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Evaluating amphibian biobanking and reproduction for captive breeding programs according to the Amphibian Conservation Action Plan objectives.
Della Togna G
,
Howell LG
,
Clulow J
,
Langhorne CJ
,
Marcec-Greaves R
,
Calatayud NE
.
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The Amphibian Conservation Action Plan (ACAP), published in 2007, is a formal document of international significance that proposed eleven relevant actions for global amphibian conservation. Action seven of the ACAP document addresses the use of amphibian captive programs as a conservation tool. Appendix material under this action explores the potential use of Genome Resource Banking (biobanking) as an urgently needed tool for these captive programs. ACAP proposed twelve objectives for Genome Resource Banking which exhibit little emphasis on reproduction as a vital underlying science for amphibian Captive Breeding Programs (CBP's). Here we have reassessed the original twelve ACAP objectives for amphibian reproduction and biobanking for CBP's as a contribution to future ACAP review processes. We have reviewed recent advances since the original objectives, as well as highlighted weaknesses and strengths for each of these objectives. We make various scientific, policy and economic recommendations based on the current reality and recent advances in relevant science in order to inform future ACAP towards new global objectives. The number of amphibian CBP'S has escalated in recent years and reproductive success is not always easily accomplished. Increases in applied and fundamental research on the natural history and reproductive biology of these species, followed by the appropriate development and application of artificial reproductive technologies (ART's) and the incorporation of genome resource banks (GRB's), may turn CBP's into a more powerful tool for amphibian conservation.
Fig. 1. Recommended steps for the systematic development of hormonal stimulation protocols for amphibian gamete collection. The process includes selection of hormones and concentrations, administration method, gamete collection, quality and morphology assessments, and determination of quality and concentration peaks (only for sperm). Following these steps would result in the nonempirical selection of the best stimulation treatment on the basis of hormone efficacy, gamete quality and concentration. FPM (forward progressive motility), PFA (paraformaldehyde) and NBF (neutral buffered formalin).
Fig. 2. Morphology assessment of Strabomantis bufoniformis eggs. Freshly laid eggs were collected and fixed in 4% PFA for 24 h, sectioned (A,B), stained with Coomassie Blue for 90 s (C,D) and washed with ARS. Staining revealed three jelly layers (E,F,G) when observed under the microscope (200X) (Della Togna, unpublished). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
Fig. 3. Steps for the systematic development of sperm pre-freezing assessment. After sample collection, either by hormonal stimulation or testicular excision, samples should be assessed for quality and concentration. Samples yielding concentration and motility parameters equal to or higher than 106 and 70% respectively, should be considered for freezing. PS collection time: poststimulation collection time, FPM (forward progressive motility), PFA (paraformaldehyde) and NBF (neutral buffered formalin).
Fig. 4. Steps to determine the cryotoxic effect of CPA’s. Selection of an effective CPA solution for sperm freezing involves choosing the most appropriate penetrating and non-penetrating CPA’s and their concentrations (in bold the most commonly used), addition rate and equilibration time on the basis of sample quality and morphology.
Fig. 5. Recommended steps to determine the most appropriate sperm freezing and thawing conditions. The process requires selection of the freezing container, freezing mode, thawing conditions and sample activation on the basis of post-thaw sperm quality and morphology recovery.
Fig. 6. Predicted cost trends across the extended lifecycle of amphibian genome resource banking infrastructure.
