Low temperature requirement for embryonic development of Itasenpara bitterling Acheilognathus longipinnis

The Itasenpara bitterling has an embryonic period up to 7 months, when the embryo experiences large seasonal temperature changes. We examined the temperature requisites for normal development during the embryonic stage. Fertilized eggs reared under any of the constant temperatures ranging from 5 degrees C to 30 degrees C did not achieve complete embryogenesis, and none reached the swim-up stage. The optimum temperature for normal embryonic development was found to be stage-dependent: 10-30 degrees C for fertilization, 15-25 degrees C for hatching, 5 degrees C for the requisite low temperature, 10-15 degrees C for eye pigmentation, and 20-30 degrees C for swim-up. These temperatures correlated well with the embryo's natural environmental conditions. Embryos raised at these temperatures sequentially grew normally, with 70% of the fertilized eggs achieving complete embryogenesis and, for the first time, developed to the swim-up stage. These results indicate that the low temperature, as required by the bitterling embryo, is an essential factor and correlates well with the embryo's natural ambient temperatures. Since the populations of Itasenpara bitterlings have been declining in Japan, this study is the first to provide additional information for successful artificial breeding of this endangered species.