A breeding program for the critically endangered Panamanian golden frog (Atelopus zeteki) was established in 2001 with the goal of forming a genetically sustainable reassurance population. Breeding in select AZA institutions has been increasingly successful; however, the prolonged amplexus that is observed in this species can result in decreasing condition and higher morbidity and mortality during the breeding season in both sexes. Therefore assisted reproductive technology was employed to aid in the release of ova and sperm in order to decrease mortality, produce eggs from genetically valuable females experiencing dystocia, and identify methods for the collection of ova and sperm for future in vitro fertilization and gamete banking studies.
Females (n=154) in the study were administered 4 µg luteinizing hormone releasing hormone (LHRH) intracoelomically every 2 days for a maximum of four doses. During the 6 yr of the study, 66.9% of the female frogs that received LHRH successfully laid eggs. The highest success of ovipositioning was noted in November–December of each year and in frogs >3 yr. The majority of frogs laid eggs after one dose of LHRH (63.2%), with 28.3% laying after two doses.
In the second part of the study, male frogs were administered LHRH intracoelomically, and urine was collected for semen analysis. Frogs that received 1 µg or 4 µg LHRH yielded spermic urine at 3 and 6 hr, with the highest sperm concentrations and highest yield of spermic urine noted with administration of 4 µg LHRH and urine collection 3 hr later.