Samples were
taken by cardiac puncture (1 ml), centrifuged at 15,000g and genomic
DNA was obtained from the buffy coat using the protocol mentioned
above. The infection was determined by MDE and MHCT
parasitological methods, and by PCR using the ITS1, 21/22-mer,
ESAG6/7 and TBR1/2 primers and 100 ng of DNA as template. In order
to compare all the methods used, a percentage (%) of detection
was calculated dividing the number of positive animals by the
experimentally infected animals, and then multiplying by 100.