Tetraselmis chuii Edible Microalga as a New Source of Neuroprotective Compounds Obtained Using Fast Biosolvent Extraction.

Tetraselmis chuii is an EFSA-approved novel food and dietary supplement with increasing use in nutraceutical production worldwide. This study investigated the neuroprotective potential of bioactive compounds extracted from T. chuii using green biobased solvents (ethyl acetate, AcOEt, and cyclopentyl methyl ether, CPME) under pressurized liquid extraction (PLE) conditions and supercritical fluid extraction (SFE). Response surface optimization was used to study the effect of temperature and solvent composition on the neuroprotective properties of the PLE extracts, including anticholinergic activity, reactive oxygen/nitrogen species (ROS/RNS) scavenging capacity, and anti-inflammatory activity. Optimized extraction conditions of 40 °C and 34.9% AcOEt in CPME resulted in extracts with high anticholinergic and ROS/RNS scavenging capacity, while operation at 180 °C and 54.1% AcOEt in CPME yielded extracts with potent anti-inflammatory properties using only 20 min. Chemical characterization revealed the presence of carotenoids (neoxanthin, violaxanthin, zeaxanthin, α- and ß-carotene) known for their anti-cholinesterase, antioxidant, and anti-inflammatory potential. The extracts also exhibited high levels of omega-3 polyunsaturated fatty acids (PUFAs) with a favorable ω-3/ω-6 ratio (>7), contributing to their neuroprotective and anti-inflammatory effects. Furthermore, the extracts were found to be safe to use, as cytotoxicity assays showed no observed toxicity in HK-2 and THP-1 cell lines at or below a concentration of 40 µg mL-1. These results highlight the neuroprotective potential of Tetraselmis chuii extracts, making them valuable in the field of nutraceutical production and emphasize the interest of studying new green solvents as alternatives to conventional toxic solvents.

TetraSOD®, a Unique Marine Microalgae Ingredient, Promotes an Antioxidant and Anti-Inflammatory Status in a Metabolic Syndrome-Induced Model in Rats.

Increased oxidative stress has been linked to the pathogenic process of obesity and can trigger inflammation, which is often linked with the risk factors that make up metabolic syndrome (MetS), including obesity, insulin resistance, dyslipidaemia and hypertension. TetraSOD®, a natural marine vegan ingredient derived from the microalgae Tetraselmis chuii that is high in the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) has recently demonstrated in vitro increased activity of these key antioxidant enzymes. In the present study, the potential bioactive effects of three dietary dosages of TetraSOD® in enhancing antioxidant and anti-inflammatory mechanisms to combat the metabolic disturbances that compose MetS were assessed in rats given a cafeteria (CAF) diet. Chronic supplementation with 0.17, 1.7, and 17 mg kg-1 day-1 of TetraSOD® for 8 weeks ameliorated the abnormalities associated with MetS, including oxidative stress and inflammation, promoting endogenous antioxidant defence mechanisms in the liver (GPx and GSH), modulating oxidative stress and inflammatory markers in plasma (NOx, oxLDL and IL-10), and regulating genes involved in antioxidant, anti-inflammatory and immunomodulatory pathways in the liver, mesenteric white adipose tissue (MWAT), thymus, and spleen. Overall, TetraSOD® appears to be a potential therapeutic option for the management of MetS.

A PMAxxTM qPCR Assay Reveals That Dietary Administration of the Microalgae Tetraselmis chuii Does Not Affect Salmonella Infantis Caecal Content in Early-Treated Broiler Chickens.

Salmonella enterica serovars cause infections in humans. S. enterica subsp. enterica serovar Infantis is considered relevant and is commonly reported in poultry products. Evaluating innovative approaches for resisting colonization in animals could contribute to the goal of reducing potential human infections. Microalgae represent a source of molecules associated with performance and health improvement in chickens. Tetraselmis chuii synthesizes fermentable polysaccharides as part of their cell wall content; these sugars are known for influencing caecal bacterial diversity. We hypothesized if its dietary administration could exert a positive effect on caecal microbiota in favor of a reduced S. Infantis load. A total of 72 one-day-old broiler chickens (COBB 500) were randomly allocated into three groups: a control, a group infected with bacteria (day 4), and a group challenged with S. Infantis but fed a microalgae-based diet. Caecal samples (n = 8) were collected two days post-infection. A PMAxxTM-based qPCR approach was developed to assess differences regarding bacterial viable load between groups. The inclusion of the microalga did not modify S. Infantis content, although the assay proved to be efficient, sensitive, and repeatable. The utilized scheme could serve as a foundation for developing novel PCR-based methodologies for estimating Salmonella colonization.

Improving the Nutritional, Structural, and Sensory Properties of Gluten-Free Bread with Different Species of Microalgae.

Microalgae are an enormous source of nutrients that can be utilized to enrich common food of inherently low nutritional value, such as gluten-free (GF) bread. Addition of the algae species: Tetraselmis chuii (Tc), Chlorella vulgaris (Cv), and Nannochloropsis gaditana (Ng) biomass led to a significant increase in proteins, lipids, minerals (Ca, Mg, K, P, S, Fe, Cu, Zn, Mn), and antioxidant activity. Although, a compromise on dough rheology and consequential sensory properties was observed. To address this, ethanol treatment of the biomass was necessary to eliminate pigments and odor compounds, which resulted in the bread receiving a similar score as the control during sensory trials. Ethanol treatment also resulted in increased dough strength depicted by creep/recovery tests. Due to the stronger dough structure, more air bubbles were trapped in the dough resulting in softer breads (23-65%) of high volume (12-27%) vs. the native algae biomass bread. Breads baked with Ng and Cv resulted in higher protein-enrichment than the Tc, while Tc enrichment led to an elevated mineral content, especially the Ca, which was six times higher than the other algae species. Overall, Ng, in combination with ethanol treatment, yielded a highly nutritious bread of improved technological and sensory properties, indicating that this species might be a candidate for functional GF bread development.

Administration of Dietary Microalgae Ameliorates Intestinal Parameters, Improves Body Weight, and Reduces Thawing Loss of Fillets in Broiler Chickens: A Pilot Study.

This pilot investigation aimed at studying the feasibility of using a low dose (0.2%) of dietary microalgae as a means of improving intestinal morphometry, body weight, and selected meat quality parameters in broilers. A total of 72 one-day-old ROSS 308 male chicks were randomly separated into four groups; three experimental pens in which the birds were fed with biomass from Tysochrysis lutea, Tetraselmis chuii, and Porphyridium cruentum over 30 days and a control group. T. chuii and P. cruentum had a positive effect with regard to body weight. In treated animals, duodenal and ileal sections showed characteristic tall and thin villi, with serrated surfaces and goblet cell differentiation. In both sections, values of the villus-height-to-crypt-depth ratio were increased by microalgae ingestion. The thawing weight loss of fillets was reduced in T. chuii-fed animals. The positive effects exerted by T. chuii and P. cruentum on intestinal architecture were associated with the improved body weight. Arguably, these outcomes exhibit the potential of using these species to enhance growth performance in broiler chickens by promoting gut homeostasis and thus nutrient absorption.

Special Issue: Rheology and Quality Research of Cereal-Based Food.

New trends in the cereal industry deal with a permanent need to develop new food products that are adjusted to consumer demands and, in the near future, the scarcity of food resources. Sustainable food products as health and wellness promoters can be developed redesigning traditional staple foods, using environmentally friendly ingredients (such as microalgae biomass or pulses) or by-products (e.g., tomato seeds) in accordance with the bioeconomy principles. These are topics that act as driving forces for innovation and will be discussed in the present special issue. Rheology always was the reference discipline to determine dough and bread properties. A routine analysis of cereal grains includes empirical rheology techniques that imply the use of well-known equipment in cereal industries (e.g., alveograph, mixograph, extensograph). Their parameters determine the blending of the grains and are crucial on the technical sheets that determine the use of flours. In addition, the structure of gluten-free cereal-based foods has proven to be a determinant for the appeal and strongly impacts consumers' acceptance. Fundamental rheology has a relevant contribution to help overcome the technological challenges of working with gluten-free flours. These aspects will also be pointed out in order to provide a prospective view of the relevant developments to take place in the area of cereal technology.

Update of the list of QPS-recommended biological agents intentionally added to food or feed as notified to EFSA 11: suitability of taxonomic units notified to EFSA until September 2019.

Qualified presumption of safety (QPS) was developed to provide a generic safety evaluation for biological agents to support EFSA's Scientific Panels. The taxonomic identity, body of knowledge, safety concerns and antimicrobial resistance are assessed. Safety concerns identified for a taxonomic unit (TU) are where possible to be confirmed at strain or product level, reflected by 'qualifications'. No new information was found that would change the previously recommended QPS TUs and their qualifications. The list of microorganisms notified to EFSA was updated with 54 biological agents, received between April and September 2019; 23 already had QPS status, 14 were excluded from the QPS exercise (7 filamentous fungi, 6 Escherichia coli, Sphingomonas paucimobilis which was already evaluated). Seventeen, corresponding to 16 TUs, were evaluated for possible QPS status, fourteen of these for the first time, and Protaminobacter rubrum, evaluated previously, was excluded because it is not a valid species. Eight TUs are recommended for QPS status. Lactobacillus parafarraginis and Zygosaccharomyces rouxii are recommended to be included in the QPS list. Parageobacillus thermoglucosidasius and Paenibacillus illinoisensis can be recommended for the QPS list with the qualification 'for production purposes only' and absence of toxigenic potential. Bacillus velezensis can be recommended for the QPS list with the qualification 'absence of toxigenic potential and the absence of aminoglycoside production ability'. Cupriavidus necator, Aurantiochytrium limacinum and Tetraselmis chuii can be recommended for the QPS list with the qualification 'production purposes only'. Pantoea ananatis is not recommended for the QPS list due to lack of body of knowledge in relation to its pathogenicity potential for plants. Corynebacterium stationis, Hamamotoa singularis, Rhodococcus aetherivorans and Rhodococcus ruber cannot be recommended for the QPS list due to lack of body of knowledge. Kodamaea ohmeri cannot be recommended for the QPS list due to safety concerns.

Improved Extraction Efficiency of Antioxidant Bioactive Compounds from Tetraselmis chuii and Phaedoactylum tricornutum Using Pulsed Electric Fields.

Pulsed electric fields (PEF) is a promising technology that allows the selective extraction of high-added value compounds by electroporation. Thus, PEF provides numerous opportunities for the energy efficient isolation of valuable microalgal bioactive substances (i.e., pigments and polyphenols). The efficiency of PEF-assisted extraction combined with aqueous or dimethyl sulfoxide (DMSO) solvents in recovering pigments and polyphenols from microalgae Tetraselmis chuii (T. chuii) and Phaeodactylum tricornutum (P. tricornutum) was evaluated. Two PEF treatments were applied: (1 kV/cm/400 pulses, 3 kV/cm/45 pulses), with a specific energy input of 100 kJ/kg. The total antioxidant capacity (TAC) was positively influenced by the use of DMSO. The highest TAC in the T. chuii culture was achieved at a lower extraction time and electric field than for P. tricornutum. The use of DMSO only improved the polyphenols' extraction for P. tricornutum, whereas the PEF and extraction time were more important for T. chuii. Carotenoids and chlorophyll a were more efficiently extracted using DMSO, while chlorophyll b levels were higher following aqueous extraction for both microalgae. In P. tricornutum, the TAC and pigment extraction efficiency were in general higher at lower extraction times. It can be concluded that PEF may be a promising alternative for the enhancement of the selective extraction of antioxidant bioactive compounds from microalgae.

Tetraselmis chuii as a Sustainable and Healthy Ingredient to Produce Gluten-Free Bread: Impact on Structure, Colour and Bioactivity.

The objective of this work is to increase the nutritional quality of gluten-free (GF) bread by addition of Tetraselmis chuii microalgal biomass, a sustainable source of protein and bioactive compounds. The impact of different levels of T. chuii (0%-Control, 1%, 2% and 4% w/w) on the GF doughs and breads' structure was studied. Microdough-Lab mixing tests and oscillatory rheology were conducted to evaluate the dough´s structure. Physical properties of the loaves, total phenolic content (Folin-Ciocalteu) and antioxidant capacity (DPPH and FRAP) of the bread extracts were assessed. For the low additions of T. chuii (1% and 2%), a destabilising effect is noticed, expressed by lower dough viscoelastic functions (G' and G'') and poor baking results. At the higher level (4%) of microalgal addition, there was a structure recovery with bread volume increase and a decrease in crumb firmness. Moreover, 4% T. chuii bread presented higher total phenolic content and antioxidant capacity when compared to control. Bread with 4% T. chuii seems particularly interesting since a significant increase in the bioactivity and an innovative green appearance was achieved, with a low impact on technological performance, but with lower sensory scores.

Safety assessment of a lyophilized biomass of Tetraselmis chuii (TetraSOD®) in a 90 day feeding study.

TetraSOD® is a powder of the lyophilized biomass of Tetraselmis chuii strain CCFM03, a marine microalga with a history of use as feed in the aquaculture industry. Recently, algae including T. chuii have been investigated for their potential use in human food. However, published toxicology studies addressing the safety of T. chuii as a food ingredient are not available. To address this issue, the toxicity of TetraSOD® was evaluated using a 90-day oral toxicology study in rats following the Organisation for Economic Co-operation and Development (OECD) test guideline 408. No treatment-related mortality or clinical signs were noted with TetraSOD® at doses of 625, 1667, or 2500 mg/kg/day. Additionally, no adverse effects on haematology, blood biochemistry, organ weights, gross or histopathology were observed. The Non Observed Adverse Effect Level (NOAEL) for TetraSOD® is greater than the highest tested dose of 2500 mg/kg/day.

Are gold nanoparticles and microplastics mixtures more toxic to the marine microalgae Tetraselmis chuii than the substances individually?

The widespread use of microplastics and nanomaterials resulting in environmental contamination is of high concern. Microplastics have been found to modulate the toxicity of other environmental contaminants. Thus, the hypothesis that microplastics increase the toxicity of gold nanoparticles to the marine microalgae Tetraselmis chuii was tested. In a laboratory bioassay, T. chuii cultures were exposed for 96 h to ∼5 nm diameter gold nanoparticles (AuNP) and to virgin 1-5 µm diameter microplastics (MP), alone and in mixture. The treatments were: control; citrate-control; AuNP alone (0.1, 0.3 and 3 mg/L); MP alone (0.3, 0.9 and 4 mg/L) and mixture of the two substances in three different concentrations (0.1 mg/L AuNP + 0.3 mg/L MP; 0.3 mg/L AuNP + 0.9 mg/L MP; 3 mg/l AuNP + 4 mg/L MP). The effect criterion was the inhibition of the average specific growth rate. AuNP alone and MP alone did not cause significant decrease of T. chui average specific growth rate up to 3 mg/L and 4 mg/L, respectively. The mixture containing 3 mg/L AuNP + 4 mg/L MP significantly reduced the average specific growth rate of the microalgae. Therefore, this mixture was more toxic to T. chuii than its components individually. Overall, the results of the present study indicated that the MP and AuNP tested have a relatively low toxicity to T. chuii, but the toxicity increases when they are in mixtures containing high concentrations of both substances. These proof-of-concept findings stress the need of more research on the toxicity of mixtures containing microplastics and nanomaterials.