• 1

    Uilenberg G. International collaborative research: significance of tick-borne hemoparasitic diseases to world animal health. Vet Parasitol 1995;57:1941.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 2

    Dumler JS, Barbet AF, Bekker CPJ, et al. Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and ‘HGE agent’ as subjective synonyms of Ehrlichia phagocytophila. Int J Syst Evol Microbiol 2001;51:21452165.

    • Search Google Scholar
    • Export Citation
  • 3

    Kocan KM, de laFuente J, Guglielmone AA, et al. Antigens and alternatives for control of Anaplasma marginale infection in cattle. Clin Microbiol Rev 2003;16:698712.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 4

    Office International des Épizooties (OIE) Web site. Terrestrial Animal Health Code. Bovine anaplasmosis. Chapter 2.3.7. Available at: www.oie.int/eng/normes/mcode/code2006_back/ en_chapitre_2.3.7.htm. Accessed Mar 27, 2007.

    • Search Google Scholar
    • Export Citation
  • 5

    Kuttler KL, Simpson JE. Relative efficacy of two oxytetracycline formulations and doxycycline in the treatment of acute anaplasmosis in splenectomized calves. Am J Vet Res 1978;39:347349.

    • Search Google Scholar
    • Export Citation
  • 6

    Stewart CG, Immelman A, Grimbeek P, et al. The use of a short and long acting oxytetracycline for the treatment of Anaplasma marginale in splenectomized calves. J S Afr Vet Assoc 1979;50:8385.

    • Search Google Scholar
    • Export Citation
  • 7

    Eriks IS, Palmer GH, McGuire TC, et al. Detection and quantitation of Anaplasma marginale in carrier cattle by using a nucleic acid probe. J Clin Microbiol 1989;27:279284.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 8

    Reeves JD, Swift BL. Iatrogenic transmission of Anaplasma marginale in beef cattle. Vet Med Small Anim Clin 1977;72:911912.

  • 9

    Futse JE, Ueti MW, Knowles DP, et al. Transmission of Anaplasma marginale by Boophilus microplus: retention of vector competence in the absence of vector-pathogen interaction. J Clin Microbiol 2003;41:38293834.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 10

    Torioni de Echaide S, Knowles DP, McGuire TC, et al. Detection of cattle naturally infected with Anaplasma marginale by nested PCR and a competitive enzyme-linked immunosorbent assay using recombinant major surface protein 5. J Clin Microbiol 1998;36:777782.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 11

    Bradway DS, Torioni deEchaide S, Knowles DP, et al. Sensitivity and specificity of the complement fixation test for detection of cattle persistently infected with Anaplasma marginale. J Vet Diagn Invest 2001;13:7981.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 12

    Office International des Épizooties (OIE) Web site. Manual of standards for diagnostic tests and vaccines. Chapter 2.3.7. 5th ed. 2004. Available at: www.oie.int/eng/normes/mmanual/ a_00058. Accessed Oct 26, 2006.

    • Search Google Scholar
    • Export Citation
  • 13

    Veterinary Medical Research & Development (VMRD) Web site. Anaplasma antibody test kit, cELISA. Available at: www.vmrd. com/docs/tk/Anaplasma/Anaplasma_2_and_5-plate_circular_060505.pdf. Accessed Mar 27, 2007.

    • Search Google Scholar
    • Export Citation
  • 14

    Knowles DP, Torioni deEchaide S, Palmer GH, et al. Antibody against an Anaplasma marginale MSP5 epitope common to tick and erythrocyte stages identified persistently infected cattle. J Clin Microbiol 1996;34:22252230.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 15

    Coetzee JF, Apley MD, Kocan KM, et al. Comparison of three oxytetracycline regimens for the treatment of persistent Anaplasma marginale infections in beef cattle. Vet Parasitol 2005;127:6173.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 16

    Blouin EF, Barbet AF, Jooyoung Yi, et al.Establishment and characterization of an Oklahoma isolate of Anaplasma marginale in cultured Ixodes scapularis cells. Vet Parasitol 2000;87:301313.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 17

    Blouin EF, Kocan KM, de laFuente J, et al. Effect of tetracycline on development of Anaplasma marginale in cultured Ixodes scapularis cells. Vet Parasitol 2002;107:115126.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 18

    Martin WH, Richie WN. A microtiter technique for the complement fixation test for anaplasmosis. Proc Annu Meet U S Anim Health Assoc 1973;77:582592.

    • Search Google Scholar
    • Export Citation
  • 19

    USDA. A microtiter technique for the complement fixation test for anaplasmosis. Beltsville, Md: USDA, APHIS, Veterinary Services, 1974.

  • 20

    Thompson JR, Kersting KW, Wass WM, et al. Splenectomy in cattle via transthoracic approach. Am J Vet Res 1992;53:143144.

  • 21

    Agresti A, Coull BA. Approximate is better than “exact” for interval estimation of binomial proportions. Am Stat 1998;52:119126.

  • 22

    Le CT. Probability and probability models. In:Introductory biostatistics. Hoboken, NJ: Wiley-Interscience, 2003;118119.

  • 23

    Dohoo I, Martin W, Stryhn H. Screening and diagnostic tests. In:Veterinary epidemiologic research. Charlottetown, PE, Canada: AVC Inc, 2003;9192.

    • Search Google Scholar
    • Export Citation
  • 24

    Morris DD. Normal values for erythron data in ruminants and the horse. In:Smith BP, ed.Large animal internal medicine. 2nd ed. St Louis: Mosby-Year Book Inc, 1996;1996

    • Search Google Scholar
    • Export Citation
  • 25

    Goff WL, Stiller D, Roeder RA, et al. Comparison of a DNA probe, complement-fixation and indirect immunofluorescence tests for diagnosing Anaplasma marginale in suspected carrier cattle. Vet Microbiol 1990;24:381390.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 26

    Jones EW, Brock WE. Bovine anaplasmosis: its diagnosis, treatment, and control. J Am Vet Med Assoc 1966;149:16241633.

  • 27

    Jones EW, Kliewer IO, Norman BB, et al. Anaplasma marginale infection in young and aged cattle. Am J Vet Res 1968;29:535544.

  • 28

    Potgieter FT, Stoltsz WH. Anaplasmosis. In:Coetzer JAW, Tustin RC, ed.Infectious diseases of livestock. 2nd ed. Cape Town, South Africa: Oxford University Press, 2004:594610.

    • Search Google Scholar
    • Export Citation
  • 29

    Gale KR, Leatch G, DeVos AJ, et al. Anaplasma marginale: effect of challenge of cattle with varying doses of infected erythrocytes. Int J Parasitol 1996;26:14171420.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 30

    Visser ES, McGuire TC, Palmer GH, et al. The Anaplasma marginale msp5 gene encodes a 19-kilodalton protein conserved in all recognized Anaplasma species. Infect Immun 1992;60:51395144.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 31

    Dreher UM, Hofmann-Lehmann R, Meli ML, et al. Seroprevalence of anaplasmosis among cattle in Switzerland in 1998 and 2003: no evidence of an emerging disease. Vet Microbiol 2005;107:7179.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 32

    Munodzana D, McElwain TF, Knowles DP, et al. Conformational dependence of Anaplasma marginale major surface protein 5 surface-exposed B-cell epitopes. Infect Immun 1998;66:26192624.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 33

    Hofmann-Lehmann R, Meli ML, Dreher UM, et al. Concurrent infections with vector-borne pathogens associated with fatal hemolytic anemia in a cattle herd in Switzerland. J Clin Microbiol 2004;42:37753780.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 34

    Lin Q, Rikihisa Y, Feleks, et al.Anaplasma phagocytophilum has a functional msp2 gene that is distinct from p44. Infect Immun 2004;72:38833889.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 35

    de la Fuente J, Lew A, Lutz H, et al. Genetic diversity of Anaplasma species major surface proteins and implications for anaplasmosis serodiagnosis and vaccine development. Anim Health Res Rev 2005;6:7589.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 36

    Alderink FJ, Dietrick RA. Economic and epidemiological implications of anaplasmosis in Texas cattle herds. Proc Annu Meet U S Anim Health Assoc 1982;86:6675.

    • Search Google Scholar
    • Export Citation
  • 37

    de la Fuente J, Van DenBussche RA, Kocan KM. Molecular phylogeny and biogeography of North American isolates of Anaplasma marginale (Rickettsiaceae: Ehrlichieae). Vet Parasitol 2001;97:6576.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 38

    de la Fuente J, Blouin EF, Kocan KM. Infection of ticks with the intracellular rickettsia, Anaplasma marginale excludes infection with other genotypes. Clin Diagn Lab Immunol 2003;10:182184.

    • Search Google Scholar
    • Export Citation
  • 39

    Kocan KM, de laFuente J, Blouin EF, et al. Anaplasma marginale (Rickettsiales: Anaplasmataceae): recent advances in defining host-pathogen adaptations of a tick-borne rickettsia. Parasitology 2004;129 (suppl):S285S300.

    • Crossref
    • Search Google Scholar
    • Export Citation

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Comparison of the complement fixation test and competitive ELISA for serodiagnosis of Anaplasma marginale infection in experimentally infected steers

Johann F. CoetzeeDepartment of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011

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Peggy L. SchmidtDepartment of Production Medicine and Epidemiology, College of Veterinary Medicine, Western University of Health Sciences, Pomona, CA 91766-1854

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Michael D. ApleyDepartment of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011

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James B. ReinboldDepartment of Clinical Sciences, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506-5606

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Katherine M. KocanDepartment of Pathobiology, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK 74078-2007

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Abstract

Objective—To compare sensitivity of a complement fixation (CF) test and competitive ELISA (cELISA) for detection of Anaplasma marginale in experimentally infected steers.

Animals—40 crossbred (Angus-Simmental) steers.

Procedures—Steers were inoculated with 2.6 × 109 A marginale–infected erythrocytes (day 0). Blood samples were collected on days 9, 13, 20, 28, 34, 41, 61, 96, 126, and 156 days after inoculation. The percentage of parasitized erythrocytes (PPE) was determined by microscopic examination of stained blood films, and sera were evaluated with the CF test and cELISA by use of USDA-approved methods. Sensitivity and agreement (κ statistic) between the 2 methods were determined. Persistent infections were confirmed by inoculation of blood obtained from infected steers into susceptible, splenectomized calves.

Results—9 days after inoculation, sensitivity of the cELISA was 47.5%, whereas the CF test failed to identify seropositive steers. After day 13, sensitivity of the cELISA and CF test was 100% and 20%, respectively. During peak parasitemia (day 20), sensitivity of the cELISA and CF test was 100%. Thereafter, sensitivity of the CF test fluctuated between 7.5% and 37.5%, whereas sensitivity of the cELISA remained at 100%. Overall sensitivity of the cELISA and CF test was 94.8% and 26.5%, respectively (κ statistic, 0.039).

Conclusions and Clinical Relevance—The cELISA had superior sensitivity for serologic detection of A marginale.The CF test and cELISA each had a high percentage of false-negative results during the prepatent period. These findings are relevant for export certification and anaplasmosis prevention or eradication programs.

Abstract

Objective—To compare sensitivity of a complement fixation (CF) test and competitive ELISA (cELISA) for detection of Anaplasma marginale in experimentally infected steers.

Animals—40 crossbred (Angus-Simmental) steers.

Procedures—Steers were inoculated with 2.6 × 109 A marginale–infected erythrocytes (day 0). Blood samples were collected on days 9, 13, 20, 28, 34, 41, 61, 96, 126, and 156 days after inoculation. The percentage of parasitized erythrocytes (PPE) was determined by microscopic examination of stained blood films, and sera were evaluated with the CF test and cELISA by use of USDA-approved methods. Sensitivity and agreement (κ statistic) between the 2 methods were determined. Persistent infections were confirmed by inoculation of blood obtained from infected steers into susceptible, splenectomized calves.

Results—9 days after inoculation, sensitivity of the cELISA was 47.5%, whereas the CF test failed to identify seropositive steers. After day 13, sensitivity of the cELISA and CF test was 100% and 20%, respectively. During peak parasitemia (day 20), sensitivity of the cELISA and CF test was 100%. Thereafter, sensitivity of the CF test fluctuated between 7.5% and 37.5%, whereas sensitivity of the cELISA remained at 100%. Overall sensitivity of the cELISA and CF test was 94.8% and 26.5%, respectively (κ statistic, 0.039).

Conclusions and Clinical Relevance—The cELISA had superior sensitivity for serologic detection of A marginale.The CF test and cELISA each had a high percentage of false-negative results during the prepatent period. These findings are relevant for export certification and anaplasmosis prevention or eradication programs.

Contributor Notes

Drs. Coetzee and Apley's present address is Department of Clinical Sciences, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506-5606.

Supported in part by the Alberta Beef Producers, Alberta Cattle Feeders Association, Alberta Livestock Industry Development Fund, Merial Canada Incorporated, Norbrook Laboratories Limited, and the Oklahoma Agricultural Experiment Station (project No. 1669). Dr. Kocan was supported by the Walter R. Sitlington Endowed Chair for Food Animal Research. Dr. Coetzee was supported by the Lloyd Endowed Professorship.

The authors thank Dr. Doug Ensley for technical assistance during the animal phase of the study; Drs. Jo Fisher, Karl Kersting, and Jim Thompson for surgical assistance; and Erin Rienstra, Joy Yoshioka, and Dollie Clawson for technical laboratory assistance.

Address correspondence to Dr. Coetzee.