Search Results (58)

Brewer’s spent grain (BSG), the primary byproduct generated by the brewing industry, holds significant potential as an ingredient in aquafeeds. However, its high content of non-starch polysaccharides (NSP) restricts the amount that can be incorporated into fish diets. To address these limitations, various pretreatment methods (physical, chemical, or enzymatic) can be applied prior to its inclusion in feed formulations. The objectives of the present study were (i) to optimize the conditions for enzymatic or microwave pretreatments to enhance the nutritional and functional profile of BSG, (ii) to determine the bioaccessibility and availability of specific nutrients and antioxidant compounds in diets including a high amount of BSG (pretreated or not) using an in vitro assay simulating the digestion of the gilthead seabream (Sparus aurata), and (iii) to evaluate the effect of such diet pretreatments on growth, metabolism, intestinal microbiota, and oxidative status in live fish. For this study, three experimental diets were formulated: one containing 20% untreated BSG (C) and two containing the same amount of BSG pretreated either enzymatically (H) or with microwave heating (MW). Each diet was administered ad libitum to triplicate groups of 22 juvenile sea bream (mean weight 60 g) over a period of three months. The results indicated that the microwave-treated BSG led to improved growth (0.69 ± 0.02%) and feed efficiency (0.80 ± 0.02 weight gain/total feed intake) and enhanced immune status (alkaline phosphatase activity = 11,811.68 ± 3426.92 U/mg SP), compared to the control diet (SGR = 0.59 ± 0.06%; FE: 0.68 ± 0.03 weight gain/total feed intake; alkaline phosphatase activity = 8590.29 ± 3663.44 U/mg SP). Moreover, fish fed on both pretreated BSG diets exhibited significant differences in metabolic parameters and functional profile of their intestinal microbiota when compared to the control group. Consequently, the findings suggest that the pretreatment of BSG, whether by enzymatic or microwave methods, results in notable differences in its nutritional value and the bioavailability of functional components, which, in turn, have a substantial impact on the growth and metabolism of gilthead seabream. Full article

Peripheral blood has been always used as the mirror of physiological status in a minimally invasive manner. In homeothermic vertebrates, different blood parameters have been correlated to a variety of biochemical and physiological processes and the establishment of physiological values have rendered them valuable indicators for research, diagnosis, and welfare status. Despite the volume of research in fish physiology and teleost significance for food security, information about the physiological values of minimally invasive biomarkers is still fragmented while there is a great need for such biomarkers for monitoring fish health, welfare, and factors that can impact their homeostasis. For the first time, we have focused on determining the physiological ranges of cholesterol, triglycerides, and total proteins in the plasma of gilthead sea bream (Sparus aurata) and detecting how size, diet, environment, and genetic background may influence these levels. A large heterogeneous dataset of 5318 healthy individuals of two different size classes (1–100 g; 101–200 g), dietary status (10 feeds and feeding regimes), and different genetic background (125 families), in combination with different geographic locations (GxE), was produced over a period of three years and analyzed to ensure a comprehensive representation in establishing the physiological range for these biomarkers. Dietary status and GxE introduced significant variations in the range of the physiological values of all biomarkers in both size classes. Triglyceride, total protein and cholesterol values did differentiate with age/size. Interestingly, all parameters responded to diet in a size-specific way, revealing their potential as biomarkers of dietary status. Full article

This study explores the impact of geographical origin, harvest time, and cooking on the volatile organic compound (VOC) profiles of wild and reared seabream from the Adriatic and Tyrrhenian Seas. A Proton Transfer Reaction–Time of Flight–Mass Spectrometry (PTR-ToF-MS) allowed for VOC profiling with high sensitivity and high throughput. A total of 227 mass peaks were identified. Principal component analysis (PCA) showed a clear separation between cooked and raw samples, with cooking causing a significant increase in 64% of VOCs, especially hydrogen sulphide, methanethiol, and butanal. A two-way ANOVA revealed significant effects of origin, time, and their interaction on VOC concentration, with 102 mass peaks varying significantly based on all three factors. Seasonal effects were also notable, particularly in reared fish from the Adriatic Sea, where compounds like monoterpenes and aromatics were higher during non-breeding months, likely due to environmental factors unique to that area. Differences between wild and reared fish were influenced by lipid content and seasonal changes, impacting the VOC profile of seabream. These findings provide valuable insights into how cooking, geographical origin, and seasonality interact to define the flavour profile of seabream, with potential applications in improving quality control and product differentiation in seafood production. Full article

A confinement stress test with 75% tank space reduction and behavioural monitoring through tri-axial accelerometers externally attached to the operculum was designed. This procedure was validated by demonstrating the less pronounced stress response in gilthead sea bream than in European sea bass (950–1200 g). Our study aimed to assess habituation to high stocking densities with such procedure in gilthead sea bream. Animals (420–450 g) were reared (June–August) in a flow-through system at two stocking densities (CTRL: 10–15 kg/m³; HD: 18–24 kg/m³), with natural photoperiod and temperature (21–29 °C), and oxygen levels at 5.2–4.2 (CTRL) and 4.2–3.2 ppm (HD). At the end, blood and muscle were sampled for haematology and transcriptomic analyses, and external tissue damage was assessed by image-based scoring. Four days later, fish underwent a 45 min confinement stress test over two consecutive days. HD fish showed reduced feed intake, growth rates and haematopoietic activity. Muscle transcriptome changes indicated a shift from systemic to local growth regulation and a primed muscle regeneration over protein accretion in HD animals with slight external injuries. After stress testing, HD fish exhibited a decreased recovery time in activity and respiration rates, which was shorter after a second stressor exposure, confirming habituation to high densities. Full article

Plastic pollution is a global problem affecting all ecosystems, and it represents most of the marine litter. Offshore aquaculture is a sector particularly vulnerable to this issue. To investigate this concern, the present study employed videography to monitor macroplastics at an offshore fish farm on Madeira Island (Portugal) and analysis of fish gut content to evaluate macroplastic ingestion by farmed sea bream Sparus aurata. Our analysis revealed that the majority of identified plastic debris originated from domestic use (66.66%) and fisheries/aquaculture activities (24.99%). While the number of dead fish suitable for sampling was limited (1.05% of the total mortality), macroplastic debris ingestion was identified in 5.15% of the total mortalities and reported for the first time in species in offshore farming conditions. Fish ingested fragmented plastic sheets, with the amount positively correlated with fish weight (r = 0.621, p = 0.031, n = 12). Notably, the stretched length of these fragments exceeded 50% of the standard length of most fish. Inconsistencies were observed in the number of samples collected per cage and per week. To ensure robust results, these discrepancies should be rectified in future studies. Additionally, extending the sampling period to encompass all seasons would be beneficial for a more comprehensive understanding of seasonal variations in plastic occurrence. Full article

DNA methylation clocks provide information not only about chronological but also biological age, offering a high-resolution and precise understanding of age-related pathology and physiology. Attempts based on transcriptomic and epigenetic approaches arise as integrative biomarkers linking the quantification of stress responses with specific fitness traits and may help identify biological age markers, which are also considered welfare indicators. In gilthead sea bream, targeted gene expression and DNA methylation analyses in white skeletal muscle proved sirt1 as a reliable marker of age-mediated changes in energy metabolism. To complete the list of welfare auditing biomarkers, wide analyses of gene expression and DNA methylation in one- and three-year-old fish were combined. After discriminant analysis, 668 differentially expressed transcripts were matched with those containing differentially methylated (DM) regions (14,366), and 172 were overlapping. Through enrichment analyses and selection, two sets of genes were retained: 33 showing an opposite trend for DNA methylation and expression, and 57 down-regulated and hypo-methylated. The first set displayed an apparently more reproducible and reliable pattern and 10 multifunctional genes with DM CpG in regulatory regions (sirt1, smad1, ramp1, psmd2—up-regulated; col5a1, calcrl, bmp1, thrb, spred2, atp1a2—down-regulated) were deemed candidate biological age markers for improved welfare auditing in gilthead sea bream. Full article

The growth of the aquaculture industry requires more sustainable and circular economy-driven aquafeed formulas. Thus, the goal of the present study was to assess in farmed gilthead sea bream (Sparus aurata L.) how different combinations of novel and conventional fish feed ingredients supported proper animal performance in terms of growth and physiological biomarkers of blood/liver/head kidney. A 77-day feeding trial was conducted with three experimental diets (PAP, with terrestrial processed animal protein from animal by-products; NOPAP, without processed animal protein from terrestrial animal by-products; MIX, a combination of alternative ingredients of PAP and NOPAP diets) and a commercial-type formulation (CTRL), and their effects on growth performance and markers of endocrine growth regulation, lipid metabolism, antioxidant defense and inflammatory condition were assessed at circulatory and tissue level (liver, head kidney). Growth performance was similar among all dietary treatments. However, fish fed the PAP diet displayed a lower feed conversion and protein efficiency, with intermediate values in MIX-fed fish. Such gradual variation in growth performance was supported by different biomarker signatures that delineated a lower risk of oxidation and inflammatory condition in NOPAP fish, in concurrence with an enhanced hepatic lipogenesis that did not represent a risk of lipoid liver degeneration. Full article

Fish were kept for six weeks at three different initial stocking densities and water O₂ concentrations (low-LD, 8.5 kg/m³ and 95–70% O₂ saturation; medium-MD, 17 kg/m³ and 55–75% O₂ saturation; high-HD, 25 kg/m³ and 60–45% O₂ saturation), with water temperature increasing from 19 °C to 26–27 °C. The improvement in growth performance with the decrease in stocking density was related to changes in skin and intestinal mucosal microbiomes. Changes in microbiome composition were higher in skin, with an increased abundance of Alteromonas and Massilia in HD fish. However, these bacteria genera were mutually exclusive, and Alteromonas abundance was related to a reactive behavior and systemic growth regulation via the liver Gh/Igf system, while Massilia was correlated to a proactive behavior and a growth regulatory transition towards muscle rather than liver. At the intestinal level, microbial abundance showed an opposite trend for two bacteria taxa, rendering in a low abundance of Reyranella and a high abundance of Prauserella in HD fish. This trend was correlated with up-regulated host gene expression, affecting the immune response, epithelial cell turnover, and abiotic stress response. Most of the observed responses are adaptive in nature, and they would serve to infer new welfare indicators for increased stress resilience. Full article

The skin of bony fish is the first physical barrier and is responsible for maintaining the integrity of the fish. Lesions make the skin vulnerable to potential infection by pathogens present in the aquatic environment. In this way, wound repair has barely been studied in gilthead sea bream. Thus, this study investigated the modulation of peripheral neuro-endocrine and tissue repair markers at the transcriptional level in the skin of teleost fish subjected to mechanical damage above or below the lateral line (dorsal and ventral lesions, respectively). Samples were evaluated using RT-qPCR at 2-, 4-, and 20-days post-injury. Fish with a ventral lesion presented a trend of progressive increase in the expressions of corticotropin-releasing hormone (crh), pro-opiomelanocortin-A (pomca), proenkephalin-B (penkb), cholecystokinin (cck), oxytocin (oxt), angiotensinogen (agt), and (less pronounced) somatostatin-1B (sst1b). By contrast, fish with a dorsal lesion registered no significant increase or biological trend for the genes evaluated at the different sampling times. Collectively, the results show a rapid and more robust response of neuro-endocrine and tissue repair markers in the injuries below than above the lateral line, which could be attributable to their proximity to vital organs. Full article

Nowadays, including vegetable ingredients in fish diets without growth effects is common; however, their intestinal oxidative status under these conditions is less known. Five isonitrogenous and isolipidic diets with 75% vegetable oil (VO) inclusion were formulated for juvenile gilthead sea bream (Sparus aurata). As VO, one diet contained palm oil (diet P), another rapeseed oil (diet R), and the other three included linseed oil (L) combined with the above-mentioned VOs (named PL, RL and RPL diets). After 18 weeks, pyloric caeca (PC) and proximal intestine (PI) were analyzed for oxidative stress biomarkers, lipid peroxidation (LPO), and gene expression. Dietary linseed oil diminished the superoxide dismutase activity in both intestinal regions, catalase in PC and glutathione reductase in PI; rapeseed oil reduced the glutathione peroxidase (GPx) and glutathione-S-transferase activities in PC, and palm oil upregulated GPx activity in PI. The PL diet triggered LPO levels in the PI, and RPL-fed fish showed the highest levels of LPO in the PC due to lower antioxidant activities, while RL-fed fish presented the best oxidative status. The results suggest that the dietary amount of n-6 and the unsaturated/saturated fatty acids ratio are factors to be considered in aquafeed formulation, including VOs, to improve the intestinal oxidative status in fish. Full article

The Sirtuin (SIRT1-7) family comprises seven evolutionary-conserved enzymes that couple cellular NAD availability with health, nutrition and welfare status in vertebrates. This study re-annotated the sirt3/5 branch in the gilthead sea bream, revealing three paralogues of sirt3 (sirt3.1a/sirt3.1b/sirt3.2) and two of sirt5 (sirt5a/sirt5b) in this Perciform fish. The phylogeny and synteny analyses unveiled that the Sirt3.1/Sirt3.2 dichotomy was retained in teleosts and aquatic-living Sarcopterygian after early vertebrate 2R whole genome duplication (WGD). Additionally, only certain percomorphaceae and gilthead sea bream showed a conserved tandem-duplicated synteny block involving the mammalian-clustered sirt3.1 gene (psmd13-sirt3.1a/b-drd4-cdhr5-ctsd). Conversely, the expansion of the Sirt5 branch was shaped by the teleost-specific 3R WGD. As extensively reviewed in the literature, human-orthologues (sirt3.1/sirt5a) showed a high, conserved expression in skeletal muscle that increased as development advanced. However, recent sirt3.2 and sirt5b suffered an overall muscle transcriptional silencing across life, as well as an enhanced expression on immune-relevant tissues and gills. These findings fill gaps in the ontogeny and differentiation of Sirt genes in the environmentally adaptable gilthead sea bream, becoming a good starting point to advance towards a full understanding of its neo-functionalization. The mechanisms originating from these new paralogs also open new perspectives in the study of cellular energy sensing processes in vertebrates. Full article

In this study, gilthead sea bream (Sparus aurata) fast muscle myoblasts were stimulated with two pro-growth treatments, amino acids (AA) and insulin-like growth factor 1 (Igf-1), to analyze the transcriptional response of mRNAs, microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) and to explore their possible regulatory network using bioinformatic approaches. AA had a higher impact on transcription (1795 mRNAs changed) compared to Igf-1 (385 mRNAs changed). Both treatments stimulated the transcription of mRNAs related to muscle differentiation (GO:0042692) and sarcomere (GO:0030017), while AA strongly stimulated DNA replication and cell division (GO:0007049). Both pro-growth treatments altered the transcription of over 100 miRNAs, including muscle-specific miRNAs (myomiRs), such as miR-133a/b, miR-206, miR-499, miR-1, and miR-27a. Among 111 detected lncRNAs (>1 FPKM), only 30 were significantly changed by AA and 11 by Igf-1. Eight lncRNAs exhibited strong negative correlations with several mRNAs, suggesting a possible regulation, while 30 lncRNAs showed strong correlations and interactions with several miRNAs, suggesting a role as sponges. This work is the first step in the identification of the ncRNAs network controlling muscle development and growth in gilthead sea bream, pointing out potential regulatory mechanisms in response to pro-growth signals. Full article

High-fat diets (HFDs) enhance fish growth by optimizing nutrient utilization (i.e., protein-sparing effect); however, their potential negative effects have also encouraged the search for feed additives. This work has investigated the effects of an extract rich in a polyphenolic antioxidant, hydroxytyrosol (HT), supplemented (0.52 g HT/kg feed) in a HFD (24% lipid) in gilthead sea bream (Sparus aurata). Fish received the diet at two ration levels, standard (3% of total fish weight) or restricted (40% reduction) for 8 weeks. Animals fed the supplemented diet at a standard ration had the lowest levels of plasma free fatty acids (4.28 ± 0.23 mg/dL versus 6.42 ± 0.47 in the non-supplemented group) and downregulated hepatic mRNA levels of lipid metabolism markers (ppara, pparb, lpl, fatp1, fabp1, acox1, lipe and lipa), supporting potential fat-lowering properties of this compound in the liver. Moreover, the same animals showed increased muscle lipid content and peroxidation (1.58- and 1.22-fold, respectively, compared to the fish without HT), suggesting the modulation of body adiposity distribution and an enhanced lipid oxidation rate in that tissue. Our findings emphasize the importance of considering this phytocompound as an optimal additive in HFDs for gilthead sea bream to improve overall fish health and condition. Full article

In modern aquaculture, genomics-driven breeding programs have emerged as powerful tools for optimizing fish quality. This study focused on two emblematic Mediterranean fish species, the European seabass (Dicentrarchus labrax) and the gilthead sea bream (Sparus aurata), with a primary aim of exploring the genetic basis of white muscle/fillet degradation in fresh fish following harvest. We identified 57 and 44 missense SNPs in gilthead sea bream and European seabass, respectively, located within genes encoding for endogenous proteases responsible for fillet quality. These SNPs were cherry-picked based on their strategic location within the catalytic/regulatory domains of endogenous proteases that are expressed in the white muscle. Using MassArray technology, we successfully associated differentiated enzymatic activity of those endogenous proteases post-harvest as a phenotypic trait with genetic polymorphism of six SNPs in gilthead sea bream and nine in European seabass. These findings can be valuable attributes in selective breeding programs toward the extension of freshness and shelf life of these species. The integration of MassArray technology into breeding programs offers a cost-effective strategy for harnessing the potential of these genetic variants to enhance the overall quality of the final product. Recognizing that fresh fish perishability is a challenge, extending shelf-life is pivotal in reducing losses and production costs. Full article

The administration of a single dose of chitosan nanoparticles driving the expression of sterol regulatory element-binding protein 1a (SREBP1a) was recently associated with the enhanced conversion of carbohydrates into lipids. To address the effects of the long-lasting expression of SREBP1a on the growth and liver intermediary metabolism of carnivorous fish, chitosan-tripolyphosphate (TPP) nanoparticles complexed with a plasmid expressing the N terminal active domain of hamster SREBP1a (pSG5-SREBP1a) were injected intraperitoneally every 4 weeks (three doses in total) to gilthead sea bream (Sparus aurata) fed high-protein–low-carbohydrate and low-protein–high-carbohydrate diets. Following 70 days of treatment, chitosan-TPP-pSG5-SREBP1a nanoparticles led to the sustained upregulation of SREBP1a in the liver of S. aurata. Independently of the diet, SREBP1a overexpression significantly increased their weight gain, specific growth rate, and protein efficiency ratio but decreased their feed conversion ratio. In agreement with an improved conversion of dietary carbohydrates into lipids, SREBP1a expression increased serum triglycerides and cholesterol as well as hepatic glucose oxidation via glycolysis and the pentose phosphate pathway, while not affecting gluconeogenesis and transamination. Our findings support that the periodical administration of chitosan-TPP-DNA nanoparticles to overexpress SREBP1a in the liver enhanced the growth performance of S. aurata through a mechanism that enabled protein sparing by enhancing dietary carbohydrate metabolisation. Full article