Viability of subitaneous eggs of the copepod, Acartia tonsa (Dana), following exposure to various cryoprotectants and hypersaline water

Subitaneous eggs were obtained from monocultures of the calanoid copepod Acartia tonsa (Dana), Gulf of Mexico strain. Eggs were exposed to methanol, ethylene glycol, propylene glycol, glycerine, and DMSO at 0.0, 0.1, 0.5, 1.0, 2.0, and 5.0 M and hypersaline water at 50, 75, 100, 150, and 200 g/L. Treatments were evaluated after 10 and 20 min of exposure and at 4 and 26 °C. Viability (percent hatched) was determined after 24 h of incubation in 35 g/L saltwater at 26 °C. Methanol, ethylene glycol, and glycerine had high viability up to 2M, and all experienced large decreases at 5M when the exposure temperature was 26 °C compared to 4 °C. Eggs exposed to propylene glycol had lower mean viability with greater variability at the lower concentrations although viability was greater than 81.4% at 2 M. Significant decreases in viability were observed at 5 M, and the decreases were much greater at an exposure temperature of 26 °C versus 4 °C. DMSO exposed at 26 °C produced high viability up to 1 M before significant decreases occurred, while an exposure temperature of 4 °C produced high viability up to 2 M. Viability of eggs exposed to hypersaline water of 50, 75, and 100 g/L were not significantly different from controls for all treatment combinations except the 26 °C temperature exposed for 20 min, which was significantly lower at 100 g/L. Concentrations of 150 and 200 g/L produced very few to no viable eggs. These results indicate further research is justified to investigate if viability of A. tonsa eggs can be protected by these cryoprotectants and hypersaline water after exposure to cryopreservation conditions.