Supercritical CO2 Extraction of High-Added Value Compounds from Chlorella vulgaris: Experimental Design, Modelling and Optimization.

Microalgae are well-known for their high-added value compounds and their recovery is currently of great interest. The aim of this work is the recovery of such components from Chlorella vulgaris through supercritical fluid extraction (SFE) with CO2. The effect of the extraction temperature (40-60 °C), pressure (110-250 bar), and solvent flow rate (20-40 g/min) was tested on yield, the extract's antioxidant activity, and the phenolic, chlorophyll and carotenoid content. Thus, data analysis indicated that the yield was mainly affected by temperature, carotenoids by pressure, while the extract's phenolics and antioxidant activity were affected by the synergy of temperature and pressure. Moreover, SFE's kinetic study was performed and experimental data were correlated using Sovová's mass transfer-based model. SFE optimization (60 °C, 250 bar, 40 g/min) led to 3.37% w/w yield, 44.35 mgextr/mgDPPH antioxidant activity (IC50), 18.29 mgGA/gextr total phenolic content, 35.55, 21.14 and 10.00 mg/gextr total chlorophyll, carotenoid and selected carotenoid content (astaxanthin, lutein and ß-carotene), respectively. A comparison of SFE with conventional aq. ethanol (90% v/v) extraction proved SFE's superiority regarding extraction duration, carotenoids, antioxidant activity and organoleptic characteristics of color and odor despite the lower yield. Finally, cosolvent addition (ethanol 10% w/w) at optimum SFE conditions improved the extract's antioxidant activity (19.46%) as well as yield (101.81%).

Manipulation of fatty acid profile and nutritional quality of Chlorella vulgaris by supplementing with citrus peel fatty acid.

Microalgae could be an excellent resource of functional and essential fatty acids. To achieve viable microalgal biomass production, mass cultivation of microalgae is required; however, the high cost of nutrients is the obstacle. An inexpensive and nutritious material is required to feed Chlorella vulgaris in the pharmaceutical and food sectors. Citrus peel waste with a valuable nutritional quality could be one of the promising and inexpensive candidates. In this study, the fatty acid extract from different citrus peels was used as the organic nutrient source for the cultivation of Chlorella. The proximate composition of bitter orange, sweet orange, grapefruit, and mandarin peels were determined, and their nutritional quality was evaluated. Total fatty acids from the citrus peel were prepared by acidic methanol hydrolysis and hexane extraction. Fourier transforms infrared (FT-IR) and gas chromatography-mass spectrometry (GC-MS) was used to analyze the fatty acid composition and nutrient composition. Fatty acids from the citrus peels were added to the Chlorella culture medium to study their influences on biomass, lipid production, fatty acid profile, and nutritional quality of Chlorella. The most predominant citrus peel fatty acids were linoleic, palmitic, oleic, linolenic, and stearic acids. The citrus peels contain polyunsaturated, saturated, and monounsaturated fatty acids. The most unsaturated fatty acids were omega-6, omega-3, omega-9, and omega-7. The citrus peel had acceptable atherogenicity, thrombogenicity, omega-6/omega-3, peroxidizability, hypocholesterolemic, and nutritive value indices. The major fatty acids of Chlorella were palmitic, linoleic, oleic, alpha-linolenic, gamma-linolenic, 4,7,10,13-hexadecatetraenoic, palmitoleic, 7,10-hexadecadienoic, 7,10,13-hexadecatrienoic, lauric and 5,8,11,14,17-eicosapentaenoic acids. Chlorella contains polyunsaturated, saturated, and monounsaturated fatty acids. The most unsaturated fatty acids contain omega-6, omega-3, omega-9, and omega-7. Chlorella had acceptable atherogenicity, thrombogenicity, omega-6/omega-3, hypocholesterolemic, peroxidizability, and nutritive value indices. Supplementation of Chlorella with citrus peels fatty acid increases total biomass, lipid content, and nutritional quality of Chlorella. The present research shows that citrus peels have good nutritional quality and could be used for the inexpensive cultivation of Chlorella biomass with potential utility for food application.

Preparation of the Enzymatic Hydrolysates from Chlorella vulgaris Protein and Assessment of Their Antioxidant Potential Using Caenorhabditis elegans.

This study aimed to assess the antioxidant potential of Chlorella vulgaris protein-derived enzymatic hydrolysate using Caenorhabditis elegans. Protein extraction was performed using an alkali solution after complete C. vulgaris swelling and hydrolysis using four commercial proteases (alcalase, neutrase, protamex, and flavourzyme). The results showed that the flavourzyme hydrolysates exhibited the strongest antioxidant activity both in vitro and in vivo. Under the optimum conditions of the enzymatic hydrolysis, the half-maximal effective concentration of the hydrolysates for superoxide and hydroxyl radicals was 0.323 mg/mL and 0.139 mg/mL, respectively. The hydrolysates could significantly extend the lifespan, improve the resistance to methyl viologen-induced oxidative stress, reduce the levels of reactive oxygen species, and enhance the activity of catalase and superoxide dismutase in C. elegans.

Raman Microspectroscopic Analysis of Selenium Bioaccumulation by Green Alga Chlorella vulgaris.

Selenium (Se) is an element with many commercial applications as well as an essential micronutrient. Dietary Se has antioxidant properties and it is known to play a role in cancer prevention. However, the general population often suffers from Se deficiency. Green algae, such as Chlorella vulgaris, cultivated in Se-enriched environment may be used as a food supplement to provide adequate levels of Se. We used Raman microspectroscopy (RS) for fast, reliable, and non-destructive measurement of Se concentration in living algal cells. We employed inductively coupled plasma-mass spectrometry as a reference method to RS and we found a substantial correlation between the Raman signal intensity at 252 cm-1 and total Se concentration in the studied cells. We used RS to assess the uptake of Se by living and inactivated algae and demonstrated the necessity of active cellular transport for Se accumulation. Additionally, we observed the intracellular Se being transformed into an insoluble elemental form, which we further supported by the energy-dispersive X-ray spectroscopy imaging.

Evaluation of antioxidant and anticancer activity of crude extract and different fractions of Chlorella vulgaris axenic culture grown under various concentrations of copper ions.

BACKGROUND: Chlorella vulgaris is a microalga potentially used for pharmaceutical, animal feed, food supplement, aquaculture and cosmetics. The current study aims to study the antioxidant and prooxidant effect of Chlorella vulgaris cultivated under various conc. of copper ions. METHODS: The axenic green microalgal culture of Chlorella vulgaris was subjected to copper stress conditions (0.00, 0.079, 0.158, 0.316 and 0.632 mg/L). The growth rate was measured at OD680 nm and by dry weight (DW). Moreover, the Antioxidant activity against DPPH and ABTS radical, pigments and phytochemical compounds of the crude extracts (methylene chloride: Methanol, 1:1) were evaluated. The promising Cu crude extract (0.316 mg/L) further fractionated into twenty-one fractions by silica gel column chromatography using hexane, chloroform and ethyl acetate as a mobile phase. RESULTS: The obtained results reported that nine out of these fractions exhibited more than 50% antioxidant activity and anticancer activity against Hela cancer cell lines. Based on IC50, fraction No. 7 was found to be the most effective fraction possessing a significant increase in both antioxidant and anticancer potency. Separation of active compound (s) in fraction No 7 was performed using precoated silica gel plates (TLC F254) with ethyl acetate: hexane (9:1 v/v) as mobile phase. Confirmation of active compound separation was achieved by two-dimensional TLC and visualization of the separated compound by UV lamp. The complete identification of the separated active compound was performed by UV- Vis- spectrophotometric absorption, IR, MS, H1-NMRT C13-NMR. The isolated compound ((2E,7R,11R)-3,7,11,15-Tetramethyl-2-hexadecenol) have high antioxidant activity with IC50 (10.59 µg/ml) against DPPH radical assay and comparable to the capacities of the positive controls, Butylated hydroxy toluene [BHT] (IC50 11.2 µg/ml) and Vitamin C (IC50 12.9 µg/ml). Furthermore, pure isolated compound exhibited a potent anticancer activity against Hela cell line with IC50 (4.38 µg/ml) compared to Doxorubicin (DOX) as synthetic drug (13.3 µg/ml). In addition, the interaction of the pure compound with Hela cancer cell line and gene expression were evaluated. CONCLUSIONS: The authors recommend cultivation of Chlorella vulgaris in large scale under various stress conditions for use the crude extracts and semi purified fractions for making a pharmaco-economic value in Egypt and other countries.

18O Isotope Labeling Combined with 31P Nuclear Magnetic Resonance Spectroscopy for Accurate Quantification of Hydrolyzable Phosphorus Species in Environmental Samples.

31P nuclear magnetic resonance (NMR) spectra can be biased due to the hydrolysis of labile P species during sample treatment and NMR analysis. This paper offers an approach to circumvent this problem by performing sample preparation and analysis in 18O-enriched medium. Heavy 18O isotope atoms were introduced into the resulting artificial hydrolysis products. The NMR signal of 18O-labeled P was shifted upfield relative to the unlabeled P nuclei in natural metabolites. This isotope shift enabled an immediate differentiation of artificial hydrolysis products from natural metabolites. Moreover, the hydrolysis products could be accurately quantified. Our data suggest that the extent to which artificial hydrolysis alters NMR spectra varies among different types of environmental samples. For instance, 72-84% of the detected monoesters in the organic soils of this study were actually artificially hydrolyzed diesters. By contrast, artificial hydrolysis products in the mineral soils used for this study accounted for less than 6% of the total monoesters. Polyphosphate was also hydrolyzed to yield 18O-labeled products in algal biomass.

Lipid yield from the diatom Porosira glacialis is determined by solvent choice and number of extractions, independent of cell disruption.

Cell wall disruption is necessary to maximize lipid extraction yields in conventional species of mass-cultivated microalgae. This study investigated the effect of sonication, solvent choice and number of extractions on the lipid yield, lipid class composition and fatty acid composition of the diatom Porosira glacialis. For comparison, the diatom Odontella aurita and green alga Chlorella vulgaris were included in the study. Sonication effectively disrupted P. glacialis cells, but did not increase the total lipid yield compared to physical stirring (mixing). In all three microalgae, the content of membrane-associated glyco- and phosopholipids in the extracted lipids was strongly dependent on the solvent polarity. A second extraction resulted in higher yields from the microalgae only when polar solvents were used. In conclusion, choice of solvent and number of extractions were the main factors that determined lipid yield and lipid class composition in P. glacialis.

Augmenting the expression of accD and rbcL genes using optimized iron concentration to achieve higher biomass and biodiesel in Chlorella vulgaris.

Chlorella vulgaris is a form of microalgae commonly employed as a biological source of oil for biodiesel production. Major algal cultivation strategies are focused on stimulating growth rate and lipid content. In the present study, the algal growth media was supplemented with iron (III) chloride (FeCl3), as a stimulating factor for growth and lipid production, in three iron concentrations including 90, 200, and 500 µM. The turbidity of algal cells was measured on different days, to determine the growth rate. In optimum iron concentration, this measurement experienced a 2.1-fold increase. Next, the lipid content was extracted, and the amount of lipid produced in each treatment was calculated, which demonstrated a 4.57-fold increase in lipid productivity. The expression of genes corresponding to the metabolic enzymes (i.e. acetyl-CoA carboxylase (accD) and ribulose bisphosphate carboxylase large chain (rbcL)) was evaluated using real-time PCR under different initial iron feeds. As demonstrated in the results, the initial iron feed of 90 µM was an optimum concentration that obtained the highest growth rate, more cell density, and increased lipid production. In 90 µM initial iron concentration, the expression of accD and rbcL genes showed a 4.8- and 35-fold increase, respectively, compared to that of the control genes. Based on the results, this optimum iron concentration could satisfy the industrial interest in biodiesel production from C. vulgaris as a potential stimulating factor. However, higher levels of iron (e.g. 200 and 500 µM) failed to act as positive stress for increasing biodiesel production. Finally, in this paper, different mechanisms where iron affects acetyl-CoA carboxylase (ACCase) and 1,5-ribulose bisphosphate carboxylase/oxygenase (RuBisCo) are illustrated.

Effect of Chlorella vulgaris enriched diet on growth performance, hemato-immunological responses, antioxidant and transcriptomics profile disorders caused by deltamethrin toxicity in Nile tilapia (Oreochromis niloticus).

The toxic effect of deltamethrin (DM) was documented in aquaculture. There is no obtainable data on the effect of Chlorella vulgaris against DM toxicity. The current study focused on the effect of dietary supplementation with C. vulgaris (CV) on growth performance, innate immune response, antioxidant activities, and transcriptomics disorders induced by sub-lethal dose of DM in Oreochromis niloticus. A total number of 216 O. niloticus divided into four groups with tri-replicates. The 1st control group (CT) fed a basal diet, the second group fed diet enriched with 5% CV. The third group was exposed to DM (15 µg/L), where the last group fed CV and simultaneously exposed to DM as previous-mentioned. The procedure of CV feeding and DM exposure were continued for two months. Exposures to DM revealed in stunting of the growth parameters and lessening of survival ratio of tilapias with a significant decline of the erythrogram (macrocytic hypochromic anemic picture), and leucocytes immune cells and related parameters (immunoglobulin M, lysozyme) and sever shifting in the antioxidant indicators. Sever raise was monitored in hepatic and kidney markers. Also, genes expression related to immune and antioxidant parameters were severely impacted. Where tilapias received CV showed a significant increase in the growth and immune parameters besides to an improvement of hematological, antioxidant values and their related genes expressions. The fourth group that received CV simultaneous with DM exposure showed a soothing of the previous indicators and markers toward the values of tilapias fed on basal diet (CT). In turn, CV supplementation may be presented a protective effect alongside DM toxicity in O. niloticus appeared through soothing of the immune, antioxidant and related genes expressions in addition to its hepato-renal protective effects. Therefore, the current study recommended that an incorporating of 5% CV for tilapias diet could improve their growth performance, immunity, antioxidant and transcriptomics disorders induced by deltamethrin.

Biological Activity of Hydrophilic Extract of Chlorella vulgaris Grown on Post-Fermentation Leachate from a Biogas Plant Supplied with Stillage and Maize Silage.

Algae are employed commonly in cosmetics, food and pharmaceuticals, as well as in feed production and biorefinery processes. In this study, post-fermentation leachate from a biogas plant which exploits stillage and maize silage was utilized as a culture medium for Chlorella vulgaris. The content of polyphenols in hydrophilic extracts of the Chlorella vulgaris biomass was determined, and the extracts were evaluated for their antioxidant activity (DPPH assay), antibacterial activity (against Escherichia coli, Lactobacillus plantarum, Staphylococcus aureus, Staphylococcus epidermidis) and antifungal activity (against Aspergillus niger, Candida albicans, Saccharomyces cerevisiae). The use of the post-fermentation leachate was not found to affect the biological activity of the microalgae. The aqueous extract of Chlorella vulgaris biomass was also observed to exhibit activity against nematodes. The results of this study suggest that Chlorella vulgaris biomass cultured on post-fermentation leachate from a biogas plant can be successfully employed as a source of natural antioxidants, food supplements, feed, natural antibacterial and antifungal compounds, as well as in natural methods of plant protection.

Production, extraction and characterization of Chlorella vulgaris soluble polysaccharides and their applications in AgNPs biosynthesis and biostimulation of plant growth.

Chlorella vulgaris, like a wide range of other microalgae, are able to grow mixotrophically. This maximizes its growth and production of polysaccharides (PS). The extracted polysaccharides have a complex monosaccharide composition (fructose, maltose, lactose and glucose), sulphate (210.65 ± 10.5 mg g-1 PS), uronic acids (171.97 ± 5.7 mg g-1 PS), total protein content (32.99 ± 2.1 mg g-1 PS), and total carbohydrate (495.44 ± 8.4 mg g-1 PS). Fourier Transform infrared spectroscopy (FT-IR) analysis of the extracted polysaccharides showed the presence of N-H, O-H, C-H, -CH3, >CH2, COO-1, S=O and the C=O functional groups. UV-Visible spectral analysis shows the presence of proteins, nucleic acids and chemical groups (ester, carbonyl, carboxyl and amine). Purified polysaccharides were light green in color and in a form of odorless powder. It was soluble in water but insoluble in other organic solvents. Thermogravimetric analysis demonstrates that Chlorella vulgaris soluble polysaccharide is thermostable until 240°C and degradation occurs in three distinct phases. Differential scanning calorimetry (DSC) analysis showed the characteristic exothermic transition of Chlorella vulgaris soluble polysaccharides with crystallization temperature peaks at 144.1°C, 162.3°C and 227.7°C. The X-ray diffractogram illustrated the semicrystalline nature of these polysaccharides. Silver nanoparticles (AgNPs) had been biosynthesized using a solution of Chlorella vulgaris soluble polysaccharides. The pale green color solution of soluble polysaccharides was turned brown when it was incubated for 24 hours with 100 mM silver nitrate in the dark, it showed peak maximum located at 430 nm. FT-IR analysis for the biosynthesized AgNPs reported the presence of carbonyl, -CH3, >CH2, C-H,-OH and -NH functional groups. Scanning and transmission electron microscopy show that AgNPs have spherical shape with an average particle size of 5.76. Energy-dispersive X-ray (EDX) analysis showed the dominance of silver. The biosynthesized silver nanoparticles were tested for its antimicrobial activity and have positive effects against Bacillus sp., Erwinia sp., Candida sp. Priming seeds of Triticum vulgare and Phaseolus vulgaris with polysaccharides solutions (3 and 5 mg mL-1) resulted in significant enhancement of seedling growth. Increased root length, leaf area, shoot length, photosynthetic pigments, protein content, carbohydrate content, fresh and dry biomass were observed, in addition these growth increments may be attributed to the increase of antioxidant activities.

Ameliorative effects of dietary Chlorella vulgaris and ß-glucan against diazinon-induced toxicity in Nile tilapia (Oreochromis niloticus).

The present study was carried out to investigate the toxic effects of diazinon on growth performance, hepato-renal function, antioxidant system, innate immune response and comparing the protective role of dietary Chlorella vulgaris (CV) algae and ß-glucan in intoxicated Nile tilapia (Oreochromis niloticus). One hundred and eighty healthy Nile tilapia (20 ± 6.1 g) were distributed equally into four groups; control group, DZN group (0.28 mg/L), DZN-CV group (5% CV) and DZN-ß-glucan group (0.1% ß-glucan) and treatments conducted for about 60 days. The results revealed that administration of DZN significantly increased serum liver enzymes, uric acid, creatinine, and malondialdehyde (MDA) in different tissues. Meanwhile, glutathione (GSH) and superoxide dismutase (SOD) in different tissues, as well as IgM, C-reactive protein (CRP), respiratory burst, lysozyme and bactericidal activities were significantly decreased in DZN group. In addition, expression of TNF-α gene was up-regulated and IL-10 was down-regulated in spleen of DZN intoxicated fish. The treatment of DZN exposed fish with CV and ß-glucan supplemented diets ameliorated hepatic damage and enhanced antioxidant activity and innate immune responses. Furthermore, dietary Chlorella vulgaris and ß-glucan have a potent anti-inflammatory effect as they remarkably increased the expression of IL-10 and decreased TNF-α gene expression. The results also revealed that fish in DZN-CV group had the highest survival rate, final body weight (FBW) and body weight gain (BWG). On the other hand, feed conversion ratio (FCR), specific growth rate (SGR), and protein efficiency ratio (PER) of control, DZN-CV, and DZN- ß-glucan were higher than DZN group. However, the hepatosomatic index (HSI) and spleen-somatic index (SSI) were higher in DZN group than other experimental groups. Overall, CV and ß-glucan can be recommended as a feed supplement to improve immunosuppression, oxidative damage, growth performance and hemato-biochemical alterations induced by DZN toxicity in Nile tilapia.

Effect of cell disruption methods on the extraction of bioactive metabolites from microalgal biomass.

Microalgae synthesize a variety of potentially high-value compounds. Due to their robust cell wall, cell disruption is necessary to improve extraction of these compounds. While cell disruption methods have been optimized for lipid and protein extraction, there are limited studies for other bioactive compounds. The present study investigated the effect of freeze-drying combined with sonication or ball-milling on the extraction of antioxidant and plant biostimulating compounds from Chlorella sp., Chlorella vulgaris and Scenedesmus acutus. Both cell disruption methods resulted in higher extract yields from the biomass compared to freeze-dried biomass using 50% methanol as a solvent. Antioxidant activity of Chlorella extracts was generally higher than freeze-dried extracts based on the diphenylpicrylhydrazyl (DPPH) and ß-carotene linoleic acid assays. However, the effectiveness of each treatment varied between microalgae strains. Sonication resulted in the highest antioxidant activity in Chlorella sp. extracts. Ball-milling gave the best results for C. vulgaris extracts in the DPPH assay. Both cell disruption methods decreased antioxidant activity in S. acutus extracts. Plant biostimulating activity was tested using the mung bean rooting assay. Damaging the membrane by freeze-drying was sufficient to release the active compounds using water extracts. In contrast, both cell disruption methods negatively affected the biological activity of the extracts. These results indicate that bioactive compounds in microalgae are sensitive to post-harvest processes and their biological activity can be negatively affected by cell disruption methods. Care must be taken to not only optimize yield but to also preserve the biological activity of the target compounds.

Investigation of the antifungal effects of algal extracts on apple-infecting fungi.

It is known that cell extracts of various algae have antifungal activity against microorganisms in vitro. Antifungal activities of Ulva lactuca, Chlorella vulgaris, Chlorella minutissima, and Chlorella protothecoides were investigated against: Aspergillus niger, Alternaria alternata, and Penicillium expansum fungi to present their fungicide potentials. Aspergillus niger, Alternaria sp., and Penicillium expansum are typical soft-rotting fungi and cause important loss of apple fruit in the storage. In vitro antifungal activity was evaluated by agar disc diffusion assay against pathogenic apple rot fungi. As a result, almost all of the extracts obtained from algae species were revealed to have antifungal activity against selected fungal pathogens. Free radical-scavenging activity of the extracts was determined with 1,1-diphenyl-2 picryl hydrazyl (DPPH) free radical-scavenging method. Extract of C. protothecoides was determined to have a stronger antioxidant activity than other algae extracts. This study reveals that the potential of algae should be investigated for the production of natural fungicide for pharmaceutical and food industries.

Investigation of hepatoprotective effect of some algae species on carbon tetrachloride-induced liver injury in rats.

The aim of this study was to investigate hepatoprotective effect of some algae species such as Spirulina platensis, Chlorella vulgaris, Laminaria japonica, Sargassum sp. on experimental acute hepatotoxicity model that induced with carbon tetrachloride (CCl4) in rats. Algaes at a dose of 200 mg/kg and Silymarin at a dose of 25 mg/kg were orally administered for 7 days followed by CCl4 at a single dose (0.5 ml/kg), at the 8th day to cause experimental acute hepatotoxicity. Levels of biochemical (AST, ALT etc.), lipid peroxidation (MDA), antioxidant (GSH, CAT, GPx) parameters and histopathological examination were carried out to investigate the hepatoprotective effects of algae. In Sp group ALT and ALP levels were significantly decreased compared with CCl4 (p < .05). Histological liver structures of Sp group were similar to the control group. MDA, GPx and CAT levels of Sp and La groups were significantly different compared with CCl4 (p < .05). Based on these results, algae species able to minimise the toxic effects of CCl4 and especially S. platensis could be used in the purpose of protection against chemical-induced hepatotoxicity.

Controlled synthesis of iron oxyhydroxide (FeOOH) nanoparticles using secretory compounds from Chlorella vulgaris microalgae.

FeOOH nanoparticles are commonly synthesized at very high temperature and pressure that makes the process energy consuming and non-economic. Recently, novel approaches were developed for the fabrication of these particles at room temperature. But, the main problem with these methods is that the prepared structures are aggregates of ultra-small nanoparticles where no intact separate nanoparticles are formed. In this study, for the first time, secretory compounds from Chlorella vulgaris cells were employed for the controlled synthesis of FeOOH nanoparticles at room atmosphere. Obtained particles were found to be goethite (α-FeO(OH)) crystals. Controlled synthesis of FeOOH nanoparticles resulted in uniform spherical nanoparticles ranging from 8 to 17 nm in diameter with 12.8 nm mean particle size. Fourier-transform infrared and elemental analyses were indicated that controlled synthesized nanoparticles have not functionalized with secretory compounds of C. vulgaris, and these compounds just played a controlling role over the synthesis reaction.

Photoautotrophically Grown Chlorella vulgaris Shows Genotoxic Potential but No Apoptotic Effect in Epithelial Cells.

This study investigated the effect of Chlorella vulgaris (C. vulgaris) on genotoxicity, cytotoxicity, and apoptosis in Caco-2 and HT-29 cells. C. vulgaris significantly induced DNA damage in both cell lines at a concentration of 200 µg dry matter/mL (comet tail intensity CTI: 24.6 ± 4.7% for Caco-2, 16.6 ± 0.9% for HT-29). The application of processing (sonication, ball-milling) did not affect the genotoxicity negatively and lowered the lipid peroxidation in C. vulgaris preparations. C. vulgaris-induced intracellular formation of reactive oxygen species in human cell lines and might be responsible for the genotoxic effect. A solid fraction mainly triggered the observed DNA damage (CTI: 41.5 ± 1.9%), whereas a hydrophilic (CTI: 7.9 ± 1.7%) and lipophilic (CTI: 10.2 ± 2.1%) fraction revealed a significantly lower tail intensity. C. vulgaris significantly induced DNA damage in both cell lines possibly through intracellular formation of reactive oxygen species; however, it was repaired after a 2 h recovery time or was even avoided at lower concentrations. In addition, none of the preparations indicated an adverse effect on cell proliferation or revealed apoptotic activity.

Bioavailability and Safety of Nutrients from the Microalgae Chlorella vulgaris, Nannochloropsis oceanica and Phaeodactylum tricornutum in C57BL/6 Mice.

Microalgae are rich in macronutrients and therefore, they have been proposed as a potential future food source preserving natural resources. Here, we studied safety and bioavailability of algae nutrients in mice. Three microalgae species, Chlorella vulgaris, Nannochloropsis oceanica and Phaeodactylum tricornutum, were studied after ball mill disruption at different doses (5%, 15% and 25% dry weight) for 14 days. In response to all three algae diets, we observed a weight gain similar or superior to that in response to the control diet. No substantial differences in organ weights nor gut length occurred. Protein bioavailability from the algae diets did not differ from the control diet ranging from 58% to 77% apparent biological value. Fat absorption was lower for microalgae compared to soy oil in control diets, albeit still substantial. High liver eicosapentaenoic acid levels were measured following feeding with N. oceanica, the algae richest in omega-3 fatty acids. Neither histological nor serum analyses revealed any heart, kidney or liver toxicity induced by any of the algae diets. Algae-rich diets were thus well accepted, well tolerated and suitable for the maintenance of body weight and normal organ function. No toxicological effects were observed.

In vitro evaluation of the synergistic antioxidant and anti-inflammatory activities of the combined extracts from Malaysian Ganoderma lucidum and Egyptian Chlorella vulgaris.

BACKGROUND: Since oxidative stress and inflammation are two linked factors in the pathogenesis of several human diseases. Thus identification of effective treatment is of great importance. Edible mushroom and microalgae are rich in the effective antioxidant phytochemicals. Hence, their beneficial effects on oxidative stress-associated inflammation are extremely required to be investigated. METHODS: This study evaluated the functional constituents, antioxidant and anti-inflammatory activities of Malaysian Ganoderma lucidum aqueous extract (GLE) and Egyptian Chlorella vulgaris ethanolic extract (CVE). Also, the synergistic, addictive or antagonistic activities of the combination between the two extracts (GLE-CVE) were studied. Expression of inducible nitric oxide synthase, cyclooxygenase-2, and nuclear factor-kappa B, as well as levels of nitric oxide, tumor necrosis factor (TNF)-α, lipid peroxidation, reduced glutathione and antioxidant enzymes were determined using in vitro model of lipopolysaccharide-stimulated white blood cells.

Impact of different sequences of mechanical and thermal processing on the rheological properties of Porphyridium cruentum and Chlorella vulgaris as functional food ingredients.

Microalgae are a promising and sustainable source for enhancing the nutritional value of food products. Moreover, incorporation of the total biomass might contribute to the structural properties of the enriched food product. Our previous study demonstrated the potential of Porphyridium cruentum and Chlorella vulgaris as multifunctional food ingredients, as they displayed interesting rheological properties after applying a specific combination of mechanical and thermal processing. The aim of the current study was to investigate the impact of a different sequence of high pressure homogenization (HPH) and thermal processing on the thickening and gelling potential of these microalgal biomasses in aqueous suspensions. Thermal processing largely increased the gel strength and viscosity of both microalgae, which was ascribed to larger and stronger aggregates as a result of partial solubilization of polymers, while subsequent HPH generally reduced the rheological properties. Interestingly, large amounts of intact cells were still observed for both microalgae when HPH was performed after a thermal treatment, irrespective of the applied homogenization pressure, implying that cell disruption was hindered by the preceding thermal treatment. Although thermal processing was regarded as the most effective processing technique to obtain increased rheological properties, the combination with a preceding HPH treatment should still be considered when cell disruption is desired, for instance to increase the bioavailability of intracellular components. Finally, biomass of P. cruentum showed the largest potential for use as a structuring agent, as the gel strength and viscosity in thermally treated suspensions of this microalga were about 10 times higher than for C. vulgaris.