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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.

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.

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.

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.

This work assesses the dietary use of two insect meals of Tenebrio molitor (TM) larvae reared in conventional (TM-10) or MAP-enriched substrates (MAP-TM-10) as fish meal replacements (10%) in the diets of gilthead seabream (Sparus aurata). Fish (n = 4500; 207.19 ± 1.47 g) were divided into three groups with triplicates: control (fed conventional diet), TM-10, and MAP-TM-10 groups. The fish were reared in floating cages for 12 weeks and the dietary effects on white blood cell activation, heat shock proteins, MAPKs, and apoptosis of the fish were evaluated. The MAP-TM-10 group exhibited the highest eosinophilic induction. Phosphorylated levels of p38 MAPK, p44/42 MAPK, HSP70, and HSP90 increased in the TM-10 and MAP-TM-10 groups. In terms of apoptosis, Bax levels were lower in the TM groups compared to the control, and the MAP-TM-10 group showed even lower levels than the TM-10 group. Bcl-2 levels increased in the TM-10 group compared to the control, and further increased in the MAP-TM-10 group. The Bax/Bcl-2 ratio, an apoptosis indicator, decreased in the TM groups, with the MAP-TM-10 group showing a further decrease compared to TM-10. These findings suggest that insects’ breeding substrate being enriched with MAPs modulated the effect of TM on cellular stress and apoptosis.

Mediterranean finfish aquaculture is mainly represented by the production of the gilthead sea bream (Sparus aurata) and sea bass (Dicentrarchus labrax) growing in marine cage farms. Despite the numerous benefits of fish farming, the ecological risk potentially caused by fish escapes to the wild populations is occasionally high. In the current study, an integrated review of fish escapes is presented regarding S. aurata and D. labrax escapes and their potential effect on the genetic composition of wild populations. The collected data from the literature are combined and discussed along with recorded escapes in a fish farm located in the Aegean Sea, Greece. According to these records, fish escapes present a generally stable rate, ranging between 0 and 2% in each escape event, which occur 5–8 times per year. Although for other farmed fish species worldwide, the ecological risk may be higher, this risk is probably lower in Mediterranean finfish populations, which are characterized by generally low levels of genetic differentiation. Nevertheless, the risk of genetic introgression still exists. Particularly in sea bream and sea bass farming, genetic structure and differentiation in wild populations seem to be unaffected for now. Nevertheless, suitable management measurements would be a useful strategy to avoid future negative effects of fish escapes. These strategies should be focused on further efforts towards recapture program development, reduced escapees’ survival, and proper legislation. Furthermore, more data on escapees’ survival percentage, their migration pathways, and the way these pathways relate to mortality, the type of escape event, and the season of release would be also valuable.

The possibility of simultaneous production of halophyte and euryhaline fish creates huge interest in both commercial aquaponics systems and in research. The aim of the present study was to investigate the effect of three different salinities (8, 14, and 20 ppt) on the growth performance and survival rate of sea bream (Sparus aurata) and rock samphire (Crithmum maritimum) in an experimental brackish aquaponic system. Furthermore, induction of heat shock proteins (Hsps) and phosphorylation of mitogen-activated protein kinases (MAPKs) were assessed through the sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analysis. A total number of 234 sea bream individuals were divided into nine autonomous aquaponic systems. The experiment lasted 45 days. In total, 54 individuals of rock samphire were used and were distributed into groups of six individuals per hydroponic tank using the raft method. Water quality showed stable fluctuation throughout the experiment, strongly supporting fish and plant growth performance and survival in both treatments. The results show that better growth performance for both sea bream and rock samphire (height increase) was evident in salinity 8 ppt compared to salinities 14 ppt and 20 ppt. Minimal or mild histopathological alterations were detected in gills, midgut, and liver for all three salinity groups. Exposure to different salinities modified Hsp60 and MAPKs expression in a tissue- and time-specific manner. During exposure to 8 ppt, constant Hsp60 levels and phosphorylation of MAPKs at 15 days may indicate a prominent protective role in the gills. The results show that sea bream and rock samphire can be used in brackish aquaponics systems with satisfactory growth performances, thus allowing for a range of commercial applications generating interest in their production.

Fish were kept for six weeks at three different initial stocking densities and water O 2 concentrations (low-LD, 8.5 kg/m 3 and 95-70% O 2 saturation; medium-MD, 17 kg/m 3 and 55-75% O 2 saturation; high-HD, 25 kg/m 3 and 60-45% O 2 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.

The objective of the present study was the evaluation of the effect of slurry ice, as an alternative cooling medium during harvesting and transportation, on the quality parameters (e.g., microbiological stability, sensory attributes, physicochemical changes) and shelf life of fish. The effect of seasonal variability of seawater temperature on fish preservation using the tested cooling media was also investigated. Gilthead sea bream (Sparus aurata) was slaughtered and transported in different mixtures of conventional flake ice and slurry ice for 24 h. Three mixtures of ice were tested as T: slaughtered in flake ice and transported in flake ice (control), TC: slaughtered in slurry ice and transported in flake ice, T50: slaughtered and transported in slurry ice 50%–flake ice 50%. Samples were subsequently stored isothermally at 0 °C for shelf-life evaluation. Three independent experiments were performed at three different periods, i.e., January, April, and September, referring to a sea water temperature range of 13.3–26.8 °C. Higher sea water temperatures at catching led to lower microbial growth rates and proteolytic enzyme activities and longer shelf life of refrigerated whole fish. The partial replacement of conventional flake ice with slurry ice improved the quality and extended the shelf life of fish at 0 °C by 2–7 days. The results of the study support that the use of slurry ice may enable better quality maintenance and significant shelf-life extension of whole gilthead sea bream.

Long non-coding RNAs (lncRNAs) are an emerging group of ncRNAs that can modulate gene expression at the transcriptional or translational levels. In the present work, previously published transcriptomic data were used to identify lncRNAs expressed in gilthead sea bream skeletal muscle, and their transcription levels were studied under different physiological conditions. Two hundred and ninety lncRNAs were identified and, based on transcriptomic differences between juveniles and adults, a total of seven lncRNAs showed potential to be important for muscle development. Our data suggest that the downregulation of most of the studied lncRNAs might be linked to increased myoblast proliferation, while their upregulation might be necessary for differentiation. However, with these data, as it is not possible to propose a formal mechanism to explain their effect, bioinformatic analysis suggests two possible mechanisms. First, the lncRNAs may act as sponges of myoblast proliferation inducers microRNAs (miRNAs) such as miR-206, miR-208, and miR-133 (binding energy MEF < −25.0 kcal). Secondly, lncRNA20194 had a strong predicted interaction towards the myod1 mRNA (ndG = −0.17) that, based on the positive correlation between the two genes, might promote its function. Our study represents the first characterization of lncRNAs in gilthead sea bream fast skeletal muscle and provides evidence regarding their involvement in muscle development.

The aquaculture sector has experienced rapid and important growth with the subsequent increase of feeding and nutritional issues for sustaining this activity, mainly related to the use of high quality, safe and environmentally friendly feed ingredients. The use of additives in aquafeeds has proven to be a suitable option to improve different productive indicators in farmed fish. In the present study, the effect of adding the GHRP-6 peptide, a ghrelin analog, to a commercial diet of gilthead sea bream (Sparus aurata) was studied at two proportions (100 or 500 μg/kg of feed). Both experimental diets show an increase in growth performance, as well as in feed efficiency after 97 days of experiment. The lower inclusion of GHRP-6 (100 μg/kg) results in a better aerobic metabolism, while the higher inclusion significantly increased plasma GH levels in agreement with the GH secretagogue effects of ghrelin. Similar growth outcome and differences between GHRP-6 levels in aerobic metabolism and GH stimulation suggest that improvements in culture performance by this peptide may occur through different mechanisms. Taken together, this compound can be considered as a viable dietary supplement for increasing production efficiency of sea bream aquaculture, although a better understanding of its dose-specific effects is still required.

Gill monogenean Sparicotyle chrysophrii is considered the most detrimental fish parasite to the Mediterranean aquaculture. Treatment of sparicotylosis relies on frequent gill inspections correlated with the seasonal increase in seawater temperature, application of functional feeds, and treatments with formalin baths where permitted. While the latter is bound to be banned in Europe, other synthetic anthelminthics, such as praziquantel and ivermectin, are prone to induce resistance in the parasites. Therefore, we investigated, in vitro, 14 synthetic and natural compounds against adult S. chrysophrii, developing dose–response modelsm and estimated toxicity levels at 20%, 50%, and 80% parasite mortality. Bactericidal activity of target compounds was also tested in two important aquaculture bacteria; Vibrio harveyi and V. anguillarum, while their potential host toxicity was evaluated in gilthead seabream SAF-1 cell line. Synthetic compound bithionate sodium exerted the most potent toxicity against the monogenean, no host cytotoxicity, and a medium and high potency against two bacterial pathogens. In comparison, target natural compounds were approximately 20 (cedrol) or up to 154 times (camphor) less toxic for the monogenean. Rather than completely dismissing natural compounds, we suggest that their application in combination with synthetic drugs, especially if administered in the feed, might be useful in sparicotylosis treatment.

The pressurized liquid extraction (PLE) technique was used, for the first time, to obtain protein extracts with antioxidant activity from side streams (muscle, heads, viscera, skin, and tailfins) of gilthead sea bream (Sparus aurata) in order to give added value to these underutilized matrices. Extraction conditions previously optimized for sea bass (Dicentrarchus labrax) side streams were applied. Protein recovery percentages were 22% (muscle), 33% (heads), 78% (viscera), 24% (skin), and 26% (tailfins), which represented an increase of 1.2–4.5-fold compared to control samples (extraction by stirring). The SDS-PAGE profiles revealed that PLE-assisted extraction influenced protein molecular weight distribution of the obtained extracts. PLE conditions also allowed increasing the antioxidant capacity measured by both Trolox equivalent antioxidant capacity (TEAC; 1.3–2.4 fold) and oxygen radical absorbance capacity (ORAC; 1.9–6.4) assays for all fish extracts. Inductively coupled plasma mass spectrometry (ICP-MS) and high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-qTOF-MS) were used to investigate the presence of toxic metals and mycotoxins in sea bream side streams. The levels of As, Hg, Cd, and Pb were below those established by authorities for fish muscle for human consumption (except for Cd in viscera samples). Through a nontargeted screening approach, no mycotoxins or related metabolites were detected for all sea bream side streams. This study contributes to the research on the valorization of fish processing side streams using environmentally friendly technology.

Mycobacterium pseudoshottsii, a slow-growing mycobacterium closely related to M. marinum, has been isolated only in wild fish in the United States and in Japanese fish farms to date. Here, we report cases of mortality in three farmed fish species (Dicentrarchus labrax, Sparus aurata, and Sciaenops ocellatus) caused by M. pseudoshottsii in Italy. Samples underwent necropsy, histology, and culture with pathogen identification based on PCR and sequencing of housekeeping genes (16S rRNA, hsp65, rpoB). Multifocal to coalescing granulomatous and necrotizing inflammation with acid-fast bacilli were observed in the parenchymatous organs, from which M. pseudoshottsii was isolated and identified. Phylogenetic analysis confirmed the results of gene sequencing and allowed subdivision of the isolates into three distinct groups. M. pseudoshottsii poses a potential threat for Mediterranean aquaculture. Its origin in the area under study needs to be clarified, as well as the threat to the farmed fish species.

Bisphenol A (BPA), a known endocrine disrupting chemical (EDC), was administered by diet to gilthead sea bream (Sparus aurata) in order to study its effects on the endocannabinoid system (ECS) and gonadal steroidogenesis. 2-year-old male gilthead sea bream were fed with two different concentrations of BPA (LOW at 4 and HIGH at 4000 µg/kg body weight for 21 days during the reproductive season. Exposure to 4000 µg BPA/kg bw/day (BPA HIGH) reduced sperm motility and altered the straight-line velocity (VSL) and linearity (LIN). Effects on steroidogenesis were evident, with testosterone (T) being up-regulated by both treatments and 11-ketotestosterone (11-KT) down-regulated by BPA HIGH. Plasma levels of 17β-estradiol (E2) were not affected. The Gonadosomatic Index (GSI) increased in the BPA HIGH group. Interestingly, the levels of endocannabinoids and endocannabinoid-like compounds were significantly reduced after both treatments. Unpredictably, a few changes were noticed in the expression of genes coding for ECS enzymes, while the receptors were up-regulated depending on the BPA dose. Reproductive markers in testis (leptin receptor (lepr), estrogen receptors (era, erb), progesterone receptors (pr) and the gonadotropin releasing hormone receptor (gnrhr)) were up-regulated. BPA induced the up-regulation of the hepatic genes involved in oogenesis (vitellogenin (vtg) and zona pellucida 1 (zp1)).

Insect meal derived from chironomid larvae and collected from aquatic environments was included in the feed of gilthead sea bream juveniles (75 ± 1.1 g) in a growth trial of 90 days. Three feeds, which were namely one control (L1) and two experimental diets (L2, L3), were analyzed and formulated as isonitrogenous (45%) and isolipidic (13%). In L1, the protein source was mainly soybean meal (32%), followed by fish meal (20%), wheat meal (20%), gluten corn (17%), and hemoglobin (11%). In L2, the proportion of soybean meal was increased (33.5%), followed by gluten corn (21%), wheat meal (14%), and hemoglobin (11%), whereas the fish meal source was reduced (15%) due to the inclusion of chironomids (5%). In L3, the proportion of fish meal was further reduced (8%) and that of chironomid meal was increased to 10% of the protein source. The L2 and L3 groups showed similar growth performances with respect to the L1 group. The feed conversion rate was favorable in all the groups, ranging from 1.18 (L1) to 1.22 (L3). Survival rates varied from 93.62% (L3) to 94.31% (L1). Feed palatability showed similar results for all diets. Although the inclusion of chironomid meal was used in small quantities, our results suggest a significant advantage in replacing 50% of the fish meal with the chironomid meal for growing gilthead sea bream fishes.

The molecular profiles of perchloric acid solutions extracted from the flesh of Sparus aurata fish specimens, produced according to different aquaculture systems, have been investigated. The 1H-NMR spectra of aqueous extracts are indicative of differences in the metabolite content of fish reared under different conditions that are already distinguishable at their capture, and substantially maintain the same differences in their molecular profiles after sixteen days of storage under ice. The fish metabolic profiles are studied by top-down chemometric analysis. The results of this exploratory investigation show that the fish metabolome accurately reflects the rearing conditions. The level of many metabolites co-vary with the rearing conditions and a few metabolites are quantified including glycogen (stress indicator), histidine, alanine and glycine which all display significant changes dependent on the aquaculture system and on the storage times.

Gill monogenean Sparicotyle chrysophrii is considered the most detrimental fish parasite to the Mediterranean aquaculture. Treatment of sparicotylosis relies on frequent gill inspections correlated with the seasonal increase in seawater temperature, application of functional feeds, and treatments with formalin baths where permitted. While the latter is bound to be banned in Europe, other synthetic anthelminthics, such as praziquantel and ivermectin, are prone to induce resistance in the parasites. Therefore, we investigated, in vitro, 14 synthetic and natural compounds against adult S. chrysophrii, developing dose-response modelsm and estimated toxicity levels at 20%, 50%, and 80% parasite mortality. Bactericidal activity of target compounds was also tested in two important aquaculture bacteria; Vibrio harveyi and V. anguillarum , while their potential host toxicity was evaluated in gilthead seabream SAF-1 cell line. Synthetic compound bithionate sodium exerted the most potent toxicity against the monogenean, no host cytotoxicity, and a medium and high potency against two bacterial pathogens. In comparison, target natural compounds were approximately 20 (cedrol) or up to 154 times (camphor) less toxic for the monogenean. Rather than completely dismissing natural compounds, we suggest that their application in combination with synthetic drugs, especially if administered in the feed, might be useful in sparicotylosis treatment.

Mycobacterium pseudoshottsii , a slow-growing mycobacterium closely related to M. marinum , has been isolated only in wild fish in the United States and in Japanese fish farms to date. Here, we report cases of mortality in three farmed fish species ( Dicentrarchus labrax , Sparus aurata , and Sciaenops ocellatus ) caused by M. pseudoshottsii in Italy. Samples underwent necropsy, histology, and culture with pathogen identification based on PCR and sequencing of housekeeping genes (16S rRNA, hsp65 , rpoB ). Multifocal to coalescing granulomatous and necrotizing inflammation with acid-fast bacilli were observed in the parenchymatous organs, from which M. pseudoshottsii was isolated and identified. Phylogenetic analysis confirmed the results of gene sequencing and allowed subdivision of the isolates into three distinct groups. M. pseudoshottsii poses a potential threat for Mediterranean aquaculture. Its origin in the area under study needs to be clarified, as well as the threat to the farmed fish species.