A nested approach was used to amplify a region of the Internal Transcribed Spacer 1 (ITS-1) of Trypanosoma spp. (~180-640 bp). Primers Tryp_3 (50- TGCAATTATTGGTCGCGC -30) and Tryp_4 (50- CTTTGCTGCGTTCTT -30) were used for the first round of PCR, while internal primers Tryp_1 (50- AAGCCAAGTCATCCATCG -30) and Tryp_2 (50- TAGAGGAAGCAAAAG -30) were used for the second round of PCR (Adams et al., 2006). PCR was performed with Taqpolymerase (TopBio, Prague, Czech Republic) using the following program: 1 min at 95 C, 35 cycles of 1 min at 94 C, 1 min at 54 C, 30 s at 72 C, and 5 min at 72C, and PCR products were resolved in ethidium-bromide stained agarose gels.