Differentiation of the Organoleptic Volatile Organic Compound Profile of Three Edible Seaweeds.

The inclusion of seaweeds in daily-consumption food is a worthy-of-attention challenge due to their high nutritional value and potential health benefits. In this way, their composition, organoleptic profile, and toxicity must be assessed. This work focuses on studying the volatile organic compounds (VOCs) emitted by three edible seaweeds, Grateloupia turuturu, Codium tomentosum, and Bifurcaria bifurcata, with the aim of deepening the knowledge regarding their organoleptic profiles. Nine samples of each seaweed were prepared in glass vials, and the emitted headspace was analyzed, for the first time, with a gas chromatography-ion mobility spectrometry device, a highly sensitive technology. By statistically processing the collected data through PCA, it was possible to accurately differentiate the characteristic patterns of the three seaweeds with a total explained variance of 98%. If the data were pre-processed through PLS Regression, the total explained variance increased to 99.36%. The identification of 13 VOCs was accomplished through a developed database of compounds. These outstanding values in addition to the identification of the main emissions of VOCs and the utilization of a never-before-used technology prove the capacity of GC-IMS to differentiate edible seaweeds based solely on their volatile emissions, increase the knowledge regarding their organoleptic profiles, and provide an important step forward in the inclusion of these highly nutritional ingredients in the human diet.

Proximate Composition, Physicochemical and Microbiological Characterization of Edible Seaweeds Available in the Portuguese Market.

BACKGROUND: The aim of this work was the study of the proximate composition and profile of fatty acids, minerals, and some microbiological aspects of four edible seaweed species (Chondrus crispus, Palmaria palmata, Porphyra sp., and Ulva sp.) available in the Portuguese market for food consumption, and produced in a national Integrated Multi-Trophic System (IMTA). METHODS: Moisture, ash, and total lipids were determined gravimetrically. Crude protein was analysed by Duma's combustion procedures. The total carbohydrate content was assayed by the phenol/sulphuric acid method. The assessment of the fatty acids methyl esters (FAMEs) was determined through GC-MS. Characterization of elemental analysis was performed by ICP-AES. Different standard microbiological methods were applied for microorganisms. Statistics were performed using the non-parametric Mann-Whitney U test to assess significant differences between samples. RESULTS: Lipid contents (n = 3) were very low (1.6-2.3%), particularly in Palmaria palmata, and Chondrus crispus (1.6-1.7%). The protein content (n = 4) varied from 14.4% in P. palmata to 23.7% in Porphyra sp. Carbohydrates (n = 3) were the major constituent of most seaweeds (31-34%), except in Porphyra sp., with higher content in proteins than carbohydrates. Regarding the fatty acid content (n = 4), in general, saturated fatty acids (SFAs) were the most abundant followed by polyunsaturated fatty acids (PUFAs) and monounsaturated fatty acids (MUFAs). Among macro and trace elements (n = 3), Chondrus crispus shows the highest average content in Zn (71.1 mg ⋅ kg - 1 D.W.), Palmaria palmata the highest average content in K (124.8 g ⋅ kg - 1 D.W.), Porphyra sp. the highest average content in P (2.1 g ⋅ kg - 1 D.W.), and Ulva sp. the highest average content of Ca (5.5 g ⋅ kg - 1 D.W.), Mg (55.8 g ⋅ kg - 1 D.W.), and Fe (336.3 mg ⋅ kg - 1 D.W.). In general, Na and K were the most abundant elements among analysed seaweed. Additionally, the microbiological results (n = 4) comply with the Portuguese guidelines (subgroup 2D) on the application of general principles of food hygiene in ready-to-eat foods. CONCLUSIONS: Overall, the results highlight the potential of using these seaweeds as an alternative and sustainable source of elements and bioactive compounds to produce enriched food products with a beneficial potential for human nutrition.

Salinity shapes the stress responses and energy reserves of marine polychaetes exposed to warming: From molecular to functional phenotypes.

Estuarine systems are critical transition zones influenced by sea, land and freshwater. An array of human activities impacts these areas leading to multiple-stressor interactions. Temperature and salinity are among the most relevant drivers in estuaries, shaping species growth, reproduction and distribution. However, few studies provide an overview of cellular rewiring processes under multiple-stressor environments. Here, we tested how salinity could shape the response of ragworms Hediste diversicolor, an important bioindicator and commercial species, to elevated temperature. We exposed polychaetes to three temperatures for a month, simulating control, ocean warming and heatwave conditions (24, 27 and 30 °C, respectively) combined with two salinities (20 and 30). We quantified whole-organism performance (wet weight gain and survival), along with cellular stress response (CSR) and energy reserves of worms after 14 and 28 days of exposure. Significant three-way interactions between temperature, salinity and exposure time show the non-linearity of molecular responses. Worms at a salinity of 20 were more sensitive to warming than worms exposed to a salinity of 30. The combination of high temperature and low salinity can act synergistically to induce oxidative stress and macromolecular damage in worm tissues. This finding was supported by an induction of the CSR, with a concomitant decrease of energy reserves, pointing towards a metabolic compensation strategy. However, under a higher salinity (30), the need for a CSR upon thermal challenge was reduced and energy content increased with temperature, which suggests that environmental conditions were within the optimum range. Heatwaves striking low-salinity areas of estuaries can therefore negatively impact the cellular physiology of H. diversicolor, with greater metabolic costs. However, extreme stress levels were not reached as worms incremented wet weight and survival was high under all conditions tested. Our findings are important for the optimization of ragworm aquaculture and adaptive conservation strategies of estuarine systems.

Living in a multi-stressors environment: An integrated biomarker approach to assess the ecotoxicological response of meagre (Argyrosomus regius) to venlafaxine, warming and acidification.

Pharmaceuticals, such as the antidepressant venlafaxine (VFX), have been frequently detected in coastal waters and marine biota, and there is a growing body of evidence that these pollutants can be toxic to non-target marine biota, even at low concentrations. Alongside, climate change effects (e.g. warming and acidification) can also affect marine species' physiological fitness and, consequently, compromising their ability to cope with the presence of pollutants. Yet, information regarding interactive effects between pollutants and climate change-related stressors is still scarce. Within this context, the present study aims to assess the differential ecotoxicological responses (antioxidant activity, heat shock response, protein degradation, endocrine disruption and neurotoxicity) of juvenile fish (Argyrosomus regius) tissues (muscle, gills, liver and brain) exposed to VFX (via water or feed), as well as to the interactive effects of warming (ΔT °C = +5 °C) and acidification (ΔpCO2 ~ +1000 µatm, equivalent to ΔpH = -0.4 units), using an integrated multi-biomarker response (IBR) approach. Overall, results showed that VFX toxicity was strongly influenced by the uptake pathway, as well as by warming and acidification. More significant changes (e.g. increases surpassing 100% in lipid peroxidation, LPO, heat shock response protein content, HSP70/HSC70, and total ubiquitin content, Ub,) and higher IBR index values were observed when VFX exposure occurred via water (i.e. average IBR = 19, against 17 in VFX-feed treatment). The co-exposure to climate change-related stressors either enhanced (e.g. glutathione S-transferases activity (GST) in fish muscle was further increased by warming) or attenuated the changes elicited by VFX (e.g. vitellogenin, VTG, liver content increased with VFX feed exposure acting alone, but not when co-exposed with acidification). Yet, increased stress severity was observed when the three stressors acted simultaneously, particularly in fish exposed to VFX via water (i.e. average IBR = 21). Hence, the distinct fish tissues responses elicited by the different scenarios emphasized the relevance of performing multi-stressors ecotoxicological studies, as such approach enables a better estimation of the environmental hazards posed by pollutants in a changing ocean and, consequently, the development of strategies to mitigate them.

Seasonal changes in stress biomarkers of an exotic coastal species - Chaetopleura angulata (Polyplacophora) - Implications for biomonitoring.

Knowledge on baseline values of stress biomarkers in natural conditions is urgent due to the need of reference values for monitoring purposes. Here we assessed the cellular stress response of the chiton Chaetopleura angulata in situ. Biomarkers commonly used in environmental monitoring (heat shock protein 70kDa, total ubiquitin, catalase, glutathione-S-transferase, superoxide-dismutase, lipid peroxidation) were analyzed in the digestive system, gills and muscle of C. angulata, under spring and summer conditions in order to assess seasonal tissue-specific responses. Season had an effect on all targeted organs, especially affecting the digestive system which displayed clear seasonal clusters. The respective Integrated Biomarker Response (IBR) showed a 7.2-fold seasonal difference. Muscle and gills showed similar IBRs between seasons making them appropriate organs to monitor chemical pollution as they were less responsive to seasonal variation. The most stable biomarkers in these organs were ubiquitin and superoxide-dismutase thus being reliable for monitoring purposes.

Synergy of environmental variables alters the thermal window and heat shock response: an experimental test with the crab Pachygrapsus marmoratus.

The intertidal zone is an extremely variable habitat, imposing stressful conditions on its inhabiting communities. Tolerance towards extremes of temperature, salinity and pH are crucial in these habitats. Despite the vast literature on stress tolerance, few studies have focused on the synergistic effects of several variables on thermal tolerance and HSP70 (heat shock protein 70 kDa) levels. In this work, the crabs were exposed to three experimental conditions 1) thermal ramp at standard pH (8) and saline conditions (35‰) (named T), 2) thermal ramp at standard pH (8) and hyposaline conditions (15‰) (named T plus HypoS), and 3) thermal ramp at lower pH (7) and standard saline conditions (35‰) (named T plus A). Two physiological parameters (Critical Thermal Maximum - CTMax, and osmolality) and a stress biomarker (HSP70) were chosen for this analysis. These parameters were measured in all of the aforementioned conditions. CTMax for each set of conditions was reached by exposing the organisms to a rate of temperature increase of 1 °C h(-1) until loss of equilibrium. Haemolymph samples were taken every 2 °C to quantify HSP70 and osmolality. Results showed that CTMax did not differ between crabs solely exposed to T stress and crabs exposed to T plus HypoS stress. However, HSP70 production was impaired in T plus HypoS stress. When crabs were exposed to T plus A stress, they showed a significantly higher CTMax, suggesting that short-term exposure to acidified conditions may alter the thermal window of this species. Nevertheless, in T plus A conditions HSP70 production was impaired as well. Regarding osmolality it decreased according to temperature increase in all tested stress conditions. This study showed that the heat stress response is altered by the synergistic effect of variables. Physiological end-points (i.e. CTMax) may vary and the expression of stress proteins such as HSP70 may be impaired.