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.

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.

Inorganic carbon limitation decreases ammonium removal and N2O production in the algae-nitrifying bacteria symbiosis system.

Ammonium removal by a symbiosis system of algae (Chlorella vulgaris) and nitrifying bacteria was evaluated in a long-term photo-sequencing batch reactor under varying influent inorganic carbon (IC) concentrations (15, 10, 5 and 2.5 mmol L-1) and different nitrogen loading rate (NLR) conditions (270 and 540 mg-N L-1 d-1). The IC/N ratios provided were 2.33, 1.56, 0.78 and 0.39, respectively, for an influent NH4+-N concentration of 90 mg-N L-1 (6.43 mmol L-1). The results confirmed that both ammonium removal and N2O production were positively related with IC concentration. Satisfactory ammonium removal efficiencies (>98 %) and rates (29-34 mg-N gVSS-1 h-1) were achieved regardless of NLR levels under sufficient IC of 10 and 15 mmol L-1, while insufficient IC at 2.5 mmol L-1 led to the lowest ammonium removal rates of 0 mg-N gVSS-1 h-1. The ammonia oxidation process by ammonia oxidizing bacteria (AOB) played a predominant role over the algae assimilation process in ammonium removal. Long-time IC deficiency also resulted in the decrease in biomass and pigments of algae and nitrifying bacteria. IC limitation led to the decreasing N2O production, probably due to its negative effect on ammonia oxidation by AOB. The optimal IC concentration was determined to be 10 mmol L-1 (i.e., IC/N of 1.56, alkalinity of 500 mg CaCO3 L-1) in the algae-bacteria symbiosis reactor, corresponding to higher ammonia oxidation rate of ∼41 mg-N gVSS-1 h-1 and lower N2O emission factor of 0.13 %. This suggests regulating IC concentrations to achieve high ammonium removal and low carbon emission simultaneously in the algae-bacteria symbiosis wastewater treatment process.

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.

Molecular mechanism of dissolvable metal nanoparticles-enhanced CO2 fixation by algae: Metal-chlorophyll synthesis.

Algae-driven photosynthetic CO2 fixation is a promising strategy to mitigate global climate changes and energy crises. Yet, the presence of metal nanoparticles (NPs), particularly dissolvable NPs, in aquatic ecosystems introduces new complexities due to their tendency to release metal ions that may perturb metabolic processes related to algal CO2 fixation. This study selected six representative metal NPs (Fe3O4, ZnO, CuO, NiO, MgO, and Ag) to investigate their impacts on CO2 fixation by algae (Chlorella vulgaris). We discovered an intriguing phenomenon that bivalent metal ions released from the metal NPs, especially from ZnO NPs, substituted Mg2+ within the porphyrin ring. This interaction led to 81.8% and 76.1% increases in Zinc-chlorophyll and Magnesium-chlorophyll contents within algal cells at 0.01 mM ZnO NPs, respectively. Integrating metabolomics and transcriptomics analyses revealed that ZnO NPs mainly promoted the photosynthesis-antenna protein pathway, porphyrin and chlorophyll metabolism, and carbon fixation pathway, thereby mitigating the adverse effects of Zn2+ substitution in light harvesting and energy transfer for CO2 fixation. Ultimately, the genes encoding Rubisco large subunit (rbcL) responsible for CO2 fixation were upregulated to 2.60-fold, resulting in a 76.3% increase in carbon fixation capacity. Similar upregulations of rbcL expression (1.13-fold) and carbon fixation capacity (76.1%) were observed in algal cells even at 0.001 mM ZnO NPs, accompanied by valuable lipid accumulation. This study offers novel insights into the molecular mechanism underlying NPs on CO2 fixation by algae and potentially introduces strategies for global carbon sequestration.

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.

Chlorella alleviates the intestinal damage of tilapia caused by microplastics.

Polyethylene microplastics (MPs) of the different sizes may result in different response in fish. Studies showed microorganisms adhered to the surface of MPs have toxicological effect. Juveniles tilapia (Oreochromis niloticus, n = 600, 26.5 ± 0.6 g) were dispersed into six groups: the control group (A), 75 nm MP exposed group (B), 7.5 µm group (C) and 750 (D) µm group, 75 nm + 7.5 µm+750 µm group (E) and 75 nm + Chlorella vulgaris group (F), and exposed for 10 and 14 days. The intestinal histopathological change, enzymic activities, and the integrated "omics" workflows containing transcriptomics, proteomics, microbiota and metabolomes, have been performed in tilapia. Results showed that MPs were distributed on the surface of goblet cells, Chlorella group had severe villi fusion without something like intestinal damage, as in other MPs groups. The intestinal Total Cholesterol (TC, together with group E) and Tumor Necrosis Factor α (TNFα, except for group B) contents in group F were significantly increased, cytochrome p450 1a1 (EROD, group B and E) significantly increased, adenosine triphosphate (ATP), lipoprotein lipase (LPL) and caspase 3 (except group B) also significantly increased at 14 d. At 14 days, group E saw considerably higher regulation of the actin cytoskeleton, focal adhesion, insulin signaling pathway, and AGE-RAGE signaling pathway in diabetes complications. Whereas, chlorella enhanced the focal adhesion, cytokine-cytokine receptor interaction, and MAPK signaling pathways. PPAR signaling pathway has been extremely significantly enriched via the proteomics method. Candidatus latescibacteria, C. uhrbacteria, C. abyssubacteria, C. cryosericota significantly decreased caused by MPs of different particle sizes. Carboxylic acids and derivatives, indoles and derivatives, organooxygen compounds, fatty acyls and organooxygen compounds significantly increased with long-term duration, especially PPAR signaling pathway. MPs had a size-dependent long-term effect on histopathological change, gene and protein expression, and gut microbial metabolites, while chlorella alleviates the intestinal histopathological damage via the integrated "omics" workflows.

Interactions of monolayer molybdenum disulfide sheets with metalloid antimony in aquatic environment: Adsorption, transformation, and joint toxicity.

The tremendous application potentiality of transitional metal dichalcogenides (TMDs), such as molybdenum disulfide (MoS2) nanosheets, will unavoidably lead to increasing release into the environment, which could influence the fate and toxicity of co-existed contaminants. The present study discovered that 59.8 % of trivalent antimony [Sb(III)] was transformed by MoS2 to pentavalent Sb [Sb(V)] in aqueous solutions under light illumination, which was due to hole oxidation on the nanosheet surfaces. A synergistic toxicity between MoS2 and Sb(III, V) to algae (Chlorella vulgaris) was observed, as demonstrated by the lower median-effect concentrations of MoS2 + Sb(III)/Sb(V) (13.1 and 20.9 mg/L, respectively) than Sb(III)/Sb(V) (38.8 and 92.5 mg/L, respectively) alone. Particularly, MoS2 at noncytotoxic doses notably increased the bioaccumulation of Sb(III, V) in algae, causing aggravated oxidative damage, photosynthetic inhibition, and structural alterations. Metabolomics indicated that oxidative stress and membrane permeabilization were primarily associated with down-regulated amino acids involved in glutathione biosynthesis and unsaturated fatty acids. MoS2 co-exposure remarkably decreased the levels of thiol antidotes (glutathione and phytochelatins) and aggravated the inhibition on energy metabolism and ATP synthesis, compromising the Sb(III, V) detoxification and efflux. Additionally, extracellular P was captured by the nanosheets, also contributing to the uptake of Sb(V). Our findings emphasized the nonignorability of TMDs even at environmental levels in affecting the ecological hazard of metalloids, providing insight into comprehensive safety assessment of TMDs.

Bioadsorption, bioaccumulation and biodegradation of antibiotics by algae and their association with algal physiological state and antibiotic physicochemical properties.

Bioadsorption, bioaccumulation and biodegradation processes in algae, play an important role in the biomagnification of antibiotics, or other organic pollutants, in aquatic food chains. In this study, the bioadsorption, bioaccumulation and biodegradation of norfloxacin [NFX], sulfamethazine [SMZ] and roxithromycin [RTM]) is investigated using a series of culture experiments. Chlorella vulgaris was exposed to these antibiotics with incubation periods of 24, 72, 120 and 168 h. Results show the bioadsorption concentration of antibiotics in extracellular matter increases with increasing alkaline phosphatase activity (AKP/ALP). The bioaccumulation concentrations of NFX, SMZ and RTM within cells significantly increase after early exposure, and subsequently decrease. There is a significant positive antibiotics correlation to superoxide dismutase (SOD), the photosynthetic electron transport rate (ETR) and maximum fluorescence after dark adaptation (Fv/Fm), while showing a negative correlation to malondialdehyde (MDA). The biodegradation percentages (Pb) of NFX, SMZ and RTM range from 39.3 - 97.2, 41.3 - 90.5, and 9.3 - 99.9, respectively, and significantly increase with increasing Fv/Fm, density and chlorophyll-a. The accumulation of antibiotics in extracellular and intracellular substances of C. vulgaris is affected by antibiotic biodegradation processes associated with cell physiological state. The results succinctly explain relationships between algal growth during antibiotics exposure and the bioadsorption and bioaccumulation of these antibiotics in cell walls and cell matter. The findings draw an insightful understanding of the accumulation of antibiotics in algae and provide a scientific basis for the better utilization of algae treatment technology in antibiotic contaminated wastewaters. Under low dose exposures, the biomagnification of antibiotics in algae is affected by bioadsorption, bioaccumulation and biodegradation.

Quantitative modelling reservoir microalgae proliferation in response to water-soluble anions and cations influx.

Atmospheric precipitation deposits acid-forming substances into surface water. However, the effects of water-soluble components on microalgae proliferation are poorly understood. This study analysed the growth characteristics of three microalgae bioindicators of water quality: Scenedesmus quadricauda, Chlorella vulgaris, and Scenedesmus obliquus, adopting on-site monitoring, culture experiments simulating 96 types of water by supplementing anions and cations, and predictive modelling. The result quantified pH > 3.0 rain with dominant Ca2+, Mg2+, and K+ cations, together with anions of NO3- and SO42-. The presence of Ca2+ of up to 0.1 mM and Mg2+ concentrations (>0.5 mM) suppressed Scenedesmus quadricauda growth. Soluble ions, luminosity, and pH had significant impacts (p ≤ 0.01) on increased microalgae proliferation. A newly proposed microalgae growth model predicted a 10.7-fold increase in cell density six days post-incubation in the case of rainfall. The modelling supports algal outbreaks and delays prediction during regional water cycles.

Apoptosis induction in colon cancer cells (SW480) by BiFe2O4@Ag nanocomposite synthesized from Chlorella vulgaris extract and evaluation the expression of CASP8, BAX and BCL2 genes.

BACKGROUND: The use of nanomaterials in cancer diagnosis and treatment has received considerable interest. Preparation of nanoscale complex molecules could be considered to improve the efficacy and minimize toxicity of the product. This work aimed to biosynthesize BiFe2O4@Ag nanocomposite using the Chlorella vulgaris extract and its cytotoxic effect on colon cancer cell line. METHODS: The physicochemical properties of the bioengineered BiFe2O4 @Ag were investigated by Transmission Electron Microscopy (TEM), Field Emission Scanning Electron Microscopy (FE-SEM), Zeta potential, Dynamic Light Scattering (DLS), Fourier Transform Infrared Spectroscopy (FT-IR), Energy Dispersive X-ray Spectroscopy (EDX), Vibrating-sample Magnetometer (VSM) and X-ray Diffraction Analysis (XRD). The cytotoxic potential of BiFe2O4 @Ag was evaluated by MTT assay against SW480 colon cancer cell line. The expression levels of apoptotic genes including BAX, BCL2 and CASP8 were determined by Real-time PCR. The rate of apoptosis and necrosis of the cancer cells as well as the cell cycle analysis were evaluated by flow cytometry. RESULTS: Physicochemical assays indicated the nanoscale synthesis (10-70 nm) and functionalization of BiFe2O4 nanoparticles by Ag atoms. The VSM analysis revealed the magnetism of BiFe2O4 @Ag nanocomposite. According to the MTT assay, colon cancer cells (SW480) were considerably more sensitive to BiFe2O4 @Ag nanocomposite than normal cells. Apoptotic cell percentage increased from 1.93% to 73.66%, after exposure to the nanocomposite. Cell cycle analysis confirmed an increase in the number of the cells in subG1 and G0/G1 phases among nanocomposite treated cells. Moreover, treating the colon cancer cells with BiFe2O4 @Ag caused an increase in the expression of CASP8, BAX, and BCL2 genes by 3.1, 2.6, and 1.2 folds, respectively. Moreover, activity of Caspase-3 protein increased by 2.4 folds and apoptotic morphological changes appeared which confirms that exposure to the nanocomposite induces extrinsic pathway of apoptosis in colon cancer cells. CONCLUSION: The considerable anticancer potential of the synthesized BiFe2O4 @Ag nanocomposite seems to be related to the induction of oxidative stress which leads to inhibit cell cycle progression and cell proliferation. This study reveals that the BiFe2O4 @Ag is a potent compound to be used in biomedical fields.

Eco-friendly approach to decrease the harmful effects of untreated wastewater on growth, yield, biochemical constituents, and heavy metal contents of carrot (Daucus carota L.).

Here, the impact of irrigation using untreated wastewater (WW) on carrots (Daucus carota L.) was examined. We hypothesized that the addition of ethylenediaminetetraacetic acid (EDTA), dry algal powder (Spirulina platensis or Chlorella vulgaris), and Salix alba leaves powder would function as chelators for harmful contaminants in wastewater. The findings showed that irrigation of carrot plants with the sampled untreated wastewater led to significant decreases in the shoot lengths, fresh, dry weights of shoots and roots at stage I, the diameter of roots, pigment content, carotenoids, total soluble carbohydrate content, and soluble protein content. Furthermore, a significantly increased level of proline, total phenols, and the activities of polyphenol oxidase (PPO), peroxidase (POX), superoxide dismutase (SOD), and catalase (CAT) was identified in stage I samples. In contrast to the stage I, the length of the roots, the number of leaves on each plant, wet and dry weights of the stage II roots were all greatly enhanced. In spite of the increased yield due to the wastewater irrigation, carrot roots irrigated with wastewater had significantly more cadmium (Cd), nickel (Ni), cobalt (Co), and lead (Pb) than is considered safe. Our data clearly show that the application of Spirulina platensis, Chlorella vulgaris, EDTA, and leaves powder of salix was able to alleviate the toxicity of wastewater on carrot plants. For example, we recorded a significant decrease in the accumulation of carrot's Cd, Ni, Co, and Pb contents. We conclude that the treatments with Spirulina platensis and Chlorella vulgaris can be utilized as eco-friendly tools to lessen the damaging effects of wastewater irrigation on carrot plants.

Impact of Chlorella vulgaris, Nannochloropsis salina, and Arthrospira platensis as bio-stimulants on common bean plant growth, yield and antioxidant capacity.

The use of bio-stimulants in agriculture has emerged as a promising strategy to improve crop growth and yield. One type of bio-stimulant that has gained attention is microalgae extracts, which are known for their high metabolic activity, bioactive compounds, and ability to enhance plant growth and development. To investigate their effectiveness, a pot experiment was conducted at the Experimental Farm of Helwan University in Egypt during the 2022 season. The experiment aimed to evaluate the efficacy of Chlorella vulgaris, Nannochloropsis salina, and Arthrospira platensis (Spirulina platensis) extracts as bio-stimulants, applied through foliar spray at concentrations ranging from 0.25 to 2.0%, on common bean plants. Analysis of algal extract showed that . N. salina had the highest content of promotive growth hormones gibberellins (GA3) (74.85 ± 2.7mg100 g-1 d.wt). and auxins (IAA) (34.57 ± 2.7µg 100 g-1 d.wt.) compared to Chlorella and Arthrospira..The results revealed that the application of C. vulgaris, N. salina, and A. platensis extracts at concentrations up to 1.0% significantly improved various growth parameters, such as root, and shoot length, number of leaves and flowers per plant, leaf area, and total fresh and dry weight per plant. These extracts also positively affected yield attributes, including the number and fresh weight of pods per plant, seed index, seed yield per plant, and per feddan [a unit of land area]. Furthermore, the application of these extracts increased the chlorophyll content index with the maximum values of CCI (17.95. and 17.81%) was obtained at 0.50% N. salina, followed by 0.50% C.vulgaris. In addition to increase in the capacity of both non-enzymatic antioxidants [such as total antioxidant capacity, phenolics, and flavonoids] and enzymatic antioxidants [including catalase and ascorbic oxidase]. The most promising results were observed with the application of N. salina, and C. vulgaris extracts at a concentration of 0.5%. Additionally, the extracts significantly reduced the content of oxidative stress markers, such as malondialdehyde, percentage of electrolyte leakage, and hydrogen peroxide, in common bean plants compared to the control group. Contrarily, the measured parameters were reduced, while the levels of oxidative stress markers and some antioxidants including peroxidase, ascorbic peroxidase, superoxide dismutase, glutathione peroxidase, and glutathione transferase were increased by three algal extracts at a concentration of 2.0%, compared to control plants. Additionally, the application of these microalgae extracts improved the quality parameters, proximate composition, seed energy, and mineral contents of the harvested seeds, with the most significant positive impact was observed at 0.5% concentration of algal extract. These findings demonstrate the successful and safe utilization of extracts from C. vulgaris, N. salina, and A. platensis at concentrations up to 1.0% as bio-stimulants to enhance common bean yields and improve the nutritional quality of dried beans for consumers.

Insight into the roles of hematite iron oxide nanoparticles on microalgae growth, urban wastewater treatment and bioproducts generation: Gompertz simulation, nutrient mass balance and gene expression.

In this study, the effect of α-Fe2O3 nanoparticles spiking in urban wastewater (UWW) on growth rate, wastewater treatment ability and bioproducts generation of C. vulgaris and Spirulina was investigated and compared with pure cultivation system. The biomass concentration of C. vulgaris and Spirulina improved by 20 % and 39 % at 10 and 15 mg/L α-Fe2O3, respectively while the both microalgae growth pattern fitted better with Gompertz simulation after treatment with α-Fe2O3. The nutrients mass balance revealed that 1 g of treated C. vulgaris and Spirulina could uptake more COD, TN and TP in comparison to the untreated cells. The lipid generation increased remarkably (C. vulgaris: 45 % and Spirulina: 72 %) after α-Fe2O3 treatment. While, the addition of α-Fe2O3 showed no significant impact on the protein and carbohydrate productivity. Overall, this study evangelize the role of nanoparticles on promoting microalgae applications as a sustainable approach for UWW treatment and promising feedstock for biofuel production.

Application of response surface Methodology coupled with Artificial Neural network and genetic algorithm to model and optimize symbiotic interactions between Chlorella vulgaris and Stutzerimonas stutzeri strain J3BG for chlorophyll accumulation.

Research on microalgae has surged due to its diverse biotechnological applications and capacity for accumulating bioactive compounds. Despite considerable advancements, microalgal cultivation remains costly, prompting efforts to reduce expenses while enhancing productivity. This study proposes a cost-effective approach through the coculture of microalgae and bacteria, exploiting mutualistic interactions. An engineered consortium of Chlorella vulgaris and Stutzerimonas stutzeri strain J3BG demonstrated biofilm-like arrangements, indicative of direct cell-to-cell interactions and metabolite exchange. Strain J3BG's enzymatic characterization revealed amylase, lipase, and protease production, sustaining mutual growth. Employing Response Surface Methodology (RSM), Artificial Neural Network (ANN), and Genetic Algorithm (GA) in a hybrid modeling approach resulted in a 2.1-fold increase in chlorophyll production. Optimized conditions included a NaNO3 concentration of 128.52 mg/l, a 1:2 (Algae:Bacteria) ratio, a 6-day cultivation period, and a pH of 5.4, yielding 10.92 ± 0.88 mg/l chlorophyll concentration.

Apoptosis Induction by ZnFe2O4-Ag Biosynthesized by Chlorella vulgaris in MCF-7 Breast Cancer Cell Line.

The incidence and mortality of breast cancer are growing which indicates the inefficiency of the current chemotherapy drugs. Due to the anticancer potential of Zn and Ag and the magnetic feature of iron oxide, in this work, we synthesized ZnFe2O4-Ag nanocomposite using Chlorella vulgaris and investigated its anticancer effect on breast cancer cell line. Physicochemical characterization was performed by FT-IR, XRD, SEM, TEM, VSM, EDS mapping, UV, and zeta potential assays. Cell cytotoxicity and apoptosis frequency were studied by the MTT and flow cytometry assays. Also, cell cycle analysis, Hoechst staining, and measuring ROS (reactive oxygen species) level were performed. The synthesized particles were almost spherical with a size range of 14-52 nm. The FT-IR and XRD assays confirmed the proper synthesis of the particles and VSM analysis showed that particles had magnetic property and the maximum saturation magnetization was 0.8 Emu/g. Also, the EDS mapping of the nanocomposite showed the Zn, Fe, O, and Ag elements. The MTT assay showed that the 50% inhibitory concentration (IC50) of ZnFe2O4-Ag for breast cancer and normal cells were 28 and 154 µg/mL, respectively, and the nanocomposite had stronger anticancer activity than cisplatin (IC50 = 84 µg/mL). Flow cytometry analysis showed that the exposure to the nanocomposite induced cell apoptosis by 77.5% and significantly induced ROS generation. Also, treating breast cancer cells with the nanocomposite induced cell cycle arrest and apoptotic features, including chromatin condensation and fragmentation. In conclusion, ZnFe2O4-Ag nanocomposite synthesized by C. vulgaris could suppress the proliferation of breast cancer cells by the generation of oxidative stress, apoptosis induction, and cell cycle arrest.

Continuous microalgae cultivation for wastewater treatment - Development of a process strategy during day and night.

Conventional wastewater treatment (WWT) is not able to recycle nutrients from the wastewater (WW) directly. Microalgae integrate the valuable nutrients nitrogen and phosphorus within their biomass very efficiently, making them predestined for an application in WWT. Nevertheless, microalgae-based processes are driven by natural sunlight as energy source, making a continuous process mode during day and night difficult. The aim of this study was therefore to investigate metabolic activities of the continuously cultivated microalgae Chlorella vulgaris at light and dark periods (16 h,8 h) with focus on nutrient uptake during night from a synthetic WW. Varying the dilution rate D (D = 0.0-1.0 d-1 in 0.1 d-1-steps) causes different limitations for algae growth. Nutrient limitations at low D's cause maximum accumulation of intracellular storage components (sum of carbohydrates and lipids) of ~70 % of dry biomass, starch is converted to lipids at the absence of light. From middle to high D's, the growth rate is determined by light limitation, reducing the intracellular storage components to ~20 % of dry biomass. Complete nutrient uptake is measurable up to D = 0.5 d-1, marking the maximum operating point for wastewater purification. At that point, cells are characterised by high protein (up to 57%DBM) and pigment (up to 6.9%DBM) quotas. During the night, the build-up of proteins at the degradation of intracellular storage components is furthermore visible. Applying the concept of active biomass (cells without storage components), a constant cellular protein (~68%ABM) and nitrogen quota (11.94%ABM) was revealed. A nitrogen spiking experiment clearly showed nitrogen uptake and proliferation during the night period. Based on the experimental data, a window of operation for a continuous WWT process was designed, allowing the hypothesis that continuous WWT using microalgae during day and night operation is possible without the supplementation of artificial light. This revealed the system's capacity to treat WW throughout 24 h applying cell recycling and storage of carbohydrate-rich biomass. At the end of the night, protein-rich biomass is available for further valorisation.

Mechanism of the adverse outcome of Chlorella vulgaris exposure to diethyl phthalate: Water environmental health reflected by primary producer toxicity.

As a ubiquitous contaminant in aquatic environments, diethyl phthalate (DEP) is a major threat to ecosystems because of its increasing utilization. However, the ecological responses to and toxicity mechanisms of DEP in aquatic organisms remain poorly understood. To address this environmental concern, we selected Chlorella vulgaris (C. vulgaris) as a model organism and investigated the toxicological effects of environmentally relevant DEP concentrations at the individual, physiological, biochemical, and molecular levels. Results showed that the incorporation of DEP significantly inhibited the growth of C. vulgaris, with inhibition rates ranging from 10.3 % to 83.47 %, and disrupted intracellular chloroplast structure at the individual level, while the decrease in photosynthetic pigments, with inhibition rates ranging from 8.95 % to 73.27 %, and the imbalance of redox homeostasis implied an adverse effect of DEP at the physio-biochemical level. Furthermore, DEP significantly reduced the metabolic activity of algal cells and negatively altered the cell membrane integrity and mitochondrial membrane potential. In addition, the apoptosis rate of algal cells presented a significant dose-effect relationship, which was mainly attributed to the fact that DEP pollutants regulated Ca2+ homeostasis and further increased the expression of Caspase-8, Caspase-9, and Caspase-3, which are associated with internal and external pathways. The gene transcriptional expression profile further revealed that DEP-mediated toxicity in C. vulgaris was mainly related to the destruction of the photosynthetic system, terpenoid backbone biosynthesis, and DNA replication. Overall, this study offers constructive understandings for a comprehensive assessment of the toxicity risks posed by DEP to C. vulgaris.

RNA-sequencing analysis reveals the co-biodegradation performance of crude oil by marine Chlorella vulgaris under norfloxacin stress.

In this study, the microalgal growth and crude oil (CRO) biodegradation by marine Chlorella vulgaris (C. vulgaris) were assessed under norfloxacin (NFX) stress. The presence of NFX negatively affected the bio-removal of CRO within 5 days, as the NFX concentration increased from 100 to 1600 µg/L, due to its toxicity as an antibiotic. However, its negative impact on the final degradation capabilities of C. vulgaris was less significant (P-value <0.05). After 9 days of cultivation, CRO bio-removal efficiencies still exceeded 90 %, while NFX bio-removal efficiencies maintained over 47 %. RNA-seq analysis revealed that the degradation of CRO and NFX was attributed to the combined action of functional genes involved in scavenging reactive oxygen species. The production of pigments and the bio-removal performance of C. vulgaris in CRO, NFX, and CRO & NFX coexistence media were consistent with the changes in the number of differentially expressed genes in these samples.

Biodegradation of benzo[a]pyrene by a marine Chlorella vulgaris LH-1 with heterotrophic ability.

In this study, a microalga, Chlorella vulgaris LH-1, with heterotrophic ability to degrade BaP was explored. The effect of BaP concentration on microalga growth was investigated, and the possible biodegradation mechanism of BaP was proposed. Results showed that low BaP concentration (<5 mg/L) had less negative influence on the growth of this microalga under mixotrophic condition, but high BaP concentration (>5 mg/L) had a significant inhibitory effect on its growth. During heterotrophic cultivation, low BaP concentration (<20 mg/L) promoted the growth of C. vulgaris LH-1, whereas high BaP concentration (>20 mg/L) inhibited its growth significantly. The degradation rates of mixotrophic and heterotrophic C. vulgaris LH-1 were 62.56 %-74.13 % and 52.07 %-71.67 %, respectively, when the BaP concentration ranged from 0.5 mg/L to 2 mg/L. The expression of functional enzyme genes of C. vulgaris LH-1 such as phenol 2-monooxygenase activity, protocatechuate 3,4-dioxygenase activity, catechol 1,2-dioxygenase activity, styrene degradation, and benzoate degradation were upregulated in the process of BaP degradation. C. vulgaris LH-1 may degrade BaP by monooxygenase and dioxygenase simultaneously. The degradation of BaP by this microalga under mixotrophic condition goes through the degradation pathway of phthalic acid, whereas it goes through the degradation pathway of benzoic acid under heterotrophic condition.