Two young (ages: 15 and 16 yr;; studbook # 947 and # 939, respectively) parous female gorillas were initially primed with daily oral monophasic estrogen/progesterone treatment (Ovcon 35: Bristol-Myers Squibb Co., Princeton, NJ, USA) for 3 mo to synchronize their menstrual cycles. After withdrawing treatment, urine was tested daily for occult blood (Hemastix;; Baxter Healthcare Corp., Deerfield, IL, USA). On days 3–10 following the onset of menses (Day 1), ovarian activity was stimulated with 225 IU human FSH im (Repronex;; Ferring Pharmaceuticals, New South Wales, Australia). Then on Days 6–8 this treatment was combined with 25mg GnRH antagonist im (Antagon;; Organon, West Orange, NJ, USA) to prevent premature endogenous LH release. Final oocyte maturation was stimulated 36h after the last FSH/GnRH treatment with 10,000IU hCG im (Profasi;; Serono Lab., Hingham/Rockland, MA, USA) and oocyte retrieval was performed 36h post-hCG administration in sternal recumbency (knee-chest) using a 3–6mHz probe, 17-ga needles and 87mmHg (VMAR-5000 Regulated Vacuum Pump;; Cook Veterinary Products). Both gorillas displayed a thickened endometrium, and approximately 6 (# 947) to 10 (# 939) maturing follicles (10–15mm) were detected in each animal. In female # 947, one oocyte was collected but peritoneal fluid and pathology (hydrosalpinx) were also diagnosed and the right ovary showed no follicular development. A total of 3 oocytes were recovered in highly viscous follicular fluid containing massive amounts of cumulus cells. They were transported in a HEPES-buffered transport medium in a portable incubator (CryoLogic, Mulgrave, Victoria, Australia) at 37°C by airline immediately from Brownsville to Dallas, TX, USA, and within 6-h post-retrieval were fertilized by ICSI using cryopreserved sperm collected by rectal probe electrostimulation (age: est. 40yr ; studbook # 268). The fertilized oocytes were cultured in Gardner’s Sequential Medium at 37°C in 6% CO2 all three cleaved and developed to blastocysts by 115h post-ICSI. One high-quality expanding blastocyst was transported back to Brownsville, TX, and transferred transcervically into the oocyte donor (studbook # 939), and two fair-quality blastocysts were transported overnight to Omaha, NE, and transferred into a synchronized gorilla recipient (age: 29yr; studbook # 543/91). Weekly urine samples from the two embryo transfer recipients are being tested for pregnancy diagnosis using OvuQuick test strips (Quidel, San Diego, CA, USA). Acknowledgments: This research was supported in part by the Morris Animal Foundation (Ruth Morris Keesling Animal Health Study).
The safe use of the transvaginal ultrasound probe for transabdominal oocyte retrieval in patients with vaginally inaccessible ovaries