Raceways ponds are the microalgal production systems most commonly used at industrial scale. In this work, two different raceway configurations were tested under the same processing conditions to compare their performance on the production of Nannochloropsis oceanica. Biomass productivity, biochemical composition of the produced biomass, and power requirements to operate those reactors were evaluated. Water depths of 0.20 and 0.13 m, and culture circulation velocities of 0.30 and 0.15 m s⁻¹ were tested. A standard configuration, which had a full channel width paddlewheel, proved to be the most energy efficient, consuming less than half of the energy required by a modified configuration (had a half channel width paddlewheel). The later showed to have slightly higher productivity, not enough to offset the large difference in energetic consumption. Higher flow velocity (0.30 m s⁻¹) led to a 1.7 g m⁻² d⁻¹ improvement of biomass productivity of the system, but it increased the energy consumption twice as compared to the 0.15 m s⁻¹ flow velocity. The latter velocity showed to be the most productive in lipids. A water depth of 0.20 m was the most suitable option tested to cultivate microalgae, since it allowed a 54% energy saving. Therefore, a standard raceway pond using a flow velocity of 0.3 m s⁻¹ with a 0.20 m water depth was the most efficient system for microalgal cultivation. Conversely, a flow velocity of 0.15 m s⁻¹ was the most suitable to produce lipids.

Bioprospection of novel autochthonous strains is key to the successful industrial-scale production of microalgal biomass. A novel Chlorococcum strain was recently isolated from a pond inside the industrial production facility of Allmicroalgae (Leiria, Portugal). Phylogenetic analysis based on 18S ribosomal ribonucleic acid (rRNA) gene sequences suggests that this isolate is a novel, free-living Oophila amblystomatis strain. However, as our phylogenetic data strongly suggests that the aforementioned taxon belongs to the genus Chlorococcum, it is here proposed to rename this species as Chlorococcum amblystomatis. In order to characterize the biotechnological potential of this novel isolate, growth performance and biochemical composition were evaluated from the pilot (2.5-m³) to industrial (10-m³) scale. The highest maximum areal productivity (36.56 g·m⁻²·day⁻¹) was reached in a 10-m³ tubular photobioreactor (PBR), as compared to that obtained in a 2.5-m³ PBR (26.75 g·m⁻²·day⁻¹). Chlorococcum amblystomatis displayed high protein content (48%–56% dry weight (DW)) and moderate levels of total lipids (18%–31% DW), carbohydrates (6%–18% DW) and ashes (9%–16% DW). Furthermore, the lipid profile was dominated by polyunsaturated fatty acids (PUFAs). The highest pigment contents were obtained in the 2.5-m³ PBR, where total chlorophylls accounted for 40.24 mg·g⁻¹ DW, followed by lutein with 5.37 mg·g⁻¹ DW. Overall, this free-living Chlorococcum amblystomatis strain shows great potential for nutritional applications, coupling a promising growth performance with a high protein content as well as relevant amounts of PUFAs, chlorophyll, and carotenoids.

Microalgae are currently considered to be a promising feedstock for biodiesel production. However, significant research efforts are crucial to improve the current biomass and lipid productivities under real outdoor production conditions. In this context, batch, continuous and semi-continuous operation regimes were compared during the Spring/Summer seasons in 2.6 m³ tubular photobioreactors to select the most suitable one for the production of the oleaginous microalga Nannochloropsis oceanica. Results obtained revealed that N. oceanica grown using the semi-continuous and continuous operation regimes enabled a 1.5-fold increase in biomass volumetric productivity compared to that cultivated in batch. The lipid productivity was 1.7-fold higher under semi-continuous cultivation than that under a batch operation regime. On the other hand, the semi-continuous and continuous operation regimes spent nearly the double amount of water compared to that of the batch regime. Interestingly, the biochemical profile of produced biomass using the different operation regimes was not affected regarding the contents of proteins, lipids and fatty acids. Overall, these results show that the semi-continuous operation regime is more suitable for the outdoor production of N. oceanica, significantly improving the biomass and lipid productivities at large-scale, which is a crucial factor for biodiesel production.

Thraustochytrids have gained increasing relevance over the last decades, due to their fast growth and outstanding capacity to accumulate polyunsaturated fatty acids (PUFAs), particularly docosahexaenoic acid (DHA). In this context, the present work aimed to optimize the growth performance and DHA yields by improving the culture medium of Aurantiochytrium sp. AF0043. Accordingly, two distinct culture media were optimized: (i) an inorganic optimized medium (IOM), containing only monosodium glutamate and glucose as nitrogen and carbon sources, respectively; and (ii) an organic and sustainable waste-based optimized medium (WOM), containing corn steep powder and glycerol, added in fed-batch mode, as nitrogen and carbon sources, respectively. Overall, the lab-scale optimization allowed to increase the biomass yield 1.5-fold and enhance DHA content 1.7-fold using IOM. Moreover, WOM enabled a 2-fold increase in biomass yield and a significant improvement in lipid contents, from 22.78% to 31.14%. However, DHA content was enhanced almost 3-fold, from an initial content of 10.12% to 29.66% of total fatty acids contained in the biomass. Therefore, these results strongly suggest, not only that the production pipeline was significantly improved but also confirmed the potential use of Aurantiochytrium sp. AF0043 as a source of DHA.

Microalgal biomass has gained increasing attention in the last decade for various biotechnological applications, including human nutrition. Certified organic products are currently a growing niche market in which the food industry has shown great interest. In this context, this work aimed at developing a certified organic culture medium for the production of autotrophic Chlorella vulgaris biomass. A preliminary assay in 2 L bubble column photobioreactors was performed in order to screen different commercial organic substrates (OS) at a normalized concentration of N (2 mmol L⁻¹). The highest growth performance was obtained using EcoMix4 and Bioscape which showed similar biomass concentrations compared to the synthetic culture medium (control). In order to meet the nutrient needs of Chlorella, both OS underwent elemental analyses to assess their nutrient composition. The laboratory findings allowed the development of a final organic culture medium using a proportion of Bioscape/EcoMix4 (1:1.2, m/m). This organic culture medium was later validated outdoors in 125 L flat panel and 10 m³ tubular flow through photobioreactors. The results obtained revealed that the developed organic medium led to similar microalgal growth performance and biochemical composition of produced biomass, as compared to the traditional synthetic medium. Overall, the formulated organic medium was effective for the autotrophic production of organic C. vulgaris biomass.

Microalgae attract interest worldwide due to their potential for several applications. Scenedesmus is one of the first in vitro cultured algae due to their rapid growth and handling easiness. Within this genus, cells exhibit a highly resistant wall and propagate both auto- and heterotrophically. The main goal of the present work is to find scalable ways to produce a highly concentrated biomass of Scenedesmus rubescens in heterotrophic conditions. Scenedesmus rubescens growth was improved at the lab-scale by 3.2-fold (from 4.1 to 13 g/L of dry weight) through medium optimization by response surface methodology. Afterwards, scale-up was evaluated in 7 L stirred-tank reactor under fed-batch operation. Then, the optimized medium resulted in an overall productivity of 8.63 g/L/day and a maximum biomass concentration of 69.5 g/L. S. rubescens protein content achieved approximately 31% of dry weight, similar to the protein content of Chlorella vulgaris in heterotrophy.