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- Sargassum muticum and Osmundea pinnatifida enzymatic extracts: chemical, structural, and cytotoxic characterizationPublication . Rodrigues, Dina; Costa-Pinto, Ana R.; Sousa, Sérgio; Vasconcelos, Marta W.; Pintado, Manuela M.; Pereira, Leonel; Rocha-Santos, Teresa A. P.; Costa, João P. da; Silva, Artur M. S.; Duarte, Armando C.; Gomes, Ana M. P.; Freitas, Ana C.Seaweeds, which have been widely used for human consumption, are considered a potential source of biological compounds, where enzyme-assisted extraction can be an efficient method to obtain multifunctional extracts. Chemical characterization of Sargassum muticum and Osmundea pinnatifida extracts obtained by Alcalase and Viscozyme assisted extraction, respectively, showed an increment of macro/micro elements in comparison to the corresponding dry seaweeds, while the ratio of Na/K decreased in both extracts. Galactose, mannose, xylose, fucose, and glucuronic acid were the main monosaccharides (3.2–27.3 mg/glyophilized extract) present in variable molar ratios, whereas low free amino acids content and diversity (1.4–2.7 g/100gprotein) characterized both extracts. FTIR-ATR and 1H NMR spectra confirmed the presence of important polysaccharide structures in the extracts, namely fucoidans from S. muticum or agarans as sulfated polysaccharides from O. pinnatifida. No cytotoxicity against normal mammalian cells was observed from 0 to 4 mglyophilized extract/mL for both extracts. The comprehensive characterization of the composition and safety of these two extracts fulfils an important step towards their authorized application for nutritional and/or nutraceutical purposes.
- Biodegradation of polyethylene microplastics by the marine fungus zalerion maritimumPublication . Paço, Ana; Duarte, Kátia; Costa, João P. da; Santos, Patrícia S.M.; Pereira, R.; Pereira, M. E.; Freitas, Ana C.; Duarte, Armando C.; Rocha-Santos, Teresa A. P.Plastic yearly production has surpassed the 300milliontons mark and recycling has all but failed in constituting a viable solution for the disposal of plastic waste. As these materials continue to accumulate in the environment, namely, in rivers and oceans, in the form of macro-, meso-, micro- and nanoplastics, it becomes of the utmost urgency to find new ways to curtail this environmental threat. Multiple efforts have been made to identify and isolate microorganisms capable of utilizing synthetic polymers and recent results point towards the viability of a solution for this problem based on the biodegradation of plastics resorting to selected microbial strains. Herein, the response of the fungus Zalerion maritimum to different times of exposition to polyethylene (PE) pellets, in a minimum growth medium, was evaluated, based on the quantified mass differences in both the fungus and the microplastic pellets used. Additionally, molecular changes were assessed through attenuated total reflectance Fourier transform Infrared Spectroscopy (FTIR-ATR) and Nuclear Magnetic Resonance (NMR). Results showed that, under the tested conditions, Z. maritimum is capable of utilizing PE, resulting in the decrease, in both mass and size, of the pellets. These results indicate that this naturally occurring fungus may actively contribute to the biodegradation of microplastics, requiring minimum nutrients.
- Chemical and structural characterization of Pholiota nameko extracts with biological propertiesPublication . Rodrigues, Dina; Freitas, Ana C.; Sousa, Sérgio; Amorim, Manuela; Vasconcelos, Marta W.; Costa, João P. da; Silva, Artur M. S.; Rocha-Santos, Teresa A. P.; Duarte, Armando C.; Gomes, Ana M. P.Edible mushrooms including Pholiota nameko are excellent sources of extractable bioactive compounds with much to explore. Enzymatic extractions with Cellulase and Viscozyme were responsible for highest extraction yields (67-77%). No strong antioxidant activity was observed although extracts were able to scavenge ABTS(+) and OH(+). Potential prebiotic activity was observed in all extracts, some increasing 1.4-2 Log cycles of Lactobacillus acidophilus La-5 and Bifidobacterium animalis BB12. 30-50% α-glucosidase inhibition was observed in ultrasound, Flavourzyme and Cellulase extracts. Flavourzyme and Cellulase extracts are richer in macro (Mg, K and P) and micro elements (Zn, Mn and Fe) in comparison to mushroom. Monosaccharides content and profile varied slightly among both extracts with predominance of glucose, galactose and mannose with no uronic acids detection; Flavourzyme extract reported higher free amino acids content. Presence of α and β-glycosidic structures such as glucans and glucan-protein complexes are among the polysaccharides found in both extracts.
- Carbon nanotube field effect transistor biosensor for the detection of toxins in seawaterPublication . Marques, Inês; Costa, João Pinto da; Justino, Celine; Santos, Patrícia; Duarte, Kátia; Freitas, Ana; Cardoso, Susana; Duarte, Armando; Rocha-Santos, TeresaDisposable field effect transistors (FET) biosensors (bio-FET) based on carbon nanotubes were fabricated for detection of domoic acid (DA), which belongs to the group of biotoxins associated with the amnesic shellfish poisoning. The analytical results obtained with the bio-FET were compared with those obtained with a traditional methodology (enzyme-linked immunosorbent assay) in order to validate the bio-FET for DA detection. Standard solutions of DA with concentrations between 10 and 500 ng L−1 were tested in order to construct the calibration curve, where five bio-FET were used for reproducibility estimation and two analytical measurements were performed for each bio-FET for repeatability estimation. Ten spiked artificial seawater samples were used to validate the bio-FET. The obtained reproducibility (0.52–1.43%), repeatability (0.57–1.27%), limit of detection (10 ng L−1) and recovery range (92.3–100.3%) reveal an adequate analytical performance of the bio-FET for the detection of DA in environmental samples such as seawater samples.
- Salinity induced effects on the growth rates and mycelia composition of basidiomycete and zygomycete fungiPublication . Venâncio, C.; Pereira, R.; Freitas, A. C.; Rocha-Santos, T. A. P.; Costa, J. P. da; Duarte, A. C.; Lopes, I.Soil salinization, as the combination of primary and secondary events, can adversely affect organisms inhabiting this compartment. In the present study, the effects of increased salinity were assessed in four species of terrestrial fungi: Lentinus sajor caju, Phanerochaete chrysosporium, Rhizopus oryzae and Trametes versicolor. The mycelial growth and biochemical composition of the four fungi were determined under three exposure scenarios: 1) exposure to serial dilutions of natural seawater (SW), 2) exposure to serial concentrations of NaCl (potential surrogate of SW); and 3) exposure to serial concentrations of NaCl after a period of pre-exposure to low levels of NaCl. The toxicity of NaCl was slightly higher than that of SW, for all fungi species: the conductivities causing 50% of growth inhibition (EC50) were within 14.9 and 22.0 mScm−1 for NaCl and within 20.2 and 34.1 mScm−1 for SW. Phanerochaete chrysosporium showed to be the less sensitive species, both for NaCl and SW. Exposure to NaCl caused changes in the biochemical composition of fungi, mainly increasing the production of polysaccharides. When fungi were exposed to SW this pattern of biochemical response was not observed. Fungi pre-exposed to low levels of salinity presented higher EC50 than fungi non-pre-exposed, though 95% confidence limits overlapped, with the exception of P. chrysosporium. Pre-exposure to low levels of NaCl also induced changes in the biochemical composition of the mycelia of L. sajor caju and R. oryzae, relatively to the respective control. These results suggest that some terrestrial fungi may acquire an increased tolerance to NaCl after being pre-exposed to low levels of this salt, thus, suggesting their capacity to persist in environments that will undergo salinization. Furthermore, NaCl could be used as a protective surrogate of SW to derive safe salinity levels for soils, since it induced toxicity similar or higher than that of SW.