2.6 DNA sequencing and analysis
The PCR products that were E. bieneusi positive underwent bidirectional sequencing (Sangon Biotech Co., Ltd., China). Further PCR products were sequenced as necessary. Basic Local Alignment Search Tool (BLAST) and ClustalX 1.83 were used for genotyping of the E. bieneusi isolates by comparing the identified nucleotide sequence with the published GenBank sequences. The genotypes were labeled following established nomenclature based on the 243 bp of the ITS region of E. bieneusi .