Gilthead Seabream Sparus Aurata Research Articles

Assessment of the Genetic Diversity of the Monogenean Gill Parasite Lamellodiscus echeneis (Monogenea) Infecting Wild and Cage-Reared Populations of Sparus aurata (Teleostei) from the Mediterranean Sea.

The diplectanid monogenean Lamellodiscus echeneis (Wagener, 1857) is a specific and common gill parasite of the gilthead seabream Sparus aurata Linnaeus, 1758, in the Mediterranean Sea. Few isolated molecular studies of this monogenean have been conducted, and its population structure and genetic diversity are poorly understood. This study represents the first analysis of the population genetics of L. echeneis, isolated from wild and cage-reared gilthead seabream from fifteen localities in both the Southern (Tunisia) and Northern (Italy and Spain) regions of the Mediterranean Sea, using nuclear ITS rDNA markers and a partial fragment of the mitochondrial gene cytochrome oxidase subunit I (COI). The phylogenetic trees based on the newly obtained dataset and the previously published sequences of L. echeneis corroborated the spread of only a single species throughout the Mediterranean Sea. The star-like haplotypes network, inferred by COI sequences, suggested a recent population expansion of L. echeneis. This is supported by the observed high haplotype diversity (Hd = 0.918) and low nucleotide diversity (Pi = 0.01595). Population structure-based AMOVA for two groups (the Adriatic Sea and the rest of the Mediterranean Sea) attributed 35.39% of the total variation to differences within populations, 16.63% to differences among populations within groups, and 47.99% to differences among groups. Fixation indices were significant, with a high FST value (0.64612), likely related to the divergence of the parasite populations from the Adriatic Sea and other Mediterranean regions. Phylogenetic analyses grouped all samples into the main clade corresponding to L. echeneis from several localities. This study provides insight into the genetic variation between L. echeneis populations, and did not show a clear genetic structure between populations of L. echeneis throughout Tunisian, Italian, and Spanish localities, which can be attributed to the considerable gene flow between the populations favoured by the potential for host dispersion within the Mediterranean Sea. Finally, haplotypes shared between wild and cage-reared hosts provided evidence for the potential for cross-infection between wild and farmed hosts in the Mediterranean Sea.

Polystyrene nanoplastics as an ecotoxicological hazard: cellular and transcriptomic evidences on marine and freshwater in vitro teleost models

The contamination of marine and freshwater environments by nanoplastics is considered a global threat for aquatic biota. Taking into account the most recent concentration range estimates reported globally and recognizing a knowledge gap in polystyrene nanoplastics (PS-NPs) ecotoxicology, the present work investigated the harmful effects of 20 nm and 80 nm PS-NPs, at increasing biological complexity, on the rainbow trout Oncorhynchus mykiss RTG-2 and gilthead seabream Sparus aurata SAF-1 cell lines. Twenty nm PS-NPs exerted a greater cytotoxicity than 80 nm ones and SAF-1 were approximately 4-fold more vulnerable to PS-NPs than RTG-2. The engagement of PS-NPs with plasma membranes was accompanied by discernible uptake patterns and morphological alterations along with a nuclear translocation already within a 30-min exposure. Cells were structurally damaged only by the 20 nm PS-NPs in a time-dependent manner as indicated by distinctive features of the execution phase of the apoptotic cell death mechanism such as cell shrinkage, plasma membrane blebbing, translocation of phosphatidylserine to the outer leaflet of the cell membrane and DNA fragmentation. At last, functional analyses unveiled marked transcriptional impairment at both sublethal and lethal doses of 20 nm PS-NPs, with the latter impacting the “Steroid biosynthesis”, “TGF-beta signaling pathway”, “ECM-receptor interaction”, “Focal adhesion”, “Regulation of actin cytoskeleton” and “Protein processing in endoplasmic reticulum” pathways. Overall, a distinct ecotoxicological hazard of PS-NPs at environmentally relevant concentrations was thoroughly characterized on two piscine cell lines. The effects were demonstrated to depend on size, exposure time and model, emphasizing the need for a comparative evaluation of endpoints between freshwater and marine ecosystems.

Subcellular effects and lipid metabolism alterations in the gilthead seabream Sparus aurata fed on ovatoxins-contaminated mussels.

The marine microalgae Ostreopsis cf. ovata are a well-known producer of palytoxin (PlTXs) analogues, i.e. ovatoxins (OVTXs) among others, which arouse concern for animal and human health. Both in field and laboratory studies, presence of OVTXs, detected in species directly feeding on O. cf. ovata, was frequently correlated with impairment on organisms’ physiology, development and behaviour, while similar knowledge is still lacking for animals feeding on contaminated preys. In this study, transfer and toxicity of OVTXs were evaluated in an exposure experiment, in which gilthead seabream Sparus aurata was fed with bivalve mussel Mytilus galloprovincialis, contaminated by a toxic strain of O. cf. ovata. Mussels exposed to O. cf. ovata for 21 days accumulated meanly 188 ± 13 μg/kg OVTXs in the whole tissues. Seabreams fed with OVTX-contaminated mussels started to reject the food after 6 days of contaminated diet. Although no detectable levels of OVTXs were measured in muscle, liver, gills and gastro-intestinal tracts, the OVTX-enriched diet induced alterations of lipid metabolism in seabreams livers, displaying a decreased content of total lipid and fatty acid, together with overexpression of fatty acid biosynthetic genes, downregulation of β-oxidation genes and modulation of several genes related to lipid transport and regulation. Results from this study would suggest the hypothesis that OVTXs produced by O. cf. ovata may not be subject to bioaccumulation in fish fed on contaminated preys, being however responsible of significant biological effects, with important implications for human consumption of seafood products.

Tissue explants as tools for studying the epigenetic modulation of the GH-IGF-I axis in farmed fish.

Somatic growth in vertebrates is mainly controlled by the growth hormone (GH)/insulin-like growth factor I (IGF-I) axis. The role of epigenetic mechanisms in regulating this axis in fish is far from being understood. This work aimed to optimize and evaluate the use of short-term culture of pituitary and liver explants from a farmed fish, the gilthead seabream Sparus aurata, for studying epigenetic mechanisms involved in GH/IGF-I axis regulation. Our results on viability, structure, proliferation, and functionality of explants support their use in short-term assays. Pituitary explants showed no variation in gh expression after exposure to the DNA methylation inhibitor decitabine (5-Aza-2'-deoxycytidine; DAC), despite responding to DAC by changing dnmt3bb and tet1 expression, and TET activity, producing an increase in overall DNA hydroxymethylation. Conversely, in liver explants, DAC had no effects on dnmt s and tet s expression or activity, but modified the expression of genes from the GH-IGF-I axis. In particular, the expression of igfbp2a was increased and that of igfbp4, ghri and ghrii was decreased by DAC as well as by genistein, which is suggestive of impaired growth. While incubation of liver explants with S-adenosylmethionine (SAM) produced no clear effects, it is proposed that nutrients must ensure the methylation milieu within the liver in the fish to sustain proper growth, which need further in vivo verification. Pituitary and liver explants from S. aurata can be further used as described herein for the screening of inhibitors or activators of epigenetic regulators, as well as for assessing epigenetic mechanisms behind GH-IGF-I variation in farmed fish.

Weight prediction of intensively reared gilthead seabream Sparus aurata from morphometric traits measured in images

The objective of this study was to establish an accurate body weight (BW) prediction model for gilthead seabream Sparus aurata of 50–1000 g. Three thousand three hundred twelve (3312) fish were individually weighed and photographed. Traits measured from the images were total body length (TBL), fork body length (FBL), standard body length (SBL), body height (BH), head length (HL), eye diameter (ED), body area (BA, without fins), head area (HA), and eye area (EA). SBL, BH, BA, BA/SBL, and BA/BH showed a strong association with BW (correlation coefficients, r: 0.96–0.99). These traits were selected to proceed with the regression analysis. Simple, multiple linear, and 2nd-order polynomial regressions were applied to the whole data set and three BW subgroups of interest during gilthead seabream rearing (i.e., 50–100 g, 100–500 g, 500–1000 g). The prediction of BW from the whole data set was more accurate than from each BW subgroup. The models with the highest coefficient of determination (R2) and the lowest errors (mean absolute percentage error, MAPE) were either the power regression of BW with BA (R2: 99.0%, MAPE: 5.8%) or the multiple linear regression of BW with SBL, BA, BA/SBL, and BA/BH (R2: 98.6%, MAPE: 5.1%) as predictors. The accuracy of the two models is considered quite similar, and for reasons of simplicity, the power regression is advantageous, requiring only one trait to be measured (BA). The models identified in the present study can help to further develop the accuracy of machine vision-based systems for gilthead seabream BW measurement.

Functional Characterization of the Almstn2 Gene and Its Association with Growth Traits in the Yellowfin Seabream Acanthopagrus latus (Hottuyn, 1782).

Myostatin (mstn), also known as GDF8, is a growth and differentiation factor of the transforming growth factor-β (TGF-β) superfamily and plays a key inhibitory effect in the regulation of skeletal muscle development and growth in vertebrates. In the present study, to comprehend the role of the mstn2 gene of the yellowfin seabream Acanthopagrus latus (Almstn2b), the genomic sequence of Almstn2b is 2359 bp, which encodes 360 amino acids and is composed of three exons and two introns, was obtained. Two typical regions, a TGF-β propeptide and TGF-β domain, constitute Almstn2b. The topology indicated that Almstn2 was grouped together with other Perciformes, such as the gilthead seabream Sparus aurata. Moreover, Almstn2b was mainly expressed in the brain, fins, and spleen. Furthermore, five SNPs, one in the exons and four in the introns, were identified in the Almstn2b gene. The allele and genotype frequencies of SNP-Almstn2b +1885 A/G were significantly related to the total weight, interorbital distance, stem length, tail length, caudal length, caudal height, body length, and total length (p < 0.05). The allele and genotype frequencies of SNP-Almstn2b +1888 A/G were significantly related to the weight, interorbital distance, long head behind the eyes, body height, tail length, caudal length, and body length. Additionally, the relationship between the SNP-Almstn2b +1915 A/G locus and weight and long head behind the eyes was significant (p < 0.05). Furthermore, the other two SNPs were not significantly associated with any traits. Thus, the SNPs identified in this study could be utilized as candidate SNPs for breeding and marker-assisted selection in A. latus.

Residency, home range and inter-annual fidelity of three coastal fish species in a Mediterranean coastal lagoon

In the Gulf of Lion (NW Mediterranean), fish species such as gilthead sea bream Sparus aurata, European seabass Dicentrarchus labrax, and salema Sarpa salpa show seasonal occupation of coastal lagoons, presumably as feeding grounds during their adult life stage. The role of the lagoons in these species' life cycles remains unknown, particularly with respect to their residency, space-use, and inter-annual fidelity. Using acoustic telemetry, the movements of 72 seabream, 58 seabass, and 81 salema were monitored over four years within Prévost Lagoon (Hérault Department, Occitania Region), to characterize (1) the main seasonal patterns of space use inside the lagoon and (2) their annual migrations between the lagoon and the sea. Overall, all three species were highly resident in the lagoon during the spring/summer foraging season; seabass was the only species that also displayed high residency to the lagoon throughout the winter breeding season. The three species showed differences in their space use, although they all mainly inhabited the deep lagoon centre and adjacent shellfish farms, with very small individual home ranges (mean ± SD, 0.12 ± 0.06 km2 over all species). All species showed some inter-annual fidelity to the lagoon (>43% at minimum for seabream) although these fidelity rates were probably underestimated due to fishing mortality, which is probably high during the winter breeding season. Overall, this study reveals that coastal lagoons are key foraging habitats for these species in the Gulf of Lion. The high residency and inter-annual fidelity suggest that any increase of anthropogenic pressure within the lagoon could negatively impact these fish populations. Therefore, protection of such productive habitats could be beneficial for long-term management of emblematic coastal species and the fisheries that they support.

Farming Sparus aurata (Teleostei: Sparidae) in marsh ponds: trophic characterization and trace metal accumulation

Considering the overexploitation of fishing on most of the world coasts, the ingestion of fish and shellfish will depend mostly on aquaculture production. Since intensive mariculture usually involves environmental impact, developing sustainable cultures is a priority. In this sense, salt marshes can provide ecosystem services and incorporate both conservation and extensive aquaculture activities. In the present study we compared gilthead seabream Sparus aurata Linnaeus, 1758 cultured in extensive and semi-intensive marsh ponds with wild conspecifics from surrounding coastal areas, using trophic characterization (diet analysis and stable isotopes) and trace metal accumulation. Stomach content analysis revealed different feeding habits among gilthead seabream from different origin. Although wild specimens had the most diverse diet, results of stable isotopes showed that extensive diet had the wider isotopic niche and revealed the highest similarities between wild and extensively cultured gilthead seabream. A similar trace element signature was also measured in wild and extensive culture, whereas the semi-intensive culture showed different concentrations for several elements. Cr, Fe and Mn showed the highest concentrations in semi-intensive cultured fish, while As and Zn showed the lowest values in this group. In any case, average values measured in both extensive and semi-intensive culture were, in general terms, below the hazardous limits provided by the standards recommended for trace metals by national and international regulations. Therefore, marsh ponds provide a suitable environment where the cultured fish, especially extensive, should be promoted.

Citizen Science and Expert Judgement: A Cost-Efficient Combination to Monitor and Assess the Invasiveness of Non-Indigenous Fish Escapees

Mapping the distribution and evaluating the impacts of marine non-indigenous species (NIS) are two fundamental tasks for management purposes, yet they are often time consuming and expensive. This case study focuses on the NIS gilthead seabream Sparus aurata escaped from offshore farms in Madeira Island in order to test an innovative, cost-efficient combined approach to risk assessment and georeferenced dispersal data collection. Species invasiveness was screened using the Aquatic Species Invasiveness Screening Kit (AS-ISK), and revealed a high invasion risk. Occurrences of S. aurata were assessed involving citizens in GIS participatory mapping and data from recreational fishing contests. A probability map showed that S. aurata is well dispersed around Madeira Island. This assessment proved to be a cost-efficient early warning method for detecting NIS dispersal, highlighting the urgent need for additional surveys that should search for sexually mature individuals and assess the direct and indirect impacts in the native ecosystem.

Occurrence of two myxosporean parasites in the gall bladder of white seabream Diplodus sargus (L.) (Teleostei, Sparidae), with the morphological and molecular description of Ceratomyxa sargus n. sp.

Myxosporeans are widespread cnidarian parasites that usually parasitize fish as part of their complex life cycle, thus constituting a potential threat for the aquaculture industry. White seabream Diplodus sargus (L.) is a commercially valuable sparid fish reared in Southern European aquacultures. Nonetheless, knowledge on myxosporean infections potentially harming the sustainable production of this fish is extremely limited. In this study, a myxosporean survey was conducted on D. sargus specimens reared in two Southern Portuguese fish farms. Two coelozoic myxosporeans were detected infecting the gall bladder, and are herein reported based on microscopic and molecular procedures: Ceratomyxa sargus n. sp. and Zschokkella auratis Rocha et al., 2013, previously described from reared stocks of gilthead seabream Sparus aurata in the same geographic locality. Ceratomyxa sargus n. sp. is the 12th species of the genus to be reported from Southern European sparids, reinforcing a substantial radiation of Ceratomyxa within this fish family and geographic region. SSU rRNA-based Bayesian inference and maximum likelihood analyses revealed C. sargus n. sp. positioned separately from other sparid-infecting Ceratomyxa spp. reported from Southern European countries, demonstrating that this species does not share a more immediate common ancestor with its closest relatives based on host affinity and geography. The recognition of a novel sparid-infecting lineage within the Ceratomyxa clade strengthens the contention that this genus entered sparid fish multiple times, namely in the Southern European region. The identification of Zschokkella auratis infections in D. sargus demonstrates that host shift has occurred among sparids reared in the Southern Portuguese coast. This agrees with the broad host specificity that is usually attributed to this genus, and that may be suggested to be the outcome of the capacity of the Zschokkella morphotype to undergo host shift/switch based on our findings and the limited molecular data available for this genus. Thus, a better understanding of Zschokkella host-associated diversification and dispersal mechanisms requires the increasing availability of molecular data from infections of the same species occurring in multiple hosts and geographical locations.

Morphological and molecular characterization of Encotyllabe vallei Monticelli, 1907 (Monopisthocotylea, Monogenea) from the gilthead seabream Sparus aurata Linnaeus (Teleostei, Sparidae) from the southwestern Mediterranean and notes on host specificity of the genus Encotyllabe Diesing, 1850.

Incomplete original descriptions, the unavailability or poor conditions of specimens and the lack of detailed redescriptions have caused the validity of several species of the genus Encotyllabe Diesing, 1850 to be questioned. To date, seven of the recognized species were described upon one or two specimens, hindering study of intraspecific variations. This was made worse by considering few morphoanatomical differences sufficient to erect new species. Among Encotyllabe spp. occurring in Mediterranean waters, E. vallei was first described from the gilt-head bream Sparus aurata (Sparidae) off Italy. Although beautifully illustrated for a paper from that century, morphometric data for E. vallei from the type-host S. aurata remain unavailable. Previous records of E. vallei provided either morphometrical or molecular data, and its validity was questioned. We provide a redescription of E. vallei based on newly collected specimens from the S. aurata from the southwestern Mediterranean (off Algeria) using integrative taxonomy. Analysis of cox1 sequences of E. vallei from S. aurata, compared to sequences from other sparid hosts, mainly Pagellus bogaraveo, revealed a divergence not exceeding 2%, suggesting a stenoxenic specificity for this monogenean. Given that P. bogaraveo is the type-host for Encotyllabe pagelli, we were tempted to suggest a synonymy between E. vallei and E. pagelli. We refrained from doing so because E. pagelli was first described from the Atlantic coast off Brest, France. Morphological data for Encotyllabe from P. bogaraveo are warranted assessing the host specificity of E. vallei and whether there might be a species complex within individual sparid fish species.

Bottlenose dolphins share fish farm areas while maintaining sexual segregation: Investigating group memberships through spatially and temporally explicit parameters

Abstract Group membership is a key attribute of animal societies and central to the study of social structure in several taxa. However, social structure analyses are sensitive to the way data are collected and associations defined. In this study, a time–space method was used to investigate the social structure of common bottlenose dolphins Tursiops truncatus observed and photographed across 7 years in the semi‐enclosed Gulf of Corinth, Greece. Instead of adopting traditional group definitions, individuals were considered as being members of the same group if photographed within a specific time and space window. This approach can be applied post hoc across studies and can offer advantages under challenging sampling conditions (e.g. when dealing with groups spread over vast areas or when group membership is otherwise hard to assess). Dolphins were mostly found around coastal cage aquaculture facilities farming European sea bass Dicentrarchus labrax and gilthead seabream Sparus aurata. Dolphins formed clusters largely or entirely composed of individuals of the same sex, suggestive of sex‐based homophily. Habitat partitioning was not detected: there was substantial spatial overlap among dolphin clusters, with all individuals using a relatively small area in the northern portion of the Gulf, where most of the productive fish farms were located. Associations between females were stronger than those between males, and daughters tended to stay in the group of their mothers. Sex‐based social clustering may allow females and calves to limit interactions with potentially aggressive males, while individuals of both sexes benefit from prey concentrated around fish farms. Adaptation to foraging around farms can result in trade‐offs between the costs and benefits of nourishment and social interaction. This may have both positive or negative effects on the animals that should be considered in the context of ensuring their favourable conservation status.

Genetic variation of Sparicotyle chrysophrii (Monogenea: Microcotylidae) from the gilthead sea bream Sparus aurata (Teleostei: Sparidae) in the Mediterranean Sea.

The gill monogenean Sparicotyle chrysophrii (Van Beneden & Hesse, 1863) Mamaev, 1984 is a specific and common parasite of wild and cultured gilthead sea bream Sparus aurata Linnaeus, 1758, able to cause disease and mortality in aquaculture systems. Few molecular studies have been carried out on this monogenean, and its population structure and genetic diversity are barely known. This study provides the first contribution to the population genetic variation of S. chrysophrii, based on two molecular markers-the structural ribosomal RNA (rRNA) for the large subunit (28S) and the cytochrome c oxidase subunit I (COI) gene. Samples were collected from the gills of farmed and wild S. aurata from Italy and the Spanish Mediterranean. The analysis included previously published sequences. The 28S rDNA analysis was consistent with previous studies of specimens isolated from S. aurata and confirmed the presence of only one species on the gills of this host in the Mediterranean Sea. The COI sequences analysis suggested that the samples isolated in a previous study from a different host species, wild Boops boops (Linnaeus, 1758) in the Adriatic Sea, may represent a new undescribed sister species of S. chrysophrii. The low nucleotide diversity of S. chrysophrii isolated only from S. aurata versus the high haplotype diversity revealed small differences between haplotypes. The haplotypes shared between wild and farmed hosts from Spain provided the first molecular evidence of the possible transfer of S. chrysophrii between wild and farmed populations of S. aurata. The mtDNA COI analysis did not show a clear genetic structure, probably the result of several factors including coevolution, wild and farmed host interactions, and host population structure in space and time.

Pelvetia canaliculata as an Aquafeed Supplement for Gilthead Seabream Sparus aurata: A Biorefinery Approach for Seaweed Biomass Valorisation

For sustainable and economically viable aquaculture, it is necessary to search for alternative sources of aquafeeds. Algae have been studied because of their bioactive compounds with several activities such as antioxidants. The direct incorporation of the macroalgae Pelvetia canaliculata in sunflower oil to increase oxidative stability and biological value results in waste with high nutritional value that may be used as an ingredient in aquaculture feed. This study aimed to evaluate the effect of incorporating algae powder (PEL 1%, PEL 10%) and algae waste obtained after sunflower oil supplementation (WO 1%, WO 10%) in aquafeeds for gilthead seabream. We studied the growth performance, haematological profile, oxidative stress and metabolic parameters, and intestine histomorphology. Experimental diets did not influence growth performance or somatic indexes, and barely affected the haematological profile. Catalase showed higher activity in seabream fed with PEL10 than with control diet. Total glutathione had a higher activity in fish fed with both WO diets. Plasmatic levels of cholesterol were higher in PEL1 and WO10. Triglyceride levels were higher in WO1 and total lipids were higher in both WO diets. The histomorphology of the intestine was slightly modulated by experimental diets but was not affected negatively. In general, supplementation with Pelvetia powder and algal waste oil may be used as an aquafeed for gilthead seabream according to the results obtained for growth, some haematological parameters, catalase and total glutathione, intestinal villi length, and the number of total and acid goblet cells.

Sparus aurata and Lates calcarifer skin microbiota under healthy and diseased conditions in UV and non-UV treated water

BackgroundThe welfare of farmed fish is influenced by numerous environmental and management factors. Fish skin is an important site for immunity and a major route by which infections are acquired. The objective of this study was to characterize bacterial composition variability on skin of healthy, diseased, and recovered Gilthead Seabream (Sparus aurata) and Barramundi (Lates calcarifer). S. aurata, which are highly sensitive to gram-negative bacteria, were challenged with Vibrio harveyi. In addition, and to provide a wider range of infections, both fish species (S. aurata and L. calcarifer) were infected with gram-positive Streptococcus iniae, to compare the response of the highly sensitive L. calcarifer to that of the more resistant S. aurata. All experiments also compared microbial communities found on skin of fish reared in UV (a general practice used in aquaculture) and non-UV treated water tanks.ResultsSkin swab samples were taken from different areas of the fish (lateral lines, abdomen and gills) prior to controlled infection, and 24, 48 and 72 h, 5 days, one week and one-month post-infection. Fish skin microbial communities were determined using Illumina iSeq100 16S rDNA for bacterial sequencing. The results showed that naturally present bacterial composition is similar on all sampled fish skin sites prior to infection, but the controlled infections (T1 24 h post infection) altered the bacterial communities found on fish skin. Moreover, when the naturally occurring skin microbiota did not quickly recover, fish mortality was common following T1 (24 h post infection). We further confirmed the differences in bacterial communities found on skin and in the water of fish reared in non-UV and UV treated water under healthy and diseased conditions.ConclusionsOur experimental findings shed light on the fish skin microbiota in relation to fish survival (in diseased and healthy conditions). The results can be harnessed to provide management tools for commercial fish farmers; predicting and preventing fish diseases can increase fish health, welfare, and enhance commercial fish yields.

Selection for high growth improves reproductive performance of gilthead seabream Sparus aurata under mass spawning conditions, regardless of the dietary lipid source

Genetic selection programmes in gilthead seabream mainly focus on traits related to growth, disease resistance, skeletal anomalies, or fillet quality. However, the effect of selection for growth on the reproductive performance of seabream broodstock has not received much attention. The present study aimed to determine the effect of selection for growth traits, high (HG) or low (LG) growth, and broodstock feeding with fish oil (FO diet) or rapeseed oil (RO diet) as main lipid sources, on reproductive performance of gilthead seabream. For the first part of the spawning season (Phase I) HG and LG broodstock were fed a commercial diet and the HG broodstock produced a higher number of larvae and higher viable eggs, hatching and larval survival rates than LG broodstock, affecting egg fatty acid profiles. For the second part of the study (Phase II) broodstock were fed one of the two diets containing FO or RO. Fecundity in terms of viable eggs, hatchlings, and larvae produced, as well as fertilization rates, were improved in HG broodstock. Some fatty acids such as 18:0, 20:2n-6, 20:3n-3 or EPA/ARA were also affected by the growth selection. According to the two-way ANOVA analysis, feeding the RO diet did not significantly affect fecundity parameters, but slightly reduced fertilization and hatching rates in HG broodstock. Nevertheless, HG broodstock showed better spawning quality parameters than LG broodstock, even when they were fed the RO diet. Egg fatty acid profiles reflected diet composition, although DHA contents were not affected. In conclusion, broodstock selected for high growth had a positive effect on broodstock performance, and FO replacement by RO did not markedly affect reproduction providing that fatty acid contents were sufficient to fulfill the essential fatty acid requirements of gilthead seabream broodstock.

Effect of Dietary Plant Feedstuffs and Protein/Carbohydrate Ratio on Gilthead Seabream (Sparus aurata) Gut Health and Functionality

This study investigated, for the first time, the integrated effects of dietary protein source and protein/carbohydrate (P/CH) ratio on gilthead seabream gut histomorphology, microbiota composition, digestive enzymes activity, and immunological and oxidative stress-related gene expressions. Four isolipidic diets: two fishmeal-based (FM) and two plant feedstuff (PF)-based diets, with P/CH ratios of 50/10 or 40/20 each (FM-P50/CH10; FM-P40/CH20; PF-P50/CH10; PF-P40/CH20), were tested. PF-based diets lead to more histomorphological alterations than FM-based diets. P/CH ratio had no relevant effect on gut histomorphology. Gut mucosa of fish fed PF-based diets presented a higher number of operational taxonomic units, and richness and diversity indices, while the P/CH ratio did not affect those parameters. The α-amylase activity was lower in fish fed with PF-based diets and in fish fed the P40/CH20 diets. Regarding the immune-related genes, only cyclooxygenase-2 was affected, being higher in fish fed the P50/CH10 diets than the P40/CH20 diets. Fish fed the FM-based diets presented higher expression of glutathione reductase and glutathione peroxidase, while fish fed the P50/CH10 diet had higher expression of superoxide dismutase. In conclusion, PF-based diets can compromise gut absorptive and digestive metabolism, but decreasing the dietary P/CH ratio had little effect on the parameters measured.

Feed Supplementation with the GHRP-6 Peptide, a Ghrelin Analog, Improves Feed Intake, Growth Performance and Aerobic Metabolism in the Gilthead Sea Bream Sparus aurata

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.

Tenebrio molitor larvae meal inclusion affects hepatic proteome and apoptosis and/or autophagy of three farmed fish species

Herein, the effect of dietary inclusion of insect (Tenebrio molitor) meal on hepatic pathways of apoptosis and autophagy in three farmed fish species, gilthead seabream (Sparus aurata), European seabass (Dicentrarchus labrax) and rainbow trout (Oncorhynchus mykiss), fed diets at 25%, 50% and 60% insect meal inclusion levels respectively, was investigated. Hepatic proteome was examined by liver protein profiles from the three fish species, obtained by two-dimensional gel electrophoresis. Although cellular stress was evident in the three teleost species following insect meal, inclusion by T. molitor, D. labrax and O. mykiss suppressed apoptosis through induction of hepatic autophagy, while in S. aurata both cellular procedures were activated. Protein abundance showed that a total of 30, 81 and 74 spots were altered significantly in seabream, European seabass and rainbow trout, respectively. Insect meal inclusion resulted in individual protein abundance changes, with less number of proteins altered in gilthead seabream compared to European seabass and rainbow trout. This is the first study demonstrating that insect meal in fish diets is causing changes in liver protein abundances. However, a species-specific response both in the above mentioned bioindicators, indicates the need to strategically manage fish meal replacement in fish diets per species.

Behavioural Stress Propagation in Benthic Invertebrates Caused by Acute pH Drop-Induced Metabolites

Studies on pH stress in marine animals typically focus on direct or species-specific aspects. We here test the hypothesis that a drop to pH = 7.6 indirectly affects the intra- and interspecific interactions of benthic invertebrates by means of chemical communication. We recorded fitness-relevant behaviours of small hermit crabs Diogenes pugilator, green shore crabs Carcinus maenas, and harbour ragworms Hediste diversicolor in response to short-term pH drop, and to putative stress metabolites released by conspecifics or gilt-head sea bream Sparus aurata during 30 min of acute pH drop. Not only did acute pH drop itself impair time to find a food cue in small hermit crabs and burrowing in harbour ragworms, but similar effects were observed under exposure to pH drop-induced stress metabolites. Stress metabolites from S. aurata, but not its regular control metabolites, also induced avoidance responses in all recipient species. Here, we confirm that a short-term abrupt pH drop, an abiotic stressor, has the capacity to trigger the release of metabolites which induce behavioural responses in conspecific and heterospecific individuals, which can be interpreted as a behavioural cost. Our findings that stress responses can be indirectly propagated through means of chemical communication warrant further research to confirm the effect size of the behavioural impairments caused by stress metabolites and to characterise their chemical nature.