Cultivation and nutritional characteristics of Chlorella vulgaris cultivated using Martian regolith and synthetic urine.

Long-term spatial missions will require sustainable methods for biomass production using locally available resources. This study investigates the feasibility of cultivating Chlorella vulgaris, a high value microalgal specie, using a leachate of Martian regolith and synthetic human urine as nutrient sources. The microalga was grown in a standard medium (BBM) mixed with 0, 20, 40, 60, or 100 % Martian medium (MM). MM did not significantly affect final biomass concentrations. Total carbohydrate and protein contents decreased with increasing MM fractions between 0 % and 60 %, but biomass in the 100% MM showed the highest levels of carbohydrates and proteins (25.2 ± 0.9 % and 37.1 ± 1.4 % of the dry weight, respectively, against 19.0 ± 1.7 % and 32.0 ± 2.7 % in the absence of MM). In all MM-containing media, the fraction of the biomass represented by total lipids was lower (by 3.2 to 4.5%) when compared to BBM. Conversely, total carotenoids increased, with the highest value (97.3 ± 1.5 mg/100 g) measured with 20% MM. In a three-dimensional principal component analysis of triacylglycerols, samples clustered according to growth media; a strong impact of growth media on triacylglycerol profiles was observed. Overall, our findings suggest that microalgal biomass produced using regolith and urine can be used as a valuable component of astronauts' diet during missions to Mars.

Engineered Chlorella vulgaris improves bioethanol production and promises prebiotic application.

Microalgal biomass for biofuel production, integration into functional food, and feed supplementation has generated substantial interest worldwide due to its high growth rate, non-competitiveness for agronomic land, ease of cultivation in containments, and presence of several bioactive molecules. In this study, genetic engineering tools were employed to develop transgenic lines of freshwater microalga Chlorella vulgaris with a higher starch content, by up-regulating ADP-glucose pyrophosphorylase (AGPase), which is a rate-limiting enzyme in starch biosynthesis. Expression of the Escherichia coli glgC (AGPase homolog) gene in C. vulgaris led to an increase in total carbohydrate content up to 45.1% (dry cell weight, DCW) in the transgenic line as compared to 34.2% (DCW) in the untransformed control. The starch content improved up to 16% (DCW) in the transgenic alga compared to 10% (DCW) in the control. However, the content of total lipid, carotenoid, and chlorophyll decreased differentially in the transgenic lines. The carbohydrate-rich biomass from the transgenic algal line was used to produce bioethanol via yeast fermentation, which resulted in a higher ethanol yield of 82.82 mg/L as compared to 54.41 mg/L from the untransformed control. The in vitro digestibility of the transgenic algal starch revealed a resistant starch content of up to 7% of total starch. Faster growth of four probiotic bacterial species along with a lowering of the pH of the growth medium indicated transgenic alga to exert a positive prebiotic effect. Taken together, the study documents the utilization of genetically engineered C. vulgaris with enriched carbohydrates as bioethanol feedstock and functional food ingredients.

N-acyl homoserine lactones (AHLs) enhanced removal of cadmium and other pollutants by algae-bacteria consortia.

Signal transduction is an important mode of algae-bacteria interaction, in which bacterial quorum sensing (QS) may affect microalgal growth and metabolism. Currently, little is known whether acyl homoserine lactones (AHLs) released by bacteria can affect the pollutant removal by algae-bacteria consortia (ABC). In this study, we constructed ABC using Chlorella vulgaris (Cv) with two AHLs-producing bacteria and investigated their performance in the removal of multiple pollutants, including chemical oxygen demand (COD), total nitrogen (TN), phosphorus (P), and cadmium (Cd). The AHLs-producing bacteria, namely Agrobacterium sp. (Ap) and Ensifer adherens (Ea), were capable of forming a symbiosis with C. vulgaris. Consortia of Cv and Ap with ratio of 2:1 (Cv2-Ap1) showed the optimal growth promotion and higher removal of Cd, COD, TN, and P compared to the C. vulgaris monoculture. Cv2-Ap1 ABC removed 36.1-47.5% of Cd, 94.5%-94.6% COD, 37.1%-56.0% TN, and 90.4%-93.5% P from the culture medium. In addition, increase of intracellular neutral lipids and extracellular protein, as well as the types of functional groups on cell surface contributed to Cd removal and tolerance in the Cv2-Ap1 ABC. Six AHLs were detected in the Cv2-Ap1 culture. Among these, 3OC8-HSL and 3OC12-HSL additions promoted the ABC growth and enhanced their Cd accumulation. These findings may contribute to further understanding of AHL-mediated communication between algae and bacteria and provide support bioremediation efforts of metal-containing wastewater.

Towards a circular economy - Repurposing side streams from the potato processing industry by Chlorella vulgaris.

Common wastewater treatment strategies in the food industry do not include efficient remediation strategies for nitrogen, phosphorous and organic carbon. Incorporating microalgae in water treatment plants is rising in popularity because of their high nutrient and trace element uptake driven by light. In this study, four different side streams from an Austrian potato processing company have been screened for their applicability of microalgal cultivation. The side streams were assessed for Chlorella vulgaris growth and their requirement of any additional pretreatment or media supplementation. One side stream specifically, called blanching water II, a stream generated by boiling the potatoes for ease of peeling, turned out very useful to cultivate Chlorella vulgaris and concomitantly remedy the wastewater. Compared to a state-of-the-art cultivation in BG11, cultivating Chlorella vulgaris in blanching water II led to a 45 % increase in specific growth rate of 1.29 day-1 and a 48% increase in biomass productivity to 294.6 mg/L/day, while all nitrogen and phosphate present in the side stream were metabolized. Overall, the results demonstrate that the water remediation process for blanching water II shows vast potential in regard to water purification and waste to value approaches.

Cultivation of Chlorella vulgaris in wastewater: biodiesel potential and wastewater remediation.

The present investigation has evaluated the use of effluents from a secondary municipal wastewater treatment plant for biomass production and potential of the biomass for biodiesel production. Cultivations of Chlorella vulgaris using wastewater, wastewater with supplementation, and WC medium were carried out. Effect of wastewater collected in different months on biomass productivity (BP) and lipid composition was studied. Methods based on NMR and GC-MS techniques were applied for determining the composition of the lipids and their fatty acid profile including poly unsaturated fatty acids (PUFAs). Lipids extracted are comprised of both neutral (tri acyl glycerides, TAG; free fatty acids, FFA) and polar (glyco glycero/phospho) lipids. The TAG content of the extracted lipids was determined in the range of 22.5-41.3% w/w. The NMR and GC-MS compositional results of microalgal lipids of biomasses cultivated in wastewater without nutrient supplementation, collected in different months, showed potential for biodiesel production. The fatty acid profiles of neutral and polar lipids, which are mainly comprised of saturated and unsaturated long alkyl chain (C16-C22) fatty acids, are potential sources for the biodiesel and food industry. The concentration of nitrates (45-78 mg L-1) in wastewater without supplementation, collected in different months, was found to be optimum to enable cultivation of biomasses with reasonably good BP of 21.5-28.1 mg L-1 day-1. Similar results have been obtained in the present work as well as reported in the literature in the case of WC medium (nitrate, 69 mg L-1) with BP of 25.5-28.2 mg L-1 day-1, thus highlighted the significance of the presented work.

Enhancing immobilized Chlorella vulgaris growth with novel buoyant barium alginate bubble beads.

Microalgae immobilization in alginate beads shows promise for biomass production and water pollution control. However, carrier instability and mass transfer limitations are challenges. This study introduces buoyant barium alginate bubble beads (BABB), which offer exceptional stability and enhance Chlorella vulgaris growth. In just 12 days, compared to traditional calcium alginate beads, BABB achieved a 20 % biomass increase while minimizing cell leakage and simplifying harvesting. BABB optimization involved co-immobilization with BG-11 medium, enrichment of CO2 in internal bubbles, and the integration of Fe nanoparticles (FeNPs). In the open raceway pond reactor, these optimizations resulted in a 39 % increase in biomass over 7 days compared to the unoptimized setup in closed flasks. Furthermore, enhancements in pigment and organic matter production were observed, along with improved removal of ammonia nitrogen and phosphate. These results highlight the overall advantages of BABB for microalgae immobilization, offering a scientific foundation for their effective utilization.

Microcystin-LR (MC-LR) inhibits green algae growth by regulating antioxidant and photosynthetic systems.

Microcystins release from bloom-forming cyanobacteria is considered a way to gain competitive advantage in Microcystis populations, which threaten water resources security and aquatic ecological balance. However, the effects of microcystins on microalgae are still largely unclear. Through simulated culture experiments and the use of UHPLC-MS-based metabolomics, the effects of two microcystin-LR (MC-LR) concentrations (400 and 1,600 µg/L) on the growth and antioxidant properties of three algae species, the toxic Microcystis aeruginosa, a non-toxic Microcystis sp., and Chlorella vulgaris, were studied. The MC-LR caused damage to the photosynthetic system and activated the protective mechanism of the photosynthetic system by decreasing the chlorophyll-a and carotenoid concentrations. Microcystins triggered oxidative stress in C. vulgaris, which was the most sensitive algae species studied, and secreted more glycolipids into the extracellular compartment, thereby destroying its cell structure. However, C. vulgaris eliminated reactive oxygen species (ROS) by secreting terpenoids, thereby resisting oxidative stress. In addition, two metabolic pathways, the vitamin B6 and the sphingolipid pathways, of C. vulgaris were significantly disturbed by microcystins, contributing to cell membrane and mitochondrial damage. Thus, both the low (400 µg/L) and the high (1,600 µg/L) MC-LR concentration inhibited algae growth within 3 to 7 days, and the inhibition rates increased with the increase in the MC-LR concentration. The above results indicate that the toxin-producing Microcystis species have a stronger toxin tolerance under longer-term toxin exposure in natural water environments. Thus, microcystins participates in interspecific interaction and phytoplankton population regulation and creates suitable conditions for the toxin-producing M. aeruginosa to become the dominant species in algae blooms.

Production and Evaluation of Set-Type Yoghurt Fortified with Chlorella vulgaris and Moringa oleifera Powder.

Considering the growing popularity of functional foods, fortifying yoghurt with natural ingredients with various flavours and appearances could improve its nutritional and health potential. The current study aimed to evaluate the effect of Chlorella vulgaris (0.3 and 0.5%) and Moringa oleifera (0.3 and 0.5%) on the fermentation kinetics, apparent viscosity, antioxidant activity, microbiological, sensorial, and FTIR properties of yoghurt during storage. The results demonstrated that the incorporation of Chlorella vulgaris and Moringa oleifera into yoghurt increased acidification rate and decreased fermentation time (p < 0.05). Moringa oleifera (0.5%) improved the growth and survival of lactic acid bacteria as well as the phenolic and antioxidant properties of yoghurt. However, Chlorella vulgaris, at a concentration of 0.5% reduced the viability of lactic acid bacteria, viscosity, total phenolic, and antioxidant properties of yoghurt. In conclusion, it was found that Chlorella vulgaris, at 0.3%, and Moringa oleifera improved the phenolic, antioxidant properties, and acidification rate of yoghurt.

A Comparative Analysis Assessing Growth Dynamics of Locally Isolated Chlorella sorokiniana and Chlorella vulgaris for Biomass and Lipid Production with Biodiesel Potential.

Prior research has emphasized the potential of microalgae in biodiesel production, driven by their ability to replace fossil fuels. However, the significant costs associated with microalgae cultivation present a major obstacle to scaling up production. This study aims to develop an eco-friendly microalgae cultivation system by integrating carbon dioxide from flue gas emissions with an affordable photobioreactor, providing a sustainable biomass production. The research evaluates the growth performance of Chlorella sorokiniana and Chlorella vulgaris across this integrated system for biomass and lipid production. Results indicate substantial biomass yields of 1.97 and 1.84 g/L, with lipid contents of 35 % and 41 % for C. sorokiniana and C. vulgaris, respectively. The macrobubble photobioreactor demonstrates high potential for microalgae biomass and lipid production, yielding quality fatty acid methyl esters such as palmitic, linoleic and stearic. This study presents an environmentally friendly system for efficient microalgae cultivation, generating lipid-rich biomass suitable for biodiesel production.

Exploring the potential nutritional benefits of Arthrospira maxima and Chlorella vulgaris: A focus on vitamin B12, amino acids, and micronutrients.

Arthrospira (Limnospira) maxima (A. maxima) and Chlorella vulgaris (Ch. vulgaris) are among the approved microalgae and cyanobacteria (MaC) in the food industry that are known to be safe for consumption. However, both organisms are controversial regarding their vitamin B12 content, due to the possible occurrence of pseudo-cobalamin. Concurrently, their nutrition profiles remain understudied. The main purpose of the present study was to identify their nutrition profiles, focusing mainly on vitamin B12, amino acids, and micronutrients under iron-induced hormesis (10 mg/L Fe in treated samples). Our findings indicate a higher B12 content in A. maxima compared to Ch. vulgaris (both control and treated samples). Using liquid chromatography with tandem mass spectrometry (LC-MS/MS), the cyanocobalamin content was determined as 0.42 ± 0.09 µg/g dried weight (DW) in the A. maxima control and 0.55 ± 0.02 µg/g DW in treated A. maxima, resulting in an insignificant difference. In addition, the iron-enriched medium increased the amount of iron in both tested biomasses (p < 0.01). However, a more pronounced (approximately 100×) boost was observed in Ch. vulgaris, indicating a better absorption capacity (control Ch. vulgaris 0.16 ± 0.01 mg/g Fe, treated Ch. vulgaris 15.40 ± 0.34 mg/g Fe). Additionally, Ch. vulgaris also showed a higher micronutrient content. Using both tested microalgae, meeting the sufficient recommended daily mineral allowance for an adult is possible. By combining biomass from A. maxima and Ch. vulgaris in a ratio of 6:1, we can fulfill the recommended daily allowance of vitamin B12 and iron by consuming 6 tablets/6 g. Importantly, iron hormesis stimulated amino acid composition in both organisms. The profile of amino acids may suggest these biomasses as promising potential nutrition sources.

Modeling and optimization of biomass productivity of Chlorella vulgaris using response surface methodology, analysis of variance and machine learning for carbon dioxide capture.

This study explores bioremediation's effectiveness in reducing carbon emissions through the use of microalgae Chlorella vulgaris, known for capturing carbon dioxide and producing biomass. The impact of temperature and light intensity on productivity and carbon dioxide capture was investigated, and cultivation conditions were optimized in a photobioreactor using response surface methodology (RSM), analysis of variance (ANOVA), and deep neural networks (DNN). The optimal conditions determined were 28.74 °C and 225 µmol/m2/s with RSM, and 29.55 °C and 226.77 µmol/m2/s with DNN, closely aligning with literature values (29 °C and 225 µmol/m2/s). DNN demonstrated superior performance compared to RSM, achieving higher accuracy due to its capacity to process larger datasets using epochs and batches. The research serves as a foundation to further in this field by demonstrating the potential of utilizing diverse mathematical models to optimize bioremediation conditions, and offering valuable insights to improve carbon dioxide capture efficiency in microalgae cultivation.

Removal of polycyclic aromatic hydrocarbons (PAHs) from produced water using the microalgae Chlorella vulgaris cultivated in mixotrophic and heterotrophic conditions.

Chlorella vulgaris was cultivated for 15 days in 10 different treatments under mixotrophic and heterotrophic conditions, using wastewater from oil and poultry industries as the culture medium. The blends were made with produced water (PW), sterilized produced water (PWs), sterilized poultry wastewater (PoWs), sterilized seawater (SWs), and the addition of sodium nitrate to evaluate cell growth in treatments and the removal of PAHs. The heterotrophic condition showed more effective removal, having an initial concentration of 3.93 µg L-1 and a final concentration of 0.57 µg L-1 of total PAHs reporting 83%, during phycoremediation of (PW) than the mixotrophic condition, with an initial concentration of 3.93 µg L-1 and a final concentration of 1.96 and 43% removal for the PAHs. In the heterotrophic condition, the blend with (PWs + SWs) with an initial concentration of 0.90 µg L-1 and a final concentration of 0.32 µg L-1 had 64% removal of total PAHs compared to the mixotrophic condition with 37% removal having an initial concentration of 0.90 µg L-1 and a final concentration of 0.56 µg L-1. However, the best result in the mixotrophic condition was obtained using a blend of (PWs + PoWs) that had an initial cell concentration of 1.18 × 105 cells mL-1 and reached a final cell concentration of 4.39 × 105 cells mL-1, an initial concentration of 4.76 µg L-1 and a final concentration of 0.37 µg L-1 having a 92% total removal of PAHs. The biostimulation process increased the percentage of PAHs removal by 45% (PW) in the mixotrophic condition. This study showed that it is possible to allow an environmental remediation strategy that significantly reduces effluent toxicity and generates high value-added biomass in contaminated effluents rich in nutrients and carbon, based on a circular bioeconomy model.

Impact of organic matter molecular weight on hexavalent chromium enrichment in green microalgae.

In lightly polluted water containing heavy metals, organic matter, and green microalgae, the molecular weight of organic matter may influence both the growth of green microalgae and the concentration of heavy metals. This study elucidates the effects and mechanisms by which different molecular weight fractions of fulvic acid (FA), a model dissolved organic matter component, facilitate the bioaccumulation of hexavalent chromium (Cr(VI)) in a typical green alga, Chlorella vulgaris. Findings show that the addition of FA fractions with molecular weights greater than 10 kDa significantly enhances the enrichment of total chromium and Cr(VI) in algal cells, reaching 21.58%-31.09 % and 16.17 %-22.63 %, respectively. Conversely, the efficiency of chromium enrichment in algal cells was found to decrease with decreasing molecular weight of FA. FA molecular weight within the range of 0.22 µm-30 kDa facilitated chromium enrichment primarily through the algal organic matter (AOM) pathway, with minor contributions from the algal cell proliferation and extracellular polymeric substances (EPS) pathways. However, with decreasing FA molecular weight, the AOM and EPS pathways become less prominent, whereas the algal cell proliferation pathway becomes dominant. These findings provide new insights into the mechanism of chromium enrichment in green algae enhanced by medium molecular weight FA.

Concentration-dependent effects of spinetoram on nontarget freshwater microalgae: A comparative study on Chlorella vulgaris and Microcystis aeruginosa.

The rising global demand for agricultural products is leading to the widespread application of pesticides, such as spinetoram, resulting in environmental pollution and ecotoxicity to nontarget organisms in aquatic ecosystems. This research focused on assessing the toxicity of spinetoram at various concentrations (0, 0.01, 0.1, 0.5, 1.0, and 3.0 mg L-1) on two common freshwater microalgae, Chlorella vulgaris and Microcystis aeruginosa, to shed light on the ecotoxicological effects of insecticides. Our findings demonstrate that M. aeruginosa is more sensitive to spinetoram than is C. vulgaris, with a concentration-dependent reduction in the growth rate observed for M. aeruginosa, whereas only the highest concentration of spinetoram adversely affected C. vulgaris. At a concentration of 0.01 mg L-1, the growth rate of M. aeruginosa unexpectedly increased beginning on day 7, indicating a potential hormetic effect. Although initial exposure to spinetoram improved the photosynthetic efficiency of both microalgae strains at all concentrations, detrimental effects became apparent at higher concentrations and with prolonged exposure. The photosynthetic efficiency of C. vulgaris recovered, in contrast to that of M. aeruginosa, which exhibited limited recovery. Spinetoram more significantly inhibited the effective quantum yield of PSII (EQY) in M. aeruginosa than in C. vulgaris. Although spinetoram is not designed to target phytoplankton, its toxicity can disrupt primary productivity and modify phytoplankton-consumer interactions via bottom-up control mechanisms. This study enhances our understanding of spinetoram's ecotoxicity and potential effects on aquatic ecosystems.

Enhancing bioremediation potential of microalgae Chlorella vulgaris and Scenedesmus acutus by NaCl for pyrene degradation.

Microalgae are increasingly recognized as promising organisms for bioremediation of organic pollutants. This study investigates the potential of enhancing the bioremediation efficiency of pyrene (PYR), a polycyclic aromatic hydrocarbon (PAH), through NaCl induced physiological and biochemical alterations in two microalgae species, Chlorella vulgaris and Scenedesmus acutus. Our findings reveal significant improvement in PYR removal when these microalgae were cultivated in the presence of 0.1% NaCl where PYR removal increased from 54 to 74% for C. vulgaris and from 26 to 75% for S. acutus. However, it was observed that NaCl induced stress had varying effects on the two species. While C. vulgaris exhibited increased PYR removal, it experienced reduced growth and biomass production, as well as lower photosynthetic efficiency when exposed to PYR and PYR + NaCl. In contrast, S. acutus displayed better growth and biomass accumulation under PYR + NaCl conditions, making it a more efficient candidate for enhancing PYR bioremediation in the presence of NaCl. In addition to assessing growth and biochemical content, we also investigated stress biomarkers, such as lipid peroxidation, polyphenol and proline contents. These findings suggest that S. acutus holds promise as an alternative microalgae species for PYR removal in the presence of NaCl, offering potential advantages in terms of bioremediation efficiency and ecological sustainability. This study highlights the importance of understanding the physiological and biochemical responses of microalgae to environmental stressors, which can be harnessed to optimize bioremediation strategies for the removal of organic pollutants like PYR.

View on a mechanistic model of Chlorella vulgaris in incubated shake flasks.

Kinetic growth models are a useful tool for a better understanding of microalgal cultivation and for optimizing cultivation conditions. The evaluation of such models requires experimental data that is laborious to generate in bioreactor settings. The experimental shake flask setting used in this study allows to run 12 experiments at the same time, with 6 individual light intensities and light durations. This way, 54 biomass data sets were generated for the cultivation of the microalgae Chlorella vulgaris. To identify the model parameters, a stepwise parameter estimation procedure was applied. First, light-associated model parameters were estimated using additional measurements of local light intensities at differ heights within medium at different biomass concentrations. Next, substrate related model parameters were estimated, using experiments for which biomass and nitrate data were provided. Afterwards, growth-related model parameters were estimated by application of an extensive cross validation procedure.

Simulating the Interplay between the Uptake of Inorganic Phosphate and the Cell Phosphate Metabolism under Phosphorus Feast and Famine Conditions in Chlorella vulgaris.

Using a mathematical simulation approach, we studied the dynamics of the green microalga Chlorella vulgaris phosphate metabolism response to shortage and subsequent replenishing of inorganic phosphate in the medium. A three-pool interaction model was used to describe the phosphate uptake from the medium, its incorporation into the cell organic compounds, its storage in the form of polyphosphates, and culture growth. The model comprises a system of ordinary differential equations. The distribution of phosphorous between cell pools was examined for three different stages of the experiment: growth in phosphate-rich medium, incubation in phosphate-free medium, and phosphate addition to the phosphorus-starving culture. Mathematical modeling offers two possible scenarios for the appearance of the peak of polyphosphates (PolyP). The first scenario explains the accumulation of PolyP by activation of the processes of its synthesis, and the decline in PolyP is due to its redistribution between dividing cells during growth. The second scenario includes a hysteretic mechanism for the regulation of PolyP hydrolysis, depending on the intracellular content of inorganic phosphate. The new model of the dynamics of P pools in the cell allows one to better understand the phenomena taking place during P starvation and re-feeding of the P-starved microalgal cultures with inorganic phosphate such as transient PolyP accumulation. Biotechnological implications of the observed dynamics of the polyphosphate pool of the microalgal cell are considered. An approach enhancing the microalgae-based wastewater treatment method based on these scenarios is proposed.

Contribution of vitamin B12 to biogas upgrading and nutrient removal by different microalgae-based technology.

The algae-based technology has a positive effect on the treatment of biogas slurry and the purification of biogas, while vitamin B12 (VB12) is one of the important regulatory substances in the algae-based cultivation system. In this study, different concentrations of VB12 were used in three microalgal treatment technologies to assess their effect on simultaneous removal of nutrients from biogas slurry and removal of CO2 from raw biogas. Results showed that Chlorella vulgaris exhibited higher growth rate, mean daily productivity, chlorophyll a content, carbonic anhydrase activity and better photosynthetic properties when co-cultivated with Ganoderma lucidum, rather than when co-cultivated with activated sludge or under mono-cultivation. Maximum mean chemical oxygen demand, total nitrogen, total phosphorus and CO2 removal efficiencies were found to be 84.29 ± 8.28%, 83.27 ± 8.14%, 85.27 ± 8.46% and 65.71 ± 6.35%, respectively when microalgae were co-cultivated with Ganoderma lucidum under 100 ng L-1 of VB12. This study shows the potential of microalgae and fungi co-cultivation supplemented with VB12 for the simultaneous upgradation of biogas production as well as for the purification of biogas slurry.

Feedback between bottom-up and top-down control of stream biofilm mediated through eutrophication effects on grazer growth.

Algal biofilms in streams are simultaneously controlled by light and nutrient availability (bottom-up control) and by grazing activity (top-down control). In addition to promoting algal growth, light and nutrients also determine the nutritional quality of algae for grazers. While short-term experiments have shown that grazers increase consumption rates of nutrient-poor algae due to compensatory feeding, nutrient limitation in the long run can constrain grazer growth and hence limit the strength of grazing activity. In this study, we tested the effects of light and phosphorus availability on grazer growth and thus on the long-term control of algal biomass. At the end of the experiment, algal biomass was significantly affected by light, phosphorus and grazing, but the interactive effects of the three factors significantly changed over time. At both high light and phosphorus supply, grazing did not initially reduce algal biomass, but the effect of grazing became stronger in the final three weeks of the experiment. Snail growth was enhanced by light, rather than phosphorus, suggesting that algal quantity rather than quality was the main limiting factor for grazer growth. Our results highlight the role of feedback effects and the importance of long-term experiments in the study of foodweb interactions.

High-throughput screening for heterotrophic growth in microalgae using the Biolog Plate assay.

Microalgae produce a broad range of organic compounds that are increasingly being recognised for their value in novel product production and biotechnological applications. Most microalgae are photoautotrophic, but some are capable of either mixotrophy or heterotrophy. Reported enhanced biomass yields or contrasting metabolite profiles compared to autotrophic growth improve the economics of large-scale production of microalgae, which currently limits industrial applications. Here, the potential of a high-throughput method for the rapid screening of microalgal metabolism was assessed against 95 different carbon sources, using the cost-effective Biolog plate. Of the 5 microalgae tested, Desmodesmus communis (30 carbon sources) and Chlorella vulgaris (19 carbon sources) had the highest number of positive responses to carbon sources, whereas Chlorella sorokiniana had the most negative (toxic) response to the various carbon sources (77 carbon sources). Comparison of Biolog plate results with traditional culture techniques showed good agreement. Species with a high number of positive responses on the Biolog plate exhibited the highest biomass yield under heterotrophic conditions, whilst those with low number of positive responses exhibited the highest biomass yield under autotrophic conditions, using traditional culturing techniques. While the use of these plates is limited to obtaining axenic lines of microalgal species, the method provided a high-throughput assessment of carbon source metabolism, without the expense of undertaking large, laborious traditional culturing assessments. Such high-throughput assessments can be regarded as useful tools for progressing species selection, metabolic capacity and optimal culture conditions for microalgal biotechnology applications.