ABSTRACT
Reproductive technologies (RTs) can assist integrated conservation breeding programs to attain propagation targets and manage genetic diversity more effectively. While the application of RTs to enhance the conservation management of threatened amphibians has lagged behind that of other taxonomic groups, a recent surge in research is narrowing the divide. The present study reports on the first application of RTs (hormone-induced spawning, hormone-induced sperm-release, and sperm cryopreservation) to the critically endangered Baw Baw frog, Philoria frosti. To determine the effect of hormone therapy on spawning success, male-female pairs were administered either 0 µg/g gonadotropin-releasing hormone agonist (GnRHa), 0.5 µg/g GnRHa, or 0.5 µg/g GnRHa + 10 µg/g metoclopramide (MET) (n = 6-7 pairs/treatment), and the number of pairs ovipositing, total eggs, and percent fertilisation success were quantified. To determine the effect of hormone therapy on sperm-release and to establish the peak time to collect sperm post-hormone administration, males were administered 0 IU/g (n = 4), or 20 IU/g hCG (n = 16). Total sperm, sperm concentration, and percent viability were quantified at 0, 2, 4, 6, 8, 10, and 12 h post-hormone administration. Overall, the percentage of pairs ovipositing was highest in the GnRHa + MET treatment, with 71% of pairs ovipositing, compared to 57% and 33% of pairs in the GnRHa and control treatments, respectively. The quantity of sperm released from males in response to hCG peaked at 4 h post-hormone administration, though it remained high up to 12 h. The percent sperm viability also peaked at 4 h post-administration (94.5%), exhibiting a steady decline thereafter, though viability remained above 77% throughout the 12 h collection period. The remaining sperm samples (n = 22) were cryopreserved using established protocols and biobanked for long-term storage and future conservation applications. The mean post-thaw sperm viability was 59%, and the percent total motility was 17%. The results from this preliminary study will direct further applications of RTs to the critically endangered Baw Baw frog to assist with species recovery.
ABSTRACT
Temperature and food availability are known to independently trigger phenotypic change in ectotherms, but the interactive effects between these factors have rarely been considered. This study investigates the independent and interactive effects of water temperature and food availability on larval growth and development of the critically endangered Baw Baw frog, Philoria frosti. Larvae were reared at low (12°C) or high (17°C) water temperature in the absence or presence of substrate that controlled food availability, and body size and time to metamorphosis were quantified. Growth and development of larvae was influenced by the individual effects of temperature and food availability; time to metamorphosis was shorter in warm water treatment groups and in the presence of substrate and increased food. Unexpectedly, however, water temperature and food availability did not have an interactive effect on either time to metamorphose or body size at metamorphosis. Under all treatment groups, metamorphic onset occurred once a developmental size threshold was reached, indicating that growth rate and body size are key factors controlling the metamorphic process in Baw Baw frogs (consistent with the Wilbur-Collins model for ectotherm development). From an applied perspective, our findings have implications for amphibian conservation because they indicate that simple manipulations of temperature and food availability can be used to increase the rate of frog production in conservation breeding programs.
