The bioremediation of digestate using microalgae presents a solution to the current eutrophication issue in Northwest Europe, where the use of digestate as soil fertiliser is limited, thus resulting in an excess of digestate. Ammonium is the main nutrient of interest in digestate for microalgal cultivation, and improving its availability and consequent uptake is crucial for optimal bioremediation. This work aimed to determine the influence of pH on ammonium availability in cultures of two green microalgae, additionally screened for their growth performances on three digestates produced from different feedstocks, demonstrating the importance of tailoring a microalgal strain and digestate for bioremediation purposes. Results showed that an acidic pH of 6–6.5 resulted in a better ammonium availability in the digestate media, translated into better growth yields for both S. obliquus (GR: 0.099 ± 0.001 day⁻¹; DW: 0.23 ± 0.02 g L⁻¹) and C. vulgaris (GR: 0.09 ± 0.001 day⁻¹; DW: 0.49 ± 0.012 g L⁻¹). This result was especially true when considering larger-scale applications where ammonium loss via evaporation should be avoided. The results also demonstrated that digestates from different feedstocks resulted in different growth yields and biomass composition, especially fatty acids, for which, a digestate produced from pig manure resulted in acid contents of 6.94 ± 0.033% DW and 4.91 ± 0.3% DW in S. obliquus and C. vulgaris, respectively. Finally, this work demonstrated that the acclimation of microalgae to novel nutrient sources should be carefully considered, as it could convey significant advantages in terms of biomass composition, especially fatty acids and carbohydrate, for which, this study also demonstrated the importance of harvesting time.

The use of microalgae as a sustainable source of n-3 long-chain polyunsaturated fatty acids (LC-PUFA) as an alternative to fish oils from small pelagic fish (e.g., anchovy, sardine) has received growing interest in the past few years. The present study aimed to: (i) produce Aurantiochytrium mangrovei biomass by heterotrophic fermentation using a medium containing anaerobic digestion liquid effluent, and (ii) evaluate a biomass rich in n-3 LC-PUFA and good quality proteins as a feed ingredient for sea bass juveniles and larvae. Two 800 L bioreactors were used to produce Aurantiochytrium biomass in non-axenic conditions. Biomass was then filtered through a crossflow filtration system (300 Kda ceramic membrane) and freeze-dried. Sea bass juveniles (32.7 ± 4.2 g) were fed both a control diet and a diet containing 15% of freeze-dried A. mangrovei biomass for 38 days. Juvenile survival percentage was 90% on average in both dietary conditions. Similar growth was observed between fish fed with both diets, demonstrating the feasibility to replace 15% of a standard fish feed by Aurantiochytrium biomass. The liver of sea bass juveniles fed with the A. mangrovei diet contained significantly higher proportions of 22:6n-3, 22:5n-6, and 20:4n-6 than those fed with the control diet, while the proportions of 16:0, 16:1n-7, and 18:1n-9 were significantly lower. The secondary oxidation, as measured by malonylaldehyde (MDA) content, in the liver and muscle of juveniles fed with the microalgae diet tended to be higher than in fish fed the control diet, but the differences were not statistically significant. Although the larvae survival percentage was low for all the tanks after 41 days of rearing, the inclusion of 15% of hydrolyzed A. mangrovei biomass in the larvae micro-diet did not impair the development of sea bass larvae and only marginally affected their lipid composition. In the future, we have to further optimize a sustainable workflow between Aurantiochytrium cultivation and fish feed production and confirm the zootechnical and biochemical results.

The production of non-fish based docosahexaenoic acid (DHA) for feed and food has become a critical need in our global context of over-fishing. The industrial-scale production of DHA–rich Thraustochytrids could be an alternative, if costs turned out to be competitive. In order to reduce production costs, this study addresses the feasibility of the non-axenic (non-sterile) cultivation of Aurantiochytrium mangrovei on industrial substrates (as nitrogen and mineral sources and glucose syrup as carbon and energy sources), and its scale-up from laboratory (250 mL) to 500 L cultures. Pilot-scale reactors were airlift cylinders. Batch and fed-batch cultures were tested. Cultures over 38 to 62 h achieved a dry cell weight productivity of 3.3 to 5.5 g.L⁻¹.day⁻¹, and a substrate to biomass yield of up to 0.3. DHA productivity ranged from 10 to 0.18 mg.L⁻¹.day⁻¹. Biomass productivity appears linearly related to oxygen transfer rate. Bacterial contamination of cultures was low enough to avoid impacts on fatty acid composition of the biomass. A specific work on microbial risks assessment (in supplementary files) showed that the biomass can be securely used as feed. However, to date, there is a law void in EU legislation regarding the recycling of nitrogen from digestate from animal waste for microalgae biomass and its usage in animal feed. Overall, the proposed process appears similar to the industrial yeast production process (non-axenic heterotrophic process, dissolved oxygen supply limiting growth, similar cell size). Such similarity could help in further industrial developments.