Post-thaw Addition of Caffeine and/or Pentoxifylline Affect Differently Motility of Horse and Donkey-Cryopreserved Spermatozoa

To increase sperm motility, several molecules have been tested in mammals. Methylxanthines have shown effects on sperm motility, capacitation, and on in vitro fertilization processes. The aim of the study was to evaluate if the post-thaw addition of caffeine and/or pentoxifylline changes motility parameters of cryopreserved stallion and donkey spermatozoa. Straws derived from 14 horses and 7 donkeys were thawed and diluted in a milk-based extender to obtain the following final concentrations: CTR (control, no additives), CAF 5 (5 mM caffeine), CAF 10 (10 mM caffeine), PTX 5 (5 mM pentoxifylline), PTX 10 (10 mM pentoxifylline), CAF-PTX (5 mM caffeine and 5 mM pentoxifylline). Samples were evaluated immediately post-thaw and after 60 and 120 minutes of incubation at 37°C. In horses, overall total motility was significantly lower in CTR than in to CAF5, CAF-PTX, PTX5, PTX10, whereas progressive motility increased only in CAF5 and PTX5 (P <.05). No differences between control and treatments were seen for donkey semen. In CTR, during the first hour of incubation horses’ sperm cells showed a larger decrease than donkeys’ ones in all parameters (P <.05), except for lateral sperm head displacement. Thus, post-thaw motility and velocity decreased more sharply in horses than in donkeys. Caffeine and pentoxifylline-added post-thaw were able to increase the proportion of motile spermatozoa only for stallions and not for donkeys. Whether the improvement in post-thaw motility of equine spermatozoa may have an effect on in vivo or in vitro pregnancy rates remains to be determined.