This study extends the findings of prior studies proposing and validating nutrient recycling for the heterotrophic microalgae, Thraustochytrium sp. (T18), grown in optimized fed-batch conditions. Sequential nutrient recycling of enzymatically-derived hydrolysate in fermentors succeeded at growing the tested thraustochytrid strain, with little evidence of inhibition or detrimental effects upon culture health. The average maximum biomass obtained in the recycled hydrolysate was 63.68 +/- 1.46 g L-1 in 90 h the first recycle followed by 65.27 +/- 1.15 g L-1 in 90 h in the subsequent recycle of the same material. These compared to 58.59 g L-1 and 64.92 g L-1 observed in fresh media in the same time. Lipid production was slightly impaired, however, with a maximum total fatty acid content of 62.2 +/- 0.30% in the recycled hydrolysate compared to 69.4% in fresh control media. (C) 2016 Elsevier Ltd. All rights reserved.