Abstract
Viability of spermatozoa in honey bee semen from individual drones was highly variable, based on an assay using dual fluorescent staining. Temperature, physical manipulation and the saline used during artificial insemination were evaluated for their impact on sperm viability. Temperature of the assay buffer in which semen was collected had much less effect on viability than the way in which the semen was collected. Washing semen into the buffer did less harm than sucking it up into a syringe. The highest viability levels were seen when semen was collected directly from seminal vesicles. Given that obtaining semen from the seminal vesicles is difficult, collection of semen with the highest viability is best done by washing semen directly from an ejaculated drone into a buffer, rather than using a syringe. The sodium chloride/antibiotic solution used for prophylactic purposes during artificial insemination improved sperm viability slightly.