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Bovine babesiosis is caused by
Babesia bovis and
B. bigemina in Argentina. These protozoans are prevalent north of parallel 30°S, where their natural vector
Rhipicephalus (
Boophilus)
microplus is widespread. To prevent babesiosis outbreaks in endemic areas, an increasing population of 4–10-month-old calves are vaccinated with low virulence
B. bovis R1A (BboR1A) and
B. bigemina S1A (BbiS1A) strains. In non-endemic areas, an additional calf population is also vaccinated and boostered as adults, before they are relocated to
R. microplus-endemic areas of the country.
Serological tests are currently utilized not only to determine the status of natural
Babesia spp. infections, but also to confirm the infection caused by vaccine strains. For this purpose, an indirect enzyme immunoassay (ELISA) based on the recombinant major surface antigen-2c (rMSA-2c) of
B. bovis expressed in
Escherichia coli, was standardized using sera from
Babesia spp. experimentally infected cattle. ELISA
rMSA-2c was validated using sera obtained weekly during 336 days from steers primed and boostered with BboR1A and/or BbiS1A on days 0 and 154, then compared with the immunofluorescent-antibody test (IFAT). Western blot (WB) protein analysis was used to confirm the specificity of the immune response to rMSA-2c.
The sensitivity and specificity for ELISA
rMSA-2c were 92 and 96% after the
Babesia spp. priming and 88 and 73% after the boostering immunization, respectively. The sensitivity and specificity for IFAT were 99 and 90% after priming and 92 and 98% after boostering, respectively. Unlike IFAT, ELISA
rMSA-2c detected a remarkable delayed booster response and a significant drop in specificity between 35 and 84 days after the booster immunization. Simultaneously, 87.5% of cattle boostered with
B. bigemina showed cross-reactions in the ELISA
rMSA-2c, particularly between 63 and 77 days after the inoculation. A reaction against
E. coli was observed, since bands of approximately 40 and/or 42
kDa were detected using sera from cattle before and after
Babesia spp. inoculations. ELISA
rMSA-2c showed to be useful between 42 and 98 days after priming with
Babesia spp. live vaccine to evaluate the success of infecting cattle. However, after boostering the test showed low specificity.