title: The impact of ovarian stimulation protocol on oocyte quality, subsequent in vitro embryo development, and pregnancy after transfer to recipients in Eld's deer (Rucervus eldii thamin).
abstract: Propagating genetically valuable individuals through oocyte collection, in vitro fertilization (IVF) and embryo transfer is critical to maintain sustainable populations of the endangered Eld's deer. The objectives of this study were to assess the impact of exogenous FSH injections on (1) the number and in vitro competence of oocytes collected and (2) the developmental potential of resulting IVF embryos after transfer into recipients during the breeding season (February-April). In a pilot experiment, estrus synchronization was conducted in three surplus females (using intravaginal progesterone-releasing devices, CIDRG for 14 days and injections of buserelin (a GnRH agonist). Five days after CIDR removal, ovaries were excised, minced and a total of 133 oocytes were recovered. Following in vitro maturation (IVM) and IVF, 63% of the oocytes formed embryos but only 5% reached the blastocyst stage. In a subsequent study, follicle numbers and diameters were compared between synchronized does stimulated with 0 or 80 mg FSH (-FSH and +FSH; n = 8 does in each group) and oocytes collected either by laparoscopic ovum pick-up or ovariectomy. FSH stimulation increased the main follicular diameter from 2-3 mm to 4-5 mm (P < 0.05) but not the oocyte number (∼20/donor) or the percentage of good quality oocytes (57%) regardless of the treatment. FSH stimulation did not either affect the percentage of cleaved embryos after IVF (25-35%; P > 0.05). Lastly, embryos at the 2-to 8-cell stage (from either + FSH or -FSH groups) were transferred into the oviducts of 11 synchronized recipients. With the +FSH embryos, three pregnancies failed between 90 and 120 days of gestation and two fawns that were born preterm (Days 215 and 224 of gestation) died at birth. In the -FSH group one healthy female fawn was born on Day 234 of gestation. This is the first report of a successful in vitro embryo production and subsequent birth of a live Eld's deer fawn. Further investigations are required to improve IVM/IVF success and the developmental potential of the embryos.
authors.affiliation: Wildlife Reproductive Innovation Center, Bureau of Conservation and Research, Zoological Park Organization under the Royal Patronage of H.M. the King, Bangkok, 10300, Thailand.
authors.forename: Ampika
authors.initials: A
authors.lastname: Thongphakdee
authors.affiliation: AgResearch, Ltd., Ruakura Research Centre, Hamilton, New Zealand. Electronic address: debra.berg@agresearch.co.nz.
authors.forename: Debra K
authors.initials: DK
authors.lastname: Berg
authors.affiliation: Faculty of Veterinary Science, Chulalongkorn University, Bangkok, 10130, Thailand.
authors.forename: Theerawat
authors.initials: T
authors.lastname: Tharasanit
authors.affiliation: Faculty of Veterinary Medicine, Kasetsart University, Nakhon Pathom, 73140, Thailand.
authors.forename: Nikorn
authors.initials: N
authors.lastname: Thongtip
authors.affiliation: Wildlife Reproductive Innovation Center, Bureau of Conservation and Research, Zoological Park Organization under the Royal Patronage of H.M. the King, Bangkok, 10300, Thailand.
authors.forename: Wanlaya
authors.initials: W
authors.lastname: Tipkantha
authors.affiliation: Khao Kheow Open Zoo, 235, Bang Pra, Sri Racha, Chonburi, 20110, Thailand.
authors.forename: Chainarong
authors.initials: C
authors.lastname: Punkong
authors.affiliation: Khao Kheow Open Zoo, 235, Bang Pra, Sri Racha, Chonburi, 20110, Thailand; Faculty of Veterinary Science, Rajamangala University of Technology Tawan-ok, 43 Bang Pra, Sri Racha, Chonburi, 20110, Thailand.
authors.forename: Daraka
authors.initials: D
authors.lastname: Tongthainan
authors.affiliation: Khao Kheow Open Zoo, 235, Bang Pra, Sri Racha, Chonburi, 20110, Thailand.
authors.forename: Sakhon
authors.initials: S
authors.lastname: Noimoon
authors.affiliation: Khao Kheow Open Zoo, 235, Bang Pra, Sri Racha, Chonburi, 20110, Thailand.
authors.forename: Umaporn
authors.initials: U
authors.lastname: Maikeaw
authors.affiliation: Wildlife Reproductive Innovation Center, Bureau of Conservation and Research, Zoological Park Organization under the Royal Patronage of H.M. the King, Bangkok, 10300, Thailand.
authors.forename: Nudthakamol
authors.initials: N
authors.lastname: Kajornklin
authors.affiliation: Wildlife Reproductive Innovation Center, Bureau of Conservation and Research, Zoological Park Organization under the Royal Patronage of H.M. the King, Bangkok, 10300, Thailand.
authors.forename: Boripat
authors.initials: B
authors.lastname: Siriaroonrat
authors.affiliation: Smithsonian Conservation Biology Institute, National Zoological Park, Washington, DC, USA.
authors.forename: Pierre
authors.initials: P
authors.lastname: Comizzoli
authors.affiliation: Wildlife Reproductive Innovation Center, Bureau of Conservation and Research, Zoological Park Organization under the Royal Patronage of H.M. the King, Bangkok, 10300, Thailand.
authors.forename: Sumate
authors.initials: S
authors.lastname: Kamolnorranath
mesh_terms: D000818:Animals; D003670:Deer / Q000196:embryology*; D004624:Embryo Transfer / Q000662:veterinary; D047108:Embryonic Development / Q000187:drug effects*; D004973:Estrus Synchronization; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D059471:In Vitro Oocyte Maturation Techniques / Q000662:veterinary; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology; D010062:Ovulation Induction / Q000379:methods / Q000662:veterinary*
chemical_list: D005640:Follicle Stimulating Hormone
keywords: Eld's deer; Embryo development; Embryo transfer; IVF; Oocyte quality; Ovarian stimulation