2.5 Genotyping of E. bieneusi
Nested PCR amplification of the ITS region was done to identify and genotype E. bieneusi using TaKaRa TaqDNA Polymerase along with genotype BEB6 DNA from deer (positive control) and 2 μL distilled water (negative control). Buckholt and colleagues designed the primers and cycle parameters (Buckholt et al., 2002). The PCR products were analyzed using 1.5% agarose gel electrophoresis, followed by visualization by DNAGREEN staining (Tiandz, Inc., China).