Cryopreservation of different strains of the euryhaline rotifer Brachionus plicatilis embryos

Abstract

Cryopreserevation of different strains of Brachionus plicatilis symmetrical stage embryos was conducted. One S-type strain, namely Yashima-S (YS), and three L-type strains, namely Yashima-L (YL), Nagasaki (NG), and Hamana (HA), were used. Symmetrical stage embryos in 10% DMSO-28‰ sea water solution were frozen at -196°C using a two-step freezing procedure; the embryos were first cooled slowly from -5°C to -20°C at a rate of 0.3°C/min and then cooled rapidly by direct transfer to liquid nitrogen. Frozen samples were thawed in a gently stirred water bath at room temperature.

A mean survival rate of 36% after 30 days storage in liquid nitrogen of HA strain was significantly lower than YS (55%), YL (58%), and NG (54%), and is due likely to its relatively larger embryo volume. There was no significant difference in the intrinsic rate of increase (r) between strains among clones taken from the cryopreservation or unfrozen control group. The results also indicate that various strains of B. plicatilis embryos can be cryopreserve without seriously altering their reproductive capability. The freezing method described could be of great potential in aquaculture and in future studies on rotifer genetics.