Publications by authors named "Jaume Pérez-Sánchez"

90 Publications

Physiological Effects of Water Flow Induced Swimming Exercise in Seabream .

Front Physiol 2020 7;11:610049. Epub 2020 Dec 7.

Future Genomics Technologies B.V., Leiden, Netherlands.

A longer on-land rearing period of Gilthead seabream before transfer to sea-cages would allow the farmer to benefit from exercise-enhanced growth, resilience, and robustness as induced by increasing water flow in the tanks. In this study, the physiological effects of flow-conditioning were investigated by subjecting large groups of experimental fish to minimal flow or to flow regimes inducing swimming exercise at 1 or 2 body length (BL) s for a period of 8 months (February-October) in 1,500 L tanks. Fish representing the three treatment groups were then used for: (1) a stress challenge netting test and plasma cortisol measurement (baseline, peaking, and recovery levels), (2) blood plasma measurements of glucose, triglycerides, lactate, cholesterol, growth hormone (GH), and insulin-like growth factor 1 (IGF1), and (3) heart and muscle gene expression of the GH and IGF1 receptors and the muscle transcriptome by deep RNA sequencing (RNAseq). Fish size after 8 months of flow conditioning was 92 ± 27 g body weight (BW) for fish under minimal flow, 106 ± 24 g BW (+15%) at 1 BL s, and 125 ± 27 g BW (+36%) at 2 BL s. Flow conditioning at 1 BL s provided optimal conditions for growth and uniformity, but also stress (lowest baseline plasma cortisol), robustness (higher condition factor and larger hearts), and energy mobilization (increased plasma glucose). Although flow enhanced growth linearly with swimming speed, also the percentage of lordotic fish increased with exercise, particularly high for swimming at 2 BL s. The absence of important differences in plasma GH and IGF1, and expression levels of their receptors in heart and white skeletal muscle, indicated that other factors may be involved in growth enhancement. RNAseq of the white skeletal muscle showed upregulated expression of genes involved in muscle contraction, muscle development and its molecular regulation, and immune genes that may play a role in the muscle repair mechanism. An exercise regime of swimming at 1 BL s can be considered as optimal for farming robust seabream although the increase of skeletal deformities should be avoided.
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http://dx.doi.org/10.3389/fphys.2020.610049DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7750471PMC
December 2020

Genetic selection for growth drives differences in intestinal microbiota composition and parasite disease resistance in gilthead sea bream.

Microbiome 2020 11 23;8(1):168. Epub 2020 Nov 23.

Nutrigenomics and Fish Endocrinology Group, Instituto de Acuicultura Torre de la Sal (IATS-CSIC), Castellón, Spain.

Background: The key effects of intestinal microbiota in animal health have led to an increasing interest in manipulating these bacterial populations to improve animal welfare. The aquaculture sector is no exception and in the last years, many studies have described these populations in different fish species. However, this is not an easy task, as intestinal microbiota is composed of very dynamic populations that are influenced by different factors, such as diet, environment, host age, and genetics. In the current study, we aimed to determine whether the genetic background of gilthead sea bream (Sparus aurata) influences the intestinal microbial composition, how these bacterial populations are modulated by dietary changes, and the effect of selection by growth on intestinal disease resistance. To that aim, three different groups of five families of gilthead sea bream that were selected during two generations for fast, intermediate, or slow growth (F3 generation) were kept together in the same open-flow tanks and fed a control or a well-balanced plant-based diet during 9 months. Six animals per family and dietary treatment were sacrificed and the adherent bacteria from the anterior intestinal portion were sequenced. In parallel, fish of the fast- and slow-growth groups were infected with the intestinal parasite Enteromyxum leei and the disease signs, prevalence, intensity, and parasite abundance were evaluated.

Results: No differences were detected in alpha diversity indexes among families, and the core bacterial architecture was the prototypical composition of gilthead sea bream intestinal microbiota, indicating no dysbiosis in any of the groups. The plant-based diet significantly changed the microbiota in the intermediate- and slow-growth families, with a much lower effect on the fast-growth group. Interestingly, the smaller changes detected in the fast-growth families potentially accounted for more changes at the metabolic level when compared with the other families. Upon parasitic infection, the fast-growth group showed significantly lower disease signs and parasite intensity and abundance than the slow-growth animals.

Conclusions: These results show a clear genome-metagenome interaction indicating that the fast-growth families harbor a microbiota that is more flexible upon dietary changes. These animals also showed a better ability to cope with intestinal infections. Video Abstract.
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http://dx.doi.org/10.1186/s40168-020-00922-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7686744PMC
November 2020

Effects of genetics and early-life mild hypoxia on size variation in farmed gilthead sea bream (Sparus aurata).

Fish Physiol Biochem 2021 Feb 13;47(1):121-133. Epub 2020 Nov 13.

Nutrigenomics and Fish Growth Endocrinology, Institute of Aquaculture Torre de la Sal (IATS-CSIC), 12595, Ribera de Cabanes, Castellón, Spain.

The present study evaluated, in an 18-month gilthead sea bream trial, the time course effects of genetics on individual size variation and growth compensation processes in families selected by heritable growth in the PROGENSA breeding program. Families categorized as fast, intermediate, and slow growing had different growth trajectories with a more continuous growth in fast growth families. This feature was coincident with a reduced size variation at the beginning of the trial that clustered together the half-sib families sharing the same father. Regression analysis evidenced that the magnitude of compensatory growth was proportional to the initial size variation with no rescaling of families at this stage. By contrast, the finishing growth depensation process can mask, at least partially, the previous size convergence. This reflects the different contribution across the production cycle of genetics in growth. How early-life experiences affect growth compensation at juvenile stages was also evaluated in a separate cohort, and intriguingly, a first mild-hypoxia pulse at 60-81 days post-hatching (dph) increased survival rates by 10%, preventing growth impairment when fish were exposed to a second hypoxia episode (112-127 dph). The early hypoxia experience did not have a negative impact on growth compensatory processes at juvenile stages. By contrast, a diminished capacity for growth compensation was found with repeated or late hypoxia experiences. All this reinforces the use of size variation as a main criterion for improving intensive fish farming and selective breeding.
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http://dx.doi.org/10.1007/s10695-020-00899-1DOI Listing
February 2021

SeqEditor: an application for primer design and sequence analysis with or without GTF/GFF files.

Bioinformatics 2020 Oct 18. Epub 2020 Oct 18.

Biotechvana, Parc Científic Universitat de València, Valencia, Spain.

SeqEditor is a cross-platform desktop application for the analysis of nucleotide and protein sequences. It is managed through a Graphical User Interface (GIU) and can work either as a graphical sequence browser or as a fasta task manager for multi-fasta files. SeqEditor has been optimized for the management of large sequences, such as contigs, scaffolds or even chromosomes, and includes a GTF/GFF viewer to visualize and manage annotation files. In turn, this allows for content mining from reference genomes and transcriptomes with similar efficiency to that of command line tools. SeqEditor also incorporates a set of tools for singleplex and multiplex PCR primer design and pooling that uses a newly optimized and validated search strategy for target and species-specific primers. All these features make SeqEditor a flexible application that can be used to analyses complex sequences, design primers in PCR assays oriented for diagnosis, and/or manage, edit, and personalize reference sequence data sets.

Availability: SeqEditor was developed in Java using Eclipse Rich Client Platform and is publicly available at https://gpro.biotechvana.com/download/SeqEditor as binaries for Windows, Linux and Mac OS. The user manual and tutorials are available online at https://gpro.biotechvana.com/tool/seqeditor/manual.

Supplementary Information: Supplementary data are available at Bioinformatics online.
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http://dx.doi.org/10.1093/bioinformatics/btaa903DOI Listing
October 2020

Local DNA methylation helps to regulate muscle sirtuin 1 gene expression across seasons and advancing age in gilthead sea bream ().

Front Zool 2020 15;17:15. Epub 2020 May 15.

Nutrigenomics and Fish Growth Endocrinology Group, Institute of Aquaculture Torre de la Sal, IATS-CSIC, 12595 Ribera de Cabanes s/n, Castellón, Spain.

Background: Sirtuins (SIRTs) are master regulators of metabolism, and their expression patterns in gilthead sea bream (GSB) reveal different tissue metabolic capabilities and changes in energy status. Since little is known about their transcriptional regulation, the aim of this work was to study for the first time in fish the effect of age and season on gene expression, correlating expression patterns with local changes in DNA methylation in liver and white skeletal muscle (WSM).

Methods: Gene organization of the seven was analyzed by BLAT searches in the IATS-CSIC genomic database (www.nutrigroup-iats.org/seabreamdb/). The presence of CpG islands (CGIs) was mapped by means of MethPrimer software. DNA methylation analyses were performed by bisulfite pyrosequencing. A PCR array was designed for the simultaneous gene expression profiling of and related markers (, , , and ) in the liver and WSM of one- and three-year-old fish during winter and summer.

Results: The occurrence of CGIs was evidenced in the and promoters. This latter CGI remained hypomethylated regardless of tissue, age and season. Conversely, DNA methylation of at certain CpG positions within the promoter varied with age and season in the WSM. Among them, changes at several SP1 binding sites were negatively correlated with the decrease in expression in summer and in younger fish. Changes in regulation match well with variations in feed intake and energy metabolism, as judged by the concurrent changes in the analyzed markers. This was supported by discriminant analyses, which identified as a highly responsive element to age- and season-mediated changes in energy metabolism in WSM.

Conclusions: The gene organization of is highly conserved in vertebrates. GSB family members have CGI- and non-CGI promoters, and the presence of CGIs at the promoter agrees with its ubiquitous expression. Gene expression analyses support that , especially , are reliable markers of age- and season-dependent changes in energy metabolism. Correlation analyses suggest the involvement of DNA methylation in the regulation of expression, but the low methylation levels suggest the contribution of other putative mechanisms in the transcriptional regulation of .
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http://dx.doi.org/10.1186/s12983-020-00361-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7227224PMC
May 2020

No transfer of the non-regulated mycotoxins, beauvericin and enniatins, from feeds to farmed fish reared on plant-based diets.

Food Chem 2020 Apr 10;323:126773. Epub 2020 Apr 10.

Institute of Aquaculture, University of Stirling, Stirling FK9 4LA, United Kingdom.

Concern about the risk of exposure to emerging plant-derived mycotoxins such as beauvericin and enniatins has been addressed by the European Commission who requested the European Food Safety Authority for a scientific opinion on their risk to human and animal health. The studied mycotoxins were found in feeds with enniatin B and beauvericin at average concentrations of 19.9 μg/kg and 30 μg/kg, respectively. In all cases, concentrations of all the mycotoxins analyzed were below quantification limits (<0.1 μg/kg) in fish samples (n = 82). The present work provides comprehensive and traceable data of emerging mycotoxins in plant-based aquafeeds and fish reared on the feeds, responding to increasing concerns about safety of farmed fish fed on sustainable feeds. On the basis of data reported, there was no transfer of the emerging mycotoxins, beauvericin and enniatins, from feeds to fish and so, no risk for human consumption.
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http://dx.doi.org/10.1016/j.foodchem.2020.126773DOI Listing
April 2020

Effects of Dietary Lipid Composition and Fatty Acid Desaturase 2 Expression in Broodstock Gilthead Sea Bream on Lipid Metabolism-Related Genes and Methylation of the Gene Promoter in Their Offspring.

Int J Mol Sci 2019 Dec 11;20(24). Epub 2019 Dec 11.

Aquaculture Research Group (GIA), IU-ECOAQUA, Universidad de Las Palmas de Gran Canaria, Crta. Taliarte s/n, 35214 Telde, Spain.

Polyunsaturated fatty acids (PUFA) in parental diets play a key role in regulating LC-PUFA metabolism of the offspring. However, it is not clear whether this metabolic regulation is driven by the precursors presented in the diet or by the parental ability to synthesize them. To elucidate this, broodstocks of gilthead sea bream with different blood expression levels of , which encodes for the rate-limiting enzyme in the LC-PUFA synthesis pathway, were fed either a diet supplemented with alpha-linolenic acid (ALA, 18:3) or a control diet. The progenies obtained from these four experimental groups were then challenged with a low LC-PUFA diet at the juvenile stage. Results showed that the offspring from parents with high expression presented higher growth and improved utilization of low LC-PUFA diets compared to the offspring from parents with low expression. Besides, an ALA-rich diet during the gametogenesis caused negative effects on the growth of the offspring. The epigenetic analysis demonstrated that methylation in the promoter of of the offspring was correlated with the parental expression levels and type of the broodstock diet.
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http://dx.doi.org/10.3390/ijms20246250DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6940931PMC
December 2019

Stearoyl-CoA desaturase () is epigenetically regulated by broodstock nutrition in gilthead sea bream ().

Epigenetics 2020 05 6;15(5):536-553. Epub 2019 Dec 6.

Nutrigenomics and Fish Growth Endocrinology Group, Institute of Aquaculture Torre de la Sal, IATS-CSIC, Castellón, Spain.

The aim of this study was to generate new knowledge on fish epigenetics, assessing the effects of linolenic acid (ALA) conditioning of broodstock in the offspring of the marine fish . Attention was focused on gene organization, methylation signatures and gene expression patterns of fatty acid desaturase 2 () and stearoyl-CoA desaturase 1a (). Blat searches in the genomic IATS-CSIC database (www.nutrigroup-iats.org/seabreamdb) highlighted a conserved exon-intron organization, a conserved PUFA response region, and CG islands at the promoter regions of each gene. The analysed CpG positions in the promoter were mostly hypomethylated and refractory to broodstock nutrition. The same response was achieved after conditioning of juvenile fish to low water oxygen concentrations, thus methylation susceptibility at individual CpG sites seems to be stringently regulated in fish of different origin and growth trajectories. Conversely, the promoter was responsive to broodstock nutrition and the offspring of parents fed the ALA-rich diet shared an increased DNA-methylation, mainly in CpG sites neighbouring SP1 and HNF4α binding sites. Cytosine methylation at these sites correlated inversely with the hepatic expression of the offspring. Co-expression analyses supported that the HNF4α-dependent regulation of is affected by DNA methylation. The phenotypic output is a regulated liver fat deposition through changes in expression, which would also allow the preservation of fatty acid unsaturation levels in fish fed reduced levels of LC-PUFA. Collectively, these findings reveal a reliable mechanism by which parent's nutrition can shape gene expression in the fish offspring.
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http://dx.doi.org/10.1080/15592294.2019.1699982DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7188398PMC
May 2020

Sex, Age, and Bacteria: How the Intestinal Microbiota Is Modulated in a Protandrous Hermaphrodite Fish.

Front Microbiol 2019 31;10:2512. Epub 2019 Oct 31.

Nutrigenomics and Fish Growth Endocrinology Group, Institute of Aquaculture Torre de la Sal (CSIC), Castellón, Spain.

Intestinal microbiota is key for many host functions, such as digestion, nutrient metabolism, disease resistance, and immune function. With the growth of the aquaculture industry, there has been a growing interest in the manipulation of fish gut microbiota to improve welfare and nutrition. Intestinal microbiota varies with many factors, including host species, genetics, developmental stage, diet, environment, and sex. The aim of this study was to compare the intestinal microbiota of adult gilthead sea bream () from three groups of age and sex (1-year-old males and 2- and 4-year-old females) maintained under the same conditions and fed exactly the same diet. Microbiota diversity and richness did not differ among groups. However, bacterial composition did, highlighting the presence of and starting at 2 years of age (females) and a higher presence of and in 1-year-old males. The core microbiota was defined by 14 Operational Taxonomic Units (OTUs) and the groups that showed more OTUs in common were 2- and 4-year-old females. Discriminant analyses showed a clear separation by sex and age, with bacteria belonging to the phyla Firmicutes, Proteobacteria and Actinobacteria driving the separation. Pathway analysis performed with the inferred metagenome showed significant differences between 1-year-old males and 4-year-old females, with an increase in infection-related pathways, nitrotoluene degradation and sphingolipid metabolism, and a significant decrease in carbohydrate metabolism pathways with age. These results show, for the first time, how intestinal microbiota is modulated in adult gilthead sea bream and highlight the importance of reporting age and sex variables in these type of studies in fish.
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http://dx.doi.org/10.3389/fmicb.2019.02512DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6834695PMC
October 2019

Protective effects of seaweed supplemented diet on antioxidant and immune responses in European seabass (Dicentrarchus labrax) subjected to bacterial infection.

Sci Rep 2019 11 6;9(1):16134. Epub 2019 Nov 6.

CIIMAR - Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto, Av. General Norton de Matos s/n, 4450-208, Matosinhos, Portugal, Portugal.

European seabass (Dicentrarchus labrax) production is often hampered by bacterial infections such as photobacteriosis caused by Photobacterium damselae subsp. piscicida (Phdp). Since diet can impact fish immunity, this work investigated the effect of dietary supplementation of 5% Gracilaria sp. aqueous extract (GRA) on seabass antioxidant capacity and resistance against Phdp. After infection, mortality was delayed in fish fed GRA, which also revealed increased lysozyme activity levels, as well as decreased lipid peroxidation, suggesting higher antioxidant capacity than in fish fed a control diet. Dietary GRA induced a down-regulation of hepatic stress-responsive heat shock proteins (grp-78, grp-170, grp-94, grp-75), while bacterial infection caused a down-regulation in antioxidant genes (prdx4 and mn-sod). Diet and infection interaction down-regulated the transcription levels of genes associated with oxidative stress response (prdx5 and gpx4) in liver. In head-kidney, GRA led to an up-regulation of genes associated with inflammation (il34, ccr9, cd33) and a down-regulation of genes related to cytokine signalling (mif, il1b, defb, a2m, myd88). Additionally, bacterial infection up-regulated immunoglobulins production (IgMs) and down-regulated the transcription of the antimicrobial peptide leap2 in head kidney. Overall, we found that GRA supplementation modulated seabass resistance to Phdp infection.
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http://dx.doi.org/10.1038/s41598-019-52693-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6834676PMC
November 2019

Effects of Dietary Bisphenol A on the Reproductive Function of Gilthead Sea Bream () Testes.

Int J Mol Sci 2019 Oct 10;20(20). Epub 2019 Oct 10.

Dipartimento Scienze della Vita e dell'Ambiente, Università Politecnica delle Marche, Via Brecce Bianche, 60131 Ancona, Italy.

Bisphenol A (BPA), a known endocrine disrupting chemical (EDC), was administered by diet to gilthead sea bream () 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 (E) 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 (), estrogen receptors (, ), progesterone receptors () and the gonadotropin releasing hormone receptor ()) were up-regulated. BPA induced the up-regulation of the hepatic genes involved in oogenesis (vitellogenin () and zona pellucida 1 ()).
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http://dx.doi.org/10.3390/ijms20205003DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6835794PMC
October 2019

Disruption of gut integrity and permeability contributes to enteritis in a fish-parasite model: a story told from serum metabolomics.

Parasit Vectors 2019 Oct 16;12(1):486. Epub 2019 Oct 16.

Associated Unit of Marine Ecotoxicology (IATS-IUPA), Castellon, Spain.

Background: In the animal production sector, enteritis is responsible for serious economic losses, and intestinal parasitism is a major stress factor leading to malnutrition and lowered performance and animal production efficiency. The effect of enteric parasites on the gut function of teleost fish, which represent the most ancient bony vertebrates, is far from being understood. The intestinal myxozoan parasite Enteromyxum leei dwells between gut epithelial cells and causes severe enteritis in gilthead sea bream (Sparus aurata), anorexia, cachexia, growth impairment, reduced marketability and increased mortality.

Methods: This study aimed to outline the gut failure in this fish-parasite model using a multifaceted approach and to find and validate non-lethal serum markers of gut barrier dysfunction. Intestinal integrity was studied in parasitized and non-parasitized fish by immunohistochemistry with specific markers for cellular adhesion (E-cadherin) and tight junctions (Tjp1 and Cldn3) and by functional studies of permeability (oral administration of FITC-dextran) and electrophysiology (Ussing chambers). Serum samples from parasitized and non-parasitized fish were analyzed using non-targeted metabolomics and some significantly altered metabolites were selected to be validated using commercial kits.

Results: The immunodetection of Tjp1 and Cldn3 was significantly lower in the intestine of parasitized fish, while no strong differences were found in E-cadherin. Parasitized fish showed a significant increase in paracellular uptake measured by FITC-dextran detection in serum. Electrophysiology showed a decrease in transepithelial resistance in infected animals, which showed a diarrheic profile. Serum metabolomics revealed 3702 ions, from which the differential expression of 20 identified compounds significantly separated control from infected groups in multivariate analyses. Of these compounds, serum inosine (decreased) and creatine (increased) were identified as relevant and validated with commercial kits.

Conclusions: The results demonstrate the disruption of tight junctions and the loss of gut barrier function, a metabolomic profile of absorption dysfunction and anorexia, which further outline the pathophysiological effects of E. leei.
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http://dx.doi.org/10.1186/s13071-019-3746-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6796429PMC
October 2019

Tissue-Specific Orchestration of Gilthead Sea Bream Resilience to Hypoxia and High Stocking Density.

Front Physiol 2019 10;10:840. Epub 2019 Jul 10.

Nutrigenomics and Fish Growth Endocrinology, Institute of Aquaculture Torre de la Sal (IATS-CSIC), Castellón, Spain.

Two different O levels (normoxia: 75-85% O saturation; moderate hypoxia: 42-43% O saturation) and stocking densities (LD: 9.5, and HD: 19 kg/m) were assessed on gilthead sea bream () in a 3-week feeding trial. Reduced O availability had a negative impact on feed intake and growth rates, which was exacerbated by HD despite of the improvement in feed efficiency. Blood physiological hallmarks disclosed the enhancement in O-carrying capacity in fish maintained under moderate hypoxia. This feature was related to a hypo-metabolic state to cope with a chronic and widespread environmental O reduction, which was accompanied by a differential regulation of circulating cortisol and growth hormone levels. Customized PCR-arrays were used for the simultaneous gene expression profiling of 34-44 selected stress and metabolic markers in liver, white skeletal muscle, heart, and blood cells. The number of differentially expressed genes ranged between 22 and 19 in liver, heart, and white skeletal muscle to 5 in total blood cells. Partial Least-Squares Discriminant Analysis (PLS-DA) explained [R2Y(cum)] and predicted [Q2Y(cum)] up to 95 and 65% of total variance, respectively. The first component (R2Y = 0.2889) gathered fish on the basis of O availability, and liver and cardiac genes on the category of energy sensing and oxidative metabolism (α, α, β, s -----), antioxidant defense and tissue repair (, and ) and oxidative phosphorylation (, and ) highly contributed to this separation. The second component (R2Y = 0.2927) differentiated normoxic fish at different stocking densities, and the white muscle clearly promoted this separation by a high over-representation of genes related to GH/IGF system (, and ). The third component (R2Y = 0.2542) discriminated the effect of stocking density in fish exposed to moderate hypoxia by means of hepatic fatty acid desaturases (, and and muscle markers of fatty acid oxidation (). All these findings disclose the different contribution of analyzed tissues (liver ≥ heart > muscle > blood) and specific genes to the hypoxic- and crowding stress-mediated responses. This study will contribute to better explain and understand the different stress resilience of farmed fish across individuals and species.
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http://dx.doi.org/10.3389/fphys.2019.00840DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6635561PMC
July 2019

Dietary tryptophan supplementation induces a transient immune enhancement of gilthead seabream (Sparus aurata) juveniles fed fishmeal-free diets.

Fish Shellfish Immunol 2019 Oct 13;93:240-250. Epub 2019 Jul 13.

Centro Interdisciplinar de Investigação Marinha e Ambiental (CIIMAR), Universidade do Porto, Terminal de Cruzeiros do Porto de Leixões, Avenida General Norton de Matos, S/N, 4450-208, Matosinhos, Portugal; Instituto de Ciências Biomédicas Abel Salazar (ICBAS-UP), Universidade do Porto, Rua de Jorge Viterbo Ferreira nº 228, 4050-313, Porto, Portugal. Electronic address:

European aquaculture is an industry with a high sustainability profile contributing to the supply of safe seafood. However, several diseases can affect farmed fish and it is imperative to find alternatives for chemotherapeutic treatments when disease outbreaks occur. Maintenance of health through nutrition is well-establish in modern animal farming, and amino acids (AA) are promising candidates as functional additives to improve fish health. Therefore, the goal of this research is to provide a better understanding of the influence of tryptophan supplementation on nutritional condition and immune mechanisms in fish. Triplicate groups of fish (13.3 ± 0.3g) previously fed with a fishmeal-based diet were either fed a control diet with an extreme formulation (0% fishmeal) but meeting the AA requirements (CTRL), or the SUP diet, formulated as the CTRL with an increase in tryptophan (TRP) content. After 2 and 13 weeks of feeding, head-kidney (HK), liver (L) and white skeletal muscle (WSM) were collected for gene expression, whereas plasma was suited for humoral immune parameters. A holistic approach using transcriptomic, humoral and zootechnical parameters was undertaken. The expression of 29-31 genes for WSM, L or HK confirms an effect due to the treatment across time. A two-way ANOVA analysis revealed that 15-24 genes varied significantly depending on the tissue, and the multivariate analysis by means of PLS-DA explained (R) and predicted (Q) with four components up to 93% and 78% of total variance, respectively. Component 1 (R = 50.06%) represented the time effects, whereas components 2 (24.36%) and 3 (13.89%) grouped fish on the basis of dietary treatment, at early sampling. The HK results in particular suggest that fish fed SUP diet displayed an immunostimulated state at 2 weeks. No major differences were observed in plasma humoral parameters, despite an increase in antiprotease and peroxidase activities after 13 weeks regardless of dietary treatment. These results suggest that tryptophan supplementation may improve the seabream immune status after 2 weeks. Hence, the use of functional feeds is especially relevant during a short-term feeding period before a predictable stressful event or disease outbreak, considering that these putative advantageous effects seem to disappear after a 13 weeks feeding period.
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http://dx.doi.org/10.1016/j.fsi.2019.07.033DOI Listing
October 2019

Ultra-Low Power Sensor Devices for Monitoring Physical Activity and Respiratory Frequency in Farmed Fish.

Front Physiol 2019 29;10:667. Epub 2019 May 29.

Nutrigenomics and Fish Growth Endocrinology Group, Institute of Aquaculture Torre de la Sal, Consejo Superior de Investigaciones Científicas (CSIC), Castellón, Spain.

Integration of technological solutions aims to improve accuracy, precision and repeatability in farming operations, and biosensor devices are increasingly used for understanding basic biology during livestock production. The aim of this study was to design and validate a miniaturized tri-axial accelerometer for non-invasive monitoring of farmed fish with re-programmable schedule protocols. The current device (AE-FishBIT v.1s) is a small (14 mm × 7 mm × 7 mm), stand-alone system with a total mass of 600 mg, which allows monitoring animals from 30 to 35 g onwards. The device was attached to the operculum of gilthead sea bream () and European sea bass () juveniles for monitoring their physical activity by measurements of movement accelerations in - and -axes, while records of operculum beats (-axis) served as a measurement of respiratory frequency. Data post-processing of exercised fish in swimming test chambers revealed an exponential increase of fish accelerations with the increase of fish speed from 1 body-length to 4 body-lengths per second, while a close relationship between oxygen consumption (MO) and opercular frequency was consistently found. Preliminary tests in free-swimming fish kept in rearing tanks also showed that device data recording was able to detect changes in daily fish activity. The usefulness of low computational load for data pre-processing with on-board algorithms was verified from low to submaximal exercise, increasing this procedure the autonomy of the system up to 6 h of data recording with different programmable schedules. Visual observations regarding tissue damage, feeding behavior and circulating levels of stress markers (cortisol, glucose, and lactate) did not reveal at short term a negative impact of device tagging. Reduced plasma levels of triglycerides revealed a transient inhibition of feed intake in small fish (sea bream 50-90 g, sea bass 100-200 g), but this disturbance was not detected in larger fish. All this considered together is the proof of concept that miniaturized devices are suitable for non-invasive and reliable metabolic phenotyping of farmed fish to improve their overall performance and welfare. Further work is underway for improving the attachment procedure and the full device packaging.
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http://dx.doi.org/10.3389/fphys.2019.00667DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6548888PMC
May 2019

A long-term growth hormone treatment stimulates growth and lipolysis in gilthead sea bream juveniles.

Comp Biochem Physiol A Mol Integr Physiol 2019 06 15;232:67-78. Epub 2019 Mar 15.

Department of Cell Biology, Physiology and Immunology, Faculty of Biology, University of Barcelona, 08028 Barcelona, Spain. Electronic address:

The enhancement of the endocrine growth hormone (GH)/insulin-like growth factor I (IGF-I) system by the treatment with a sustained release formulation of a recombinant bovine GH (rBGH), is a good strategy to investigate growth optimization in aquaculture fish species. To further deepen into the knowledge of rBGH effects in fish and to estimate the growth potential of juveniles of gilthead sea bream, the present work evaluated rBGH injection on growth, GH/IGF-I axis and lipid metabolism modulation, and explored the conservation of GH effects provoked by the in vivo treatment using in vitro models of different tissues. The rBGH treatment increased body weight and specific growth rate (SGR) in juveniles and potentiated hyperplastic muscle growth while reducing circulating triglyceride levels. Moreover, the results demonstrated that the in vivo treatment enhanced also lipolysis in both isolated hepatocytes and adipocytes, as well as in day 4 cultured myocytes. Furthermore, these cultured myocytes extracted from rBGH-injected fish presented higher gene expression of GH/IGF-I axis-related molecules and myogenic regulatory factors, as well as stimulated myogenesis (i.e. increased protein expression of a proliferation and a differentiation marker) compared to Control fish-derived cells. These data, suggested that cells in vitro can retain some of the pathways activated by in vivo treatments in fish, what can be considered an interesting line of applied research. Overall, the results showed that rBGH stimulates somatic growth, including specifically muscle hyperplasia, as well as lipolytic activity in gilthead sea bream juveniles.
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http://dx.doi.org/10.1016/j.cbpa.2019.03.012DOI Listing
June 2019

Acting locally - affecting globally: RNA sequencing of gilthead sea bream with a mild Sparicotyle chrysophrii infection reveals effects on apoptosis, immune and hypoxia related genes.

BMC Genomics 2019 Mar 11;20(1):200. Epub 2019 Mar 11.

Fish Pathology Group, Institute of Aquaculture Torre de la Sal (IATS-CSIC), Ribera de Cabanes, Castellón, Spain.

Background: Monogenean flatworms are the main fish ectoparasites inflicting serious economic losses in aquaculture. The polyopisthocotylean Sparicotyle chrysophrii parasitizes the gills of gilthead sea bream (GSB, Sparus aurata) causing anaemia, lamellae fusion and sloughing of epithelial cells, with the consequent hypoxia, emaciation, lethargy and mortality. Currently no preventive or curative measures against this disease exist and therefore information on the host-parasite interaction is crucial to find mitigation solutions for sparicotylosis. The knowledge about gene regulation in monogenean-host models mostly comes from freshwater monopysthocotyleans and almost nothing is known about polyopisthocotyleans. The current study aims to decipher the host response at local (gills) and systemic (spleen, liver) levels in farmed GSB with a mild natural S. chrysophrii infection by transcriptomic analysis.

Results: Using Illumina RNA sequencing and transcriptomic analysis, a total of 2581 differentially expressed transcripts were identified in infected fish when compared to uninfected controls. Gill tissues in contact with the parasite (P gills) displayed regulation of fewer genes (700) than gill portions not in contact with the parasite (NP gills) (1235), most likely due to a local silencing effect of the parasite. The systemic reaction in the spleen was much higher than that at the parasite attachment site (local) (1240), and higher than in liver (334). NP gills displayed a strong enrichment of genes mainly related to immune response and apoptosis. Processes such as apoptosis, inflammation and cell proliferation dominated gills, whereas inhibition of apoptosis, autophagy, platelet activation, signalling and aggregation, and inflammasome were observed in spleen. Proteasome markers were increased in all tissues, whereas hypoxia-related genes were down-regulated in gills and spleen.

Conclusions: Contrasting forces seem to be acting at local and systemic levels. The splenic down-regulation could be part of a hypometabolic response, to counteract the hypoxia induced by the parasite damage to the gills and to concentrate the energy on defence and repair responses. Alternatively, it can be also interpreted as the often observed action of helminths to modify host immunity in its own interest. These results provide the first toolkit for future studies towards understanding and management of this parasitosis.
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http://dx.doi.org/10.1186/s12864-019-5581-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6416957PMC
March 2019

Effects of diisononyl phthalate (DiNP) on the endocannabinoid and reproductive systems of male gilthead sea bream (Sparus aurata) during the spawning season.

Arch Toxicol 2019 03 2;93(3):727-741. Epub 2019 Jan 2.

Dipartimento Scienze Della Vita e dell'Ambiente, Università Politecnica Delle Marche, Via Brecce Bianche, 60131, Ancona, Italy.

Diisononyl phthalate (DiNP) is a plasticizer used to improve plastic performance in a large variety of items which has been reported as an endocrine-disrupting chemical (EDC) in several organisms. The endocannabinoid system (ECS) is a cellular signaling system, whose functionality is tightly involved with reproductive function. The aim of the present study was the assessment of the effects of DiNP on the gonadal ECS and on the reproductive function of male gilthead sea bream Sparus aurata, an important marine aquacultured species in Europe, during the reproductive season. Fish were fed for 21 days with two diets contaminated with different nominal concentrations of DiNP (DiNP LOW at 15 µg DiNP kg bw day and DiNP HIGH at 1500 µg DiNP kg bw day), based on the tolerable daily intake (TDI) ruled by the European Food Safety Authority for humans. The transcription of several genes related to the ECS was affected by the DiNP. Specifically, DiNP reduced the levels of endocannabinoids and endocannabinoid-like mediators, concomitant with the increase of fatty acid amide hydrolase (FAAH) activity. At the histological level, DiNP LOW induced the highest occurrence of individuals with regressed testes. Steroidogenesis was affected significantly, since plasma 11-ketotestosterone (11-KT), the main active androgen in fish, was significantly decreased by the DiNP HIGH treatment, while plasma 17β-estradiol (E) levels were raised, associated with an increase of the gonadosomatic index (GSI). Additionally, the level of testosterone (T) was significantly increased in the DiNP LOW group, however, the same DiNP concentration reduced the levels of 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P). The production of sperm was in general not affected, since spermiation index, sperm density, survival and the duration of forward motility did not exhibit any changes compared to controls. However, computer-assisted sperm analysis (CASA) showed that DiNP reduced the percentage of motile cells. The results clearly suggest a negative effect of DiNP via the diet on the male endocrine system of gilthead sea bream during the reproductive season.
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http://dx.doi.org/10.1007/s00204-018-2378-6DOI Listing
March 2019

Somatotropic Axis Regulation Unravels the Differential Effects of Nutritional and Environmental Factors in Growth Performance of Marine Farmed Fishes.

Front Endocrinol (Lausanne) 2018 27;9:687. Epub 2018 Nov 27.

Nutrigenomics and Fish Growth Endocrinology, Institute of Aquaculture Torre de la Sal (IATS-CSIC), Castellón, Spain.

The Gh/Prl/Sl family has evolved differentially through evolution, resulting in varying relationships between the somatotropic axis and growth rates within and across fish species. This is due to a wide range of endogenous and exogenous factors that make this association variable throughout season and life cycle, and the present minireview aims to better define the nutritional and environmental regulation of the endocrine growth cascade over precisely defined groups of fishes, focusing on Mediterranean farmed fishes. As a result, circulating Gh and Igf-i are revitalized as reliable growth markers, with a close association with growth rates of gilthead sea bream juveniles with deficiency signs in both macro- or micro-nutrients. This, together with other regulated responses, promotes the use of Gh and Igf-i as key performance indicators of growth, aerobic scope, and nutritional condition in gilthead sea bream. Moreover, the sirtuin-energy sensors might modulate the growth-promoting action of somatotropic axis. In this scenario, transcripts of and receptors mirror changes in plasma Gh and Igf-i levels, with the expression ratio mostly unaltered over season. However, this ratio is nutritionally regulated, and enriched plant-based diets or diets with specific nutrient deficiencies downregulate hepatic , decreasing the / ratio. The same trend, due to a increase, is found in skeletal muscle, whereas impaired growth during overwintering is related to increase in the / and / ratios in liver and skeletal muscle, respectively. Overall, expression of insulin receptors and receptors is less regulated, though the expression quotient is especially high in the liver and muscle of sea bream. Nutritional and environmental regulation of the full Igf binding protein 1-6 repertoire remains to be understood. However, tissue-specific expression profiling highlights an enhanced and nutritionally regulated expression of the clade in liver, whereas the clade is overexpressed and regulated in skeletal muscle. The somatotropic axis is, therefore, highly informative of a wide-range of growth-disturbing and stressful stimuli, and multivariate analysis supports its use as a reliable toolset for the assessment of growth potentiality and nutrient deficiencies and requirements, especially in combination with selected panels of other nutritionally regulated metabolic biomarkers.
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http://dx.doi.org/10.3389/fendo.2018.00687DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6277588PMC
November 2018

Gene expression analysis of Atlantic salmon gills reveals mucin 5 and interleukin 4/13 as key molecules during amoebic gill disease.

Sci Rep 2018 09 12;8(1):13689. Epub 2018 Sep 12.

Fish Pathology Group, Instituto de Acuicultura Torre de la Sal (IATS-CSIC), Castellón, 12595, Spain.

Amoebic gill disease (AGD) is one of the main diseases affecting Atlantic salmon (Salmo salar L.) mariculture. Hallmarks of AGD are hyperplasia of the lamellar epithelium and increased production of gill mucus. This study investigated the expression of genes involved in mucus secretion, cell cycle regulation, immunity and oxidative stress in gills using a targeted 21-gene PCR array. Gill samples were obtained from experimental and natural Neoparamoeba perurans infections, and sampling points included progressive infection stages and post-freshwater treatment. Up-regulation of genes related to mucin secretion and cell proliferation, and down-regulation of pro-inflammatory and pro-apoptotic genes were associated with AGD severity, while partial restoration of the gill homeostasis was detected post-treatment. Mucins and Th2 cytokines accoun ted for most of the variability observed between groups highlighting their key role in AGD. Two mucins (muc5, muc18) showed differential regulation upon disease. Substantial up-regulation of the secreted muc5 was detected in clinical AGD, and the membrane bound muc18 showed an opposite pattern. Th2 cytokines, il4/13a and il4/13b2, were significantly up-regulated from 2 days post-infection onwards, and changes were lesion-specific. Despite the differences between experimental and natural infections, both yielded comparable results that underline the importance of the studied genes in the respiratory organs of fish, and during AGD progression.
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http://dx.doi.org/10.1038/s41598-018-32019-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6135806PMC
September 2018

Ghrelin and Its Receptors in Gilthead Sea Bream: Nutritional Regulation.

Front Endocrinol (Lausanne) 2018 30;9:399. Epub 2018 Jul 30.

Department of Cell Biology, Physiology and Immunology, Faculty of Biology, University of Barcelona, Barcelona, Spain.

Ghrelin is involved in the regulation of growth in vertebrates through controlling different functions, such as feed intake, metabolism, intestinal activity or growth hormone (Gh) secretion. The aim of this work was to identify the sequences of and Ghrelin receptors (), and to study their responses to different nutritional conditions in gilthead sea bream () juveniles. The structure and phylogeny of was analyzed, and a tissue screening was performed. The effects of 21 days of fasting and 2, 5, 24 h, and 7 days of refeeding on plasma levels of Ghrelin, Gh and Igf-1, and the gene expression of and members of the Gh/Igf-1 system were determined in key tissues. and the receptors are well conserved, being expressed mainly in stomach, and in the pituitary and brain, respectively. Twenty-one days of fasting resulted in a decrease in growth while Ghrelin plasma levels were elevated to decrease at 5 h post-prandial when pituitary expression was minimum. Gh in plasma increased during fasting and slowly felt upon refeeding, while plasma Igf-1 showed an inverse profile. Pituitary expression augmented during fasting reaching maximum levels at 1 day post-feeding while liver expression and that of its splice variants decreased to lowest levels. Liver Gh receptors expression was down-regulated during fasting and recovered after refeeding. This study demonstrates the important role of Ghrelin during fasting, its acute down-regulation in the post-prandial stage and its interaction with pituitary Ghsrs and Gh/Igf-1 axis.
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http://dx.doi.org/10.3389/fendo.2018.00399DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6077198PMC
July 2018

Hints on T cell responses in a fish-parasite model: Enteromyxum leei induces differential expression of T cell signature molecules depending on the organ and the infection status.

Parasit Vectors 2018 Jul 31;11(1):443. Epub 2018 Jul 31.

Fish Pathology Group, Institute of Aquaculture Torre de la Sal (IATS-CSIC), Ribera de Cabanes, Castellón, Spain.

Backgroud: Enteromyxum leei is a myxozoan parasite that produces a slow-progressing intestinal disease. This parasite invades the paracellular space of the intestinal epithelium and progresses from the posterior to the anterior intestine. The aim of the present study was to gain insights into fish T cell responses in the gilthead sea bream-E. leei infection model using a PCR-array with 30 signature molecules for different leukocyte responses in head kidney, spleen, anterior and posterior intestine.

Results: The PCR-array results suggest that E. leei induced migration of T cells from head kidney to intestines where T, CTL and T profiles were activated and kept in balance by the upregulation of regulatory cytokines. These results were partially validated by the use of cross-reacting antibodies and BrdU immunostaining to monitor proliferation. Zap70 immunostaining supported the increased number of T cells in the anterior intestine detected by gene expression, but double staining with BrdU did not show active proliferation of this cell type at a local level, supporting the migration from lymphohaematopoietic tissues to the site of infection. Global analyses of the expression profiles revealed a clear separation between infected and exposed, but non-infected fish, more evident in the target organ. Exposed, non-infected animals showed an intermediate phenotype closer to the control fish.

Conclusions: These results evidence a clear modulation of the T cell response of gilthead sea bream upon E. leei infection. The effects occurred both at local and systemic levels, but the response was stronger and more specific at the site of infection, the intestine. Altogether, this research poses a promising basis to understand the response against this important parasite and establish effective preventive or palliative measures.
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http://dx.doi.org/10.1186/s13071-018-3007-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6069777PMC
July 2018

Endocrine disruptors in the diet of male Sparus aurata: Modulation of the endocannabinoid system at the hepatic and central level by Di-isononyl phthalate and Bisphenol A.

Environ Int 2018 10 19;119:54-65. Epub 2018 Jun 19.

Dipartimento Scienze della Vita e dell'Ambiente, Università Politecnica delle Marche, Via Brecce Bianche, 60131 Ancona, Italy. Electronic address:

The increasing manufacture of plastics and their mismanagement has turned plastic into a ubiquitous waste in the marine environment. Among all the substances conforming the plastic items, the effects of a dietary Bisphenol A (BPA) and Di-isononyl phthalate (DiNP) have been evaluated in adult male gilthead sea bream, focusing on their effects in the modulation of the Endocannabinoid System (ECS). In zebrafish, the ECS has been recently chosen as a new target for the activity of some Endocrine Disrupting Chemicals (EDC), since it represents a complex lipid signaling network essential for the well-being of the organisms. The results obtained in gilthead seabream showed that BPA and DiNP altered the structure and the biochemical composition of liver, increasing the presence of lipids and triglycerides and decreasing the glycogen and phospholipids. Moreover, the addition of BPA or DiNP in the gilthead sea bream diet altered the levels of endocannabinoids (EC) and EC-like mediators in the liver. These alterations were also associated to changes at the transcriptomic level of genes involved in lipid biosynthesis and ECS metabolism. At the central level, both BPA and DiNP reduced the expression of the endocannabinoid receptor type I (cnr1) and the neuropeptide Y (npy) as well as the levels of the endocannabinoid Anandamide (AEA), suggesting a downregulation of appetite. The results herein reported highlighted the negative effects of chronic dietary exposure to DiNP or BPA on ECS functions and lipid metabolism of male gilthead sea bream liver, showing a similar disruptive activity of these contaminants at metabolic level. Moreover, the novelty of the biomarkers used evidenced possible innovative endpoints for the development of novel OEDCS test guidelines.
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http://dx.doi.org/10.1016/j.envint.2018.06.011DOI Listing
October 2018

Co-expression Analysis of Sirtuins and Related Metabolic Biomarkers in Juveniles of Gilthead Sea Bream () With Differences in Growth Performance.

Front Physiol 2018 5;9:608. Epub 2018 Jun 5.

Nutrigenomics and Fish Growth Endocrinology, Institute of Aquaculture Torre de la Sal-CSIC, Castellón, Spain.

Sirtuins (SIRTs) represent a conserved protein family of deacetylases that act as master regulators of metabolism, but little is known about their roles in fish and livestock animals in general. The present study aimed to assess the value of SIRTs for the metabolic phenotyping of fish by assessing their co-expression with a wide-representation of markers of energy and lipid metabolism and intestinal function and health in two genetically different gilthead sea bream strains with differences in growth performance. Fish from the fast-growing strain exhibited higher feed intake, feed efficiency and plasma IGF-I levels, along with higher hepatosomatic index and lower mesenteric fat (lean phenotype). These observations suggest differences in tissue energy partitioning with an increased flux of fatty acids from adipose tissue toward the liver. The resulting increased risk of hepatic steatosis may be counteracted in the liver by reduced lipogenesis and enhanced triglyceride catabolism, in combination with a higher and more efficient oxidative metabolism in white skeletal muscle. These effects were supported by co-regulated changes in the expression profile of SIRTs (liver, skeletal muscle, adipose tissue, ) and markers of oxidative metabolism (α), mitochondrial respiration uncoupling () and fatty acid and triglyceride metabolism (αγ) that were specific to each strain and tissue. The anterior intestine of the fast-growing strain was better suited to cope with improved growth by increased expression of markers of nutrient absorption (), epithelial barrier integrity () and immunity (β, ), which were correlated with low expression levels of and markers of fatty acid oxidation (). In the posterior intestine, the fast-growing strain showed a consistent up-regulation of and concurrently with increased expression levels of markers of cell proliferation (), oxidative metabolism () and immunity (). Together, these findings indicate that SIRTs may play different roles in the regulation of metabolism, inflammatory tone and growth in farmed fish, arising as powerful biomarkers for a reliable metabolic phenotyping of fish at the tissue-specific level.
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http://dx.doi.org/10.3389/fphys.2018.00608DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5996159PMC
June 2018

Under control: how a dietary additive can restore the gut microbiome and proteomic profile, and improve disease resilience in a marine teleostean fish fed vegetable diets.

Microbiome 2017 12 28;5(1):164. Epub 2017 Dec 28.

Nutrigenomics and Fish Growth Endocrinology Group, Instituto de Acuicultura Torre de la Sal (IATS-CSIC), Castellón, Spain.

Background: The constant increase of aquaculture production and wealthy seafood consumption has forced the industry to explore alternative and more sustainable raw aquafeed materials, and plant ingredients have been used to replace marine feedstuffs in many farmed fish. The objective of the present study was to assess whether plant-based diets can induce changes in the intestinal mucus proteome, gut autochthonous microbiota and disease susceptibility of fish, and whether these changes could be reversed by the addition of sodium butyrate to the diets. Three different trials were performed using the teleostean gilthead sea bream (Sparus aurata) as model. In a first preliminary short-term trial, fish were fed with the additive (0.8%) supplementing a basal diet with low vegetable inclusion (D1) and then challenged with a bacteria to detect possible effects on survival. In a second trial, fish were fed with diets with greater vegetable inclusion levels (D2, D3) and the long-term effect of sodium butyrate at a lower dose (0.4%) added to D3 (D4 diet) was tested on the intestinal proteome and microbiome. In a third trial, the long-term effectiveness of sodium butyrate (D4) to prevent disease outcome after an intestinal parasite (Enteromyxum leei) challenge was tested.

Results: The results showed that opposed forces were driven by dietary plant ingredients and sodium butyrate supplementation in fish diet. On the one hand, vegetable diets induced high parasite infection levels that provoked drops in growth performance, decreased intestinal microbiota diversity, induced the dominance of the Photobacterium genus, as well as altered the gut mucosal proteome suggesting detrimental effects on intestinal function. On the other hand, butyrate addition slightly decreased cumulative mortality after bacterial challenge, avoided growth retardation in parasitized fish, increased intestinal microbiota diversity with a higher representation of butyrate-producing bacteria and reversed most vegetable diet-induced changes in the gut proteome.

Conclusions: This integrative work gives insights on the pleiotropic effects of a dietary additive on the restoration of intestinal homeostasis and disease resilience, using a multifaceted approach.
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http://dx.doi.org/10.1186/s40168-017-0390-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5745981PMC
December 2017

Sodium salt medium-chain fatty acids and -based probiotic strategies to improve growth and intestinal health of gilthead sea bream ().

PeerJ 2017 4;5:e4001. Epub 2017 Dec 4.

Nutrigenomics and Fish Growth Endocrinology Group, Institute of Aquaculture Torre de la Sal, CSIC, Castellón, Spain.

Background: The increased demand for fish protein has led to the intensification of aquaculture practices which are hampered by nutritional and health factors affecting growth performance. To solve these problems, antibiotics have been used for many years in the prevention, control and treatment against disease as well as growth promoters to improve animal performance. Nowadays, the use of antibiotics in the European Union and other countries has been completely or partially banned as a result of the existence of antibiotic cross-resistance. Therefore, a number of alternatives, including enzymes, prebiotics, probiotics, phytonutrients and organic acids used alone or in combination have been proposed for the improvement of immunological state, growth performance and production in livestock animals. The aim of the present study was to evaluate two commercially available feed additives, one based on medium-chain fatty acids (MCFAs) from coconut oil and another with a -based probiotic, in gilthead sea bream (GSB, ), a marine farmed fish of high value in the Mediterranean aquaculture

Methods: The potential benefits of adding two commercial feed additives on fish growth performance and intestinal health were assessed in a 100-days feeding trial. The experimental diets (D2 and D3) were prepared by supplementing a basal diet (D1) with MCFAs in the form of a sodium salt of coconut fatty acid distillate (DICOSAN; Norel, Madrid, Spain), rich on C-12, added at 0.3% (D2) or with the probiotic CECT 5940, added at 0.1% (D3). The study integrated data on growth performance, blood biochemistry, histology and intestinal gene expression patterns of selected markers of intestinal function and architecture.

Results: MCFAs in the form of a coconut oil increased feed intake, growth rates and the surface of nutrient absorption, promoting the anabolic action of the somatotropic axis. The probiotic (D3) induced anti-inflammatory and anti-oxidant effects with changes in circulating cortisol, immunoglobulin M, leukocyte respiratory burst, and mucosal expression levels of cytokines, lymphocyte markers and immunoglobulin T.

Discussion: MCFA supplementation showed positive effects on GSB growth and intestinal architecture acting mainly in the anterior intestine, where absorption takes place. The probiotic CECT 5940 exhibited key effects in the regulation of the immune status inducing anti-inflammatory and anti-oxidant effects which can be potentially advantageous upon infection or exposure to other stressors. The potential effects of these feed additives in GSB are very promising to improve health and disease resistance in aquaculture.
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http://dx.doi.org/10.7717/peerj.4001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5719961PMC
December 2017

Gene expression profiling of whole blood cells supports a more efficient mitochondrial respiration in hypoxia-challenged gilthead sea bream ().

Front Zool 2017 6;14:34. Epub 2017 Jul 6.

Nutrigenomics and Fish Growth Endocrinology Group, Institute of Aquaculture Torre de la Sal, Consejo Superior de Investigaciones Científicas (IATS-CSIC), Ribera de Cabanes, E-12595 Castellón, Spain.

Background: Acclimation to abiotic challenges, including decreases in O availability, requires physiological and anatomical phenotyping to accommodate the organism to the environmental conditions. The retention of a nucleus and functional mitochondria in mature fish red blood cells makes blood a promising tissue to analyse the transcriptome and metabolic responses of hypoxia-challenged fish in an integrative and non-invasive manner.

Methods: Juvenile gilthead sea bream () were reared at 20-21 °C under normoxic conditions (> 85% O saturation) followed by exposure to a gradual decrease in water O concentration to 3.0 ppm (41-42% O saturation) for 24 h or 1.3 ppm (18-19% O saturation) for up to 4 h. Blood samples were collected at three different sampling points for haematological, biochemical and transcriptomic analysis.

Results: Blood physiological hallmarks remained almost unaltered at 3.0 ppm, but the haematocrit and circulating levels of haemoglobin, glucose and lactate were consistently increased when fish were maintained below the limiting oxygen saturation at 1.3 ppm. These findings were concurrent with an increase in total plasma antioxidant activity and plasma cortisol levels, whereas the opposite trend was observed for growth-promoting factors, such as insulin-like growth factor I. Additionally, gene expression profiling of whole blood cells revealed changes in upstream master regulators of mitochondria ( and ), antioxidant enzymes ( and ), outer and inner membrane translocases ( and ), components of the mitochondrial dynamics system ( and ), apoptotic factors (), uncoupling proteins () and oxidative enzymes of fatty acid β-oxidation ( and ), the tricarboxylic acid cycle () and the oxidative phosphorylation pathway. The overall response is an extensive reduction in gene expression of almost all respiratory chain enzyme subunits of the five complexes, although mitochondrial-encoded catalytic subunits and nuclear-encoded regulatory subunits of Complex IV were primarily increased in hypoxic fish.

Conclusions: Our results demonstrate the re-adjustment of mitochondrial machinery at transcriptional level to cope with a decreased basal metabolic rate, consistent with a low risk of oxidative stress, diminished aerobic ATP production and higher O-carrying capacity. Taken together, these results suggest that whole blood cells can be used as a highly informative target tissue of metabolic condition.
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http://dx.doi.org/10.1186/s12983-017-0220-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5501551PMC
July 2017

Recombinant bovine growth hormone (rBGH) enhances somatic growth by regulating the GH-IGF axis in fingerlings of gilthead sea bream (Sparus aurata).

Gen Comp Endocrinol 2018 02 27;257:192-202. Epub 2017 Jun 27.

Departament de Biologia Cel·lular, Fisiologia i Immunologia, Facultat de Biologia, Universitat de Barcelona, 08028 Barcelona, Spain. Electronic address:

The growth hormone (GH)/insulin-like growth factors (IGFs) endocrine axis is the main growth-regulator system in vertebrates. Some authors have demonstrated the positive effects on growth of a sustained-release formulation of a recombinant bovine GH (rBGH) in different fish species. The aim of this work was to characterize the effects of a single injection of rBGH in fingerlings of gilthead sea bream on growth, GH-IGF axis, and both myogenic and osteogenic processes. Thus, body weight and specific growth rate were significantly increased in rBGH-treated fish respect to control fish at 6weeks post-injection, whereas the hepatosomatic index was decreased and the condition factor and mesenteric fat index were unchanged, altogether indicating enhanced somatic growth. Moreover, rBGH injection increased the plasma IGF-I levels in parallel with a rise of hepatic mRNA from total IGF-I, IGF-Ic and IGF-II, the binding proteins IGFBP-1a and IGFBP-2b, and also the receptors IGF-IRb, GHR-I and GHR-II. In skeletal muscle, the expression of IGF-Ib and GHR-I was significantly increased but that of IGF-IRb was reduced; the mRNA levels of myogenic regulatory factors, proliferation and differentiation markers (PCNA and MHC, respectively), or that of different molecules of the signaling pathway (TOR/AKT) were unaltered. Besides, the growth inhibitor myostatin (MSTN1 and MSTN2) and the hypertrophic marker (MLC2B) expression resulted significantly enhanced, suggesting altogether that the muscle is in a non-proliferative stage of development. Contrarily in bone, although the expression of most molecules of the GH/IGF axis was decreased, the mRNA levels of several osteogenic genes were increased. The histology analysis showed a GH induced lipolytic effect with a clear decrease in the subcutaneous fat layer. Overall, these results reveal that a better growth potential can be achieved on this species and supports the possibility to improve growth and quality through the optimization of its culture conditions.
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http://dx.doi.org/10.1016/j.ygcen.2017.06.019DOI Listing
February 2018

Dietary supplementation of heat-treated and seaweeds enhanced acute hypoxia tolerance in gilthead sea bream ().

Biol Open 2017 Jun 15;6(6):897-908. Epub 2017 Jun 15.

Nutrigenomics and Fish Growth Endocrinology Group, Institute of Aquaculture Torre de la Sal (CSIC), Ribera de Cabanes, Castellón 12595, Spain.

Intensive aquaculture practices involve rearing fish at high densities. In these conditions, fish may be exposed to suboptimal dissolved O levels with an increased formation of reactive O species (ROS) in tissues. Seaweeds (SW) contain biologically active substances with efficient antioxidant capacities. This study evaluated the effects of dietary supplementation of heat-treated SW (5% or 5% ) on stress bioindicators in sea bream subjected to a hypoxic challenge. 168 fish (104.5 g average weight) were distributed in 24 tanks, in which eight tanks were fed one of three experimental diets for 34 days: (i) a control diet without SW supplementation, (ii) a control diet supplemented with , or (iii) a control diet with Thereafter, fish from 12 tanks (=4 tanks/dietary treatment) were subjected to 24 h hypoxia (1.3 mg O l) and subsequent recovery normoxia (8.6 mg O l). Hypoxic fish showed an increase in hematocrit values regardless of dietary treatment. Dietary modulation of the O-carrying capacity was conspicuous during recovery, as fish fed SW supplemented diets displayed significantly higher haemoglobin concentration than fish fed the control diet. After the challenge, survival rates in both groups of fish fed SW were higher, which was consistent with a decrease in hepatic lipid peroxidation in these groups. Furthermore, the hepatic antioxidant enzyme activities were modulated differently by changes in environmental O condition, particularly in sea bream fed the diet. After being subjected to hypoxia, the gene expression of antioxidant enzymes and molecular chaperones in liver and heart were down regulated in sea bream fed SW diets. This study suggests that the antioxidant properties of heat-treated SW may have a protective role against oxidative stress. The nature of these compounds and possible mechanisms implied are currently being investigated.
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http://dx.doi.org/10.1242/bio.024299DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5483021PMC
June 2017

Olive oil bioactive compounds increase body weight, and improve gut health and integrity in gilthead sea bream (Sparus aurata).

Br J Nutr 2017 02 1;117(3):351-363. Epub 2017 Mar 1.

3Department of Marine Species Biology, Culture and Pathology,Nutrigenomics and Fish Growth Endocrinology Group,Institute of Aquaculture Torre de la Sal,12595,Castellón,CSIC,Spain.

An olive oil bioactive extract (OBE) rich in bioactive compounds like polyphenols, triterpenic acids, long-chain fatty alcohols, unsaturated hydrocarbons, tocopherols and sterols was tested (0, 0·08, 0·17, 0·42 and 0·73 % OBE) in diets fed to sea bream (Sparus aurata) (initial weight: 5·4 (sd 1·2) g) during a 90-d trial (four replicates). Fish fed diets containing 0·17 and 0·42 % OBE were 5 % heavier (61·1 (sd 1·6) and 60·3 (sd 1·1) g, respectively) than those of the control group (57·0 (sd 0·7) g), although feed conversion ratio and specific feed intake did not vary. There were no differences in lipid peroxidation (LPO) levels, catalase, glutathione reductase and glutathione S-transferase activities in the intestine and liver, although there was a tendency of lower intestinal and hepatic LPO levels in fish fed OBE diets. No differences in villus size were found among treatments, whereas goblet cell density in the control group was on average14·3 % lower than in fish fed OBE diets. The transcriptomic profiling of intestinal markers, covering different biological functions like (i) cell differentiation and proliferation, (ii) intestinal permeability, (iii) enterocyte mass and epithelial damage, (iv) IL and cytokines, (v) pathogen recognition receptors and (vi) mitochondria function, indicated that among the eighty-eight evaluated genes, twenty-nine were differentially expressed (0·17 % OBE diet), suggesting that the additive has the potential of improving the condition and defensive role of the intestine by enhancing the maturation of enterocytes, reducing oxidative stress, improving the integrity of the intestinal epithelium and enhancing the intestinal innate immune function, as gene expression data indicated.
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http://dx.doi.org/10.1017/S0007114517000228DOI Listing
February 2017