Establishing an astaxanthin-rich live feed strain of Pseudodiaptomus annandalei

This study aimed to establish an astaxanthin-rich strain of the calanoid copepod Pseudodiaptomus annandalei, through selective breeding based on RGB (red, green and blue) value, a parameter indicating color intensity. We evaluated the RGB value frequency distributions of the copepod populations, and selected individuals with the highest 10% and the lowest 10% RGB value over six generations. The RGB value, nauplii production, clutch interval and clutch number were assessed, and the genetic gain was calculated across generations (G₀-G₅). Two strains of copepods were selected and defined as dark body copepod strain (DBS) and light body copepod strain (LBS) at the end of experiment. Results revealed significantly lower RGB values (male: 121.5 ± 14.1; female: 108.8 ± 15) in the G₅ DBS population compared to the G₀ (male: 163.9 ± 13.1; female: 162.2 ± 14.6), with higher genetic gains of RGB values during G₀ to G₂. While DBS females exhibited longer clutch intervals in the G₃ and G₄, there was no significant difference in nauplii production between the two strains across all generations. Significantly higher astaxanthin content was found in the DBS copepods (0.04 μg/ ind.) compared to the LBS copepods (0.01 μg/ ind.) and the non-selective copepods (0.02 μg/ ind.) 20 months post selective breeding, validating the stability of the desired trait in the DBS strain. This study successfully established an astaxanthin-rich strain of P. annandalei, which provides implications for enhancing marine and brackish larviculture production.