Development of alternative and sustainable methodologies in laboratory research on sea urchin gametes

The aim of the present work is to develop a laboratory-scaled methodology for an on-demand supply of semen from the sea urchin Paracentrotus lividus. Firstly, sea urchin specimens were acclimatized to the long-term rearing in a recirculating aquaculture system and gonad maturation was obtained under controlled conditions. Semen samples were then collected from mature sea urchins and cryopreserved. Finally, post-thawing motility was evaluated, to verify whether the cryopreserved semen had maintained enough viability to be used in laboratory activities. The post-thawing motility parameters remained quite unchanged for up to 60 min after activation; moreover, the semen even retained the ability of motility activation for 60 min after thawing. This motility pattern makes the use of cryopreserved semen a feasible option in spermiotoxicity bioassays. The preliminary ecotoxicity test, carried out using motility parameters as endpoints, showed sensitivity levels to cadmium falling in the same order of magnitude as those recorded for fresh sea urchin semen and for cryopreserved sea bream semen. . Therefore, semen samples produced and stored according to the developed methodology described in this paper, can be considered a promising and sustainable alternative to those collected from mature sea urchins harvested in the field.