First successful intra-uterine artificial insemination in a cownose ray (Rhinoptera javanica) using chilled semen.

Reproduction of endangered wildlife, including elasmobranchs, is a fundamental mission of modern aquariums. Captive breeding programs necessitate artificial insemination to better utilize the limited stock in each facility. A cownose ray (Rhinoptera javanica) was successfully inseminated producing a live offspring. The semen was collected under chemical restraint, extended with a custom-made solution and kept chilled at 6°C for 24 hr. The quality of the semen was assessed at collection and just prior to insemination, noting color, volume, subjective sperm motility, sperm viability and concentration. Intrauterine insemination was carried out under chemical restraint. The monitoring of pregnancy was performed by serial ultrasound scans of the fetus assessing gilling and heartbeat. A healthy female pup was born after a gestation of 429 days (61 weeks and 2 days). Genetic microsatellite analysis confirmed that the pup was not a product of parthenogenesis and identified the donor male as the sire. This is the first report of a successful artificial insemination in an aplacental viviparous ray, suggesting these techniques can be applied to other species in this clade.