Determination of the rates of fertilization, hatching and larval survival in the Japanese eel, Anguilla japonica, using tissue culture microplates

A method was developed for assessing fertility, hatchability and larval survival in the Japanese eel, Anguilla japonica. Fertilized eggs were stocked in 24-, 48-, and 96-well plates at one egg to one well and maintained at 23 °C without removing dead larvae, changing the water or feeding. Upon hatching, the larvae were often trapped on the surface of the water where they died (surface death). The remaining larvae then showed rather stable survival in the three types of plates beyond the yolk-sac stage, which was completed by day 8 after hatching (DAH). Polyethylene glycol 6000 (PEG 6000) added to the rearing water at 1 or 10 μg/ml markedly suppressed surface death without apparent effect on hatchability, the survival profile or abnormality. These results indicated that the survival rate can be determined easily by stocking the fertilized eggs individually in the wells of microplates with water containing PEG 6000 and by rearing the hatched larvae until the completion of yolk absorption, with little management. By this method, a series of parameters for evaluating the egg quality, rates of fertilization, hatching, survival and abnormality were determined in the artificially maturation-induced Japanese eel: unstable fertility and frequent abnormal development were revealed to be impediments to constant production of larvae at first feeding.