Fertilizing ability of canine spermatozoa cryopreserved with skim milk-based extender in a retrospective study

We previously reported that skim milk (SM) is an effective cryoprotectant for cryopreservation of canine spermatozoa instead of egg yolk (EY), which is the conventional cryoprotectant. In this study, the fertilizing ability and practical use of frozen canine spermatozoa prepared with SM were evaluated by transcervical insemination. Frozen-thawed spermatozoa were inseminated one to four times on days 2-9 after the LH surge. In SM group, a single transcervical insemination (TCI) on Day 5 led to higher delivery rate (83%) than any other days (33%-50%) post-LH surge. In EY group, delivery rate in double TCI on days 5 and 6 (71%) was higher compared to any other experimental groups (0%-44%). Regardless of single or double, TCI on Day 5 or Day 6 led to higher litter sizes in SM or EY groups, respectively. The breeding efficiency and litter size of single TCI on Day 5 (4.2) and double TCI on Day 5 and Day 6 (3.7) were significantly higher than in the other experimental groups in SM and EY groups, respectively (p < .05). These findings suggest that skim milk is a suitable alternative to egg yolk for cryopreservation of canine spermatozoa, and the suitable timing for insemination might be on Day 5 post-LH surge.