Preliminary studies on cryopreservation of common tench (Tinca tinca) embryos (work in progress)

Vitrification could provide a promising tool for the cryopreservation of fish embryos. However, to achieve cryopreservation using vitrification, chilling sensitivity and cryoprotectants toxicity were determined using tench embryos at four developmental stages (11, 17, 23 and 29 h). Embryos treated with alcalase (2 ml/998 ml, 2 min at 22 degrees C) were exposed to chilling with/without warming. Other embryos were exposed to methanol and glycerol at the concentration of 10% and 20% for periods of 20 min. At last, embryos were incubated at special incubator cages where hatching rates were counted. Regarding chilling sensitivity and exposure to chilling followed by warming, the hatching rates of embryos decreased significantly (p < 0.001) after exposure to 0 degrees C at all developmental stages except the 29-h stage compared with the controls. The embryo stage most sensitive to chilling was 11-h stage. The 29-h stage exhibited the least sensitivity to low temperature while 17-h and 23-h stages were intermediate in their sensitivity to chilling. The toxicity of methanol increased significantly (p < 0.001) with developmental stage for 11, 17 and 23-h stages. The highest hatching rates of tench embryos were obtained with 29-h embryos using various concentrations of methanol. The hatching rates of tench embryos exposed to glycerol concentrations were approximately similar to those embryos exposed to methanol concentrations except for 11-h embryos that showed no hatching. Unfortunately, we could not obtain living embryos in any of the conditions examined after vitrification. In conclusion, it was quite difficult to vitrify the tench embryos during this study using various vitrifying solutions and the method reported by Chen & Tian (2005) and further studies are needed to achieve successful cryopreservation.