Two sets of primers were designed based on the target sequences of the corresponding group of hemoparasites.These primer sets were termed BH18S and EA16S. First,forward and reverse primers were designed to bind to the particular regions capable of discriminating hemoparasites, and the amplified products from these primers were subsequently used to design the sequencing primers. All primers were designed, and all quantitative parameters were evaluated for pyrosequencing with PyroMarkTMQ96 ID software version 2.0. The sequences and alignments of these 2 primer sets arepresented in Table 1 and Fig. 1.