Research on the Application of Automatic Apparatus for Chromosome Manipulation in FishTechnology from established platforms in the first year of the project was utilized to study the application of a patented automatic system for chromosome manipulation in cyprinid loach. Large amount of eggs obtained by induced maturation and fertilized with cryopreserved or fresh sperm were used for a series of experiment on chromosome manipulation in order to evaluate the best parameter combination and to construct the feasible protocol program in the software of the self-designed automatic apparatus. It was found that heat shock (HS) using 40 or 42℃ to manipulate chromosome resulted in comparatively non-acceptable overall outcome including low rates of fertilization, embryo formation, growth of fingerlings and survival. However, cold shock (CS) using 1, 3, or 5℃ to manipulate chromosome was much better in terms of satisfactory fertilization, embryo formation, and integrated propagation index. Especially, the 30-min cold shock duration gave much better results than that of 40 min. At water temperature of 24±1℃, chemical treatment with 0.5 mg/1000 ml cytochalasin B (CB) starting at 5 min after artificial fertilization and lasting for 10- 15 min also resulted in similar overall performance. At water temperature of 26±1℃, chemical treatment with 0.5 mg/1000 ml cytochalasin B (CB) starting at 8-10 min after artificial fertilization and lasting for 10- 15 min resulted in ideal overall performance. Parameter combination of cold shock and chemical treatment methods were collected and keyed into the software of the automatic apparatus.Judging the ploidy of mixed cells from cell suspension of crushed and ground larvae or erythrocyte of young fish using flowcytometer gave the index integrated with fertilization and embryo formation rates. From these widely distributed data, high-ranking index was found mainly with properly designed cold shock and chemical treatment groups.As for research on practical application of the automatic apparatus, the advantages in cold shock for chromosome manipulation included simultaneous exposure of the selected cold temperature in mass manipulation, quick-in and quick-out to and from manipulation temperature, favorable fertilization and embryo formation rates, and triploidy percentage, as proven in this study. The advantages in chemical treatment for chromosome manipulation included avoiding direct contact with trans-cell-membrane CB by manipulators, precise timing and sufficient exposure during ten procedures such as sperm addition, sperm rinsing-out, CB addition, CB rinsing-out by DMSO, water addition, and three times DMSO rinsing. Generally speaking, overall efficiency to conduct the cold shock in loach using the automatic apparatus leads to a satisfactory integrated index of embryo formation rate and triploidy rate, far above that of chemical treatment. However, further studies on both methods of chromosome manipulation using the automatic apparatus are needed. It is found that more parameters were closely involved with artificial chromosome manipulation in finfish than in shellfish, although both with potentially high feasibility.