| In this study, fertilized egg of Chinese medaka (Oryzias latiipessinensis) at28°C was continuously observed. We confirmed thedevelopmental stages and the elapsed time of each developmental stage,describing the development status of each period. Two solutions for threeeyed period, the16muscle feasts periods and notochord vacuolizationwere designed. Four ways to preserve in low temperature (0-4°C) for15days were carried out and embryo survival rates were recorded. Thenmedaka embryo was incubated at28°C under constant temperature andhatchability was recorded. Finally, compare the total number of embryosin the number of samples of normal larvae hatched, obtaining aftercryopreservation for15days the normal larvae rate of28°C incubator, Thestudy includes:1. Development of China medaka embryo (28°C)In this study development of fertilized eggs of China medaka, and thedevelopmental stages of segmentation and description were continuously observed. The results showed that fertilized egg of China medakaexperienced six periods (the period of cleavage, blastocyst formationduring the gastrulation period, neurulation period of organ formationduring the incubation period),32specific developmental stages beforehatching the larvae.2Trial of four ways to preserve medaka embryo in cryopreservation (4°C)In this trial, three different developmental stages (eyed period,16sarcomere of notochord vacuolization completion date) of medakaembryos were stored at low temperature for15days, the results of whichshowed that: only using povidone-iodine protective solution, the averageembryo survival rates of three periods were75.0%,71.1%and62.5%.Adding peat povidone-iodine protective solution, the average embryosurvival rates of three periods were86.0%,84.3%and74.5%. The use ofmalachite green protective solution, the average embryo survival rates ofthree periods were74.5%,68.3%and72.7%. The use of malachite greenprotective solution with peat, the average embryo survival rates of threeperiods were84.3%,79.0%and80.5%.3Trial of Cryopreservation thermostat incubator medaka embryoThe trial was after the last part, in which survival medaka embryocryopreservation were put out from sample bags and then put in waterbath equipped with a28°C for incubation in a Petri dish of pure water until hatching a film recorded hatching results. Embryo hatching rates ofcryopreservation with only three periods of povidone-iodine protectivesolution were73.7%,73.0%and79.1%. Average embryos hatching rates ofcryopreservation with peat soil protection solution of povidone-iodine ofthe three periods were79.5%,74.0%and79.1%. Embryo hatching rate s ofcryopreservation only of three periods using malachite green protectivesolution were73.3%,73.8%and66.2%. Embryo hatching rate of addingpeat malachite green protective solution cryopreservation of three periodswere74.5%,78.1%and69.3%.4Trial of hatching normal larvae of medaka embryosAfter28℃incubator in part2.2, observe body and internal organs of theembryo developmental status under a stereo microscope, recordingnormal development of larvae results, the results of which showed that:The normal embryonic larvae rates of cryopreservation using only theprotective solution of povidone-iodine in three periods were54.0%,48.2%and48.3%. The normal embryonic larvae rates of cryopreservation ofpovidone-iodine protection solution adding peat in three periods were67.1%,59.8%and56.8%. Normal embryonic larvae rates ofcryopreservation using only the malachite green protective solution forthree periods were51.6%,45.5%and45.0%. Normal embryonic larvaerates of cryopreservation adding peat malachite green protective solution in three periods were61.2%,57.7%and52.7%, respectively. |
