Freezability of Dog Semen after Collection in Field Conditions and Cooled Transport

SIMPLE SUMMARY: Dog semen freezing is gaining popularity, but it has to be performed in equipped facilities, which can be far from the place where the stud dog lives. To avoid animal movement, it seemed interesting to investigate whether freezing dog semen after 24 or 48 h of cooled transport to an equipped laboratory was possible when semen collection was performed in the field such as in local breeding kennels. The influence of two pre-freezing holding times (i.e., 24 or 48 h) and two holding diluents (solutions used to dilute semen before freezing) was evaluated. Post-thaw morphofunctional sperm features, such as motility, morphological integrity, and ability to bind female gametes, were assessed. No differences between times or diluents were observed, but motility tended to decrease in the samples frozen at 48 h. Since the insemination dose was based on the number of motile spermatozoa, a shorter pre-freezing time is advisable. Yet, considering that the rest of the morphofunctional parameters remained comparable between samples frozen after collection or after 24/48 h of transport, freezing after cooled transport is a good option for avoiding animal stress and for promoting a greater diffusion of semen cryopreservation. ABSTRACT: Dog semen freezing is gaining popularity, but it has to be performed in equipped facilities, which can be far from the place where the stud dog lives. The aim of this study was to evaluate whether freezing dog semen after 24 or 48 h of cooled transport to an equipped laboratory was possible when semen collection was performed in the field such as in local breeding kennels. Single ejaculates from different dogs (mixed breeds and ages) were collected. In Experiment I, 10 ejaculates were conventionally frozen using the Uppsala method or frozen after 24 or 48 h of storage in a Styrofoam transport box cooled by icepacks. In Experiment II, 10 ejaculates were used to assess the influence of two extenders (Uppsala chilling extender or freezing extender 1) used for semen dilution during the 24 or 48 h storage. Motility, morphology, membrane, and acrosome integrity were analyzed as well as spermatozoa zona-binding ability. No significant differences were observed among the frozen groups, regardless of freezing time (Experiment I) or extender (Experiment II). Motility at thawing, however, decreased in absolute value at 48 h. Freezing of freshly collected semen is the gold standard, but the results obtained in this study prompt the application of freezing after cooled transport for the long-term preservation of dog semen, especially if the transport can be organized in 24 h.