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- Comparative lipidomic analysis of Chlorella stigmatophora and Hemiselmis cf. andersenii in response to nitrogen-induced changesPublication . Fernandes, Tomásia; Ferreira, Artur; Cordeiro, NereidaThe current focus of algae biotechnology is the production of high-value lipids, and its improvement by employing abiotic perturbations such as nitrogen-induced changes. In the present study, the growth dynamics, nitrogen uptake, pigments, and lipid composition of Chlorellla stigmatophora and Hemiselmis cf. andersenii were studied, in response to low (LN), medium (MN) and high (HN) nitrogen supplementations. Both microalgae responded to increased nitrogen levels by increasing their nitrogen uptake rate and pigment content. However, for lipid accumulation, C. stigmatophora presented a different pattern (LN: 16.56% > MN: 11.51% > HN: 10.95%) to that of H. cf. andersenii (MN: 15.37% > HN: 13.06% > LN: 6.71%). Untargeted gas chromatography–mass spectrometry analysis allowed the visualization of the biochemical diversity of C. stigmatophora and H. cf. andersenii, as well as differences in lipid regulation upon nitrogen-induced changes among species. For instance, glycosyl sterols were only detected for C. stigmatophora samples grown under MN and HN conditions. Moreover, lipid analysis of H. cf. andersenii, before and after alkaline hydrolysis, suggests that wax esters play a key role in the response of this microalga to high nitrogen levels. The cultivation of H. cf. andersenii at MN and HN was shown to be ideal for providing a rich source of ω3 and polyunsaturated fatty acids for nutraceutical purposes. The hierarchical cluster analysis showed the differential intra- and interspecific effects of nitrogen on lipid composition. The diverse ways by which both microalgae responded to nitrogen-induced changes highlighted the influence of phylogeny on the carbon flux through metabolic networks, and accumulation.
- Microalgal-based industry vs. microplastic pollution: Current knowledge and future perspectivesPublication . Mendonça, Ivana; Faria, Marisa; Rodrigues, Filipa; Cordeiro, NereidaMicroalgae can play a crucial role in the environment due to their efficient capture of CO2 and their potential as a solution for a carbon-negative economy. Water quality is critical for the success and profitability of microalgal based industries, and understanding their response to emergent pollutants, such as microplastics (MPs), is essential. Despite the published studies investigating the impact of MPs on microalgae, knowledge in this area remains limited. Most studies have mainly focused on microalgal growth, metabolite analysis, and photosyn thetic activity, with significant discrepancies in what is known about the impact on biomass yield. Recent studies show that the yield of biomass production depends on the levels of water contamination by MPs, making it necessary to reduce the contamination levels in the water. However, present technologies for extracting and purifying water from MPs are limited, and further research and technological advancements are required. One promising solution is the use of bio-based polymer materials, such as bacterial cellulose, which offer biode gradability, cost-effectiveness, and environmentally friendly detoxifying properties. This review summarises the current knowledge on MPs pollution and its impact on the viability and proliferation of microalgae-based industries, highlights the need for further research, and discusses the potential of bio-solutions for MPs removal in microalgae-based industries.
- Effects of phosphorus-induced changes on the growth, nitrogen uptake, and biochemical composition of Pavlova pinguis and Hemiselmis cf. anderseniiPublication . Fernandes, Tomásia; Cordeiro, NereidaThe understanding of the phosphorus-induced changes in the biochemical composition of microalgae is of great importance for achieving efciency in high-value lipid production. To study the chemoplasticity of Pavlova pinguis (Haptophyceae) and Hemiselmis cf. andersenii (Cryptophyceae), their growth, carotenoid and chlorophyll a content, and their monosaccharide and lipid profles were analyzed against several phosphorus (P) regimes: low (LP), medium (MP), and high (HP). For both microalgal cultures, increasing initial P concentrations showed a positive efect on biomass productivities. Carbon-rich pools presented signifcant diferences (p<0.05) for P. pinguis against P treatments, in contrast to H. cf. andersenii. Diferential responses to P-induced changes in microalgae monosaccharide and lipid profle were observed. Hemiselmis cf. andersenii increased its proportion in galactose (up to 3 times) from LP to HP conditions, whereas P. pinguis decreased (up to 20%) its glucose proportion from LP to HP conditions. For P. pinguis, the lowest amount (13.12 mg g−1 dw) of sterols was observed at LP conditions, in contrast to its carotenoid content (4.32 mg g−1 dw). P-replete conditions were the most efective in induc ing high-value lipid accumulation. Non-targeted lipid analysis revealed which samples would need to be processed to fully exploit its high-value lipids, namely H. cf andersenii under MP and HP conditions. This study demonstrated that P played an important role in carbon allocation, nitrogen uptake, and lipid regulation on P. pinguis and H. cf. andersenii, and that P-replete conditions could be useful for optimizing high-value lipids with potential for nutraceutical and pharmaceutical felds.
- Tissue engineering scaffold material with enhanced cell adhesion and angiogenesis from soy protein isolate loaded with bio modulated micro-TiO2 prepared via prolonged sonication for wound healing applicationsPublication . Koshy, Rekha Rose; Mary, Siji K.; Reghunadhan, Arunima; Dalvi, Yogesh Bharat; Kailas, Lekshmi; Cordeiro, Nereida; Thomas, Sabu; Pothen, Laly A.Tissue engineering is a technique that promotes healing by creating an ideal environment for endogenous cells to migrate and grow into the site of injury via a scaffold, improving regeneration and reducing the time required for in vitro cell culture. In this work, the effect of the addition of sonicated TiO2 in the soy protein isolate (SPI) matrix for tissue engineering applications was studied. In comparison to adding expensive nano TiO2, this method of incorporating sonicated TiO2 into the SPI matrix will aid in achieving improved properties at a lower cost. The effect of the addition of sonicated TiO2 on the morphological, UV transmittance, mechanical, thermal, surface energy, and hydrophilicity of SPI films was investigated. The result shows that the uniformly distributed TiO2 particles successfully blocked 95% of UV light. Scanning electron microscopy revealed a significant reduction in the TiO2 agglomerate size and homogeneous distribution of the same when sonication was applied instead of mechanical dispersion. A simultaneous increase of tensile strength (from 3.16 to 4.58 MPa) and elongation at break values (from 24.25% to 95.31%) with 0.5% TiO2 was observed. The addition of 0.25% TiO2 was found to significantly enhance the elongation at break value to 120.83%. Incorporation of micro-TiO2 particles could improve the surface roughness, surface energy, and wettability of SPI films. In vitro cell adhesion studies and in vivo subcutaneous implantation studies were performed to assess the cell growth and angiogenesis of the developed film membranes. An MTT assay showed that SPI-1%TiO2 film favored cell viability up to 118%, and in vivo subcutaneous implantation studies showed enhanced cell growth and angiogenesis for SPI-1% TiO2 films. This SPI-TiO2 film with enhanced surface properties can be used as an ideal candidate for tissue engineering applications.
- Functional, rheological, and antioxidant properties of extracellular polymeric substances produced by a thermophilic cyanobacterium Leptolyngbya spPublication . Gongi, Wejdene; Cordeiro, Nereida; Pinchetti, Juan Luis Gomez; Ouada, Hatem BenExtracellular polymeric substances (EPSs) produced by the flamentous cyanobacterium identifed as Leptolyngbya sp. IkmLPT16 were isolated and characterized chemically, and their antioxidant, functional, and rheological properties were studied. The strain produces a signifcant amount of EPSs (2.15 g L−1) conjointly with a biomass production achieved at a maximum of 1.35 g L−1 after nine production days. Chemical analysis of EPSs revealed the presence of mannose (35%), arabinose (24%), glucose (15%), rhamnose (2%), and one uronic acid (8%). Fourier transformed infrared spectrum of EPSs further revealed the presence of νC-N groups indicating the presence of peptide moieties. Elemental analysis of EPSs showed the presence of sulfate groups (S=0.59%) as inorganic substituents. Functional properties of Leptolyngbya EPSs were determined based on water holding capacity, oil holding capacity, foaming ability, and metal sorption ability. Experimental results showed high water holding capacity (119%), water solubility index (97.43%), and oil holding ability (87.52%), with a strong metal sorption ability and consequent foam stability (22%). The rheological properties of EPSs were comparable with commercial xanthan gum with higher resistance to Temperature. Leptolyngbya sp. EPSs displayed an efective antioxidant activity via directly scavenging free radicals particularly DDPH• (IC50=4 mg. mL−1 against 10 mg. mL−1 for l-ascorbic acid) and •OH (IC50=10 mg. mL−1 against 20 mg. mL−1 for l-ascorbic acid) and as an iron-chelating agent (IC50=40 mg. mL−1 against 60 mg.mL−1 for EDTA). The outcomes of this study demonstrate the potential use of Leptolyngbya sp. EPSs in several food and pharmaceutical applications.
- Hemiselmis andersenii and Chlorella stigmatophora as new sources of high‐value compounds: a lipidomic approachPublication . Fernandes, Tomásia; Cordeiro, NereidaTo unlock the potential of Chlorella stigmatophora (Trebouxiophyceae, Chlorophyta) and Hemiselmis andersenii (Cryptophyceae, Cryptophyta) as natural reactors for biotechnological exploitation, their lipophilic extracts were characterized using Fourier Transform Infrared spectroscopy with Attenuated Total Reflectance (FTIR-ATR) and Gas Chromatography-Mass Spectrometry (GC-MS) before and after alkaline hydrolysis. The GC-MS analysis enabled the identification of 62 metabolites-namely fatty acids (27), aliphatic alcohols (17), monoglycerides (7), sterols (4), and other compounds (7). After alkaline hydrolysis, monounsaturated fatty acids increased by as much as 87%, suggesting that the esterified compounds were mainly neutral lipids. Hemiselmis andersenii yielded the highest Σω3/Σω6 ratio (7.26), indicating that it is a good source of ω3 fatty acids, in comparison to C. stigmatophora (Σω3/Σω6 = 1.24). Both microalgae presented significant amounts of aliphatic alcohols (6.81-10.95 mg · g dw-1 ), which are recognized by their cholesterol-lowering properties. The multivariate analysis allowed visualization of the chemical divergence among H. andersenii lipophilic extracts before and after alkaline hydrolysis, as well as species-specific differences. Chlorella stigmatophora showed to be a valuable source of essential fatty acids for nutraceuticals, whereas H. andersenii, due to its high chemical diversity, seems to be suitable for different fields of application.
- Nanofibrils vs nanocrystals bio-nanocomposites based on sodium alginate matrix: an improved-performance studyPublication . Deepa, B.; Abraham, E.; Cordeiro, Nereida; Faria, M.; Primc, G.; Pottathara, Y.; Leskovšek, M.; Gorjanc, M.; Mozetič, M.; Thomas, S.; Pothan, L.A.To develop bio-nanocomposites using natural biopolymers, nanocomposite films were prepared based on sodium alginate and kapok nanofibrils (CNFs). CNFs when subjected to TEMPO-mediated oxidation gave rise to cellulose nanocrystals (TOCNCs), with carboxyl groups at the surface ( Ka/Kb = 3.64). The differences between the two types of nanocelluloses (nanofibrils and nanocrystals) and their impact in the preparation of bio-nanocomposites, were studied. When incorporated in the matrix, the CNFs particles have the tendency to form surface aggregation ( Ka/Kb = 2.37), distorting the alginate network, creating heterogeneous films, with high surface roughness (S a = 29.37 nm), porosity (D p = 0.087 cm2/min) and vulnerability to heat. The TOCNCs present good dispersion creating a 3D network, which forms uniform (D p = 0.122 cm2/min) and homogeneous films, with smooth surface (S a = 16.83 nm). The ultrasonication treatment facilitated the dispersion improving the interfacial interaction between the reinforcing phase and the matrix. The results show the reinforcement potential of kapok nanocellulose in an industrially and medically important biopolymer, sodium alginate, especially when TOCNCs and ultrasonication were used.
- High-value lipids accumulation by Pavlova pinguis as a response to nitrogen-induced changesPublication . Fernandes, Tomásia; Cordeiro, NereidaThe challenges of exploring the potential of microalgal strains for biotechnological applications include the optimization of their cell growth and chemical composition. To overcome this, it is essential to understand the mechanisms that lead to the accumulation of desired products within microalgal cells. In this study, a gradient of nitrogen as nitrate (NO−3 − N) concentrations was used for the preparation of P. pinguis growth medium. The algal growth dynamics, pigments, nutrient uptake, and detailed lipid composition across treatments were assessed. Increasing the NO− 3 − N level led to higher lipid content (21%), a higher cell uptake rate (0.20 pg NO− 3 − N cell− 1 d− 1 ) and more accumulation of chlorophylls, carotenoids, and high-value lipids. Eicosapentaenoic acid, essential fatty acids, phytol and stigmasterol were the key high-value lipids that were positively influenced by higher NO− 3 − N levels. High NO− 3 − N conditions induced an increase of 54% in total sterol content, while low NO− 3 − N conditions resulted in increased proportions of saturated fatty acids (66% more) and decreased proportions of polyunsaturated fatty acids (14% less). The low NO− 3 − N level also led to higher amounts of monoglyceride (1.60 mg g− 1 ; 64% more). The analysis of P. pinguis lipids before and after hydrolysis provided an insight into the composition of the esterified lipids across treatments. NO− 3 − N supplementation was revealed to be an effective strategy for enhancing P. pinguis lipid composition, for nutraceutical and pharmaceutical industries (high NO− 3 − N level). A greater understanding of the NO− 3 − N uptake and the use efficiency by P. pinguis was reached, showing its potential for further biotechnological applications.
- Characterization of biodegradable films based on extracellular polymeric substances extracted from the thermophilic microalga Graesiella spPublication . Gongi, Wejdene; Pinchetti, Juan Luis Gómez; Cordeiro, Nereida; Sadok, Saloua; Ouada, Hatem BenIn this research, a new type of biodegradable film based on the extracellular polymeric substances (EPS) and isolated from the thermophilic microalga Graesiella sp., was formulated and characterized. The EPS film was 0.221 mm thick. Atomic force microscopy and scanning electron microscopy images revealed a homogeneous character with a lamellar microstructure. The EPS film displayed yellowish color, high transparency, high ul traviolet barrier properties, and low oxygen (0.008 SI), and water-vapor permeability (0.037 SI). Film tensile strength (16.24 MPa) and elongation at break (4.76%) were in the range of common biofilms and the thermal analyses showed high transition temperature (126 ◦C) and high thermal stability (up to 800 ◦C). Compared to ascorbic acid, results indicated that the EPS film shows a higher antioxidant activity, mainly as β-carotene anti bleaching (84%), DPPH- free radical scavenging ability (80%), and ferrous iron-chelating (55%). Graesiella sp., EPS film effects on beef meat packaging were studied during nine days of cold storage. Compared to polyvinylchloride-packed meat, EPS-packed meat samples showed higher stability of color (redness = 13.6) and pH (5.85) during storage and low proliferation of total viable counts (4.04 CFU⋅g− 1 ) and Pseudomonas bacteria (4.09 CFU⋅g− 1 ). They also exhibit lower drip loss (9%) and less metmyoglobin (32%), heme iron (4.87 μg⋅g− 1 ) total volatile basic nitrogen (TVB-N = 22.96 mg⋅kg− 1 ), and lipid oxidation (MDA = 0.025 mg⋅kg− 1 ). The obtained results highlight the potential for use of microalgae EPS as a new film forming material that could be applied in beef meat preservation.
- Determination of phthalic acid esters and di(2-ethylhexyl) adipate in fish and squid using the ammonium formate version of the QuEChERS method combined with gas chromatography mass spectrometryPublication . Sambolino, Annalisa; Ortega-Zamora, Cecilia; González-Sálamo, Javier; Dinis, Ana; Cordeiro, Nereida; Canning-Clode, João; Hernández-Borges, JavierIn the present study, the ammonium formate version of the QuEChERS method, considered highly advantageous in relation to instrument maintenance and other issues, was applied for the first time to extract a group of twelve phthalic acid esters (PAEs, i.e. dipropyl phthalate, DPP; diisobutyl phthalate, DIBP; dibutyl phthalate, DBP; diisopentyl phthalate, DIPP; di-n-pentyl phthalate, DNPP; dihexyl phthalate, DHP; butyl benzyl phthalate, BBP; dicyclohexyl phthalate, DCHP; di(2-ethylhexyl) phthalate, DEHP; di-n-octyl phthalate, DNOP; diisononyl phthalate, DINP; and diisodecyl phthalate, DIDP) and one adipate (di(2-ethylhexyl) adipate, DEHA) from two species of fish (Scomber colias and Katsuwonus pelamis) and one of squid (Loligo gahi). The method was validated in terms of linearity, trueness and matrix effects. Determination coefficients (R2 ) for matrix-matched calibration curves were higher than 0.99 in all cases, being the lowest calibration levels in the range 0.5–10 ng/g. Mean recovery values were between 70 and 117% with relative standard deviation values ≤20%. Matrix effects were soft (between − 20 and +20%) for most analytes and matrices, except in squid samples, which was mostly medium with a moderate ion suppression. The analysis of 10 samples of each type showed the presence of DIBP, DBP and DEHP at concentrations up to 44.2 ± 2.1 ng/g of wet weight in some of the samples and species, still not representing concerning values when considering the daily intake of such species of seafood in the human diet (tolerable daily intake -TDI- values were not exceeded). Results demonstrated that the ammonium formate version of the QuEChERS method can be applied with success for the extraction and determination of the selected PAEs and DEHA in fish and squid samples.