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Home / Equine Research Bank / Clin Vaccine Immunol / Serum antibodies from a subset of horses positive for Babesi...
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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  • Evaluation Study
  • Journal Article
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.

Summary

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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

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                              Citations

                              This article has been cited 4 times.
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