Abstract
A method of isolatingBabesia ovata merozoites from infected erythrocytes and an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-B. ovata antibodies were developed. AfterB. ovata-infected erythrocytes had been lysed using the nitrogen cavitation method, the merozoites were separated from erythrocyte components by differential centrifugation and density-gradient centrifugation. The light microscopic examination showed that the purified merozoites were morphologically intact and contained few contaminants. Sodium dodecyl sulfate-polyacrylamide electrophoretic (SDS-PAGE) analysis revealed that the merozoite fraction contained very little contamination with erythrocyte components. The merozoites thus obtained were sonicated and treated with Triton X-100 and then used as an antigen to measure anti-B. ovata antibodies in ELISA. The ELISA was more sensitive in detecting anti-B. ovata antibodies than was either the indirect fluorescent antibody test or the complement fixation test on sera from cattle infected withB. ovata.
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References
Allsopp BA, Wagner GG, Mathews KP, Kariavu CG (1977) Purification and characterization of precipitating antigens fromTheileria parva. J Gen Microbiol 100:319–328
Barry DN, Rodwell BJ, Timms P, McGregor W (1982) A microplate enzyme immunoassay for detecting and measuring antibodies toBabesia bovis in cattle serum. Aust Vet J 59:136–140
Dowben HS, Gaffey TA, Lynch PA (1968) Isolation of liver and muscle polyribosomes in high yield after cell disruption by nitrogen cavitation. FEBS Lett 2:1–3
Figueroa JV, Buning GM, Kinden DA (1990) Purification of the erythrocytic stage ofBabesia bigemina from cultures. Paraitol Res 76:675–680
Fujinaga T (1981a) Bovine babesiosis in Japan: clinical and clinicopathological studies on cattle experimentally infected withBabesia ovata. Jpn J Vet Sci 43:803–813
Fujinaga T (1981b) Effect of splenectomy and dexamethasone administration on cattle experimentally infected withBabesia ovata. Jpn J Vet Sci 44:71–88
Fujinaga T, Minami T (1981) Indirect fluorescent antibody and complement fixation tests in the diagnosis of bovine theileriosis and babesiosis in Japan. Vet Parasitol 8:115–126
Fujinaga T, Minami T, Ishihara T (1980) Serological relationship between a largeBabesia found in Japanese cattle andBabesia major, B. bigemina andB. bovis. Res Vet Sci 29:230
Gray MA, Luckins AG, Rae PF, Brown CGD (1980) Evaluation of enzyme immunoassay for serodiagnosis of infections withTheileria parva andT. annulata. Res Vet Sci 29:360–366
Hamburger J, Kreier JP (1980) Isolation of malaria parasites. In: Kreier EJP (ed) Malaria, Vol 3. Academic Press, New York, pp 1–65
Laemmli VK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Mahoney DF, Goodger BV (1981) The isolation ofBabesia parasites and their products from the blood. In: Ristic M, Kreier JP (eds) Babesiosis. Academic Press, New York, pp 323–335
Minami T, Yamabe K, Hayashi S, Ishihara T (1979) Serological relationship of JapaneseBabesia species andBabesia bigemina by the complement fixation and capillary-tube agglutination tests. Vet Parasitol 5:29–38
Nakamura Y, Shimizu S, Minami T, Ito S (1988a) Enzyme-linked immunosorbent assay using solubilised antigen for detection of antibodies toAnaplasma marginale. Trop Anim Health Prod 20:259–266
Nakamura Y, Shimizu S, Minami T, Ito S (1988b) Enzyme-linked immunosorbent assay for detection of antibodies toAnaplasma centrale. Jpn J Vet Sci 50:933–935
O'Donoghue PJ, Friedhoff KT, Vizcanio OG, Weyreter H (1985) The detection of IgM and IgG antibodies againstBabesia bigemina in bovine sera using semi-defined antigens in enzyme immunoassays. Vet Parasitol 18:1–12
Prior RB, Kreier JP (1972)Plasmodium berghei freed from host erythrocytes by a continuous-flow ultrasonic system. Exp Parasitol 32:239–243
Rodriguez SD, Buening GM, Vega CA, Carson CA (1986)Babesia bovis: purification and concentration of merozoites and infected bovine erythrocytes. Exp Parasitol 61:236–243
Sato M, Hirose T, Miyagami T, Sakurai H, Saito A, Suzuki N (1986) A rise and fall of humoral antibody in calves experimentally infected withBabesia ovata orTheileria sergenti. Jpn J Vet Sci 48:599–602
Shimizu S, Suzuki K, Nakamura K, Kadota K, Fujisaki K, Ito S, Minami T (1988) Isolation ofTheileria sergenti piroplasms from infected erythrocytes and development of an enzymelinked immunosorbent assay for serodiagnosis ofT. sergenti infections. Res Vet Sci 45:206–212
Taylor SM, Elliott CT, Kenny J (1986) Isolation of antigenic proteins from erythrocytes parasitized withBabesia divergens and comparison to their immunizing potential. Vet Parasitol 21:99–105
Wagner GG, Duffus WPH, Kimber CD, Lule M (1974) Immunological studies on East Coast fever: I. partial segregation and characterization of theTheileria parva piroplasm antigen. J Parasitol 60:854–859
Wallach DF, Conley M (1977) Altered membrane proteins of monkey erythrocytes infected with simian malaria. J Mol Med 2:119–136
Waltisbuhl DJ, Goodger BV, Wright IG, Commins MA, Mahoney DF (1987) An enzyme-linked immunosorbent assay to diagnoseBabesia bovis infection in cattle. Parasitol Res 73:126–131
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This work was supported in part by a Grant-in-Aid (Bio Media Program) from the Ministry of Agriculture, Forestry and Fisheries (BMP 92-IV-1-7)
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Shimizu, S., Shimura, K., Ito, S. et al. Babesia ovata: isolation from erythrocytes and development of an enzyme-linked immunosorbent assay for detection of antibodies. Parasitol Res 78, 684–688 (1992). https://doi.org/10.1007/BF00931521
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DOI: https://doi.org/10.1007/BF00931521