Abstract The aim of the present study was to determine the effects of the use of astaxanthin alternate feeding on rainbow trout pigmentation in term of astaxanthin serum concentration, muscle colour and astaxanthin muscle retention. Four hundred and fifty rainbow trout were fed the same basal diet supplemented with two different astaxanthin levels, 100, and 200 mg astaxanthin kg − 1 of diet, hereafter designated as AX100 and AX200, respectively. An additional astaxanthin-free (AX0) diet was used. The experimental treatments were as follows: (1) REF = AX100 diet at each meal each day, served as reference; (2) SD1 = AX100 diet at each meal the first day followed by AX0 diet at each meal every second day; (3) SD2 = AX100 diet and AX0 diet in alternate meals each day; (4) R2 = AX200 diet and AX0 diet in alternate meals each day; (5) R4 = AX200 diet at the first meal the first day followed by AX0 diet at the second meal the first day and at each meal every second day. Fish were fed the experimental feeding schedule for 42 days. At the end of the experiment there were no significant differences among fish fed the different feeding schedules in term of final mean weight, specific growth rate and feed efficiency ratio. SD2 fish group displayed the highest ( P < 0.05) astaxanthin serum concentration and the R4 fish group the lowest one. REF and R2 fish groups showed similar astaxanthin serum concentrations. Muscle chroma showed the most pronounced effect. It increased significantly for all fish groups during the experiment. At the end of the experiment REF and R2 fish groups displayed higher values than SD1 and R4 fish groups. Muscle astaxanthin concentrations increased significantly during the experiment whatever the astaxanthin feeding schedule. At the end of the experiment, the highest muscle astaxanthin concentration was recorded for R2 fish group while the lowest was noted for R4 fish group. Except for SD1 and R4 fish groups, muscle astaxanthin retention decreased significantly during the experiment. At the end of the experiment, muscle astaxanthin retention coefficients for SD2 fish group were significantly higher than those for REF fish group. The results reported here provide further evidence of the potential applicability of alternate astaxanthin feeding on rainbow trout pigmentation. Extending the optimisation of the SD2 treatment will therefore be subject for future studies. Its application could result in cost saving in the fish farming industry.