Artificial fertilisation in a terrestrial toadlet (Pseudophryne guentheri): effect of medium osmolality, sperm concentration and gamete storage
Aimee J. Silla
+ Author Affiliations
- Author Affiliations
Institute for Conservation Biology and Environmental Management, School of Biological Sciences, University of Wollongong, Wollongong, NSW 2522, Australia. Email: aimee.silla@gmx.com
Reproduction, Fertility and Development 25(8) 1134-1141 https://doi.org/10.1071/RD12223
Submitted: 11 July 2012 Accepted: 18 October 2012 Published: 23 November 2012
Abstract
Anurans exhibit a greater reproductive diversity than any other vertebrate order. However, studies investigating the effects of the external fertilisation environment on fertilisation success are limited to aquatic-breeding species. This study investigated the effects of fertilisation medium osmolality, sperm concentration and short-term oocyte storage on fertilisation success in a terrestrial-breeding anuran, Pseudophryne guentheri. Split-clutch experimental designs were used to determine optimal fertilisation conditions. To determine the effect of short-term sperm storage, sperm viability was assessed using fluorescence microscopy and percentage sperm motility and velocity quantified with a computer-assisted sperm analysis system. Fertilisation success was highest in media ranging in osmolality from 25 mOsm kg–1 to 100 mOsm kg–1, representing a broader range and higher optimal osmolality than previously reported for aquatic breeders. High rates of fertilisation (>75%) were achieved in relatively low sperm concentrations (2.5 × 104 mL–1). Oocytes stored in isotonic solutions (200 mOsm kg–1) retained fertilisation capacity (32%) after 8 h of storage, while sperm suspensions maintained motility (≥26%) for 13 days. Additional studies on terrestrial-breeding anurans will be required to ascertain whether the optimal fertilisation conditions reported reflect adaptations to achieve fertilisation in a terrestrial environment.
Additional keywords : amphibian, assisted reproductive technologies, ART, external fertilisers, fertilisation environment, fertilization, frog, in vitro fertilisation, IVF, reproduction.
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