horses to Babesia caballi by western blotting.. Int J Parasitol 1992 Aug;22(5):627-30.
                                pubmed: 1399247doi: 10.1016/0020-7519(92)90011-9google scholar: lookup
                              25. Tyler JW, Cullor JS. Titers, tests, and truisms: rational interpretation of diagnostic serologic testing.. J Am Vet Med Assoc 1989 Jun 1;194(11):1550-8.
                                pubmed: 2666372
                              26. Al-Adhami B, Scandrett WB, Lobanov VA, Gajadhar AA. Serological cross-reactivity between Anaplasma marginale and an Ehrlichia species in naturally and experimentally infected cattle.. J Vet Diagn Invest 2011 Nov;23(6):1181-8.
                                pubmed: 22362799doi: 10.1177/1040638711425593google scholar: lookup
                              27. Terkawi MA, Jia H, Gabriel A, Goo YK, Nishikawa Y, Yokoyama N, Igarashi I, Fujisaki K, Xuan X. A shared antigen among Babesia species: ribosomal phosphoprotein P0 as a universal babesial vaccine candidate.. Parasitol Res 2007 Dec;102(1):35-40.
                                pubmed: 17823817doi: 10.1007/s00436-007-0718-1google scholar: lookup
                              28. Dubey JP, Davis SW, Speer CA, Bowman DD, de Lahunta A, Granstrom DE, Topper MJ, Hamir AN, Cummings JF, Suter MM. Sarcocystis neurona n. sp. (Protozoa: Apicomplexa), the etiologic agent of equine protozoal myeloencephalitis.. J Parasitol 1991 Apr;77(2):212-8.
                                pubmed: 1901359
                              29. Marsh AE, Barr BC, Packham AE, Conrad PA. Description of a new Neospora species (Protozoa: Apicomplexa: Sarcocystidae).. J Parasitol 1998 Oct;84(5):983-91.
                                pubmed: 9794642
                              30. Sam-Yellowe TY. Rhoptry organelles of the apicomplexa: Their role in host cell invasion and intracellular survival.. Parasitol Today 1996 Aug;12(8):308-16.
                                pubmed: 15275182doi: 10.1016/0169-4758(96)10030-2google scholar: lookup
                              31. Suarez CE, Noh S. Emerging perspectives in the research of bovine babesiosis and anaplasmosis.. Vet Parasitol 2011 Aug 4;180(1-2):109-25.
                                pubmed: 21684084doi: 10.1016/j.vetpar.2011.05.032google scholar: lookup
                              32. Brayton KA, Lau AO, Herndon DR, Hannick L, Kappmeyer LS, Berens SJ, Bidwell SL, Brown WC, Crabtree J, Fadrosh D, Feldblum T, Forberger HA, Haas BJ, Howell JM, Khouri H, Koo H, Mann DJ, Norimine J, Paulsen IT, Radune D, Ren Q, Smith RK Jr, Suarez CE, White O, Wortman JR, Knowles DP Jr, McElwain TF, Nene VM. Genome sequence of Babesia bovis and comparative analysis of apicomplexan hemoprotozoa.. PLoS Pathog 2007 Oct 19;3(10):1401-13.
                                doi: 10.1371/journal.ppat.0030148pmc: PMC2034396pubmed: 17953480google scholar: lookup
                              33. Cheadle MA, Lindsay DS, Rowe S, Dykstra CC, Williams MA, Spencer JA, Toivio-Kinnucan MA, Lenz SD, Newton JC, Rolsma MD, Blagburn BL. Prevalence of antibodies to Neospora sp. in horses from Alabama and characterisation of an isolate recovered from a naturally infected horse [corrected].. Int J Parasitol 1999 Oct;29(10):1537-43.
                                pubmed: 10608440doi: 10.1016/s0020-7519(99)00140-xgoogle scholar: lookup
                              34. Mullaney T, Murphy AJ, Kiupel M, Bell JA, Rossano MG, Mansfield LS. Evidence to support horses as natural intermediate hosts for Sarcocystis neurona.. Vet Parasitol 2005 Oct 10;133(1):27-36.
                                pubmed: 15970386doi: 10.1016/j.vetpar.2005.05.016google scholar: lookup
                              35. Torian LV, Forgione LA, Punsalang AE, Pirillo RE, Oleszko WR. Comparison of Multispot EIA with Western blot for confirmatory serodiagnosis of HIV.. J Clin Virol 2011 Dec;52 Suppl 1:S41-4.
                                pubmed: 21995935doi: 10.1016/j.jcv.2011.09.017google scholar: lookup
                              36. Trevejo RT, Krause PJ, Sikand VK, Schriefer ME, Ryan R, Lepore T, Porter W, Dennis DT. Evaluation of two-test serodiagnostic method for early Lyme disease in clinical practice.. J Infect Dis 1999 Apr;179(4):931-8.
                                pubmed: 10068589doi: 10.1086/314663google scholar: lookup
                              37. Alvarez I, Gutierrez G, Ostlund E, Barrandeguy M, Trono K. Western blot assay using recombinant p26 antigen for detection of equine infectious anemia virus-specific antibodies.. Clin Vaccine Immunol 2007 Dec;14(12):1646-8.
                                pmc: PMC2168377pubmed: 17959820doi: 10.1128/CVI.00293-07google scholar: lookup

                              Citations

                              This article has been cited 4 times.
                              1. Lei R, Wang X, Zhang D, Liu Y, Chen Q, Jiang N. Rapid isothermal duplex real-time recombinase polymerase amplification (RPA) assay for the diagnosis of equine piroplasmosis. Sci Rep 2020 Mar 5;10(1):4096.
                                doi: 10.1038/s41598-020-60997-1pubmed: 32139744google scholar: lookup
                              2. Lobanov VA, Peckle M, Massard CL, Brad Scandrett W, Gajadhar AA. Development and validation of a duplex real-time PCR assay for the diagnosis of equine piroplasmosis. Parasit Vectors 2018 Mar 2;11(1):125.
                                doi: 10.1186/s13071-018-2751-6pubmed: 29499748google scholar: lookup
                              3. Ueti MW, Olafson PU, Freeman JM, Johnson WC, Scoles GA. A Virulent Babesia bovis Strain Failed to Infect White-Tailed Deer (Odocoileus virginianus). PLoS One 2015;10(6):e0131018.
                                doi: 10.1371/journal.pone.0131018pubmed: 26083429google scholar: lookup
                              4. Mendoza FJ, Pérez-Écija A, Kappmeyer LS, Suarez CE, Bastos RG. New insights in the diagnosis and treatment of equine piroplasmosis: pitfalls, idiosyncrasies, and myths. Front Vet Sci 2024;11:1459989.
                                doi: 10.3389/fvets.2024.1459989pubmed: 39205808google scholar: lookup
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