Tracing the Impact of Domestic Storage Conditions on Antioxidant Activity and Lipid Profiles in the Edible Microalgae Chlorella vulgaris and Tetraselmis chui.

The microalgae Chlorella vulgaris and Tetraselmis chui are valued for their nutrient-rich content, including lipids and polyunsaturated fatty acids (PUFA). However, little is known about how storage and processing affect their lipid quality. This study aimed to assess the impact of domestic storage and cooking practices in dried biomass of C. vulgaris and T. chui. Four conditions were tested: control (newly opened package), light (storage at room temperature and daily light regimen for three weeks), frozen (storage in the freezer at -20 °C for three weeks), and heated (three cycles of 90 min at 100 °C). Lipid extracts were analyzed by GC-MS and LC-MS, and antioxidant activity through DPPH and ABTS radical scavenging assays. Tested storage conditions promoted a decrease in fatty acid content and in diacyl/lyso lipid species ratios of phospholipid (PC/LPC, PE/LPE) and betaine lipids (DGTS/MGTS). Lipid extracts from light treatment showed the lowest antioxidant activity in C. vulgaris (ABTS, IC40: 104.9; DPPH, IC20: 187.9 ± 15.0), while heat affected the antioxidant activity of T. chui (ABTS, IC40: 88.5 ± 2.8; DPPH, IC20 209.4 ± 10.9). These findings underscore the impact of managing storage and processing conditions to optimize the nutritional and functional benefits of C. vulgaris and T. chui in food and feed applications.

Chlorella vulgaris-mediated bioremediation of food and beverage wastewater from industries in Mexico: Results and perspectives towards sustainability and circular economy.

The food and beverage industries in Mexico generate substantial effluents, including nejayote, cheese-whey, and tequila vinasses, which pose significant environmental challenges due to their extreme physicochemical characteristics and excessive organic load. This study aimed to assess the potential of Chlorella vulgaris in bioremediating these complex wastewaters while also producing added-value compounds. A UV mutagenesis treatment (40 min) enhanced C. vulgaris adaptability to grow in the effluent conditions. Robust growth was observed in all three effluents, with nejayote identified as the optimal medium. Physicochemical measurements conducted pre- and post-cultivation revealed notable reductions of pollutants in nejayote, including complete removal of nitrogen and phosphates, and an 85 % reduction in COD. Tequila vinasses exhibited promise with a 66 % reduction in nitrogen and a 70 % reduction in COD, while cheese-whey showed a 17 % reduction in phosphates. Regarding valuable compounds, nejayote yielded the highest pigment (1.62 mg·g-1) and phenolic compound (3.67 mg·g-1) content, while tequila vinasses had the highest protein content (16.83 %). The main highlight of this study is that C. vulgaris successfully grew in 100 % of the three effluents (without additional water or nutrients), demonstrating its potential for sustainable bioremediation and added-value compound production. When grown in 100 % of the effluents, they become a sustainable option since they don't require an input of fresh water and therefore do not contribute to water scarcity. These findings offer a practical solution for addressing environmental challenges in the food and beverage industries within a circular economy framework.

Acclimation mechanism of microalgal photosynthetic apparatus under low atmospheric pressures - new astrobiological perspectives in a Mars-like atmosphere.

This study reveals a new acclimation mechanism of the eukaryotic unicellular green alga Chlorella vulgaris in terms of the effect of varying atmospheric pressures on the structure and function of its photosynthetic apparatus using fluorescence induction measurements (JIP-test). The results indicate that low (400mbar) and extreme low (2 atmosphere (simulating the Mars atmosphere), reveals that the impact of extremely low atmospheric pressure on PQ mobility within the photosynthetic membrane, coupled with the low density of an almost 100% CO2 Mars-like atmosphere, results to a similar photosynthetic efficiency to that on Earth. These findings pave the way for the identification of novel functional acclimation mechanisms of microalgae to extreme environments that are vastly distinct from those found on Earth.

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.

The Biophysical Properties of Microalgal Cell Surfaces Govern Their Interactions with an Amphiphilic Chitosan Derivative Used for Flocculation and Flotation.

Microalgae show great promise for producing valuable molecules like biofuels, but their large-scale production faces challenges, with harvesting being particularly expensive due to their low concentration in water, necessitating extensive treatment. While methods such as centrifugation and filtration have been proposed, their efficiency and cost-effectiveness are limited. Flotation, involving air-bubbles lifting microalgae to the surface, offers a viable alternative, yet the repulsive interaction between bubbles and cells can hinder its effectiveness. Previous research from our group proposed using an amphiphilic chitosan derivative, polyoctyl chitosan (PO-chitosan), to functionalize bubbles used in dissolved air flotation (DAF). Molecular-scale studies performed using atomic force microscopy (AFM) revealed that PO-chitosan's efficiency correlates with cell surface properties, particularly hydrophobic ones, raising the question of whether this molecule can in fact be used more generally to harvest different microalgae. Evaluating this, we used a different strain of Chlorella vulgaris and first characterized its surface properties using AFM. Results showed that cells were hydrophilic but could still interact with PO-chitosan on bubble surfaces through a different mechanism based on specific interactions. Although force levels were low, flotation resulted in 84% separation, which could be explained by the presence of AOM (algal organic matter) that also interacts with functionalized bubbles, enhancing the overall separation. Finally, flocculation was also shown to be efficient and pH-independent, demonstrating the potential of PO-chitosan for harvesting microalgae with different cell surface properties and thus for further sustainable large-scale applications.

Metagenomic profiling of antibiotic resistance genes/bacteria removal in urban water: Algal-bacterial consortium treatment system.

Antibiotic resistance genes (ARGs) have exhibited significant ecological concerns, especially in the urban water that are closely associated with human health. In this study, with presence of exogenous Chlorella vulgaris-Bacillus licheniformis consortium, most of the typical ARGs and MGEs were removed. Furthermore, the relative abundance of potential ARGs hosts has generally decreased by 1-4 orders of magnitude, revealing the role of algal-bacterial consortium in cutting the spread of ARGs in urban water. While some of ARGs such as macB increased, which may be due to the negative impact of algicidal bacteria and algal viruses in urban water on exogenous C. vulgaris and the suppression of exogenous B. licheniformis by indigenous microorganisms. A new algal-bacterial interaction might form between C. vulgaris and indigenous microorganisms. The interplay between C. vulgaris and bacteria has a significant impact on the fate of ARGs removal in urban water.

Continuous UV-C/H2O2 and UV-C/Chlorine applied to municipal secondary effluent and nanofiltration retentate: Removal of contaminants of emerging concern, ecotoxicity, and reuse potential.

As global effects of water scarcity raise concerns and environmental regulations evolve, contemporary wastewater treatment plants (WWTPs) face the challenge of effectively removing a diverse range of contaminants of emerging concern (CECs) from municipal effluents. This study focuses on the assessment of advanced oxidation processes (AOPs), specifically UV-C/H2O2 and UV-C/Chlorine, for the removal of 14 target CECs in municipal secondary effluent (MSE, spiked with 10 µg L-1 of each CEC) or in the subsequent MSE nanofiltration retentate (NFR, no spiking). Phototreatments were carried out in continuous mode operation, with a hydraulic retention time of 3.4 min, using a tube-in-tube membrane photoreactor. For both wastewater matrices, UV-C photolysis (3.3 kJ L-1) exhibited high efficacy in removing CECs susceptible to photolysis, although lower treatment performance was observed for NFR. In MSE, adding 10 mg L-1 of H2O2 or Cl2 enhanced treatment efficiency, with UV-C/H2O2 outperforming UV-C/Chlorine. Both UV-C/AOPs eliminated the chronic toxicity of MSE toward Chlorella vulgaris. In the NFR, not only was the degradation of target CECs diminished, but chronic toxicity to C. vulgaris persisted after both UV-C/AOPs, with UV-C/Chlorine increasing toxicity due to potential toxic by-products. Nanofiltration permeate (NFP) exhibited low CECs and microbial content. A single chlorine addition effectively controlled Escherichia coli regrowth for 3 days, proving NFP potential for safe reuse in crop irrigation (<1 CFU/100 mL for E. coli; <1 mg L-1 for free chlorine). These findings provide valuable insights into the applications and limitations of UV-C/H2O2 and UV-C/Chlorine for distinct wastewater treatment scenarios.

Dietary supplementation with Chlorella vulgaris in broiler chickens submitted to heat-stress: effects on growth performance and meat quality.

Heat stress can greatly challenge growth and meat quality of broiler chickens where research is looking for sustainable ingredients, such as microalgae, that could also alleviate its negative impacts. Thus, in the present study, 576 1-D-old chicks (Ross 308) were housed until commercial slaughtering (42 D) in 36 pens in 2 rooms of a poultry house, according to a full factorial design encompassing 2 room temperatures (standard vs. high), 2 sexes (females vs. males), and 3 dietary treatments, that is, diet C0 (control diet), diet C3, and diet C6 containing 0, 3, and 6%, respectively, of C. vulgaris meal replacing the same quantities of soybean meal. The highest inclusion level of C. vulgaris decreased feed intake (P < 0.001) and body weight (P < 0.0001) compared to the control diet; it increased yellow and red indexes (P < 0.0001) of the breast muscle, besides the proportion of n3 polyunsaturated fatty acids (PUFA) (P = 0.028). Heat stress decreased feed intake (P = 0.001), breast (P = 0.001) and p. major yields (P = 0.036), and increased meat pH (P= 0.008) and cooking losses (P < 0.001), umami (P = 0.021) and brothy flavor (P < 0.001), and the proportion of n3 PUFA rates (P = 0.027), while reducing the contents of several amino acids in the breast meat (P ≤ 0.05). Compared to females, males displayed higher feed intake and growth, and more favorable feed conversion (P < 0.001). Carcass and p. major yields were greater in females (P < 0.001) which also showed a higher occurrence of spaghetti meat compared to males (P < 0.001). In conclusion, C. vulgaris can be used to replace until 3% of soybean meal in diets for broiler chickens without negative implications, while positively affecting breast meat color according to consumers' preferences. However, the microalgae inclusion did not mitigate the negative effects of a chronic heat stress on growth performance nor reduced the occurrence of any myopathies.

Effects of food quantity on the ingestion and egestion of MPs with different colors by Daphnia magna.

Aquatic organism uptake and accumulate microplastics (MPs) through various pathways, with ingestion alongside food being one of the primary routes. However, the impact of food concentration on the accumulation of different types of MPs, particularly across various colors, remains largely unexplored. To address this gap, we selected Daphnia magna as a model organism to study the ingestion/egestion kinetics and the preference for different MP colors under varying concentrations of Chlorella vulgaris. Our findings revealed that as the concentration of Chlorella increased, the ingestion of MPs by D. magna initially increased and then showed a decline. During the egestion phase within clean medium without further food supply, an increase in food concentration during the ingestion phase led to a slower rate of MP discharge; while when food was present during the egestion phase, the discharge rate accelerated for all treatments, indicating the importance of food ingestion/digestion process on the MPs bioaccumulation. Furthermore, in the presence of phytoplankton, D. magna demonstrated a preference for ingesting green-colored MPs, especially at low and medium level Chlorella supply, possibly due to the enhanced food searching activities. Beyond gut passage, we also examined the attachment of MPs to the organism's body surface, finding that the number of adhered MPs increased with increasing food concentration, likely due to the intensified filtering current during food ingestion. In summary, this study demonstrated that under aquatic environment with increasing phytoplankton concentrations, the ingestion and egestion rates, color preferences, as well as surface adherence of MPs to filter feeding zooplanktons will be significantly influenced, which may further pose ecological risks. Our results offer novel insights into the unintentional accumulation of MPs by zooplankton, highlighting the complex interactions between food availability and MPs accumulation dynamics.

Impact of the biomass pretreatment and simulated gastrointestinal digestion on the digestibility and antioxidant activity of microalgae Chlorella vulgaris and Tetraselmis chuii.

Chlorella vulgaris and Tetraselmis chuii are two microalgae species already marketed because of their richness in high-value and health-beneficial compounds. Previous studies have demonstrated the biological properties of compounds isolated from both microalgae, although data are not yet available on the impact that pre-treatment and gastrointestinal digestion could exert on these properties. The aim of the present study was to analyze the impact of the biomass pre-treatment (freeze/thaw cycles plus ultrasounds) and simulated gastrointestinal digestion in the bioaccessibility and in vitro antioxidant activity (ABTS, ORAC, Q-FRAP, Q-DPPH) of the released digests. The cell wall from microalgae were susceptible to the pre-treatment and the action of saliva and gastric enzymes, releasing bioactive peptides and phenolic compounds that contributed to the potent antioxidant activity of digests through their radical scavenging and iron reduction capacities. Our findings suggest the potential of these microalgae against oxidative stress-associated diseases at both, intestinal and systemic level.

Chlorella vulgaris algae ameliorates chlorpyrifos toxicity in Nile tilapia with special reference to antioxidant enzymes and Streptococcus agalactiae infection.

BACKGROUND: Chlorpyrifos (CPF) is a widely used pesticide in the production of plant crops. Despite rapid CPF biodegradation, fish were exposed to wastewater containing detectable residues. Recently, medicinal plants and algae were intensively used in aquaculture to replace antibiotics and ameliorate stress impacts. METHODS AND RESULTS: An indoor experiment was conducted to evaluate the deleterious impacts of CPF pollution on Nile tilapia health and the potential mitigation role of Chlorella vulgaris algae. Firstly, the median lethal concentration LC50 - 72 h of CPF was determined to be 85.8 µg /L in Nile tilapia (35.6 ± 0.5 g body weight) at a water temperature of 27.5 °C. Secondly, fish were exposed to 10% of LC50 - 72 h for six weeks, and tissue samples were collected and examined every two weeks. Also, Nile tilapia were experimentally infected with Streptococcus agalactiae. Exposed fish were immunosuppressed expressed with a decrease in gene expressions of interleukin (IL) 1ß, IL-10, and tumor necrosis factor (TNF)-α. Also, a decline was recorded in glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT) gene expression in the head kidney tissue. A high mortality rate (MR) of 100% was recorded in fish exposed to CPF for six weeks and challenged with S. agalactiae. Fish that received dietary C. vulgaris could restore gene expression cytokines and antioxidants compared to the control. After six weeks of CPF exposure, fish suffered from anemia as red blood cell count (RBCs), hemoglobin (Hb), and packed cell volume (PCV) significantly declined along with downregulation of serum total protein (TP), globulin (GLO), and albumin (ALB). Liver enzymes were significantly upregulated in fish exposed to CPF pollution, alanine aminotransferase (ALT) (42.5, 53.3, and 61.7 IU/L) and aspartate aminotransferase (AST) (30.1, 31.2, and 22.8) after 2, 4, and 6 weeks, respectively. On S. agalactiae challenge, high MR was recorded in Nile tilapia exposed to CPF (G3) 60%, 60%, and 100% in week 2, week 4, and week 6, and C. vulgaris provided a relative protection level (RPL) of 0, 14.29, and 20%, respectively. CONCLUSIONS: It was concluded that CPF pollution induces immunosuppressed status, oxidative stress, and anemic signs in Nile tilapia. In contrast, C. vulgaris at a 50 g/kg fish feed dose could partially ameliorate such withdrawals, restoring normal physiological parameters.

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.

Enhancing the biodiesel production in the green alga Chlorella vulgaris by heavy metal stress and prediction of fuel properties from fatty acid profiles.

The green microalga Chlorella vulgaris was used as a test organism during this study for evaluation of the impact of different heavy metal stress, Mn2+, Co2+, and Zn2+, on enhancing the biodiesel production. The algal cultures were grown for 13 days under heavy metal stress after which were subjected to estimation of growth, some primary metabolites, lipid, and fatty acid profiles. The maximum lipid accumulation (283.30 mg/g CDW) was recorded in the algal culture treated with 3 µM cobalt nitrate. Application of 2 mM manganese chloride; 1, 2, and 3 µM cobalt nitrate; and 0.2, 0.4, and 0.6 mM zinc sulfate caused highly significant increases in the lipid contents amounting to 183.8, 191.4, 230.6, 283.3, 176.3, 226.0, and 212.1 mg/g CDW, respectively, in comparison to control (153.4 mg/g CDW). The maximum proportion of saturated fatty acids (SFA) (64.44%) was noted in the culture treated with 6 mM MnCl2 due to the existence of palmitic acid (C16:0), stearic acid (C18:0), and pentadecylic acid (C15:0) which are represented by 53.59%, 5.96%, and 1.37%, respectively, of the total FAs. Relative increase in energy compound (REEC) showed that 1, 2, and 3 µM Co2+ lead to the highest stimulation in lipid and carbohydrate contents to 0.207, 0.352, and 0.329 × 103%, respectively. Empirical formulas were used for the assessment of biodiesel fuel properties based on FAME composition. The estimated properties met the prescribed international standard criteria.

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.

Formation and toxicity alteration of halonitromethanes from Chlorella vulgaris during UV/chloramination process involving bromide ion.

Frequent algal blooms cause algal cells and their algal organic matter (AOM) to become critical precursors of disinfection by-products (DBPs) during water treatment. The presence of bromide ion (Br-) in water has been demonstrated to affect the formation laws and species distribution of DBPs. However, few researchers have addressed the formation and toxicity alteration of halonitromethanes (HNMs) from algae during disinfection in the presence of Br-. Therefore, in this work, Chlorella vulgaris was selected as a representative algal precursor to investigate the formation and toxicity alteration of HNMs during UV/chloramination involving Br-. The results showed that the formation concentration of HNMs increased and then decreased during UV/chloramination. The intracellular organic matter of Chlorella vulgaris was more susceptible to form HNMs than the extracellular organic matter. When the Br-: Cl2 mass ratio was raised from 0.004 to 0.08, the peak of HNMs total concentration increased 33.99%, and the cytotoxicity index and genotoxicity index of HNMs increased 67.94% and 22.80%. Besides, the formation concentration and toxicity of HNMs increased with increasing Chlorella vulgaris concentration but decreased with increasing solution pH. Possible formation pathways of HNMs from Chlorella vulgaris during UV/chloramination involving Br- were proposed based on the alteration of nitrogen species and fluorescence spectrum analysis. Furthermore, the formation laws of HNMs from Chlorella vulgaris in real water samples were similar to those in deionized water samples. This study contributes to a better comprehension of HNMs formation from Chlorella vulgaris and provides valuable information for water managers to reduce hazards associated with the formation of HNMs.

Therapeutic potential of Lactobacillus casei and Chlorella vulgaris in high-fat diet-induced non-alcoholic fatty liver disease (NAFLD)-associated kidney damages: a stereological study.

BACKGROUND: The non-alcoholic fatty liver disease (NAFLD) is prevalent in as many as 25% of adults who are afflicted with metabolic syndrome. Oxidative stress plays a significant role in the pathophysiology of hepatic and renal injury associated with NAFLD. Therefore, probiotics such as Lactobacillus casei (LBC) and the microalga Chlorella vulgaris (CV) may be beneficial in alleviating kidney injury related to NAFLD. MATERIALS AND METHODS: This animal study utilized 30 C57BL/6 mice, which were evenly distributed into five groups: the control group, the NAFLD group, the NAFLD + CV group, the NAFLD + LBC group, and the NAFLD + CV + LBC group. A high-fat diet (HFD) was administered to induce NAFLD for six weeks. The treatments with CV and LBC were continued for an additional 35 days. Biochemical parameters, total antioxidant capacity (TAC), and the expression of kidney damage marker genes (KIM 1 and NGAL) in serum and kidney tissue were determined, respectively. A stereological analysis was conducted to observe the structural changes in kidney tissues. RESULTS: A liver histopathological examination confirmed the successful induction of NAFLD. Biochemical investigations revealed that the NAFLD group exhibited increased ALT and AST levels, significantly reduced in the therapy groups (p < 0.001). The gene expression levels of KIM-1 and NGAL were elevated in NAFLD but were significantly reduced by CV and LBC therapies (p < 0.001). Stereological examinations revealed reduced kidney size, volume, and tissue composition in the NAFLD group, with significant improvements observed in the treated groups (p < 0.001). CONCLUSION: This study highlights the potential therapeutic efficacy of C. vulgaris and L. casei in mitigating kidney damage caused by NAFLD. These findings provide valuable insights for developing novel treatment approaches for managing NAFLD and its associated complications.

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.

Pulsed electric field (PEF) processing of microalga Chlorella vulgaris and its digestibility in broiler feed.

Microalgae have potentially beneficial effects on animal health and nutritional value when added to feed. Crucial hereby is that intracellular bio-active molecules are released in the intestinal tract. Digestibility of Chlorella vulgaris and its impact on total digestibility of broiler feed is a first step in assessing its characteristics as feed supplement. Different methods could be used to increase the digestibility of the algae. Among other, pulsed electric field (PEF) and freezing to disrupt autotrophic (A) and heterotrophic (H) Chlorella vulgaris cells was assessed to increase their availability followed by in-vivo trials. In these trials effect of algae type (A and H) and effect of PEF-processing was evaluated on the apparent nutrient digestibility. Pulsed electric field showed to have a disruption efficiency of 83.90% and 79.20% for heterotrophic and autotrophic C. vulgaris respectively. Freezing C. vulgaris only showed efficiencies ranging from 3.86 to 11.58%. In the in-vivo trials, microscopic counting of intact C. vulgaris cells showed an increase in digested intact C. vulgaris cells of PEF-processed C. vulgaris compared to nonprocessed cells ranging from 12.16% to 15.20%. Autotrophic C. vulgaris had a higher digestibility compared to heterotrophic C. vulgaris, with an increase of 7.29, 9.44, and 17.29% in digestibility of C. vulgaris in the 1, 2, and 5% feed respectively. Feeds with PEF-processed C. vulgaris showed no significant increase in digestibility compared to nonprocessed C. vulgaris supplemented feeds. The 5% C. vulgaris feeds showed lower fat digestibility than the 1 and 2% and control feeds. Protein digestibility was lower for all C. vulgaris feeds compared to the control feed. There was a significant linear decreasing effect (P < 0.001) for all digestibility parameters. Except for crude ash digestibility, which first lowered for the 1 and 2% feeds, but then increased at 5% inclusion. Considering this study, including low dosages of 1 and 2% of C. vulgaris in broiler feed does not compromise its digestibility.

Predicting the Structure of Enzymes with Metal Cofactors: The Example of [FeFe] Hydrogenases.

The advent of deep learning algorithms for protein folding opened a new era in the ability of predicting and optimizing the function of proteins once the sequence is known. The task is more intricate when cofactors like metal ions or small ligands are essential to functioning. In this case, the combined use of traditional simulation methods based on interatomic force fields and deep learning predictions is mandatory. We use the example of [FeFe] hydrogenases, enzymes of unicellular algae promising for biotechnology applications to illustrate this situation. [FeFe] hydrogenase is an iron-sulfur protein that catalyzes the chemical reduction of protons dissolved in liquid water into molecular hydrogen as a gas. Hydrogen production efficiency and cell sensitivity to dioxygen are important parameters to optimize the industrial applications of biological hydrogen production. Both parameters are related to the organization of iron-sulfur clusters within protein domains. In this work, we propose possible three-dimensional structures of Chlorella vulgaris 211/11P [FeFe] hydrogenase, the sequence of which was extracted from the recently published genome of the given strain. Initial structural models are built using: (i) the deep learning algorithm AlphaFold; (ii) the homology modeling server SwissModel; (iii) a manual construction based on the best known bacterial crystal structure. Missing iron-sulfur clusters are included and microsecond-long molecular dynamics of initial structures embedded into the water solution environment were performed. Multiple-walkers metadynamics was also used to enhance the sampling of structures encompassing both functional and non-functional organizations of iron-sulfur clusters. The resulting structural model provided by deep learning is consistent with functional [FeFe] hydrogenase characterized by peculiar interactions between cofactors and the protein matrix.

Nutrients recovery from dairy wastewater by Chlorella vulgaris and comparison of the lipid's composition with various chlorella strains for biodiesel production.

Microalgae biomass is regarded as a promising feedstock for biodiesel production. The biomass lipid content and fatty acids composition are among the main selective criteria when screening microalgae strains for biodiesel production. In this study, three strains of Chlorella microalgae (C. kessleri, C. sorokiniana, C. vulgaris) were cultivated nutrient media with different nitrogen contents, and on a medium with the addition of dairy wastewater. Moreover, microalgae grown on dairy wastewater allowed the removal of azote and phosphorous. The removal efficiency of 90%, 53% and 95% of ammonium nitrogen, total nitrogen and phosphate ions, respectively, were reached. The efficiency of wastewater treatment from inorganic carbon was 55%, while the maximum growth of biomass was achieved. All four samples of microalgae had a similar fatty acid profile. Palmitic acid (C16:0) was the most abundant saturated fatty acid (SFA), and is suitable for the production of biodiesel. The main unsaturated fatty acids (UFA) present in the samples were oleic acid (C18:1 n9); linoleic acid (C18:2 n6) and alpha-linolenic acid (C18:3 n3), which belong to omega-9, omega-6, omega-3, respectively.