CSIRO Publishing blank image blank image blank image blank imageBooksblank image blank image blank image blank imageJournalsblank image blank image blank image blank imageAbout Usblank image blank image blank image blank imageShopping Cartblank image blank image blank image You are here: Journals > Reproduction, Fertility and Development   
Reproduction, Fertility and Development
  Vertebrate Reproductive Science & Technology
 
blank image Search
 
blank image blank image
blank image
 
  Advanced Search
   

Journal Home
About the Journal
Editorial Board
Contacts
Content
All Issues
Special Issues
Research Fronts
Sample Issue
For Authors
General Information
Instructions to Authors
Submit Article
Open Access
For Referees
Referee Guidelines
Review an Article
Annual Referee Index
For Subscribers
Subscription Prices
Customer Service
Print Publication Dates

blue arrow e-Alerts
blank image
Subscribe to our email Early Alert or RSS feeds for the latest journal papers.

red arrow Connect with us
blank image
facebook twitter youtube

red arrow Connect with SRB
blank image
facebook TwitterIcon

Affiliated Societies

RFD is the official journal of the International Embryo Transfer Society and the Society for Reproductive Biology.



Article << Previous     |     Next >>   Contents Vol 22(1)

199 IN VITRO EMBRYO PRODUCTION IN THE CLOUDED LEOPARD (NEOFELIS NEBULOSA)

C. E. Pope A, M. C. Gómez A and B. L. Dresser A

A Audubon Center for Research of Endangered Species, New Orleans, LA, USA;
B Department of Biological Sciences, University of New Orleans, New Orleans, LA, USA
   

Abstract
Export Citation
Cited by
Print
  


Abstract

The clouded leopard, a spotted, mid-sized cat native to southeast Asia, is classified as Endangered by the U.S. Fish and Wildlife Service. The Species Survival Plan (SSP) program has designated that clouded leopards in North American zoological institutions are a research population with the focus of enhancing management and developing assisted reproductive techniques. In this study, we examined (1) ovarian response to exogenous gonadotropin stimulation, (2) in vitro production of embryos by IVF and intracytoplasmic sperm injection (ICSI), and (3) in vivo developmental ability of in vitro-derived embryos. Eight females at Audubon Center for Research of Endangered Species (ACRES) in New Orleans, LA, were used as oocyte donors and embryo recipients. During non-estrual, but otherwise unknown, stages of the estrous cycle, females 4 to 10 years old at our first treatment were administered a total of 15 (n = 8) or 20 (n = 7) IU of porcine FSH (i.m; Sioux Biochemical Co., Sioux City, IA) daily for 4 days. On Day 5, 10 (n = 8) or 15 (n = 7) IU of porcine LH (i.m; Sioux Biochemical Co.) was given and laparoscopic oocyte retrieval (LOR) was done 24 h later. One, 2, or 3 LOR were performed on 3, 3, and 2 females, respectively (total n LOR = 15), 10 (67%) of which were done on females that were 8 to 12 years of age. A total of 176 pre-ovulatory oocytes (mean = 14.7; range = 3-31) were recovered from 12 LOR performed on 6 females. Two females, a 5-year-old and an 11-year-old, did not respond to gonadotropin treatment. Of 5 females with ≥2 LOR, an average of 16.2 and 14.6 oocytes were recovered at the first and second LOR, respectively. Luteal tissue was present on the ovaries at 3 of 4 LOR performed during January-April as compared with 0 of 11 LOR performed from June-December. Semen was obtained by electroejaculation of 3 males (6, 9, and 11 years of age) and used after cryopreservation [TEST-yolk (media prepared with egg yolk, TES, and Tris) + 6% glycerol] or after storage at 4°C for 24 h (cooled). In vitro production, cryopreservation, and transfer of embryos were done as described by Pope et al. (2006 Theriogenology 66, 1518-1524). Cleavage frequency was 43% overall, 55% (64/117) after IVF, and 20% after ICSI with frozen (10/43) or cooled (2/16) sperm. After IVF with frozen sperm, cleavage rate was 63% (48/76) v. 39% (16/41) with cooled sperm. On Days 5, 57, 26, and 17% of embryos were morula (> 16 cells), early morula (?16 cells) and pre-morula (< 16 cells), respectively. Forty-four Day 5 embryos were cryopreserved at a slow, controlled rate, 24 of which were thawed and transferred by laparotomy to the uteri of 3 Day 5 gonadotropin-treated recipients (7-9 embryos/female). Similarly, 28 Day 5 fresh embryos were auto-transferred to 3 Day 5 recipients (5-15 embryos/female). Most embryos transferred were morulae (83%) produced by IVF using frozen sperm (71%). None of the recipients established pregnancies, a failure that, undoubtedly, was multifaceted. Age may have been a factor because most recipients were 8 to 11 years old. Nonetheless, we have (1) shown that clouded leopard females are responsive to exogenous gonadotropins, (2) produced embryos in vitro, both by IVF and ICSI, using both cryopreserved and cooled sperm, and (3) provided further evidence of seasonality in ovarian activity.

Reproduction, Fertility and Development 22(5305) 258–258   http://dx.doi.org/10.1071/RDv22n1Ab199
Published online: 08 December 2009




 
Top  Email this page
 
   


Legal & Privacy | Contact Us | Help

CSIRO

© CSIRO 1996-2014