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Carbon-Storing Structural Material Based on Wastewater-Cultivated Chlorella Sorokiniana

Microalgae cultivation is a promising technology for carbon sequestration and wastewater remediation. It is economically and logistically critical to find viable applications for the large quantities of produced biomass. In current research, a structural material is developed by using nearly 100% Chlorella sorokiniana. The otherwise non-cohesive microalgae cells are first surface-activated and then compacted under a relatively high pressure. The resulting material is a dense solid, stronger than typical steel-reinforced concrete in flexural tests. The processing procedure is simple and fast, and the setup is scalable. This technique may be useful for next-generation green construction and provide a use-case for sustainable microalgae biomass.

Yi, Haozhe

Morphotype-resolved characterization of microalgal communities in a nutrient recovery process with ARTiMiS flow imaging microscopy

Microalgae-driven nutrient recovery represents a promising technology for phosphorus removal from wastewater while simultaneously generating biomass that can be valorized to offset treatment costs. As full-scale processes come online, system parameters including biomass composition must be carefully monitored to optimize performance and prevent culture crashes. In this study, flow imaging microscopy (FIM) was leveraged to characterize microalgal community composition in near real-time at a full-scale municipal wastewater treatment plant (WWTP) in Wisconsin, USA, and population and morphotype dynamics were examined to identify relationships between water chemistry, biomass composition, and system performance. Two FIM technologies, FlowCam and ARTiMiS, were evaluated as monitoring tools. ARTiMiS provided a more accurate estimate of total system biomass, and estimates derived from particle area as a proxy for biovolume yielded better approximations than particle counts. Deep learning classification models trained on annotated image libraries demonstrated equivalent performance between FlowCam and ARTiMiS, and convolutional neural network (CNN) classifiers proved significantly more accurate when compared to feature table-based dense neural network (DNN) models. Across a two-year study period, Scenedesmus spp. appeared most important for phosphorus removal, and were negatively impacted by elevated temperatures and increase in nitrite/nitrate concentrations. Chlorella and Monoraphidium also played an important role in phosphorus removal. For both Scenedesmus and Chlorella, smaller morphological types were more often associated with better system performance, whereas larger morphotypes likely associated with stress response(s) correlated with poor phosphorus recovery rates. Furthermore, these results demonstrate the potential of FIM as a critical technology for high-resolution characterization of industrial microalgal processes.

59 BASIC BIOLOGICAL SCIENCES

Impacts of feeding three strains of microalgae alone or in combination on growth performance, protein metabolism, and meat quality of broiler chickens

Variations in nutrient compositions, especially amino acid (AA) profiles, among microalgal species may enable a superior feeding outcome from a combined than singular supplementation in poultry diets. Therefore, a feeding trial was conducted to compare the effects of three strains of microalgal biomass supplemented alone or in combination to replace 5 % (starter) and 10 % (grower) soybean meal (on weight-to-weight basis) on growth performance, protein metabolism, and meat quality of broiler chickens. Day-old Cornish Cross male chicks (total = 180) were divided into 5 groups (6 cages/treatment, 6 birds/cage) and fed a corn-soybean meal basal diet (BD), BD + H117 (Chlorella sp., H117), BD + C985 (Tetraselmis sp., C985), BD + Nannochloropsis oceanica (NO), and BD + H117 + C985 + NO (Combination). Feeding any of the microalgae diets did not alter growth performance nor meat quality including texture, pH, color, and water holding capacity of breast and thigh meats. However, the breast weight percentages were decreased (P < 0.05) by feeding the C985, NO, and Combination diets. Compared with the BD, the 4 microalgal diets led to higher (P < 0.05) plasma uric acid and protein concentrations at weeks 3 and (or) 6. The mRNA levels of MAFbx, MURF1, FOXO1, and calpastatin in the breast and thigh muscles were altered by the microalgal diets but not those of genes associated with other quality traits. In conclusion, replacing 5 % or 10 % soybean meal with three sources of microalgae in broiler diets decreased breast weights percentage but not absolute weight. Furthermore, feeding chickens with the combination of three microalgae did not restore the breast loss and induced different expressions of genes related to muscle hypertrophy or atrophy.

59 BASIC BIOLOGICAL SCIENCES

Axenisation of oleaginous microalgal cultures via anoxic photosensitisation

Growing interest in sustainable food and biofuel research has necessitated high quality axenic oleaginous microalgal strains. Unfortunately, most strains available in culture banks contain commensal microbes such as bacteria and the default decontamination method involves antibiotic treatment which has begun to exacerbate the emergence of antibiotic resistance. To overcome this problem, anoxic photosensitisation was investigated as an alternate approach. Four oleaginous microalgal species (Tetradesmus obliquus, Desmodesmus armatus, Chlorella vulgaris and Nannochloropsis limnetica) were incubated in varying concentrations of Rose Bengal (0 μM, 1 μM, 3 μM or 9 μM) either in normal (oxic) or anoxic conditions, for 72 h under light (8.85 ± 0.4 W/m 2 ) in a specially designed heterotrophic growth complex (HGC) medium, followed by 72 h in standard Bold's Basal Medium (BBM). Commonly used antibiotics-based protocol was used as the control method. Post treatment, cell numbers and percentage populations were counted with Flow Cytometry, and viability was tested using standard plating methods using BBM and LB. Additionally, the contaminating microbes in the cultures were profiled using 16Ss rRNA sequencing. Anoxic conditions were able to significantly decrease bacterial content, albeit with an equally detrimental effect on the microalgal population. Although the responses differed between the microalgae, anoxic incubation along with Rose Bengal at 3 μM was able to completely decontaminate N. limnetica and C. vulgaris, while D. armatus and T. obliquus could be decontaminated with an additional streak-plating step. None of the cultures could be decontaminated using antibiotics treatment. These results suggest that axenisation of microalgal cultures was largely due to anoxy, that was synergistically enhanced by Rose Bengal at a concentration of ≥3 μM.

59 BASIC BIOLOGICAL SCIENCES

The Energy Costs of Dewatering Feedstock Microalgae Species Using Conventional Implementations of Ultrasonic and Crossflow Filtration Technologies

The use of microalgae as a feedstock for biofuels and other products continues to be explored, but large-scale biomass production faces persistent high costs. One major cost contributor is the energy required for harvesting and dewatering, a challenge resulting from the dilute nature of microalgae cultures as well as the properties of microalgae cells. This study investigated the energy demands of concentrating three diverse microalgae species (Nannochloropsis salina, Scenedesmus obliquus, and Chlorella luteoviridis) exhibiting different settling velocities, a key harvesting property, using conventional ultrasonic and crossflow membrane filtration. Membrane-free ultrasonic filtration removed 60-80% of the microalgae largely in steady-state operations, increasing concentrations 20-100 times, while consuming 1-6 kWh/m3 - similar to centrifugation. In contrast, crossflow membrane filtration retained nearly 100% of the microalgae, increasing concentrations over 20-fold, but consumed 158 kWh/m3 - over 10 times the energy of centrifugation. Both approaches exceeded the 10% energy target suggested in the 2010 National Algal Biofuel Technology Roadmap. This study also highlights potential sources of energy savings. For ultrasonic filtration, attention to an innovative, dynamic property of the ultrasonic chamber called the Energy Efficiency Factor, could lead to energy reductions by no more than 10-fold and improve the favorability of two of the microalgae species studied. For membrane filtration, a dead-end configuration could reduce energy consumption by no more than 1000-fold, making it favorable for all three species. This study not only highlights the need to further increase the energy content of microalgae cultures, but also the critical importance of continued development of ultrasonic filtration and other harvesting technologies to achieve low-cost, energy-sustainable algal biofuels.

09 BIOMASS FUELS

CRADA Number NFE-24-10495 with Algaeo, LLC (CRADA Final Report)

This study investigated the potential for a synthetic consortium of mutualistic terrestrial microbes— comprising the fungi Laccaria bicolor and Serendipita indica alongside bacterial Pseudomonas strains—to influence the growth and productivity of the freshwater microalgae Chlorella vulgaris. The project aimed to determine if microbial complexes engineered to enhance terrestrial plant growth could provide similar growth-promoting benefits or pathogen resistance within an aquatic algal system. Using a quantitative experimental design, C. vulgaris was co-cultured with the microbial mix under controlled laboratory conditions, with growth rates, biomass density, and metabolic activity monitored over a standard cultivation period. The results demonstrated no significant symbiotic relationship or growth enhancement between these terrestrial microbes and the microalgae, as the C. vulgaris maintained independent growth trajectories unaffected by the fungal or bacterial inoculants. We conclude that the specialized mutualisms of these fungi and bacteria are likely niche-specific to vascular plants and do not readily translate to the phycosphere of C. vulgaris. These findings are valuable to synthetic biologists and bioenergy researchers, as they define the functional boundaries of inter-kingdom microbial engineering and underscore the necessity of selecting niche-compatible species when designing consortia for industrial algal cultivation.

60 APPLIED LIFE SCIENCES

Tuning Shinkarev’s Bicycle: Separating the Parallel Cycles of Photosystem II Using Empirical Wavelet Transform

The oxygen-evolving complex (OEC) of Photosystem II (PSII) catalyzes light-driven water oxidation, a process necessary to sustain Earth’s atmospheric oxygen. Oxygen yields measured during single-turnover flash sequences exhibit period-four oscillations, which form the basis of the Joliot–Kok (S-state) model. However, when the oscillations of other processes contribute to the measured oxygen yield, fitting methods can conflate these signals and distort estimates of inefficiencies and initial S-state populations. To address this, we applied the empirical wavelet transform (EWT) as a model-independent method to separate overlapping oscillators and capture damping dynamics that are not well represented in Fourier analysis. We tested this framework on polarographic flash-oxygen traces from both our Synechocystis sp. PCC 6803 thylakoid membrane preparations and archival datasets on Chlorella and isolated chloroplasts. EWT consistently resolves the expected period-four component alongside a distinct binary oscillation. Simulations suggest that fitting this isolated period-four signal recovers VZAD parameters more accurately than analysis of raw traces, yielding different estimates for S-state distributions and transition probabilities. Notably, this binary oscillation aligns closely with semiquinone dynamics predicted solely from period-four fit parameters. These findings indicate that EWT can effectively distinguish complex signals in oxygen evolution, offering a framework potentially applicable to other spectroscopic probes of the S-state cycle.

Ferrari, Nicholas [Louisiana State Univ., Baton Ro