The sperm motility pattern in ecotoxicological tests. The CRYO-Ecotest as a case study

• Cryopreserved sea bream semen was exposed to dumpsite leachate samples.
• Motility parameters best fitting as endpoints in a bioassay were investigated.
• Multivariate analyses discriminated among “activation” and “duration” endpoints.
• CRYO-Ecotest endpoints may be each time combined according to the specific needs.
• CRYO-Ecotest is a multi-endpoint, high-plasticity innovative ecotoxicological tool.

Changes in environmental stressors inevitably lead to an increasing need for innovative and more flexible monitoring tools. The aim of this work has been the characterization of the motility pattern of the cryopreserved sea bream semen after exposure to a dumpsite leachate sample, for the identification of the best representative parameters to be used as endpoints in an ecotoxicological bioassay.Sperm motility has been evaluated either by visual and by computer-assisted analysis; parameters concerning motility on activation and those describing it in the times after activation (duration parameters) have been assessed, discerning them in terms of sensitivity, reliability and methodology of assessment by means of multivariate analyses. The EC50 values of the evaluated endpoints ranged between 2.3 and 4.5 ml/L, except for the total motile percentage (aTM, 7.0 ml/L), which proved to be the less sensitive among all the tested parameters. According to the multivariate analyses, a difference in sensitivity among “activation” endpoints in respect of “duration” ones can be inferred; on the contrary, endpoints seem to be equally informative either describing total motile sperm or the rapid sub-population, as well as the assessment methodology seems to be not discriminating. In conclusion, the CRYO-Ecotest is a multi-endpoint bioassay that can be considered a promising innovative ecotoxicological tool, characterized by a high plasticity, as its endpoints can be easy tailored each time according to the different needs of the environmental quality assessment programs.

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