Vitrification of oocytes from endangered Mexican gray wolves (Canis lupus baileyi).

Careful genetic management, including cryopreservation of genetic material, is central to conservation of the endangered Mexican gray wolf. We tested a technique, previously used to vitrify human and domestic animal oocytes, on oocytes from domestic dogs as a model and from the endangered Mexican wolf. This method provided a way to conserve oocytes from genetically valuable older female Mexican wolves as an alternative to embryos for preserving female genes. Oocytes were aspirated from ovaries of 36 female dogs in December and March (0 to 65 oocytes per female) and from six female wolves (4 to 73 per female) during their physiologic breeding season, or following stimulation with the GnRH agonist deslorelin. Oocytes from dogs were pooled; half were immediately tested for viability and the remainder vitrified, then warmed and tested for viability. All oocytes were vitrified by being moved through media of increasing cryoprotectant concentration, placed on Cryotops, and plunged into liquid nitrogen. There was no difference in viability (propidium iodide staining) between fresh and vitrified, warmed dog oocytes (65.7 and 61.0%, respectively, P = 0.27). Oocyte viability after warming was similarly assessed in a subset of wolves (4 to 15 oocytes from each of three females; total 29 oocytes). Of these, 57.1% of the post-thaw intact oocytes were viable, which was 41.4% of all oocytes warmed. These were the first oocytes from a canid or an endangered species demonstrated to have maintained viability after vitrification and warming. Furthermore, our results demonstrated that vitrification of oocytes with the Cryotop technique was an option for preserving female gametes from Mexican wolves for future use in captive breeding programs, although in vitro embryo production techniques must first be developed in canids for this technique to be used.

Ovulatory cycles and anovulatory periods in the addax (Addax nasomaculatus).

Changes in serum oestradiol and progesterone were measured to study their dynamics during ovulatory cycles in six female addax, an endangered antelope. Blood was collected three times per week, during chute restraint, for 3 months (November to February) before introduction of a male, and continued until pregnancy was diagnosed with ultrasound. Serum was analysed by enzymeimmunoassay. Mean luteal phase, interluteal phase, and cycle durations were 22.7 +/- 2.0, 8.78 +/- 0.5 and 32.3 +/- 1.7 days, respectively. Ultrasonography revealed coiled uterine horns and maximum follicle and corpus luteum diameters of 15 and 27 mm, respectively. Each female experienced an anovulatory period, during which oestradiol continued to fluctuate, but progesterone remained below 2 ng ml-1. These periods ranged from 39 to 131 days and were not synchronous; ovulatory cycles resumed spontaneously in all females. All four females placed with a male conceived. Because addax give birth all year round, they are not considered seasonal breeders. The sporadic periods of anovulation that occurred during the winter months of this study suggest a possible seasonal effect. However, systematic sampling has not been conducted during summer and early autumn and will be necessary to address this question.