Biocrude Oil Production by Integrating Microalgae Polyculture and Wastewater Treatment: Novel Proposal on the Use of Deep Water-Depth Polyculture of Mixotrophic Microalgae

Microalgae have attracted significant attention worldwide as one of the most promising feedstock fossil fuel alternatives. However, there are a few challenges for algal fuels to compete with fossil fuels that need to be addressed. Therefore, this study reviews the R&D status of microalgae-based polyculture and biocrude oil production, along with wastewater treatment. Mixotrophic algae are free to some extent from light restrictions using organic matter and have the ability to grow well even in deep water-depth cultivation. It is proposed that integrating the mixotrophic microalgae polyculture and wastewater treatment process is the most promising and harmonizing means to simultaneously increase capacities of microalgae biomass production and wastewater treatment with a low land footprint and high robustness to perturbations. A large amount of mixotrophic algae biomass is harvested, concentrated, and dewatered by combining highly efficient sedimentation through flocculation and energy efficient filtration, which reduce the carbon footprint for algae fuel production and coincide with the subsequent hydrothermal liquefaction (HTL) conversion. HTL products are obtained with a relatively low carbon footprint and separated into biocrude oil, solid, aqueous, and gas fractions. Algae biomass feedstock-based HTL conversion has a high biocrude oil yield and quality available for existing oil refineries; it also has a bioavailability of the recycled nitrogen and phosphorus from the aqueous phase of algae community HTL. The HTL biocrude oil represents higher sustainability than conventional liquid fuels and other biofuels for the combination of greenhouse gas (GHG) and energy return on investment (EROI). Deep water-depth polyculture of mixotrophic microalgae using sewage has a high potential to produce sustainable biocrude oil within the land area of existing sewage treatment plants in Japan to fulfill imported crude oil.

Freshwater dissolved oxygen dynamics: changes due to glyphosate, 2,4-D and their mixture, both under clear and turbid-organic conditions.

We performed two independent outdoor mesocosm experiments where we measured the variation of DO saturation (DO%) in freshwater after a single input of Roundup Max (G) (glyphosate-based formulation), AsiMax 50 (2,4-D) (2,4-D-based formulation) and their mixture (M). Two concentration levels were tested; 0.3 mg/L G and 0.135 mg/L 2,4-D (Low; L) and 3 mg/L G and 1.35 mg/L 2,4-D (High; H). We assayed consolidated microbial communities coming from a system in organic turbid eutrophic status and a system in clear mesotrophic status during 21 and 23 days, respectively. A sample of phytoplankton (micro+nano, pico-eukaryotes, pico-cyanobacteria), mixotrophic algae and heterotrophic bacteria was collected to determine abundances at each of four sampling dates. The clear and turbid systems showed similar, but not synchronized, patterns of daily DO% changes in relation to the controls (DO%v), after exposure to both single and combined formulations. Under glyphosate scenarios (GL, GH, ML and MH), the two types of systems showed similar DO%v but different microbial abundances, being associated to an increase in the micro+nano and pico-eukaryotic phytoplankton fractions for the clear system. In contrast, in the turbid system changes were associated with increased pico-cyanobacteria and decreased mixotrophic algae. Effects of 2,4-D were only observed in the turbid system, leading to decreased micro+nano phytoplankton abundances. Under the turbid scenario, the herbicide mixture at high concentration had a synergistic effect on DO%v and recovery was not detected by the end of the experiment. Our results revealed that herbicides inputs induced changes in phytoplankton abundances that leads to measurable DO variations.

An under-ice bloom of mixotrophic haptophytes in low nutrient and freshwater-influenced Arctic waters

The pelagic spring bloom is essential for Arctic marine food webs, and a crucial driver of carbon transport to the ocean depths. A critical challenge is understanding its timing and magnitude, to predict its changes in coming decades. Spring bloom onset is typically light-limited, beginning when irradiance increases or during ice breakup. Here we report an acute 9-day under-ice algal bloom in nutrient-poor, freshwater-influenced water under 1-m thick sea ice. It was dominated by mixotrophic brackish water haptophytes (Chrysochromulina/ Prymnesium) that produced 5.7 g C m−2 new production. This estimate represents about half the annual pelagic production, occurring below sea ice with a large contribution from the mixotrophic algae bloom. The freshwater-influenced, nutrient-dilute and low light environment combined with mixotrophic community dominance implies that phagotrophy played a critical role in the under-ice bloom. We argue that such blooms dominated by potentially toxic mixotrophic algae might become more common and widespread in the future Arctic Ocean.

From lethal to beneficial - contrasting dietary effects of freshwater mixotrophs on zooplankton

1. The importance of mixotrophic algae as key bacterivores in microbial food webs is increasingly acknowledged, but their effects on consumers is less understood, with previous studies having revealed contrasting results. In freshwater, this may be related to fundamental differences in the nutritional quality of two major mixotrophic groups. While cryptophytes are generally considered as high-quality food for zooplankton, chrysophytes (golden algae) are often referred to be toxic. 2. Using four chrysophyte species, we performed a comparative study as an attempt to generalize their dietary quality by (1) revealing their stoichiometric and biochemical profiles, and (2) quantifying their dietary effects in feeding bioassays with Daphnia longispina. We compared the observed effects to a known high-quality reference food (Cryptomonas sp.) and a starvation control as a reference for potential toxicity. 3. We found dramatic differences in survival and growth of D. longispina depending on the chrysophyte species provided as food. Even within the same genus, dietary quality ranged from deleterious to high. As this was not reflected in differences in cellular stoichiometry and fatty acid profiles, we suggest that toxicity may be the underlying mechanism. 4. Our results suggest that the dietary effects of chrysophytes cannot be generalised. Besides, the fact that a species previously reported to be deleterious turned out to be a beneficial food source suggests that toxic effects may dynamically vary depending on environmental cues, mode of nutrition or the investigated strain. 5. To fully understand the nutritional value of mixotrophic algae in aquatic food webs, representatives of multiple taxa need to be tested under a range of environmental conditions. This is also needed for a better predictive capability of climate change effects, as it may not only promote the dominance of mixotrophic algae, but also induce functional changes related to their nutritional quality.

Tracing the fate of microplastic carbon in the aquatic food web by compound-specific isotope analysis

Increasing abundance of microplastics (MP) in marine and freshwaters is currently one of the greatest environmental concerns. Since plastics are fairly resistant to chemical decomposition, breakdown and reutilization of MP carbon complexes requires microbial activity. Currently, only a few microbial isolates have been shown to degrade MPs, and direct measurements of the fate of the MP carbon are still lacking. We used compound-specific isotope analysis to track the fate of fully labelled 13C-polyethylene (PE) MP carbon across the aquatic microbial-animal interface. Isotopic values of respired CO2 and membrane lipids showed that MP carbon was partly mineralized and partly used for cell growth. Microbial mineralization and assimilation of PE-MP carbon was most active when inoculated microbes were obtained from highly humic waters, which contain recalcitrant substrate sources. Mixotrophic algae (Cryptomonas sp.) and herbivorous zooplankton (Daphnia magna) used microbial mediated PE-MP carbon in their cell membrane fatty acids. Moreover, heteronanoflagellates and mixotrophic algae sequestered MP carbon for synthesizing essential ω-6 and ω-3 polyunsaturated fatty acids. Thus, this study demonstrates that aquatic micro-organisms can produce, biochemically upgrade, and trophically transfer nutritionally important biomolecules from PE-MP.