Growth performance of myostatin knockout red sea bream Pagrus major juveniles produced by genome editing with CRISPR/Cas9

Abstract

We have reported the production of myostatin (Pm-mstn) complete knockout red sea bream (Pagrus major) using CRISPR/Cas9, and the Pm-mstn mutant exhibited a 17% increase in skeletal muscle. However, important characteristics in aquaculture production, such as the growth rate and the amount of feed required for growth have not been clarified. In this study, we conducted a feeding trial using apparent satiation feeding during the juvenile stage and compared growth performance metrics including; weight gain, feed efficiency, apparent protein and lipid retention rates of the Pm-mstn mutants with wild-type fish (WT). Experimental fish were produced via artificial insemination from single male and female broodfish of heterozygous Pm-mstn mutants and reared in one tank until the start of the trial. Genotypes of 356 full sib fish were identified, and the WT and the homozygous mutant (HM) were used for the trial. Seventeen fish from WT and HM (mean body weight 41.1 ± 0.3 g and 42.7 ± 0.3 g, respectively) were randomly distributed into each of 100 L circular tanks and set in triplicate for each genotype. Fish were fed twice daily to apparent satiation with each respective diet at 08:00 and 13:00 six days per week for 8 weeks. Weight measurement of all experimental fish was conducted bi-weekly. Five fish at day zero from each genotype and 5 fish from each tank at the final day of the trial were sampled for whole-body proximate analysis. At the end of the trial, weight gain, specific growth rate, and feed efficiency were significantly higher in the HM group than that in the WT group, and there was no significant difference in the daily feed intake. The protein efficiency and apparent protein retention were significantly higher in the HM than for the WT. These results suggest that HM fish feed similarly to that of WT fish during the juvenile stage. However, HM fish have a higher ability to convert feed efficiently and accumulate ingested protein, resulting in better overall growth.