Tisochrysis lutea F&M-M36 Mitigates Risk Factors of Metabolic Syndrome and Promotes Visceral Fat Browning through ß3-Adrenergic Receptor/UCP1 Signaling.

Pre-metabolic syndrome (pre-MetS) may represent the best transition phase to start treatments aimed at reducing cardiometabolic risk factors of MetS. In this study, we investigated the effects of the marine microalga Tisochrysis lutea F&M-M36 (T. lutea) on cardiometabolic components of pre-MetS and its underlying mechanisms. Rats were fed a standard (5% fat) or a high-fat diet (20% fat) supplemented or not with 5% of T. lutea or fenofibrate (100 mg/Kg) for 3 months. Like fenofibrate, T. lutea decreased blood triglycerides (p < 0.01) and glucose levels (p < 0.01), increased fecal lipid excretion (p < 0.05) and adiponectin (p < 0.001) without affecting weight gain. Unlike fenofibrate, T. lutea did not increase liver weight and steatosis, reduced renal fat (p < 0.05), diastolic (p < 0.05) and mean arterial pressure (p < 0.05). In visceral adipose tissue (VAT), T. lutea, but not fenofibrate, increased the ß3-adrenergic receptor (ß3ADR) (p < 0.05) and Uncoupling protein 1 (UCP-1) (p < 0.001) while both induced glucagon-like peptide-1 receptor (GLP1R) protein expression (p < 0.001) and decreased interleukin (IL)-6 and IL-1ß gene expression (p < 0.05). Pathway analysis on VAT whole-gene expression profiles showed that T. lutea up-regulated energy-metabolism-related genes and down-regulated inflammatory and autophagy pathways. The multitarget activity of T. lutea suggests that this microalga could be useful in mitigating risk factors of MetS.

Iron Speciation and Iron Binding Proteins in Arthrospira platensis Grown in Media Containing Different Iron Concentrations

Cyanobacteria are characterized by high iron content. This study investigated the effects of varying iron concentrations (1, 5, and 10 mg L-1) in the culture media on the biochemical composition and the iron bioaccumulation and speciation in Arthrospira platensis F&M-C256. Iron content measured in biomasses varied from 0.35 to 2.34 mg g-1 dry weight depending on the iron concentration in the culture media. These biomasses can be considered of interest for the production of spirulina-based supplements with low and high iron content. Iron speciation was studied using size exclusion chromatography followed by atomic absorption spectrometry and proteomic analysis. The role of C-phycocyanin as an iron binding protein was also investigated. Overall, the present results provide a better understanding of iron metabolism in cyanobacteria and a foundation for further studies.

A Comparative In Vitro Evaluation of the Anti-Inflammatory Effects of a Tisochrysis lutea Extract and Fucoxanthin.

In this study, we compared the effects of a Tisochrysis lutea (T. lutea) F&M-M36 methanolic extract with those of fucoxanthin (FX) at equivalent concentration, on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The T. lutea F&M-M36 methanolic extract contained 4.7 mg of FX and 6.22 mg of gallic acid equivalents of phenols per gram. HPLC analysis revealed the presence of simple phenolic acid derivatives. The T. lutea F&M-M36 extract exhibited a potent and concentration-dependent inhibitory activity against COX-2 dependent PGE2 production compared to FX alone. Compared to LPS, T. lutea F&M-M36 extract and FX reduced the expression of IL-6 and of Arg1 and enhanced that of IL-10 and of HO-1; T. lutea F&M-M36 extract also significantly abated the expression of NLRP3, enhanced mir-223 expression and reduced that of mir-146b, compared to LPS (p < 0.05). These findings indicate that T. lutea F&M-M36 methanolic extract has a peculiar anti-inflammatory activity against COX-2/PGE2 and NLRP3/mir-223 that might be attributable to the known anti-inflammatory effects of simple phenolic compounds found in the extract that may synergize with FX. Our data suggest that T. lutea F&M-M36 may serve as a source of anti-inflammatory compounds to be further evaluated in in vivo models of inflammation.

Fermented Biomass of Arthrospira platensis as a Potential Food Ingredient.

Lactic acid fermentation (LAF) is known to improve nutritional properties and functionality and to extend the shelf life of foods. We studied the LAF of Arthrospira platensis as the sole substrate using Lactobacillus plantarum as the starter culture. Fermented (FB) and non-fermented broth (NFB) were analysed by means of pH, lactic acid bacteria (LAB) count, lactic acid concentration, microbiological safety, and nutritional composition. Additionally, water and ethanol extracts were prepared on which total phenolic content, DPPH radical scavenging activity, and cellular antioxidant activity were determined. The maximum increase in LAB count and lactic acid concentration and drop in pH was observed in the first 24 h of fermentation. Total phenolic content and DPPH radical scavinging activity of ethanol extracts increased after fermentation compared with NFB. Ethanol extracts of FB have been shown as a potential source of antioxidants, which efficiently lowered oxidation level in the cells of yeast Saccharomyces cerevisiae, as well as the oxidative damage of lipids. Additionally, the level of non-protein nitrogen increased, indicating higher protein bioavailability, and fat content decreased in comparison with NFB. No presence of pathogenic bacteria and low pH indicate enhancement of FB microbiological stability. Therefore, inclusion of fermented A. platensis into food products could lead to added-value foods based on microalgae.

Microbes: Food for the Future.

Current projections estimate that in 2050 about 10 billion people will inhabit the earth and food production will need to increase by more than 60%. Food security will therefore represent a matter of global concern not easily tackled with current agriculture practices and curbed by the increasing scarcity of natural resources and climate change. Disrupting technologies are urgently needed to improve the efficiency of the food production system and to reduce the negative externalities of agriculture (soil erosion, desertification, air pollution, water and soil contamination, biodiversity loss, etc.). Among the most innovative technologies, the production of microbial protein (MP) in controlled and intensive systems called "bioreactors" is receiving increasing attention from research and industry. MP has low arable land requirements, does not directly compete with crop-based food commodities, and uses fertilizers with an almost 100% efficiency. This review considers the potential and limitations of four MP sources currently tested at pilot level or sold as food or feed ingredients: hydrogen oxidizing bacteria (HOB), methanotrophs, fungi, and microalgae (cyanobacteria). The environmental impacts (energy, land, water use, and GHG emissions) of these MP sources are compared with those of plant, animal, insect, and cultured meat-based proteins. Prices are reported to address whether MP may compete with traditional protein sources. Microalgae cultivation under artificial light is discussed as a strategy to ensure independence from weather conditions, continuous operation over the year, as well as high-quality biomass. The main challenges to the spreading of MP use are discussed.

Lactic Acid Fermentation of Arthrospira platensis (Spirulina) in a Vegetal Soybean Drink for Developing New Functional Lactose-Free Beverages.

The main objective of this study was to evaluate the suitability of Arthrospira platensis F&M-C256 (spirulina) biomass in a vegetal soybean drink or in water, as substrate for lactic acid fermentation by the probiotic bacterium Lactiplantibacillus plantarum ATCC 8014 (LAB8014) and to evaluate the fermented products in terms of bacteria content and organic acids content, biochemical composition, total phenolics, and phycocyanin content, in vitro digestibility, in vitro and in vivo antioxidant activity. After 72 h of fermentation, a bacterial concentration of about 10.5 log CFU mL-1 in the broths containing the soybean drink + spirulina + LAB8014 (SD + S + LAB8014) or water + spirulina + LAB8014 (W + S + LAB8014) was found. Lactic acid concentration reached similar values (about 1.7 g L-1) in the two broths, while a different acetic acid concentration between SD + S + LAB8014 and W + S + LAB8014 broths was observed (7.7 and 4.1 g L-1, respectively). A. platensis biomass was shown to be a suitable substrate for LAB8014 growth. After fermentation, both broths contained a high protein content (>50%). In both broths, total phenolics, in vitro and in vivo antioxidant activity increased after fermentation (+35, +20, and +93% on average, respectively), while phycocyanin content decreased (-40% on average). Digestibility of W + S + LAB8014 broth statistically improved after fermentation. This study highlights the potential of A. platensis F&M-C256 biomass as a substrate for the production of new functional lactose-free beverages.

Effects of Arthrospira platensis Extract on Physiology and Berry Traits in Vitis vinifera.

Several advantages on physiology, productivity, and grape quality have been reported for grapevine treated with seaweed extracts, but little is known about the importance of cyanobacterial-based biostimulants in viticulture. The purpose of this pioneering work was to analyze the broad-spectrum effects of the Arthrospiraplatensis F&M-C256 extract on Vitis vinifera L. cv. Pinot Nero grown in pots in optimal conditions and under water stress. To evaluate the effects, major physiological parameters of the plants and the quali-quantitative parameters of grape were analyzed. According to the results obtained in this study, ameliorating effects in leaf gas exchanges induced by A. platensis F&M-C256 treatments were detected in both irrigation regimes. Above all, A. platensis F&M-C256 allowed keeping stomata open without negative consequences in water potential in treated vines under water-stress conditions. In terms of berry traits, A. platensis F&M-C256-treated vines presented higher berry weight in comparison with untreated vines in both water regimes and improved berry composition in treated vines subjected to drought. The results of the present study demonstrated an A. platensis-dependent physiological response in case of abiotic stress, which prominently affects grape traits at harvest.

Effects of Ascophyllum nodosum extract on Vitis vinifera: Consequences on plant physiology, grape quality and secondary metabolism.

Seaweed-based extracts have been recently employed as sustainable tools to improve abiotic stress tolerance and increase grape quality. However, the effect of these extracts on secondary metabolism compounds, that are fundamental for grape and wine quality, is still scarce. In the present study, the effects of foliar treatments with an Ascophyllum nodosum extract on physiological and biochemical parameters of Vitis vinifera (cv. Sangiovese) were investigated. We hypothesized an enhancement in the biosynthesis of secondary metabolites in berry skins and in leaves in response to these treatments, effective in improve grape quality and help vines to cope with abiotic stresses. Gas exchanges, chlorophyll fluorescence and midday stem water potential on leaves treated with A. nodosum extract and non-treated control leaves, were monitored over two growing seasons at three phenological stages: full véraison, mid maturation and full maturation. In addition, anthocyanins, flavonols and hydroxycinnamic acids were quantified both in berry skins and in leaves. The foliar treatments with A. nodosum increased photosynthesis and stomatal conductance in treated compared to control plants. Furthermore, extract-treated vines were able to maintain the potential efficiency of photosystem II close to the optimal value even during the hottest periods. No effect of A. nodosum extract treatments was observed on stem water potential. A. nodosum applications delayed berry ripening, leading to a lower sugar content and a higher anthocyanin content in treated berry skins. Interestingly, treatments also affected the content and the partitioning of secondary metabolites in berry skins, as anthocyanins and flavonols contents were higher in treated compared to control plants, while the ratio of methoxylated to non-methoxylated anthocyanins was lower in treated than in control vines. Furthermore, A. nodosum extract-treated plants also had higher content of flavonols and hydroxycinnamic acids both in berry skins and in leaves and showed a reduction in the biosynthesis of methoxylated anthocyanins, which are usually accumulated in grapes under environmental constraints. Considering the challenges posed by climate change in the Mediterranean basin, the use of seaweed extracts might represent a sustainable tool to mitigate the increasing severity of drought, often associated to heat-waves, on the viticulture sector.

Microalgae as Functional Ingredients in Savory Food Products: Application to Wheat Crackers.

Crackers are widely consumed snack foods and there is an increasing trend in adding functional ingredients to their composition. In the present work, the dried biomasses of four microalgae strains-Arthrospira platensis F&M-C256, Chlorella vulgaris Allma, Tetraselmis suecica F&M-M33, and Phaeodactylum tricornutum F&M-M40-were used as a source of proteins, antioxidants, and other bioactive molecules in artisanal wheat crackers. Two incorporation levels were tested: 2% (w/w) and 6% (w/w). The impact of microalgae addition was evaluated in terms of physical properties, biochemical composition, antioxidant activity, in vitro digestibility, and sensory characteristics. Microalgae crackers presented stable color and texture throughout eight weeks of storage. Microalgae crackers were slightly thinner and lighter than the control but presented a similar density in agreement with scanning electron microscope images, indicating that gas retention was not greatly affected by microalgae addition. Regarding biochemical composition, 6% A. platensis and C. vulgaris crackers presented a significantly higher protein content (13.2-13.5%), for which they could be claimed to be a "source of protein" according to the Regulation (EC) No. 1924/2006. A. platensis crackers showed the highest antioxidant activity and attained better sensory analysis scores. T. suecica and P. tricornutum crackers showed high phenolic content and antioxidant activity but attained low sensory scores mainly because of their unattractive fishy off-flavor.

Development of new microalgae-based sourdough "crostini": functional effects of Arthrospira platensis (spirulina) addition.

The aim of this work was to evaluate the influence of Arthrospira platensis F&M-C256 (spirulina) incorporation on the nutritional and functional properties of "crostini", a leavened bakery product largely consumed in Italy and Europe. Sourdough was used as leavening and fermentation agent and three concentrations of A. platensis F&M-C256 were tested: 2%, 6% and 10% (w/w). Despite a lower volume increase compared to the control, the A. platensis F&M-C256 "crostini" doughs reached a technological appropriate volume after fermentation. At the end of fermentation, no significant differences in microorganisms concentrations were observed. A. platensis F&M-C256 "crostini" showed higher protein content compared to the control. Considering the European Commission Regulation on nutritional claims, "crostini" incorporated with 6% and 10% biomass can be claimed to be a "source of protein". Six and ten percent A. platensis "crostini" also presented significantly higher antioxidant capacity and phenolics. A significantly lower value of in vitro dry matter and protein digestibility between A. platensis F&M-C256 "crostini" and the control was found. The overall acceptability decreased with increasing A. platensis F&M-C256 addition. The combination of spirulina biomass addition and the sourdough technology led to the development of a novel microalgae-based bakery product with nutritional and functional features.

Safety evaluations and lipid-lowering activity of an Arthrospira platensis enriched diet: A 1-month study in rats.

Arthrospira platensis (A. platensis) is worldwide consumed as dietary supplement, but its use in the form of whole biomass for food purposes may raise toxicity concerns. The aim of this study was to preliminarily evaluate the safety of an A. platensis F&M-C256-enriched diet (20% (weight/weight) corresponding to 12g/kg body weight/day), administered to rats for 1month. A. platensis F&M-C256-enriched diet was well tolerated: behavior, body weight, food consumption and growth curves were not affected; no discomfort, no deaths and no physical signs related to the treatment were observed during the administration period; food daily consumption and apparent digestibility were comparable to those of the standard laboratory AIN-76 control diet. Daily water consumption and urine excretion were, on the contrary, significantly higher (27.18±1.24 vs 21.53±1.68ml and 12.63±0.99 vs 7.00±1.29ml respectively), probably because of a slight increase in sodium intake in rats fed A. platensis F&M-C256-enriched diet. Biochemical markers of kidney and liver function were not varied but a significant increase in cholesterol-HDL and a decreased plasma triglycerides level was observed in rats fed A. platensis F&M-C256-enriched diet. These last changes were associated with an increased fecal lipids excretion and liver PPAR-α gene expression. These results indicate that A. platensis F&M-C256 is likely safe and well tolerated even at a high dosage in rodents and suggest that it may represent a promising functional food for preventing or even for managing dyslipidemias.