Semen collection, semen analysis and artificial insemination in Columbian sharp-tailed grouse (Tympanuchus phasianellus columbianus) as part of a species conservation project.

Columbian sharp-tailed grouse (Tympanuchus phasianellus columbianus; hereafter CSTG) have experienced substantial decreases in population numbers and geographic range during the early 20th century, primarily due to habitat loss. The conservation aim of this project was to re-establish a self-sustaining population of CSTG within an unoccupied portion of their historic range in northeastern Nevada via reintroduction from source populations in Idaho, USA. Female nest initiation rates post-translocation due to low fertilization rates are believed to be one limiting factor in the establishment of some translocated CSTG populations. However, studies on semen collection and artificial insemination in this species are absent. Assisted reproduction was evaluated as an additional tool in this species conservation project in order to gain knowledge on the reproductive status of yearling and adult male CSTG, establish orientation values for semen parameters and evaluate artificial insemination procedures on female CSTG. In two consecutive breeding seasons, semen collection was attempted 51 times in 47 males using the established massage method, and a novel electro-stimulation technique. Semen collection was successful in all attempts, even in yearling grouse, which represents a novel confirmation that yearling male CSTG can produce live spermatozoa in their first breeding season. Volume, color, consistency, contamination, pH of semen, and the motility, concentration, viability and morphology of spermatozoa were analyzed. Extracted semen volume ranged between 6 and 74 µl and the mean pH was 6.9 ±â€¯0.5 (x¯ ± SD). Morphology analysis revealed that on average, 42.8% of sperm was morphologically normal, but 34.8% had malformed tails. Additionally, artificial insemination was practiced in 17 females (sham-insemination group; insemination lacking spermatozoa) and performed in 17 females. Intravenous catheters G20 1.0 mm × 32 mm enabled safe intravaginal insemination under visual control. Significant (p ≤ 0.05) differences in semen parameters between adult and yearling birds were detected. It is well established that adult males receive a majority of copulations during lekking, but our novel findings also indicate that they produce significantly more spermatozoa which is of higher quality than yearling males. This finding offers insights into male reproductive biology in a lekking grouse species.