Effect of sperm concentration, medium osmolality and oocyte storage on artificial fertilisation success in a myobatrachid frog (Limnodynastes tasmaniensis)

The present study optimised artificial fertilisation and oocyte storage conditions in Limnodynastes tasmaniensis (Myobatrachidae). Data on general reproductive biology, the effect of sperm motility and concentration, medium osmolality and oocyte storage on artificial fertilisation success are presented. Egg number was most strongly correlated with bodyweight (r = 0.819). Sperm yield was correlated with testes weight (r = 0.827), which was strongly correlated with snout - vent length (r = 0.772). Optimal artificial fertilisation occurred in 0 - 7 mOsm kg(-1) amphibian Ringer, similar to ranid, bufonid and hylid species. High fertilisation rates were achieved using spermatozoa with little forwards progressive motility at comparatively low concentrations (3x10(4) sperm cells mL(-1)) and with no relationship between percentage sperm motility and fertilisation success ( correlation of fertilisation rate with spermmotility after activation: r= - 0.145). Oocytes stored in 5 mOsm kg(-1) solutions showed no significant decline in fertilisability after 2 h, showing that swelling of the jelly surrounding the eggs does not prevent sperm from fusing with the oocyte in this species. Fertilisability of oocytes was extended to> 4 h in medium to high osmolality solutions (124 - 271 mOsm kg(-1)). These data allow for the future use of L. tasmaniensis in developing assisted reproductive technology protocols for foam-nesting myobatrachid species, many of which are now threatened with extinction in the wild.