The ability of microalgae, such as the rich in docosahexaenoic acid (DHA) heterotrophic marine microalgae Schizochytrium limacinum SR21, to utilize nutrients in their culture media derived from low-cost nutrient sources makes them a promising low-cost alternative source for the production of useful substances used in aqua feeds. The assessment of culture parameters, one of which is the pH, for every different nutrient growth medium used for microalgae cultivation is important, as they affect the biomass and secondary metabolite microalgae production. This study assessed the effect of different growth medium pH levels (6, 7, 8 and 9), at laboratory and pilot scale systems, on S. limacinum biomass productivity, lipid accumulation, proximate composition, carbon assimilation and DHA. The microalgae were cultivated in growth media containing two different alternative low-cost nutrient sources: (a) crude glycerol derived from biofuel industry as carbon source and (b) effluent digestate from biogas production of livestock decomposition as a source of nutrients and trace elements. It was found that a neutral pH (7) was the optimum level, as it enhanced biomass productivity of the lab and pilot scale cultivation systems at 44.9 g L⁻¹ and 11 g L⁻¹ and DHA content at 7.5% and 19% of the total lipid content, respectively.

The challenges to feed the world in 2050 are becoming more and more apparent. This calls for producing more with fewer inputs (most of them under scarcity), higher resource efficiency, minimum or zero effect on the environment, and higher sustainability. Therefore, increasing the circularity of production systems is highly significant for their sustainability. This study investigates the utilization of waste streams from greenhouse hydroponic drainage nutrient solutions for the cultivation of the microalgae Desmodesmus sp. The cultivation was done in an automatically controlled container-scale closed tubular Photo Bio-Reactor (PBR). The study included lab-scale open-pond system experiments and in situ container-scale experiments in the greenhouse wastewater system to assess biomass growth, optical density, nitrogen consumption, and the influence of enzymatic complexes on microalgae cell breakdown. A batch-harvesting process was followed, and the harvested microalgae biomass was pre-concentrated using FeCl₃ as a flocculant that has demonstrated efficient sedimentation and biomass recovery. Following microalgae sedimentation, the produced biomass was used for biostimulant production by means of a biocatalysis process. The enzymatic complexes, “EnzProt”, “EnzCell”, and “EnzMix” were tested for cell breakdown, with “EnzMix” at a dosage of 10% showing the most promising results. The results demonstrate successful biomass production and nitrogen uptake in the lab-scale open-pond system, with promising upscaling results within container-scale cultivation. The findings contribute to a better assessment of the needs of Desmodesmus sp. culture and highlight the importance in optimizing culture conditions and enzymatic processes for the production of biostimulants.