title: Comparison of acetamide and dimethylsulfoxide as cryoprotectants of European brown hare (Lepus europaeus) spermatozoa.
abstract: The aim of the present study was to assess the effect of dimethylsulfoxide (DMSO) and acetamide on the post-thaw properties of hare semen and to perform an AI trial with frozen-thawed semen. Semen was collected under general anaesthesia by the electroejaculation method from 6 males. Immediately after collection, the semen was diluted with an extender containing the following components: 250 mM Tris, 80 mM citric acid, 70 mM glucose, 1.0 M DMSO, egg yolk (17% v/v) and kanamycin (80 mg/l); this extender was used for Protocol I (n=17). In Protocol II (n=15), the DMSO was replaced with 1.0 M acetamide. Immediately after thawing and after incubation for 90 and 180 min at 37 C, the properties of semen were evaluated by computer-assisted semen analysis, and the percentage of viable, acrosome intact spermatozoa was evaluated using flow cytometry. During the 3-h incubation, the percentages of motile spermatozoa and spermatozoa with progressive motility were significantly higher in Protocol I (P<0.01). Immediately after thawing, path and straight velocity were significantly higher in Protocol I (P<0.01), as was the curvilinear velocity (P<0.05). The amplitude of lateral head displacement was higher after 3-h incubation in Protocol I (P<0.05), and no differences in beat cross frequency were found between Protocol I and II at any incubation time. The percentage of viable, acrosome intact spermatozoa determined with flow cytometry was higher in Protocol I (P<0.01) at all incubation times. As a result of artificial insemination with the semen frozen with DMSO as a cryoprotectant, two out of three inseminated females delivered two healthy young each. Following artificial insemination with the semen frozen with acetamide as a cryoprotectant, two out of three inseminated females delivered one healthy young each. On the basis of the results, it should be stated that DMSO ensures better post-thaw properties of hare spermatozoa than acetamide.
authors.affiliation: Department and Clinic of Obstetrics, Ruminant Diseases and Animal Health Care, Wroclaw University of Environmental and Life Sciences, Plac Grunwaldzki, Wroclaw, Poland. rkozdrowski@wp.pl
authors.forename: Roland
authors.initials: R
authors.lastname: Kozdrowski
authors.forename: Andrzej
authors.initials: A
authors.lastname: Dubiel
authors.forename: Agniaszka
authors.initials: A
authors.lastname: Noszczyk-Nowak
mesh_terms: D000081:Acetamides / Q000494:pharmacology*; D020101:Acrosome Reaction / Q000187:drug effects / Q000502:physiology; D000818:Animals; D003080:Cold Temperature; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D005260:Female; D034621:Hares / Q000502:physiology; D007315:Insemination, Artificial / Q000662:veterinary; D008297:Male; D011247:Pregnancy; D018873:Pregnancy Rate; D055101:Semen Analysis / Q000379:methods / Q000662:veterinary; D012662:Semen Preservation / Q000379:methods; D013094:Spermatozoa
chemical_list: D000081:Acetamides; D003451:Cryoprotective Agents; C030686:acetamide; D004121:Dimethyl Sulfoxide