Browsing by Author "Fuentes, Juan"
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- 16.P15. Branchial osmoregulatory response to salinity challenge in the Lusitanian toadfishPublication . Guerreiro, Pedro M; Laiz-Carrión, R.; Haond, Christophe; Modesto, Teresa; Fuentes, Juan; Mancera, J. M.; Canario, Adelino V. M.Halobatrachus didactylus, a marine teleost found in coastal lagoons and river estuaries is often exposed to important salinity changes. Despite its aglomerular kidney, it is able to survive in hypo-osmotic environments, likely via compensatory actions from gills and intestine. We aimed at evaluating the response of the branchial tissue of H. didactylus to salinity changes.
- Aflatoxicosis Dysregulates the Physiological Responses to Crowding Densities in the Marine Teleost Gilthead Seabream (Sparus aurata)Publication . Barany, Andre; Fuentes, Juan; Martínez-Rodríguez, Gonzalo; Mancera, Juan MiguelSeveral studies in fish have shown that aflatoxin B1 (AFB1) causes a disparity of speciesdependent physiological disorders without compromising survival. We studied the effect of dietary administration of AFB1 (2 mg AFB1 kg−1 diet) in gilthead seabream (Sparus aurata) juveniles in combination with a challenge by stocking density (4 vs. 40 g L−1 ). The experimental period duration was ten days, and the diet with AFB1 was administered to the fish for 85 days prior to the stocking density challenge. Our results indicated an alteration in the carbohydrate and lipid metabolites mobilization in the AFB1 fed group, which was intensified at high stocking density (HSD). The CT group at HSD increased plasma cortisol levels, as expected, whereas the AFB1-HSD group did not. The star mRNA expression, an enzyme involved in cortisol synthesis in the head kidney, presented a ninefold increase in the AFB1 group at low stocking density (LSD) compared to the CT-LSD group. Adenohypophyseal gh mRNA expression increased in the AFB1-HSD but not in the CT-HSD group. Overall, these results confirmed that chronic AFB1 dietary exposure alters the adequate endocrinological physiological cascade response in S. aurata, compromising the expected stress response to an additional stressor, such as overcrowding.
- Agouti overexpression in a transgenic model regulates integrity, permeability and electrogenic amino acid transport in zebrafish intestinePublication . Leal, Esther; Angotzi, Anna Rita; Godino-Gimeno, Alejandra; Gregorio, Silvia F.; Rotllant, Josep; Saera-Vila, Alfonso; Fuentes, Juan; Cerdá-Reverter, José MiguelOverexpression of asip1 in transgenic zebrafish disrupts dorsoventral pigment pattern in addition to increasing food intake levels and linear growth. A higher feed intake is unnecessary in transgenic fish to enable larger and heavier growth. A plausible explanation may rely on the enhanced feeding efficiency mediated by improved nutrient absorption in transgenic animals. To test this hypothesis, wide scope transcriptomic techniques were used to elucidate the potential pathways involved in the enhanced nutrient absorption and intestinal epithelium permeability/integrity. In addition, the electrogenic capacity for amino acid transport was analysed. Transcriptomic analysis reveal that amino acid, monocarboxylates, ionic and vitamin transmembrane transporters were substantially modified. Enrichment analysis also revealed an inhibition of intestinal lipid metabolism and down-regulation of KEGG pathways related to membrane integrity suggesting augmented intestinal laxity that may enhance paracellular transport. Electrophysiological experiments carried out in Ussing chambers show that asip1 overexpression decrease membraned tissue resistance (Rt), indicating a modification of the intestinal barrier function in ASIP1 transgenic animals. Similarly, paracellular permeability was higher in transgenic zebrafish. Both the decrease in Rt and the increase in permeability point to an ASIP1-dependent decrease in the tissue barrier function. Electrogenic amino acid transport was also enhanced in transgenic animals providing strong indication that ASIP1 fish can extract more amino acids from their diet at similar feeding levels. Both transcriptomic and electrophysiological results suggest that asip1-overexpressing zebrafish display improved nutrient absorption and by extension a higher feed efficiency which explains enhanced growth in the absence of augmented food intake. The enhanced growth of ASIP1 zebrafish potentially mediated by improved nutrient uptake and feed efficiency suggests that the melanocortin system, specifically asip1 overexpression, is a potential target for the development of genetically engineered fish displaying improved performance and no differential lipid accumulation.
- Alternative formulations for gilthead seabream diets: towards a more sustainable productionPublication . Aragão, Cláudia; Cabano, Miguel; Colen, R.; Fuentes, Juan; Dias, JorgeTo support the expected increase in aquaculture production during the next years, a wider range of alternative ingredients to fishmeal is needed, towards contributing to an increase in production sustainability. This study aimed to test diets formulated with non‐conventional feed ingredients on gilthead seabream (Sparus aurata) growth performance, feed utilization, apparent digestibility of nutrients and nutrient outputs to the environment. Four isonitrogenous and isoenergetic diets were formulated: a control diet (CTRL) similar to a commercial feed and three experimental diets containing, as main protein sources, plant by‐products, glutens and concentrates (PLANT); processed animal proteins (PAP); or micro/macroalgae, insect meals and yeast (EMERG). Diets were tested in triplicate during 80 days. The EMERG treatment resulted in lower fish growth performance, higher FCR and lower nutrient and energy retentions than the other treatments. The lowest protein digestibility was found for the EMERG diet, which caused increased nitrogen losses. The PLANT and PAP treatments resulted in better fish growth performance, higher nutrient and energy retentions, and lower FCR than the CTRL treatment. The significant improvement in FCR found for fish fed PLANT and PAP diets and the high protein digestibility of these diets contribute towards minimizing the environmental impacts of seabream production.
- Biotechnological treatment of microalgae enhances growth performance, hepatic carbohydrate metabolism and intestinal physiology in gilthead seabream (Sparus aurata) juveniles close to commercial sizePublication . Molina-Roque, Luis; Bárany, André; Sáez, María Isabel; Alarcón, Francisco Javier; Tapia, Silvana Teresa; Fuentes, Juan; Mancera, Juan Miguel; Perera, Erick; Martos-Sitcha, Juan AntonioThe aim of this work was to evaluate the effects on growth performance, intermediary metabolism and welfare of the inclusion of two commercial microalgae-based ingredients called LB-ChromaBream (LB-CB) and LB-ChromaBream-plus (LB-CBplus) in the diet of gilthead seabream (Sparus aurata) close to commercial size. For this purpose, fish of similar to 182 g of initial body mass were fed to satiety (ad libitum) for 41 days with three different diets: (i) CONTROL diet (CTRL), with a commercial-like formulation; (ii) LB-CB diet, with a 10 % inclusion of microalgal product; (iii) LB-CBplus diet, with a 10 % inclusion of the same product but enzymatically hydrolysed to increase the bioavailability of the nutrients. The results obtained show that the use of these microalgal products leads an overall improvement in productive parameters in terms of growth (15 % in SGR) and feed efficiency (11 %), as well as a significant reduction in circulating cortisol with the LB-CBplus diet. Observations on plasma and liver metabolites, and particularly on hepatic metabolic enzymes, collectively indicate that microalgae supplementation of feed lead to a better use of carbohydrates as a source of energy in the liver and other tissues, potentially sparing triglycerides within this tissue, and a channelling hepatic triglycerides to fuels growth. Finally, the specimens fed the supplemented diets experienced a substantial improvement in intestinal health, achieved by longer intestines, a higher transepithelial resistance and better apparent permeability measured by electrophysiological methods, especially those fed LB-CBplus, which could explain the increase in productive performance by improving nutrient assimilation. In conclusion, this study shows that the experi-mental feeds, especially the one containing biotechnologically treated microalgae, are suitable for improving some important indicators of growth performance and physiological condition of gilthead seabream, thus revealing the potential for their inclusion in new functional feeds for this species at an advanced stage of the production cycle.
- Cloning, characterisation and tissue distribution of an aquaporin-3 cDNA from fish (Sparus aurata)Publication . Santos, CRA; Fuentes, Juan; Cardoso, João CR; Estêvão, Dulce; Power, DeborahThe major intrinsic protein (MIP) family consists of several transmembrane channel proteins specific for water and neutral solutes. All proteins belonging to the MIP family evolved from two divergent bacterial paralogues, one giving rise to the CHIP group, functionally characterised as water channels and the other to the GLP group, specialised in glycerol transport. Three forms of MIP proteins belonging to the GLP group have been identified in mammals: aquaporin-3 (AQP-3), aquaporin-7 (AQP-7) and aquaporin-9 (AQP-9). We have recently cloned and characterised a GLP cDNA from the marine teleost sea bream (Sparus aurata) and studied its tissue distribution. Phylogenetic analysis revealed it was most like AQP-3 and further studies are now underway to determine its role in hydromineral balance.
- Corrigendum to ‘Dietary aflatoxin B1 (AFB1) reduces growth performance, impacting growth axis, metabolism, and tissue integrity in juvenile gilthead sea bream (Sparus aurata)’. Aquaculture, volume 533, 25 February 2021, 736189Publication . Barany, A.; Guilloto, M.; Cosano, J.; de Boevre, M.; Oliva, M.; de Saeger, S.; Fuentes, Juan; Martínez-Rodriguez, G.; Mancera, J.M.The authors regret the errors in a few table references within the text. Specifically, it should reads as follows within the following subsections/ page: 3.2. Blood analysis (page 5)
- Dietary butyrate helps to restore the Intestinal status of a marine teleost (Sparus aurata) fed extreme diets low in fish meal and fish oilPublication . Estensoro, Itziar; Ballester-Lozano, Gabriel; Benedito-Palos, Laura; Grammes, Fabian; Martos-Sitcha, Juan Antonio; Mydland, Liv-Torunn; Calduch-Giner, Josep Alvar; Fuentes, Juan; Karalazos, Vasileios; Ortiz, Alvaro; Overland, Margareth; Sitja-Bobadilla, Ariadna; Perez-Sanchez, JaumeThere is a constant need to find feed additives that improve health and nutrition of farmed fish and lessen the intestinal inflammation induced by plant-based ingredients. The objective of this study was to evaluate the effects of adding an organic acid salt to alleviate some of the detrimental effects of extreme plant-ingredient substitution of fish meal (FM) and fish oil (FO) in gilthead sea bream diet. Three experiments were conducted. In a first trial (T1), the best dose (0.4%) of sodium butyrate (BP-70 (R) NOREL) was chosen after a short (9 weeks) feeding period. In a second longer trial (T2) (8 months), four diets were used: a control diet containing 25% FM (T2-D1) and three experimental diets containing 5% FM (T2-D2, T2-D3, T2-D4). FO was the only added oil in D1, while a blend of plant oils replaced 58% and 84% of FO in T2-D2, and T2-D3 and T2-D4, respectively. The latter was supplemented with 0.4% BP-70. In a third trial (T3), two groups of fish were fed for 12 and 38 months with D1, D3 and D4 diets of T2. The effects of dietary changes were studied using histochemical, immunohistochemical, molecular and electrophysiological tools. The extreme diet (T2-D3) modified significantly the transcriptomic profile, especially at the anterior intestine, up-regulating the expression of inflammatory markers, in coincidence with a higher presence of granulocytes and lymphocytes in the submucosa, and changing genes involved in antioxidant defences, epithelial permeability and mucus production. Trans-epithelial electrical resistance (Rt) was also decreased (T3-D3). Most of these modifications were returned to control values with the addition of BP-70. None of the experimental diets modified the staining pattern of PCNA, FABP2 or ALPI. These results further confirm the potential of this additive to improve or reverse the detrimental effects of extreme fish diet formulations.
- Disruption of gut integrity and permeability contributes to enteritis in a fish-parasite model: a story told from serum metabolomicsPublication . Sitjà-Bobadilla, Ariadna; Gil-Solsona, Rubén; Estensoro, Itziar; Piazzon, M. C; Martos-Sitcha, Juan A; Picard-Sánchez, Amparo; Fuentes, Juan; Sancho, Juan V; Calduch-Giner, Josep A; Hernández, Félix; Pérez-Sánchez, JaumeBackground 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.
- Dysregulation of intestinal physiology by Aflatoxicosis in the Gilthead Seabream (Sparus aurata)Publication . Barany, Andre; Oliva, Milagrosa; Gregorio, Silvia; Martínez-Rodríguez, Gonzalo; Mancera, Juan Miguel; Fuentes, JuanAflatoxin B1 (AFB1) is a mycotoxin often present in food. This study aimed to understand the physiological effects of AFB1 on the seabream (Sparus aurata) gastrointestinal system. In a first in vitro approach, we investigated ion transport using the short-circuit current (Isc) technique in Ussing chambers in the anterior intestine (AI). Application of apical/ luminal AFB1 concentrations of 8 and 16μM to healthy tissues was without effect on tissue transepithelial electrical resistance (TER), and apparent tissue permeability (Papp) was measured using fluorescein FITC (4 kD). However, it resulted in dose-related effects on Isc. In a second approach, seabream juveniles fed with different AFB1 concentrations (1 and 2mg AFB1 kg−1 fish feed) for 85days showed significantly reduced gill Na+ /K+-ATPase (NKA) and H+-ATPase (HA) activities in the posterior intestine (PI). Moreover, dietary AFB1 modified Isc in the AI and PI, significantly affecting TER in the AI. To understand this effect on TER, we analyzed the expression of nine claudins and three occludins as markers of intestinal architecture and permeability using qPCR. Around 80% of the genes presented significantly different relative mRNA expression between AI and PI and had concomitant sensitivity to dietary AFB1. Based on the results of our in vitro, in vivo, and molecular approaches, we conclude that the effects of dietary AFB1 in the gastrointestinal system are at the base of the previously reported growth impairment caused by AFB1 in fish.