Chilling sensitivity of Steindachneridion parahybae (Siluriformes: Pimelodidae) oocytes in different cryoprotectants

Abstract

The viability of post-thaw fish oocytes can be affected by different stages of the freezing process, such as cryoprotectant toxicity, cold sensitivity, freezing curves and thawing. Therefore, these steps need to be investigated for the development of a protocol. In the present study, the aim was to investigate chilling sensitivity at different oocyte stages of Steindachneridion parahybae. Immature and mature oocytes were incubated in Hanks’ or 90% L15 solutions containing different CPAs (cryoprotectant solutions) per experiment: (1) 0.1–0.4 M sucrose + 1–2 M methanol and (2) 1–4 M methanol X 1–4 M propylene glycol X 1–4 M DMSO for mature oocytes; (3) 0.5 M sucrose or fructose + 2 M methanol or PG or DMSO and (4) 0.25–1 M fructose + 1–4 M DMSO for immature oocytes. All treatments were kept for 120 min at −5.9 ± 2.8°C. For the control treatment, only Hanks’ or 90% L15 solutions were carried out. Evaluations were made by viability tests: membrane integrity staining in 0.4% Trypan blue (TB) and fertilization rate (%F) sole for mature oocytes. Results presented that mature oocytes were the most sensitive to lower temperatures, because there was no %F. All cryoprotectants tested in the different concentrations can be used for immature oocytes, however the statistically superior cryoprotectant was CPA with fructose and DMSO, with the low concentration of this CPA being was the best statistically. This may indicate that for this species the immature stages have presented a lower chilling sensitivity than the mature stages.