Extraction of Antioxidants from Brown Macroalgae Fucus spiralis.

In this study, different extraction methods and conditions were used for the extraction of antioxidants from brown macroalgae Fucus spiralis. The extraction methodologies used were ultrasound-assisted extraction (ultrasonic bath and ultrasonic probe), extraction with a vortex, extraction with an Ultra-Turrax® homogenizer, and high-pressure-assisted extraction. The extracts were analyzed for their total phenolic content (TPC) and their antioxidant activity, and evaluated through the 2,2-difenil-1-picrilhidrazil (DPPH) free radical scavenging method and ferric reducing antioxidant power (FRAP) assay. Ultrasonic probe-assisted extraction yielded the highest values of TPC (94.78-474.16 mg gallic acid equivalents/g extract). Regarding the antioxidant activity, vortex-assisted extraction gave the best DPPH results (IC50 1.89-16 µg/mL), while the highest FRAP results were obtained using the Ultra-Turrax® homogenizer (502.16-1188.81 µmol ascorbic acid equivalents/g extract). For each extraction method, response surface methodology was used to analyze the influence of the experimental conditions "extraction time" (t), "biomass/solvent ratio" (R), "solvent" (S, water % in water/ethanol mixture), and "pressure" (P) on TPC, DPPH, and FRAP of the F. spiralis extracts. In general, higher TPC content and higher antioxidant capacity (lower IC50 and higher FRAP) were obtained with higher R, t, and P, and lower S (higher ethanol %). The model regarding the combined effects of independent variables t, R, and S on the FRAP response values for vortex-assisted extractions best fitted the experimental data (R2 0.957), with optimal extraction conditions of t = 300 s, R = 50 g, and S = 25%.

Neurosyphilis-Induced Folie À Deux: A Case of Prolonged Psychosis.

Neurosyphilis presentations can include psychiatric symptoms such as psychosis, mania, depression, or changes in personality. Therefore, it can lead to a misdiagnosis with psychiatric disorders. The diagnosis is particularly difficult in a subset of patients whose psychotic symptoms are developed as a manifestation of the disease later in life. With the aim to highlight the importance of considering neurosyphilis in the differential diagnosis of late psychosis, it is presented the clinical case of a folie à deux from a Portuguese 69-year-old man, who presented symptoms of psychosis in the emergency room, ultimately diagnosed with neurosyphilis. A selective review of the literature was made using the Pubmed database, with "neurosyphilis", "psychosis", "syphilis", and "folie à deux" as keywords. Patient consent was obtained for the use of clinical data. Neurosyphilis represents the most severe consequence of an untreated syphilis infection and there is a need in the scientific community to establish tools to enhance the precision of diagnosis and treatment.

Morphologic and Genetic Analysis of Synhimantus (Synhimantus) laticeps from a Long-Eared Owl (Asio otus).

The long-eared owl (Asio otus) is a medium-sized owl species that is well-distributed in almost all of the territories in Portugal. Nematodes were found in the oral cavity of a long-eared owl (A. otus) admitted to CRASSA (Wildlife Rehabilitation Centre of Santo André). During a physical exam and stabilization of the bird, five nematodes were collected. The worms were examined and measured under light microscopy, and photos were taken. After a morphological analysis was conducted, all the nematodes (five females) were identified as Synhimantus (Synhimantus) laticeps. Two specimens were subjected to molecular analysis, which confirmed the result. This study provides a combined morphological and genetic approach to S. laticeps. To the authors' best knowledge, this is the first report including genetic sequencing of S. laticeps in a long-eared owl (A. otus) from Portugal.

Diastolic Function and Left Atrial Strain in Young Patients with History of Aortic Coarctation Repair.

Aortic coarctation (AoCo) leads to long-term sequelae that may impair heart function. Data regarding new echocardiographic function parameters such as atrial strain, in affected patients, are scarce. This study aims to describe these parameters in AoCo patients and define their association with severity measures. 53 AoCo patients and 31 healthy controls, aged 12-40 years, were evaluated. Effectively corrected AoCo (cAoCo) was defined as aortic trans-isthmic corrected Doppler gradient (Dgrad) ≤ 20 mmHg (n = 36), and recoarctation (rAoCo) as Dgrad > 20 mmHg (n = 17). Dependent variables were: E/E'; atrial reservoir strain (Ares); and atrial conduit strain (Acd). T-tests/Mann-Whitney U tests were used to compare these among groups. Multivariable regression was used to test correlation with systolic blood pressure (SBP), indexed LV mass (ILVM), Dgrad, and the ratio between the narrowest diameter of aortic arch and aorta at diaphragm level (Aoratio). In cAoCo and rAoCo patients, E/E' was higher (p < 0.001), Ares, and Acd were lower (p < 0.001 for both) comparing with controls. Acd was higher in cAoCo than rAoCo (p = 0.045). Higher Ares was associated with higher Aoratio (p = 0.002), and lower Acd with higher Dgrad (0.014). EF and GLS were not different among groups. Young patients with effectively corrected aortic coarctation have persistent changes in diastolic function parameters (E/E' and atrial strain), and these are affected by anatomical sequelae. These patients' physiology is closer to patients with recoarctation, than to healthy individuals. This provides rationale for a stronger prevention, and treatment, of arterial dysfunction and high left ventricular afterload in these patients.

Reproduction of the Blue Jack Mackerel, Trachurus picturatus, in Western Portugal: Microscopic Gonad Analysis Reveals Indeterminate Fecundity and Skipped Spawning Patterns.

Blue jack mackerel, Trachurus picturatus, is the fifth most landed fish species in mainland Portugal, but information on its reproductive biology is scarce. From September 2018 to August 2019, 626 specimens were collected from commercial vessels to clarify the reproductive strategy of the T. picturatus population off the west coast of Portugal. The proportion and length range of males and females were similar. Only three of the specimens collected were categorized as immature, indicating that the fish caught in the fishery are primarily mature. The spawning season lasted from late January until the end of March, with gonadosomatic indices being similar for males and females. Fecundity was indeterminate, and estimated batch fecundity ranged between 6,798 (at 25.4 cm TL) and 302,358 oocytes (at 33.8 cm TL). The low number of females showing direct evidence of imminent or recent spawning suggests a low number of spawning events. In addition, 12.7% of females were considered non-reproductive due to ovary abnormalities including parasitic infection by Kudoa species, atretic structures and skipped spawning events. This study highlights the importance of accounting for skipped spawning events and ovary abnormalities in the management of species fisheries.

Seasonal study of the nutritional composition of unexploited and low commercial value fish species from the Portuguese coast.

Target species diversification is essential for fisheries sustainability and fish market revitalization. Fish discards are a widely recognized problem resulting from fisheries worldwide, and are of major concern for all sector players, from administrations, to fishermen, and scientists. However, non-target species are seldom studied, and information on nutritional profiles and seasonal changes in nutritional properties is generally lacking. This study assessed the seasonal nutritional composition of two unexploited (Serranus cabrilla, Capros aper) and three low commercial value fish species (Trachurus picturatus, Spondyliosoma cantharus, and Trigla lyra), captured on the Portuguese coast over 1 year. Significant seasonal variations were observed in the nutritional composition of all the species studied. Moisture and ash contents varied from 70% to 81% and from 5% to 13%, respectively. The maximum fat contents were 5% for C. aper and 4% for T. picturatus, allowing to classify all studied fishes as lean. The highest protein contents were recorded for C. aper (25%) and S. cantharus (20%). The unexploited and low commercial value fish species studied were shown to be good fat and protein sources, comparable to commonly consumed species, such as cod and salmon, having a great potential to become commonly consumed fish in Portugal.

Using Aerial Thermal Imagery to Evaluate Water Status in Vitis vinifera cv. Loureiro.

The crop water stress index (CWSI) is a widely used analytical tool based on portable thermography. This method can be useful in replacing the traditional stem water potential method obtained with a Scholander chamber (PMS Model 600) because the latter is not feasible for large-scale studies due to the time involved and the fact that it is invasive and can cause damage to the plant. The present work had three objectives: (i) to understand if CWSI estimated using an aerial sensor can estimate the water status of the plant; (ii) to compare CWSI from aerial-thermographic and portable thermal cameras with stem water potential; (iii) to estimate the capacity of an unmanned aerial vehicle (UAV) to calculate and spatialize CWSI. Monitoring of CWSI (CWSIP) using a portable device was performed directly in the canopy, by measuring reference temperatures (Tdry, Twet, and canopy temperature (Tc)). Aerial CWSI calculation was performed using two models: (i) a simplified CWSI model (CWSIS), where the Tdry and Twet were estimated as the average of 1% of the extreme temperature, and (ii) an air temperature model (CWSITair) where air temperatures (Tair + 7 °C) were recorded as Tdry and in the Twet, considering the average of the lowest 33% of histogram values. In these two models, the Tc value corresponded to the temperature value in each pixel of the aerial thermal image. The results show that it was possible to estimate CWSI by calculating canopy temperatures and spatializing CWSI using aerial thermography. Of the two models, it was found that for CWSITair, CWSIS (R2 = 0.55) evaluated crop water stress better than stem water potential. The CWSIS had good correlation compared with the portable sensor (R2 = 0.58), and its application in field measurements is possible.

Annual variations in the mineral element content of five fish species from the Portuguese coast.

The most consumed fish species in Portugal do not correspond to the most caught fish species on the Portuguese coast. From the point of view of ocean sustainability and economics, it is important to study unexploited or underexploited fish species that have the potential to be introduced into the food market. This potential can be reflected by their nutritional value, where mineral elements are often overlooked despite their importance in human health. Therefore, it is important to study the elemental concentration of those fish species, which should also include seasonal variations of essential mineral elements such as Ca, P, Mg, Na, K, Fe, Cu, Mn, Zn, and I, and non-essential elements such as As, Ni, Cd, and Pb. This work is part of a project that studied five fish species with the potential to be brought to the market: three species with low commercial value (Trachurus picturatus, Spondyliosoma cantharus and Trigla lyra) and two species with no commercial value (Serranus cabrilla and Capros aper), caught over a year in the coast of Portugal. Mineral elements were quantified by ICP-OES (ICP-MS for I). The results showed that the studied fish species present significant elemental concentrations, similar to those found in the most caught fish species in Portugal. Trachurus picturatus was found to be a source of Fe, and all species are sources of I. Overall, Spondyliosoma cantharus proved to be the most element-rich species. Sensorially, Cu was related to "metallic odour" and Fe to "metallic flavour", and I was related to "seaweed odour" and "seawater flavour". Additionally, toxic elements such as Cd, Pb, Ni and As were not detected. The corresponding Hazard Quotients (HQ) show the safety of the consumption of these fish species concerning the content of potentially toxic elements.

Reproductive Biology of the Sea Cucumber Holothuria mammata (Echinodermata: Holothuroidea).

Holothuria mammata is one of the most valuable species of sea cucumber, as well as one of the main target species harvested in the Mediterranean and NE-Atlantic regions. This study aims to describe the reproductive cycle of H. mammata in a coastal area of southwest Portugal. Monthly samplings were carried out for 19 months, with the concomitant collection of environmental data and biometric data. H. mammata had a sex ratio of 1:1.2 (male:female) and a size at first maturity of 142 mm for males and 167 mm for females. The gonadosomatic index (GI) peaked between April and May for both sexes. Gonad development started when days had a shorter photoperiod (9 to 13 h of sunlight) and lower seawater temperature (<15 °C), and spawning occurred later, with longer photoperiod (13 to 15 h of sunlight) and higher seawater temperature (>15 °C) and chlorophyll-a concentrations. The development of new studies to increase the biological and ecological knowledge of the populations of H. mammata is essential to create conditions for the domestication of broodstock in captivity, to allow the development of fishing regulations based on informed decisions and to create precise measures for the conservation of ecosystems.

Study of the Seasonal Variations of the Fatty Acid Profiles of Selected Macroalgae.

Due to the high consumption of fat-rich processed foods, efforts are being done to reduce their saturated fat (SFA) contents and replace it with polyunsaturated fatty acids (PUFA), creating a necessity to find alternative PUFA sources. Macroalgae, being a promising natural source of healthy food, may be such an alternative. The fatty acid (FA) profile of Fucus spiralis, Bifurcaria bifurcata, Ulva lactuca, and Saccorhiza polyschides were determined through direct transesterification and their seasonal variation was studied. F. spiralis showed the highest FA content overall, B. bifurcata presented the higher PUFA amounts, and U. lactuca and S. polyschides the higher SFA. The production of FA was shown to be influenced by the seasons. Spring and summer seemed to induce the FA production in F. spiralis and B. bifurcata while in U. lactuca the same was verified in winter. U. lactuca presented a ω6/ω3 ratio between 0.59 and 1.38 while B. bifurcata presented a ratio around 1.31. The study on the seasonal variations of the macroalgal FA profile can be helpful to understand the best season to yield FA of interest, such as ALA, EPA, and DHA. It may also provide valuable information on the best culturing conditions for the production of desired FAs.

Seasonal Evaluation of Phlorotannin-Enriched Extracts from Brown Macroalgae Fucus spiralis.

Fucus spiralis that was collected in the four seasons was submitted to an extraction with ethanol:water (crude extracts Et80), followed by a liquid-liquid fractionation with organic solvents (fraction He from n-hexane; aqueous fractions AQ1, AQ2, AQ3 and AQ4; ethyl acetate fraction EA), with the aim of obtaining phlorotannin-enriched extracts. All the extracts (Et80, He, AQ1, AQ2, AQ3, AQ4 and EA) that were obtained for the F. spiralis of the four seasons were evaluated for their antioxidant capacity and total phenolic compounds. The summer extracts presented the highest contents in polyphenols (TPC), as well as the highest ferric reducing antioxidant power (FRAP), when compared to the samples from the other seasons. The reductive percentage of the DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) compound was similar between the seasons. For all the seasons, the EA extract showed the highest polyphenol content (TPC), and the highest antioxidant capacity (highest ferric reducing power (FRAP) and lowest concentration needed to reduce 50% of the DPPH compound), which is in agreement with a phlorotannin-enriched fraction. This study revealed that the polyphenol content and antioxidant power of the F. spiralis extracts are influenced by the time of harvest, as well as by the solvents used for their extraction.

Enhancing knowledge on low-value fishing species: the distinct reproductive strategy of two gurnard species.

The depletion and overexploitation of several fish stock demands for a valorisation of non-target and discarded species. Nonetheless, such species are often poorly studied, and information on their biological parameters must be gathered for effective population management. For 1 year, the reproductive strategy of the piper gurnard Trigla lyra and the red gurnard Chelidonichthys cuculus was studied by monthly samples obtained from commercial boats operating on western Portuguese coast. Both species showed a biased sex ratio towards females, especially for larger length classes. Length at first maturity could be estimated only for red gurnard (22.1 and 19.9 cm for females and males, respectively) because all piper gurnard individuals caught were mature. Piper gurnard showed determinate fecundity and a short spawning season, from November to February with a peak in January, whereas red gurnard showed indeterminate fecundity and a wide spawning season, from late December to May. The relative annual fecundity estimated for red gurnard (1893 ± 728 oocytes × g-1 eviscerated weight [EW]) was higher than the one estimated for piper gurnard (1018 ± 250 oocytes×g-1 EW). Although important information for understanding the species dynamics is presented in this study, additional information on other life-history parameters and of species landings is required.

Recent Efforts to Recover Armeria berlengensis, an Endemic Species from Berlengas Archipelago, Portugal.

Berlengas archipelago is a UNESCO world heritage site and the only location where Armeria berlengensis is found. This species faces various threats, namely, human disturbance, the presence of Carpobrotus edulis, yellow-legged gull, common-rabbit, and black-rat populations. Thus, exclusion areas were installed, which blocked the access of most Gulls, aiming to promote the recovery of A. berlengensis. Additionally, rabbits and rats were removed from the island. After six years of surveys, there has been an increase in the number of individuals of A. berlengensis in the exclusion areas, and a clear shift in the size structure of the A. berlengensis population. Significant changes in the height and diameter of the individuals were also noted. These findings indicate that the population of A. berlengensis is changing and becoming a healthier population. Principal component analysis results show a straightforward dissimilarity between the areas with A. berlengensis and those without the species and allowed the clustering of two groups: the rupicolous species and the nitrophilous species. A. berlengensis produces few seeds (seed set 3.4%), which raises concern regarding the long-term survival of the species. Thus, further conservation efforts must be implemented, such as the control of invasive species, gulls, and ruderals, to allow for the recovery of A. berlengensis.

Global warming overrides physiological anti-predatory mechanisms in intertidal rock pool fish Gobius paganellus.

In nature, a multitude of factors influences the fitness of an organism at a given time, which makes single stressor assessments far from ecologically relevant scenarios. This study focused on the effects of water temperature and predation stress on the metabolism and body mass gain of a common intertidal rock pool fish, Gobius paganellus, addressing the following hypotheses: (1) the energy metabolism of G. paganellus under predation stress is reduced; (2) G. paganellus shows thermal compensation under heat stress; and (3) thermal stress is the dominant stressor that may override predation stress responses. Individuals were exposed to simulated predation stress and temperature increase from 20 °C to 29 °C, and both stressors combined. Physiological effects were addressed using biochemical biomarkers related with energy metabolism (isocitrate dehydrogenase, lactate dehydrogenase, energy available, energy consumption rates), oxidative stress (superoxide dismutase, catalase, DNA damage, lipid peroxidation), and biotransformation (glutathione-S-transferase). The results of this study revealed that predation stress reduced the cellular metabolism of G. paganellus, and enhanced storage of protein reserves. As hypothesized, hyperthermia decreased the aerobic mitochondrial metabolism, indicating thermal compensation mechanisms to resist against unfavourable temperatures. Hyperthermia was the dominant stressor overriding the physiological responses to predation stress. Both stressors combined might further have synergistically activated detoxification pathways, even though not strong enough to counteract lipid peroxidation and DNA damage completely. The synergistic effect of combined thermal and predation stress thus may not only increase the risk of being preyed upon, but also may indicate extra energy trade-off for the basal metabolism, which in turn may have ecologically relevant consequences for general body functions such as somatic growth and reproduction. The present findings clearly underline the ecological importance of multi-stressor assessments to provide a better and holistic picture of physiological responses towards more realistic evaluations of climate change consequences for intertidal populations.

Seasonal Sensory Evaluation of Low Commercial Value or Unexploited Fish Species from the Portuguese Coast.

Overfishing is increasing over time, and according to FAO (Food and Agriculture Organization), about one third of the world's fish stocks are now overfished. Thus, diversifying the target species is essential for fisheries sustainability contributing to improve resource-efficient processes. Non-target species can be valuable resources for the development of new food products. However, those species are scarcely studied, and it is of high importance to trace their seasonal sensory profile as a first step towards their valorisation. Therefore, in this study, seasonal influence on sensory properties of five low commercial value or unexploited fish species, namely Trachurus picturatus (blue jack mackerel), Spondyliosoma cantharus (black seabream), Trigla lyra (piper gurnard), Serranus cabrilla (comber) and Capros aper (boarfish), was assessed in order to identify the most favourable season for catching each species. Fish samples were assessed by a panel of 16 semi-trained assessors for sensory attributes previously identified. The evaluation takes place every 2 months. Statistical differences were reported between attributes and seasons for all species, except for T. lyra, which did not present any difference in its sensory attributes throughout the year.

Optimization of Extraction Conditions for Gracilaria gracilis Extracts and Their Antioxidative Stability as Part of Microfiber Food Coating Additives.

Incorporation of antioxidant agents in edible films and packages often relies in the usage of essential oils and other concentrated hydrophobic liquids, with reliable increases in antimicrobial and antioxidant activities of the overall composite, but with less desirable synthetic sources and extraction methods. Hydroethanolic extracts of commercially-available red macroalgae Gracilaria gracilis were evaluated for their antioxidant potential and phenolic content, as part of the selection of algal biomass for the enrichment of thermoplastic film coatings. The extracts were obtained through use of solid-liquid extractions, over which yield, DPPH radical reduction capacity, total phenolic content, and FRAP activity assays were measured. Solid-to-liquid ratio, extraction time, and ethanol percentages were selected as independent variables, and response surface methodology (RSM) was then used to estimate the effect of each extraction condition on the tested bioactivities. These extracts were electrospun into polypropylene films and the antioxidant activity of these coatings was measured. Similar bioactivities were measured for both 100% ethanolic and aqueous extracts, revealing high viability in the application of both for antioxidant coating purposes, though activity losses as a result of the electrospinning process were above 60% in all cases.

Stability, kinetics, and application study of phycobiliprotein pigments extracted from red algae Gracilaria gracilis.

Phycobiliprotein (PBP) pigments were extracted from red algae Gracilaria gracilis through maceration in phosphate buffer using previously optimized conditions. The stability of PBPs in the extracts was assessed by monitoring the extracts at different pHs and temperatures for 10 days. Since phycoerythrin (PE) is the main PBP present in G. gracilis, PE content was spectroscopically determined and used as a response factor. Kinetic modeling was used to describe PE degradation under different ranges of T and pH. The pigment extracts presented higher stability at pH 6.9 and -20 °C. PE was semipurified by precipitation with ammonium sulphate 65% followed by dialysis against water until a purity index of 0.7. The pigment was successfully applied as colorant in pancakes and yogurts with a pigment concentration of 0.15%. This study highlights the potential of PE pigments extracted from G. gracilis for applications in food products. PRACTICAL APPLICATION: Phycobiliprotein pigments were extracted from red algae Gracilaria gracilis through maceration in phosphate buffer. The stability of the pigment was evaluated at different pHs and temperatures, presenting higher stability at neutral pH and low temperatures. The pigment was successfully applied as colorant in pancakes and yogurts with a low pigment concentration. This study highlights the potential of phycobiliprotein pigments extracted from G. gracilis for applications in food products.

The Influence of Atrial Strain on Functional Capacity in Patients with the Fontan Circulation.

Speckle-tracking echocardiography has been used to assess atrial function. This analysis is feasible in univentricular hearts. The aim of this study was to characterize the relationship between atrial strain and functional capacity in the Fontan circulation. Apical four-chamber echocardiographic loops of 39 Fontan patients were reviewed. The dominant atrium was assessed for active (εact), conduit (εcon), and reservoir (εres) strain and εact/εres ratio. Cardiopulmonary exercise test was performed on the same day and oxygen uptake (VO2) at ventilatory threshold (VT) and peak VO2 were chosen as the dependent variables. Statistical analysis was performed using SPSS® version 23. Unpaired t test was used for binomial and continuous variable correlation; single and multivariable linear regression were used for continuous variable correlation. Statistical significance was defined as p value < 0.05. VO2 at VT as a percentage of predicted VO2 was 36.8% (SD 10.7). Peak VO2 was 64.7% (SD 18.9) of the predicted value. In univariate analysis, both were associated with age, atrioventricular regurgitation, ejection fraction, εres, εcon, and εact/εres. In multivariate regression, higher VO2 at VT and peak VO2 were associated with younger age (p = 0.003 and p = 0.001, respectively) and higher εcon (p = 0.026 and p = 0.020). Evaluation of heart function is difficult in the Fontan circulation, hindered by complex ventricular morphology and lack of normative data. VO2 provides a good surrogate. Atrial strain parameters are compromised in these patients and associated with VO2. Therefore, whenever possible, atrial strain should be measured as it may provide a new method of risk stratification.

Seaweeds' neuroprotective potential set in vitro on a human cellular stress model.

Neurodegenerative diseases, such as Parkinson's disease, represent a biggest challenge for medicine, imposing high social and economic impacts. As a result, it is of utmost importance to develop new therapeutic strategies. The present work evaluated the neuroprotective potential of seaweeds extracts on an in vitro dopamine (DA)-induced neurotoxicity cellular model. The neuroprotective effects on SH-SY5Y cells' viability were estimated by the MTT assay. Changes in mitochondrial membrane potential (MMP), caspase-3 activity, and hydrogen peroxide (H2O2) production were determined. DA (30-3000 µM; 24 h) treatment decreased SH-SY5Y cells' viability in concentration and time-dependent manner, increasing the H2O2 production, MMP depolarization, and caspase-3 activity. On the other hand, DA (1000 µM; 24 h) toxicity was reduced (10-15%) with Sargassum muticum and Codium tomentosum extracts (1000 µg/mL; 24 h). The highest neuroprotective activity was exhibited by a methanolic extract obtained from Saccorhiza polyschides, which completely blunted DA effects. Results show that the marine seaweed S. polyschides contain substances with high neuroprotective potential against the toxicity induced by DA, exhibiting anti-apoptotic effects associated with both mitochondrial protection and caspase-3 inhibition. S. polyschides reveals, therefore, to be an excellent source of bioactive molecules, for new drugs development aiming PD therapeutics.

Impact of Aqueous Extract of Arbutus unedo Fruits on Limpets (Patella spp.) Pâté during Storage: Proximate Composition, Physicochemical Quality, Oxidative Stability, and Microbial Development.

Limpets are molluscs widely used in food diet and much appreciated in many regions. The consumption of fishery products rich in polyunsaturated fatty acids has been increasing through filleted products and restructured products. Since food oxidation is the major cause of nutritional quality deterioration in fish products, the interest in the replacement of synthetic antioxidants with natural sources, namely in the preparation of restructured animal products such as burgers, sausages and pâtés, has been increasing. Phenolic compounds from fruits and vegetables have recognised antioxidant properties and are therefore currently considered as good alternatives to synthetic antioxidants in the food industry. In this study, the effects of the extracts of Arbutus unedo fruits, at two concentration levels (3% and 6%), on proximate composition, physicochemical properties, oxidative stability and safety of limpets pâté, during 90 days at refrigerated storage, were investigated. After processing, the addition of 3% and 6% of A. unedo extracts into limpets pâté contributed to an increase of 18% and 36% in the total phenolic content and 5% and 36% in the antioxidant capacity, respectively. During storage, the enriched limpets pâté with A. unedo fruit extracts at 6% was more efficient as an enhancer of oxidative stability, with 34% inhibition of lipid oxidation, highlighting the potential use of A. unedo fruits as a functional ingredient in the fish industry. Overall, the limpets pâté with 6% of A. unedo fruit extracts proved to be more efficient regarding microbial control, and had the lowest changes in the quality parameters such as in colour, texture and pH during 90 days at refrigerated storage.