We sought to further characterize this parasite by microscopy, PCR, culturing and DNA sequencing. Honey bee intestines were dissected in a sterile environment from which Crithidia mellificae was cultured. Light microscopy of these parasites enabled visualization of the flagella and motility ( Figure 6 ; Figure S6 and Figure S7 ). Fixed sample imaging facilitated DAPI visualization of the kinetoplast DNA, as well as nuclear DNA ( Figure 6 ). Previous studies describing trypanosomatids in honey bees lacked DNAsequencing data with the exception of Cox-Foster et al. (2007) who published a 715-nt sequence of 18S ribosomal RNA that was too conserved between trypanosomatids for precise taxonomic assignment [9] . Together, the features observed by microscopy (flagella and kinetoplast) and phylogenetic analysis unambiguously identify this species taxonomically. We have deposited the GADPH sequence (JF423199) for future molecular identification, and genomic sequencing of C. mellificae is underway.