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The impacts of climate change-related stressors are becoming more noticeable in the ocean, particularly in coastal marine ecosystems. Yet limited information still exists on the physiological state and ecological resilience of marine fish species, especially during their early life stages (i.e., larvae and juveniles). The present study investigated the effects of chronic exposure to seawater warming (OW; ΔT = +4 °C) and acidification (OA; ΔpH = −0.3 pH units, equivalent to pCO2~1000 µatm), acting alone or combined (OWA), on juvenile zebra seabream (Diplodus cervinus) physiological resilience, considering distinct levels of biological organization (i.e., biochemical, cell, organ and individual levels). After 60 days of exposure, both stressors, in isolation or combination, significantly decreased specific growth rate (−11% in OW, −42% in OA and −49% in OWA) and leukocyte counts (from −29% in OA and OWA up to −37% in OW) in relation to the control treatment. In addition, a decreased Fulton’s condition index (K) was observed under warming and acidification in combination (−35% in OWA). At the cell level, OW, OA and OWA triggered different biomarker responses in D. cervinus (i.e., up-regulation, down-regulation, or absence of significant effect). In general, the results are suggestive of an antagonistic effect when warming and acidification are combined. OWA yielded the highest integrated biomarker response (IBR) index value in the whole organism, muscle, brain and gills of D. cervinus juveniles, therefore suggesting that the effects of these stressors are more severe when they act together. The distinct patterns observed in each stress scenario highlight the importance of carrying out further studies adjusted to the specificities of different regions, i.e., accounting not only for the type and degree of severity of environmental stressors already felt and/or projected for that specific area, but also the physiological plasticity of species that inhabit a particular ecosystem. The gathered knowledge will allow one to determine the vulnerability of particular marine species and geographic areas and, most importantly, to draw up effective and tailor-made conservation strategies to overcome climate change impacts.

Extreme weather events, like marine heatwaves (MHWs), are becoming more frequent and severe due to climate change, posing several challenges to marine ecosystems and their services. As disease outbreaks are often prompted by these acute phenomena, it is essential to develop eco-innovative strategies that can efficiently improve farmed fish resilience, especially under sub-optimal rearing conditions, thereby ensuring a sustainable aquaculture production.This study aimed to unveil farmed juvenile white seabream (Diplodus sargus, 28.50 ± 1.10 g weight, n = 150) immune and antioxidant responses under a category II MHW in the Mediterranean Sea (+4 °C, 8 days of temperature increase plus 15 days of plateau at the peak temperature) and to investigate whether a 30 days period of prophylactic biofortification with Asparagopsis taxiformis (1.5 %, 3 % and 6 %) enhanced fish resilience to these extreme events. Several biomarkers from different organization levels (individual, cellular, biochemical and molecular) were assessed upon 30 days of biofortification (T30), exposure (after 8 days of temperature increase + 15 days at peak temperature, T53) and recovery (8 days of temperature decrease, T61) from the MHW.Results showed that MHW negatively affected the fish physiological status and overall well-being, decreasing specific growth rate (SGR) and haematocrit (Ht) and increasing erythrocyte nuclear abnormalities (ENAs) and lipid peroxidation (LPO). These adverse effects were alleviated through biofortification with A. taxiformis. Seaweed inclusion at 1.5 % was the most effective dose to minimize the severity of MHW effects, significantly improving immune responses of D. sargus (i.e. increased levels of immunoglobulin M, peroxidase activity and lysozyme expression) and modulating antioxidant responses (i.e. decreased LPO, catalase and glutathione S-transferase activity). These findings confirm that A. taxiformis is a functional ingredient of added value to the aquaculture industry, as its inclusion in marine fish diets can beneficially modulate fish immunity and resilience under optimal and adverse rearing conditions.

Coastal habitats are increasingly threatened by multiple anthropogenic-related activities, which include ocean warming and loss of structural habitat complexity. These two pressures have the potential to severely affect the structure and function of marine biodiversity. Early life stages of many fish species recruit to coastal habitats at the end of their pelagic phase, benefiting from access to food, shelter and protection. However, changes in temperature have been shown to influence ecologically relevant behaviours in post-settlement stage fish, and the loss of structural habitat complexity has been related to low recruitment and deleterious behaviours of fish in coastal habitats. Here, we evaluated the individual and interactive effects of prolonged exposure to increasing temperature and changed structural habitat complexity on routine swimming speed and escape response of post-settlement white seabream, Diplodus sargus (Linnaeus, 1758). Fish were reared under different temperatures (control 19 °C; high 22 °C) and structural habitat complexity (low and high) scenarios, in a cross-experimental design, and the routine swimming and escape responses were analyzed after 6 weeks of exposure. Change in temperature did not induce alterations at the behavioural level, but loss of structural habitat complexity increased speed and distance travelled during routine swimming, and responsiveness to a stimulus during the escape response behaviour. The interaction of the two factors did not influence performance. Determining how species are affected by changes in their environment, and the mechanisms that underlie these changes, will be critical to understanding the fish recruitment and populations’ fitness and survival.

The increasing frequency and duration of marine heatwaves (MHWs) due to climate change pose severe threats to aquaculture, causing drastic physiological and growth impairments in farmed fish, undermining their resilience against additional environmental pressures. To ensure sustainable production that meets the global seafood demand and animal welfare standards, cost-effective and eco-friendly strategies are urgently needed. This study explored the efficacy of the red macroalga Asparagopsis taxiformis on juvenile white seabream Diplodus sargus reared under optimal conditions and upon exposure to a MHW. Fish were fed with four experimental diets (0%, 1.5%, 3% or 6% of dried powdered A. taxiformis) for a prophylactic period of 30 days (T30) and subsequently exposed to a Mediterranean category II MHW for 15 days (T53). Biometric data and samples were collected at T30, T53 and T61 (8 days post-MHW recovery), to assess performance indicators, biomarker responses and histopathological alterations. Results showed that A. taxiformis supplementation improved catalase and glutathione S-transferase activities and reduced lipid peroxidation promoted by the MHW, particularly in fish biofortified with 1.5% inclusion level. No histopathological alterations were observed after 30 days. Additionally, fish biofortified with 1.5% A. taxiformis exhibited increased citrate synthase activity and fish supplemented with 1.5% and 3% showed improved digestive enzyme activities (e.g., pepsin and trypsin activities). Overall, the present findings pointed to 1.5% inclusion as the optimal dosage for aquafeeds biofortification with A. taxiformis, and confirmed that this seaweed species is a promising cost-effective ingredient with functional properties and great potential for usage in a climate-smart context.

The present work aims to evaluate the macroinvertebrate community associated with macroalgae in earthen pond systems to better understand their potential in detritus recycling and as an accessory production. Sampling took place on the settling pond of an aquaculture research station, where macroalgae permanently occurred at high densities. The results suggest differentiation between seasons but not between sites within the settling pond. Seasonal variation was observable in terms of macroinvertebrate density, biomass, and diversity. Two non-indigenous species of invertebrates were found, the crustaceans Grandidierella japonica and Paracerceis sculpta Amphipods were the most abundant group, and their high nutritional value can be exploited. Detritus and the epiphyte layer are the main food items for the invertebrates, reinforcing the advantages of these organisms being present to enhance the recycling of excess detritus and to transfer organic matter to upper trophic levels. These species, naturally present in aquaculture facilities, can improve the water quality and increase the variability of food nutrients for reared species.

The increasing demand for healthy aquatic products has contributed to the rapid growth of aquaculture. Despite the many progresses, over the years, in aquaculture production, animal welfare and reduction of the stress induced by captivity remain a challenge in this sector, and overcoming it is fundamental to improve animal growth performance and resilience to environmental threats (e.g., disease outbreaks and/or climate change impacts). To address this challenge, aquafeeds biofortified with natural ingredients with functional properties, such as macroalgae, have emerged as a promising strategy to enhance fish immunity. The present study aimed at evaluating the potential of Laminaria digitata to improve juvenile gilthead seabream (Sparus aurata) physiological status and resilience, through the modulation of immunomodulatory and antioxidant responses. To explore this hypothesis, four independent treatments were carried out in which fish were fed with different experimental aquafeeds: control feed and biofortified feeds with 1.5%, 3% and 6% of whole L. digitata (incorporated as a dried and powdered ingredient). Fish (n=9) were collected from each treatment upon 30 and 60 days of biofortification in order to assess animal growth and condition indexes, haematological parameters, innate humoral parameters (antiprotease (AP) activity, peroxidase (POD) activity and immunoglobulin M (IgM)) and oxidative stress biomarkers (lipid peroxidation (LPO), catalase (CAT) activity, glutathione S-transferase (GST) activity, superoxide dismutase (SOD) activity and total antioxidant capacity (T-AOC)). Significant differences (α = 0.05) between treatments in biomarker responses were tested through a nested-design ANOVA, followed by Tukey HSD multiple comparisons. Pearson correlations between the different endpoints were also explored. Results indicate that a dietary inclusion of 1.5% L. digitata is the most effective biofortification dose, significantly improving antioxidant responses of S. aurata, by increasing levels of CAT, GST and T-AOC, along with the reduction in LPO levels. Additionally, the inclusion of 1.5% L. digitata enhances immunity by increasing the levels of IgM and POD, while maintaining the growth performance and condition indices of the fish. These findings underscore the potential utilization of L. digitata as a functional feed ingredient in aquaculture with relevant immune and antioxidant attributes.

Understanding and controlling reproductive cycles and gonad maturation in cultivated species are crucial in aquaculture. The sea urchin Paracentrotus lividus, known for its edible gonads, requires careful maturation control for both reproduction and commercialization. This study explores the impact of temperature, photoperiod, and diet on gonad development to enhance year-round aquaculture practices. Using two independent cultivation systems, we manipulated environmental conditions to mimic different seasons. Sea urchins were exposed to natural or manipulated temperature and photoperiod conditions and fed either natural (Ulva spp.) or formulated diets. The gonadosomatic index (GI) and histological analysis were used to assess gonad development. The results revealed a clear correlation between environmental conditions, diet, and gonad maturation. Manipulated conditions accelerated maturation, with sea urchins showing advanced stages compared to natural conditions. Furthermore, sea urchins fed formulated diets exhibited higher GI values, indicating enhanced maturation. Histological analysis confirmed accelerated maturation, particularly in females. This study underscores the feasibility of controlling gonad maturation through environmental manipulation and diet, enabling year-round marketable gonad production. Providing formulated diets rich in polyunsaturated fatty acids, notably docosahexaenoic acid (DHA), enhances the commercial value of sea urchins. These findings optimize aquaculture practices for P. lividus, highlighting its adaptability to the maximal production of gonads throughout the year.

There are a high number of stressors present in aquaculture that can affect fish welfare and quality. One way of mitigating stress response is by increasing dietary tryptophan. In this study, three diets containing 0.5% (Tript1), 0.6% (Tript2), and 0.8% (Tript3) of tryptophan were tested in 32 g juvenile meagre for 56 days. At the end of the trial, survival, growth, and proximate composition were similar between treatments. Significant differences were found in the plasma parameters before and after a stress test consisting of 30 s of air exposure. Blood glucose levels were higher in the post-stress for all treatments (e.g., 63.9 and 76.7 mg/dL for Tript1 before and after the stress test), and the hemoglobin values were lower in the post-stress of Tript1 (1.9 g/dL compared to 3.0 and 2.4 g/dL for Tript2 and Tript3, respectively). In terms of behavior, three tests were carried out (novel tank diving and shoaling assays, and lateralization test), but no significant differences were found, except for the number of freezing episodes during the anxiety test (1.4 for Tript3 compared to 3.5 and 4.2 for the other treatments). This study suggests that supplementation with dietary tryptophan, particularly in higher dosage (0.8%), can reduce anxiety-like behavior in meagre exposure to acute stress (novel tank). Although the remaining results showed mild effects, they provide some clues as to the potential of this amino acid as a stress mitigator in aquaculture.

Dietary supplementation with Omega-3 fatty acids seems to promote skeletal health. Therefore, their consumption at imbalanced or excessive levels has offered less beneficial or even prejudicial effects. Fish produced in aquaculture regimes are prone to develop abnormal skeletons. Although larval cultures are usually fed with diets supplemented with Omega-3 Long Chain Polyunsaturated fatty acids (LC-PUFAs), the lack of knowledge about the optimal requirements for fatty acids or about their impact on mechanisms that regulate skeletal development has impeded the design of diets that could improve bone formation during larval stages when the majority of skeletal anomalies appear. In this study, Argyrosomus regius larvae were fed different levels of Omega-3s (2.6% and 3.6% DW on diet) compared to a commercial diet. At 28 days after hatching (DAH), their transcriptomes were analyzed to study the modulation exerted in gene expression dynamics during larval development and identify impacted genes that can contribute to skeletal formation. Mainly, both levels of supplementation modulated bone-cell proliferation, the synthesis of bone components such as the extracellular matrix, and molecules involved in the interaction and signaling between bone components or in important cellular processes. The 2.6% level impacted several genes related to cartilage development, denoting a special impact on endochondral ossification, delaying this process. However, the 3.6% level seemed to accelerate this process by enhancing skeletal development. These results offered important insights into the impact of dietary Omega-3 LC-PUFAs on genes involved in the main molecular mechanism and cellular processes involved in skeletal development.

Efforts have been made to find alternatives to fish meal (FM), as the sustainability of aquaculture depends on it. Insect meal (IM) is a potential candidate to partially replace FM, being more sustainable and economically viable. In this experimental trial, three diets were tested with different yellow mealworm incorporation: a control diet with no IM, a diet with an inclusion of 10% IM (Ins10), and a diet with an incorporation of 20% IM (Ins20). The diets were tested on 10.5 g meagre for 47 days. The results showed that an IM inclusion higher than 10% affected both growth (2.6 vs. 2.2) and FCR (1.5 vs. 1.9) of meagre juveniles. However, this reduction in growth did not result from lower protein retention or changes in muscle fibre area or density. Little differences were observed in the activity of pancreatic and intestinal enzymes except for aminopeptidase total activity which was higher in the control and Ins10 compared to Ins20 (3847 vs. 3540 mU/mg protein), suggesting no limitations in protein synthesis. Also, the alkaline phosphatase intestinal maturation index was higher in the control group compared to the IM groups (437 vs. 296). On the contrary, several differences were also found in the proteolytic activity in the hepatic and muscle tissues of meagre juveniles fed the Ins10 diet. The inclusion of IM had no impact on intestine histomorphology but changes were detected in the enterocytes of fish from control and Ins10 which showed hypervacuolization and nucleus misplacement compared to the Ins20 treatment. Nevertheless, a higher percentage of Vibrionaceae was recorded for meagre fed on the Ins20 diet. Since no signs of inflammation were observed in the distal intestine, this suggests IM incorporation could have had an important impact on intestinal health due to its antimicrobial properties. This is supported by an increase in the haematocrit in the treatments where IM was added (20 to 25%). In conclusion, incorporations of IM at percentages up to 10% do not seem to have a negative impact on meagre performance at this age but can enhance the fish immune system and protection against intestinal inflammation.

Water temperature variations affect fish growth and health, often leading to huge losses in fish production, especially during the cold season. To alleviate this constraint, fish farmers can use a water heating system driven by solar energy during daytime. This action will cause a water temperature drop during the night period, making it important to understand the physiological response of fish exposed to the resulting day/night temperature oscillations. To investigate this scenario, gilthead seabream juveniles (96.3 ± 1.0 g) were exposed to different thermal regimes for 67 days: Tconstant and Tdaily cycles. The latter group was exposed to daily water temperature oscillations between ~19 and 13 °C compared with a constant temperature of ~19 °C for the other experimental group. Temperature fluctuations compromised fish growth efficiency and reduced the proportion of fatty acids in several tissues, with implications for the whole proximate composition. Moreover, temperature oscillations influenced several blood parameters. These results favor the usage of a constant water temperature of ~19 °C for optimal gilthead seabream juvenile production instead of a day/night water temperature oscillating regime. Nevertheless, the type of energy used to warm the water will depend on the operational conditions and/or business strategy of fish farmers.

This study aimed to determine the effects of dietary inclusion of Tenebrio molitor larvae (yellow worms) meal (TM) on meagre fish (Argyrosomus regius) whole-body fatty acids (FA) profile and hepatic and intestine oxidative status. For that purpose, fish were fed for 9 weeks a fishmeal-based diet (control) or diets including 10%, 20%, or 30% TM. With the increase in dietary TM level, whole-body oleic acid, linoleic acid, monounsaturated FA, and n−6 polyunsaturated FA (PUFA) increased while saturated FA (SFA), n−3 PUFA, n−3 long chain-PUFA, SFA:PUFA ratio, n3:n6 ratio, and FA retention decreased. Hepatic superoxide dismutase (SOD), glucose-6-phosphate dehydrogenase (G6PDH), and glutathione reductase (GR) activities increased and catalase (CAT) and glutathione peroxidase (GPX) activities decreased with dietary TM inclusion. Hepatic total and reduced glutathione were lower in fish fed 20% TM. Intestinal CAT activity and oxidized glutathione increased and GPX activity decreased with dietary TM inclusion. Intestine SOD, G6PDH, and GR activities increased and malondialdehyde concentration decreased in fish fed the diets with lower TM inclusion levels. Liver and intestine oxidative stress index and liver malondialdehyde concentration were unaffected by dietary TM. In conclusion, to avoid major whole-body FA changes or antioxidant status imbalances, it is recommended to limit TM to 10% inclusion in meagre diets.

Efforts have been made to find natural, highly nutritious alternatives to replace fish meal (FM) and fish oil (FO), which can simultaneously promote fish health and improve the nutritional quality of filets for human consumption. This study evaluated the impact of biofortified diets containing microalgae (as replacement for FM and FO), macroalgae (as natural source of iodine) and selenised yeast (organic source of selenium) on gilthead seabream growth, nutrient utilization, tissue composition and gene expression. A control diet (CTRL) with 15% FM and 5.5% FO was compared with three experimental diets (AD1, AD2, and AD3), where a microalgae blend (Chlorella sp., Tetraselmis sp., and DHA-rich Schizochytrium sp.) replaced 33% of FM. Diet AD1 contained 20% less FO. Diets were supplemented with Laminaria digitata (0.4% AD1 and AD2; 0.8% AD3) and selenised yeast (0.02% AD1 and AD2; 0.04% AD3). After feeding the experimental diets for 12 weeks, growth was similar in fish fed AD1, AD2, and CTRL, indicating that microalgae meal can partially replace both FM and FO in diets for seabream. But AD3 suppressed fish growth, suggesting that L. digitata and selenised yeast supplementation should be kept under 0.8 and 0.04%, respectively. Despite lower lipid intake and decreased PUFAs bioavailability in fish fed AD3, compared to CTRL, hepatic elovl5 was upregulated resulting in a significant increase of muscle EPA + DHA. Indeed, filets of fish fed AD2 and AD3 provided the highest EPA + DHA contents (0.7 g 100 g–1), that are well above the minimum recommended values for human consumption. Fish consuming the AD diets had a higher retention and gain of selenium, while iodine gain remained similar among diets. Upregulation of selenoproteins (gpx1, selk, and dio2) was observed in liver of fish fed AD1, but diets had limited impact on fish antioxidant status. Overall, results indicate that the tested microalgae are good sources of protein and lipids, with their LC-PUFAs being effectively accumulated in seabream muscle. Selenised yeast is a good fortification vehicle to increase selenium levels in fish, but efforts should be placed to find new strategies to fortify fish in iodine.

Gilthead seabream (Sparus aurata) production is a highly valued aquaculture industry in Europe. The presence of skeletal deformities in farmed gilthead seabream represents a major bottleneck for the industry leading to economic losses, negative impacts on the consumers’ perception of aquaculture, and animal welfare issues for the fish. Although past work has primarily focused on the hatchery phase to reduce the incidence of skeletal anomalies, this work targets the successive pre-ongrowing phase in which more severe anomalies affecting the external shape often arise. This work aimed to test the effects of: (i) larger and smaller tank volumes, stocked at the same density; and (ii) higher and lower stocking densities maintained in the same water volume, on the skeleton of gilthead seabream fingerlings reared for ~63 days at a pilot scale. Experimental rearing was conducted with gilthead seabream juveniles (~6.7 ± 2.5 g), which were selected as ‘non-deformed’ based on external inspection, stocked at three different densities (Low Density (LD): 5 kg/m3; Medium Density (MD): 10 kg/m3; High Density (HD): 20 kg/m3) in both 500 L and 1000 L tanks. Gilthead seabream were sampled for growth performance and radiographed to assess the skeletal elements at the beginning and end of the experimental trial. Results revealed that (i) LD fish were significantly longer than HD fish, although there were no differences in final weights, regardless of the water volume; (ii) an increase in the prevalence of seabream exhibiting cranial and vertebral axis anomalies was found to be associated with increased density. These results suggest that farmers can significantly reduce the presence of some cranial and axis anomalies affecting pre-ongrown gilthead seabream by reducing the stocking density.

Histological techniques are a traditional and essential tool for adequately comprehending cellular morphology and physiology in several scientific fields, including ecotoxicological studies. Although bioassays generally recommend using small test organisms, based on advantages such as low maintenance costs and the possibility of a higher number of replicates, the histological process of such organisms is still challenging due to the difficulty of handling them. The present study proposes an accessible protocol for performing the histology of a biological sample of 8 mm maximum length and using histopathological biomarkers in Senegal sole larvae for ecotoxicological tests. Eight-day old larvae of Solea senegalensis obtained from breeders’ natural spawns in cultivation conditions were submitted to histological processing by an adjusted protocol comprising fixation, agarose array pre-embedding, dehydration, paraffin embedding, slides’ preparation, and image analysis by light optical microscope coupled to a digital camera. Low-cost and easy-to-use alternatives were applied in the technique execution. The histological steps were detailed, and S. senegalensis morphological features were presented. This can be considered valuable background information for ecotoxicological studies where changes in cell morphology and physiology are expected.

This study aimed to assess the effect of Hermetia illucens meal (HM) dietary inclusion on meagre oxidative status. Thus, fish were fed a fishmeal-based diet (CTR diet) and three other diets with increasing levels of HM inclusion, namely 10%, 20%, and 30% (diets HM10, HM20, and HM30, respectively). At the end of the trial, hepatic and intestine superoxide dismutase, catalase, and glucose-6-phosphate dehydrogenase activities and malondialdehyde concentration were unaffected by the diet composition. Liver glutathione peroxidase activity was higher in the fish fed the HM20 diet than in the fish fed the CTR and HM30 diets, and glutathione reductase activity linearly increased with the dietary HM level. The hepatic total glutathione and reduced glutathione contents were significantly lower in fish fed the HM20 diet than in fish fed the CTR and HM10 diets. In the intestine, the oxidized glutathione (GSSG) content and oxidative stress index linearly increased with the increase in dietary HM level, with the GSSG content of fish fed the HM20 diet being significantly higher than of fish fed the CTR diet. In conclusion, 30% HM might be included in meagre diets without negatively affecting hepatic and intestine oxidative status.

Production costs in extensive and semi-intensive fish culture in earthen ponds are often too high to offer sustainable economic activity due to the low productivity of these systems. The right combination of commercial finfish species with inorganic (primary producers) and organic extractive (bivalves) species in Integrated MultiTrophic Aquaculture (IMTA) create a balanced system with higher profitability and risk reduction. To achieve this, it is crucial to understand the role of each functional groups within the system what we did by comparing three different IMTA production three different IMTA production treatments with distinct combinations of trophic levels: • fish, filter feeders, phytoplankton and macroalgae, • fish, filter feeders and phytoplankton • fish, phytoplankton and macroalgaeEach treatment was carried out in two similar ponds under semi-intensive conditions and flow through system, in a total of 6 earthen ponds of 500 m2 surface and depth of 1.5 m.Results showed that the presence of oysters in the ponds enhanced water quality by decreasing turbidity and by controlling phytoplankton which led to regulation of dissolved oxygen levels. The enhanced water quality in these systems lead to improved fish performance and higher biomass production contributing to greater profitability. The combination of fish, oyster, phytoplankton and macroalgae was particularly good providing much more fish supply compared with the other two treatments. • Oysters enhanced water quality in the ponds by decreasing turbidity and controlling phytoplankton which regulated the dissolved oxygen levels. • The enhanced water quality in systems with oysters improve fish performance resulting in higher biomass production and greater profitability. • The combination of fish, oyster, phytoplankton and macroalgae was particularly good providing much more fish supply compared with the other two treatments. Method name: Pond IMTA, Keywords: Aquaculture, IMTA, Meagre, White seabream, Grey mullet, Japanese oyster, Phytoplankton, Ulva

The dinoflagellate Amyloodinium ocellatum is the etiological agent of a parasitic disease named amyloodiniosis. Mortalities of diseased fish are usually attributed to anoxia, osmoregulatory impairment, or opportunistic bacterial infections. Nevertheless, the phylogenetic proximity of A. ocellatum to a group of toxin-producing dinoflagellates from Pfiesteria, Parvodinium and Paulsenella genera suggests that it may produce toxin-like compounds, adding a new dimension to the possible cause of mortalities in A. ocellatum outbreaks. To address this question, extracts prepared from different life stages of the parasite were tested in vitro for cytotoxic effects using two cell lines derived from branchial arches (ABSa15) and the caudal fin (CFSa1) of the gilthead seabream (Sparus aurata), and for hemolytic effects using erythrocytes purified from the blood of gilthead seabream juveniles. Cytotoxicity and a strong hemolytic effect, similar to those observed for Karlodinium toxins, were observed for the less polar extracts of the parasitic stage (trophont). A similar trend was observed for the less polar extracts of the infective stage (dinospores), although cell viability was only affected in the ABSa15 line. These results suggest that A. ocellatum produces tissue-specific toxic compounds that may have a role in the attachment of the dinospores’ and trophonts’ feeding process.

There is growing demand for the territorial tube-building genus Marphysa (Eunicidae: Polychaete), commonly known in Portugal as “goose”, for use as fishing bait, and it is being harvested all around the world for that purpose. Effects of intraspecific density on juvenile growth were studied over a four-month period in laboratory facilities. Three polychaete densities (low, 50 worms; medium, 150 worms; and high, 250 worms) were used in a 0.25 m2 aquarium containing sandy sediment and recirculating water. Total length, dry weight and number of segments were recorded for 60% of the initial population. All polychaetes were also counted to determine mortality rate and territorial behaviour through the existence of body lesions and broken and regenerating posterior segments. The results obtained in this study showed that density had a significant effect (p

No-take areas are key instruments to promote the effectiveness of Marine Protected Areas (MPAs), particularly concerning the protection of endangered species such as the dusky grouper (Epinephelus marginatus). However, despite the establishment of no-take areas and the prohibition of catching this species in a southwestern Portuguese MPA (SACVMP—‘Sudoeste Alentejano’ and ‘Costa Vicentina’ Marine Park) since 2011, there is still no evidence of population recovery. By using acoustic biotelemetry, this work aimed to evaluate the feasibility of restocking hatchery-reared adult dusky groupers in two no-take areas within the SACVMP. In 2019 and 2021, thirty groupers were tagged with acoustic transmitters and the site attachment and movements of the groupers were assessed in the releasing sites (no-take areas). None of the tagged fish settled down in either of the areas, leaving the no-take areas mainly at dusk and night. Some individuals displayed extended movements of more than a hundred kilometers along the Portuguese coast which was rarely reported for this species. At least in some coastal stretches, those movements were performed close to the shore, which may evidence the importance of coastal MPAs to protect and promote the connectivity of species more associated with rocky reef habitats. Following studies should focus on the conditions that promote site attachment and fidelity by hatchery-reared dusky groupers so that future large-scale restocking programs can be successful in MPAs with appropriate habitats.

Background Many fish taxa produce sound in voluntary and in disturbance contexts but information on the full acoustic repertoire is lacking for most species. Yet, this knowledge is critical to enable monitoring fish populations in nature through acoustic monitoring. Methods In this study we characterized the sounds emitted during disturbance and voluntary contexts by juvenile and adult meagre, Argyrosomus regius, in laboratory conditions. Breeding sounds produced by captive adults were also compared with meagre sounds registered in the Tagus estuary (Lisbon, Portugal) from unseen fish during the breeding season. Results The present dataset demonstrates for the first time that in this species dominant frequency is inversely related to fish size, and that sounds vary according to sex, context and age. Sounds from captive breeding adults were similar to sounds recorded in the field. Discussion Our findings indicate that A. regius sound features carry information about size, sex, age and motivation. This variability could potentially be used to identify meagre in the field and to infer about ontogenetic phase (i.e., juveniles vs. adults, and variation with size) and motivation (e.g., spawning). Future studies should confirm sex differences and ascertain the influence of water temperature on acoustic features.

Passive Acoustic Monitoring (PAM) is a non-intrusive and cost-effective method capable of providing high-resolution, long-term information on the status and health of vocal populations and communities. To successfully monitor the same species over wide geographical and temporal scales, it is necessary to characterise the range of sound variability, as well as the consistency of sound features between populations. The meagre (Argyrosomus regius, Asso 1801) is an interesting case study because recent investigations suggest a wider vocal repertoire than previously described. In this study, meagre vocalizations were recorded and analysed from a variety of settings, ranging from rearing facilities to wild populations to provide a comprehensive characterisation of its vocal repertoire, while investigating the consistency of spawning sound features between populations. All sounds presented a similar acoustic structure in their basic unit (i.e. the pulse), while an important variability was found in the number of pulses; the meagre can emit sounds made of one single pulse or many pulses (up to more than 100). High level of overlap in the Principal Component Analysis made difficult to differentiate sound type clusters. Despite this, two sound types were identifiable: knocks (sounds from 1 to 3 pulses) and long grunts (sounds with more than 29 pulses). Discriminant Analysis carried out on PCA residuals showed that knock had the highest proportion of correct placement (92% of the observations correctly placed) followed by long grunts (80%). All other previously described sound types (intermediate grunt, short grunt and disturbance sounds) could not be separated and presented low levels of correct placement, suggesting that care should be taken when defining these as independent sound types. Finally, acoustic features consistency was found in meagre grunts emitted by different populations during spawning nights; statistical differences could be explained by recording settings and fish conditions. The results of this study provide important information for fostering PAM programs of wild meagre populations, while contributing to the discussion around the definition of fish sound types in vocal fish communities. Studies of this kind, which evaluate both variability and consistency of sound features, are of fundamental importance for maximising PAM efforts in the wild, at both the specific and the community level.

Water extracts from sea lavender (Limonium algarvense Erben) plants cultivated in greenhouse conditions and irrigated with freshwater and saline aquaculture effluents were evaluated for metabolomics by liquid chromatography-tandem high-resolution mass spectrometry (LC-HRMS/MS), and functional properties by in vitro and ex vivo methods. In vitro antioxidant methods included radical scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), ferric-reducing antioxidant power (FRAP), and copper and iron chelating assets. Flowers’ extracts had the highest compounds’ diversity (flavonoids and its derivatives) and strongest in vitro antioxidant activity. These extracts were further tested for ex vivo antioxidant properties by oxidative haemolysis inhibition (OxHLIA), lipid peroxidation inhibition by thiobarbituric acid reactive substances (TBARS) formation, and anti-melanogenic, anti-tyrosinase, anti-inflammation, and cytotoxicity. Extract from plants irrigated with 300 mM NaCl was the most active towards TBARS (IC50 = 81 µg/mL) and tyrosinase (IC50 = 873 µg/mL). In OxHLIA, the activity was similar for fresh- and saltwater-irrigated plants (300 mM NaCl; IC50 = 136 and 140 µg/mL, respectively). Samples had no anti-inflammatory and anti-melanogenic abilities and were not toxic. Our results suggest that sea lavender cultivated under saline conditions could provide a flavonoid-rich water extract with antioxidant and anti-tyrosinase properties with potential use as a food preservative or as a functional ingredient in herbal supplements.

This data article includes data and analyses on the effect of different agronomic techniques on the production of Polygonum maritimum L. (sea knotgrass), namely different salinity irrigation treatments (0, 100, 200, 300 and 600 mM of NaCl) and a multi-harvest regime, and their relation with the chemical profile (ultra-high-resolution mass spectrometry - UHRMS), in vitro antioxidant [radical-scavenging activity (RSA) of DPPH and ABTS, copper chelating activity and ferric reducing antioxidant power] and anti-inflammatory (nitric oxide reduction on lipopolysaccharide-stimulated macrophages) activities. For further interpretation of the data presented in this work, please see the related research article “The irrigation salinity and harvesting affect the growth, chemical profile and biological activities of Polygonum maritimum L.” (Rodrigues et al., 2019). Keywords: Antioxidant activity, Anti-inflammatory activity, Halophyte domestication, Saline agriculture

In this paper we report a case of amyloodiniosis in a sea bass (Dicentrarchus labrax) broodstock in Portugal. Microscopic examination of gill filaments showed the presence of trophonts while histological observation revealed gills epithelial hyperplasia, hypertrophy and lamellar fusion of secondary lamellae. The amplification and sequencing of the small subunit ribosomal RNA gene allowed the identification of the parasite as Amyloodinium ocellatum. It was also possible to amplify a partial sequence of ribosomal RNA from a Colpodellid, a predator of protists. Keywords: Amyloodinium ocellatum, Dicentrarchus labrax, Fish parasites, Colpodellids, Broodstock

Mercury (Hg) is globally recognized as a persistent chemical contaminant that accumulates in marine biota, thus constituting an ecological hazard, as well as a health risk to seafood consumers. Climate change-related stressors may influence the bioaccumulation, detoxification, and toxicity of chemical contaminants, such as Hg. Yet, the potential interactions between environmental stressors and contaminants, as well as their impacts on marine organisms and seafood safety, are still unclear. Hence, the aim of this work was to assess the bioaccumulation of Hg and neuro-oxidative responses on the commercial flat fish species Solea senegalensis (muscle, liver, and brain) co-exposed to dietary Hg in its most toxic form (i.e., MeHg), seawater warming (ΔT°C = +4 °C), and acidification (pCO2 = +1000 µatm, equivalent to ΔpH = −0.4 units). In general, fish liver exhibited the highest Hg concentration, followed by brain and muscle. Warming enhanced Hg bioaccumulation, whereas acidification decreased this element’s levels. Neuro-oxidative responses to stressors were affected by both climate change-related stressors and Hg dietary exposure. Hazard quotient (HQ) estimations evidenced that human exposure to Hg through the consumption of fish species may be aggravated in tomorrow’s ocean, thus raising concerns from the seafood safety perspective.

The effects of two different slipping methods on the survival, physical and physiological response of sardines, Sardina pilchardus, captured in a purse-seine fishery were investigated in southern Portugal. Sardines were collected and transferred into holding tanks onboard a commercial fishing vessel after being captured, crowded and deliberately released using two slipping procedures: standard and modified. The standard slipping procedure aggregated fish at high densities and made them "roll over" the floatline, while the modified procedure aggregated the fish at moderate densities and enabled them to escape through an opening created by adding weights to the floatline. Both slipping methods were compared with minimally harmed non-slipped sardines (sardines collected from the loose pocket of the purse seine). Survival rates were monitored in captivity over 28 days using three replicates for each treatment. The estimated survival of sardines was 43.6% for the non-slipped fish, 44.7% for the modified slipping and 11.7% for the standard slipping treatments. Scale loss indicated the level of physical impact experienced, with dead fish from the non-slipped and modified slipping technique showing significantly lower scale loss than those fish from the standard slipping treatment within the same period. Of the physiological indicators of stress measured, cortisol, glucose, lactate and osmolality attained peak values during slipping and up to the first hours after introduction to captivity. This work indicates that although delayed mortality after release may be substantial, appropriately modified slipping techniques significantly enhance survival of slipped sardines.