INDIRECT HEMAGGLUTINATION TEST FOR DETECTION OF ANTIBODIES IN CATTLE NATURALLY INFECTED WITH MYCOPLASMAS

  • 1 January 1976
    • journal article
    • research article
    • Vol. 40  (1) , 20-29
Abstract
Stable mycoplasma antigens for the indirect hemagglutination test (IHA) were prepared employing glutaraldehyde treated sheep erythrocytes sensitized with Mycoplasma agalactiae ssp. bovis and M. bovigenitalium antigens. Employing these antigens, mycoplasma antibodies were detected in sera from cattle which had mastitic symptoms due to natural infection with either M. agalactiae ssp. bovis or M. bovigenitalium. A total of 200 cows from 4 herds were examined at varying intervals for the presence of M. agalactiae ssp. bovis and for the detection of antibody using growth inhibition and IHA tests. Mycoplasmas were isolated from 37 animals. Growth inhibiting antibody was detected from 56 of the 200 animals. In the IHA tests, antibody titers .gtoreq. 1:80 were detected in 148 animals, 76 of these having antibody titers .gtoreq. 1:160, while sera of 116 normal control animals had no growth inhibiting antibody and none had IHA antibody titers greater than 1:40. M. bovigenitalium was isolated from the milk of 3 of 26 animals in a 5th herd during an outbreak of mastitis. Growth inhibiting antibodies were demonstrated in the sera of 10 of the 26 animals. However, the IHA test detected antibody titers of .gtoreq. 1:160 in 13 animals and of 1:80 in 1 of the 26 animals. To determine the specificity of the IHA tests, M. agalactiae ssp. bovis and M. bovigenitalium antigens were reacted with rabbit hyperimmune typing sera produced against 12 species of bovine mycoplasmatales. Homologous antisera showed IHA antibody titers of 1:1280 and 1:2560 against M. agalactiae ssp. bovis and M. bovigenitalium, respectively, whereas heterologous antisera showed IHA antibody titers of .ltoreq. 1:20. Eight type-specific bovine antisera were reacted with M. agalactiae ssp. bovis and M. bovigenitalium antigens in homologous and heterologous tests. Homologous reactions showed IHA antibody titers .gtoreq. 1:320, whereas heterologous reactions showed IHA titers of .ltoreq. 1:20. This IHA test promises to be useful for the detection of bovine mycoplasma antibodies in sera from cattle infected with M. agalactiae ssp. bovis or M. bovigenitalium. The test is sensitive, reproducible and specific and the technique is relatively simple and rapid. The antigens were stable for at least 7 mo.