title: Sperm distribution and fertilization after unilateral and bilateral laparoscopic artificial insemination with frozen-thawed goat semen.
abstract: Generally, laparoscopic artificial insemination (LAI) provides a higher success rate than of cervical insemination in goats. However, the sperm distribution after LAI in goats remains unknown, particularly when frozen-thawed semen is used. This study evaluated the distribution of frozen-thawed goat spermatozoa after LAI and compared the effects of sperm numbers and deposition sites (unilateral and bilateral sites) on pregnancy rate. In experiment 1, the frozen-thawed spermatozoa were stained either with CellTracker Green CMFDA (CT-Green) or CellTracker Red CMPTX (CT-Red), and in vitro evaluations of viability and motility were performed. In experiment 2, the labeled spermatozoa were deposited via LAI into the left (CT-Green) and right (CT-Red) uterine horns (n = 4). After ovariohysterectomy (6 hours after insemination), the distributions of green- and red-colored spermatozoa were assessed via tissue section, flushing, and the oviductal contents were also collected. Experiment 3 was designed to test the pregnancy rates in a group of 120 does after LAI using different numbers of spermatozoa (60 and 120 × 10(6) sperm per LAI) and different deposition sites. The results demonstrated that the fluorochromes used in this study did not impair sperm motility or viability. Frozen-thawed goat spermatozoa can migrate transuterinally after LAI, as evidenced by the observations of both CT-Green- and CT-Red-labeled spermatozoa in both uterine horns. Lower numbers of spermatozoa (60 × 10(6)) that are inseminated unilaterally (either ipsilateral or contralateral to the site of ovulation) can efficiently be used for LAI in goats (with a 56.67% pregnancy rate).
authors.affiliation: Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand.
authors.forename: Nitira
authors.initials: N
authors.lastname: Anakkul
authors.affiliation: Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand.
authors.forename: Junpen
authors.initials: J
authors.lastname: Suwimonteerabutr
authors.affiliation: Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand.
authors.forename: Theerawat
authors.initials: T
authors.lastname: Tharasanit
authors.affiliation: The Office of the Commission on Agricultural Resource Education, Chulalongkorn University, Nan, Thailand.
authors.forename: Sarawanee
authors.initials: S
authors.lastname: Khunmanee
authors.affiliation: The Office of the Commission on Agricultural Resource Education, Chulalongkorn University, Nan, Thailand.
authors.forename: Paweena
authors.initials: P
authors.lastname: Diloksumpan
authors.affiliation: Reproduction and Development Biology Group, AgResearch Ltd, Ruakura Research Center, Hamilton, New Zealand.
authors.forename: Debra K
authors.initials: DK
authors.lastname: Berg
authors.affiliation: Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand. Electronic address: mongkol.t@chula.ac.th.
authors.forename: Mongkol
authors.initials: M
authors.lastname: Techakumphu
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000662:veterinary; D005260:Female; D005456:Fluorescent Dyes; D006041:Goats; D006358:Hot Temperature; D007315:Insemination, Artificial / Q000379:methods / Q000662:veterinary*; D010535:Laparoscopy / Q000662:veterinary*; D008297:Male; D011247:Pregnancy; D018873:Pregnancy Rate; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013076:Sperm Count / Q000662:veterinary; D013081:Sperm Motility; D014599:Uterus / Q000166:cytology*
chemical_list: D005456:Fluorescent Dyes
keywords: Frozen-thawed semen; Goat; Laparoscopic artificial insemination; Sperm distribution