Browsing by Author "Engrola, Sofia"
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- A nutritional strategy to promote gilthead seabream performance under low temperaturesPublication . Teodósio, Rita; Aragão, Cláudia; Colen, R.; Carrilho, Raquel; Dias, Jorge; Engrola, SofiaGilthead seabream (Sparus aurata) is vulnerable to low water temperature, which may occur in the Southern Europe and Mediterranean region during Winter. Fish are poikilothermic animals, therefore feed intake, digestion, metabolism and ultimately growth are affected by water temperature. This study aimed to evaluate growth performance, feed utilisation, nutrient apparent digestibility, and nitrogen losses to the environment in gilthead seabream juveniles reared under low temperature (similar to 13 degrees C). Three isolipid and isoenergetic diets were formulated: a diet similar to a commercial feed (COM) that contained 44% crude protein and 27.5% fishmeal, and two experimental diets with a lower protein content of 42% (ECO and ECOSup). In both ECO diets fishmeal inclusion was reduced (10% in ECO and 7.5% in ECOSup diet) and 15% poultry meal was included. Additionally, the ECOSup diet was supplemented with a mix of feed additives intended to promote fish growth performance and feed intake. The ECO diets presented lower production costs than the COM diet, whilst incorporating more sustainable ingredients. Gilthead seabream juveniles (+/- 154.5 g initial body weight) were randomly assigned to triplicate tanks and fed the diets for 84 days. Fish fed the ECOSup diet attained a similar final body weight than fish fed the COM diet, significantly higher than fish fed the ECO diet. ECOSup fed fish presented significantly higher hepatosomatic index than COM fed fish, most likely due to higher hepatic glycogen reserves. The viscerosomatic index of ECOSup fed fish were significantly lower compared to COM fed fish, which is a positive achievement from a consumer's point of view. ECOSup diet exhibited similar nutrient digestibility than the COM diet. Moreover, feeding fish with the ECO diets resulted in lower faecal nitrogen losses when compared to COM fed fish. The results suggest that feeding gilthead seabream with an eco-friendly diet with a mix of feed additives such as the ECOSup diet, promoted growth and minimised nitrogen losses to the environment. Nutritional strategies that ultimately promote feed intake and diet utilisation are valuable tools that may help conditioning fish to sustain growth even under low temperatures.
- Alternative proteins for fish diets: implications beyond growthPublication . Aragão, Cláudia; Gonçalves, Ana Teresa; Costas, Benjamín; Azeredo, Rita; Xavier, Maria João; Engrola, SofiaAquaculture has been challenged to find alternative ingredients to develop innovative feed formulations that foster a sustainable future growth. Given the most recent trends in fish feed formulation on the use of alternative protein sources to decrease the dependency of fishmeal, it is fundamental to evaluate the implications of this new paradigm for fish health and welfare. This work intends to comprehensively review the impacts of alternative and novel dietary protein sources on fish gut microbiota and health, stress and immune responses, disease resistance, and antioxidant capacity. The research results indicate that alternative protein sources, such as terrestrial plant proteins, rendered animal by-products, insect meals, micro- and macroalgae, and single cell proteins (e.g., yeasts), may negatively impact gut microbiota and health, thus affecting immune and stress responses. Nevertheless, some of the novel protein sources, such as insects and algae meals, have functional properties and may exert an immunostimulatory activity. Further research on the effects of novel protein sources, beyond growth, is clearly needed. The information gathered here is of utmost importance, in order to develop innovative diets that guarantee the production of healthy fish with high quality standards and optimised welfare conditions, thus contributing to a sustainable growth of the aquaculture industry.
- Amino acid metabolism in gilthead seabream is affected by the dietary protein to energy ratiosPublication . Teodósio, Rita; Aragão, Cláudia; Conceicao, Luis; Dias, Jorge; Engrola, SofiaThe dietary protein to energy ratio (P/E) has proven to influence protein utilization and/or growth in several fish species. This study intended to unravel the bioavailability and metabolic fate of lysine and methionine in gilthead seabream (Sparus aurata) juveniles fed plant diets with different P/E ratios. Seabream juveniles were fed two isonitrogenous diets (45% crude protein) differing in crude lipids (20 and 14%): LowP/E (P/E ratio=20.0 mg protein kJ-1) and HighP/E (P/E ratio=21.4 mg protein kJ-1). After three weeks, fish (11.6 +/- 4.3 g) were tube-fed the respective diet labelled with C-14-protein (L-amino acid mixture), C-14-lysine, or C-14-methionine. Protein, lysine, and methionine utilization were determined based on the proportion of C-14-amino acid evacuated, retained in the free or protein-bound fraction of liver and muscle, or catabolized. This study revealed that a decrease in P/E ratio resulted in lower amino acid evacuation (p < 0.05), contributing to a more efficient amino acid uptake. Results indicate that amino acids are retained as protein in the liver and not only temporarily available in the free pool. The amount of free amino acids retained in the muscle of LowP/E fed fish was significantly higher than in HighP/E fish (p < 0.05) due to a simultaneous higher retention of lysine and methionine, without affecting the overall protein retention. Methionine catabolism was significantly lower than lysine or protein independently of the P/E ratio (p < 0.05), reinforcing that this amino acid is preferentially spared for metabolic functions and not used as energy source. In contrast, increasing the dietary P/E ratio decreased lysine catabolism and increased its availability for growth. The bioavailability and metabolism of individual amino acids should be considered when optimizing P/E ratios in diets for gilthead seabream juveniles. Formulating diets with optimum P/E ratios will improve diet utilization and fish performance.
- Amino acid supplementation in fish nutrition and welfarePublication . Aragão, Cláudia; Engrola, Sofia; Costas, BenjamínAmino acids are not only the fundamental building blocks of proteins and, consequently, essential for animal growth, but they have also gained recognition in recent years for their critical roles in regulating key metabolic processes. In aquaculture, where the demand for more sustainable production systems continues to grow, nutrition remains a central pillar. Within this context, amino acids serve dual purposes: they are essential to balance the nutritional profiles of alternative protein sources, and they also act as functional ingredients that can enhance fish health, robustness, and resilience to stressors.
- Antemortem versus postmortem methods for detection of betanodavirus in Senegalese sole (Solea senegalensis)Publication . Olveira, J. G.; Soares, F.; Engrola, Sofia; Dopazo, C. P.; Bandín, I.The suitability of nested reverse transcription polymerase chain reaction (nRT-PCR) to detect betanodavirus in blood samples from naturally infected Senegalese sole (Solea senegalensis) was evaluated in comparison with other diagnostic methods. Results indicated that histologic examination of brain lesions could be regarded as the most consistent indicator of nodavirus infection in this species. The nRT-PCR showed low to moderate levels of detection; the best values were obtained in brain samples followed by blood samples. Inoculation of SSN-1 and SAF-1 cells with fish samples did not cause cytopathic effect, although virus was detected by reverse transcription polymerase chain reaction in approximately 25% of the SSN-1 inoculated wells. The efficiency of detection of the viral genome was dramatically increased by the use of nRTPCR, reaching 90.6% of positives in brain samples and 84.4% in blood samples. The sensitivity and the negative predictive value of nRT-PCR in blood samples were slightly lower than those obtained using brain samples. Nevertheless, it is suggested that the advantage of being able to perform diagnosis on live fish adequately counterbalances the slightly lower sensitivity of nRT-PCR on blood samples. This technique is proposed as a useful tool, not only for the selection of nodavirus-free breeders but also to check the fish status during ongrowing.
- Data on European seabass fed with methionine-enriched diets obtained through label free shotgun proteomicsPublication . Farinha, Ana Paula; Schrama, Denise; Silva, Tomé; Conceição, Luís E.C.; Colen, R.; Engrola, Sofia; Rodrigues, Pedro; Cerqueira, MarcoThis data article is associated with the research article "Evaluating the impact of methionine-enriched diets in the liver of European seabass through label-free shotgun proteomics". Here it is described the data obtained from proteomic analysis of 36 European seabass juveniles (3 fish x 3 replicate tanks) after 18 days of feeding with experimental diets containing four inclusion levels of methionine (Met): 0.77%, 1%, 1.36% and 1.66% Met (w/w). We analysed this dataset and compared it with that obtained during the long-term feeding period i.e., 85 days. Fish liver proteins were digested with trypsin and purified peptides were analysed by LC-MS/MS. Proteins were identified with at least two peptides at 0.1% Decoy false discovery rate (FDR). In this dataset, we present the analysis of the differential abundant proteins (DAP) with significant differences across treatments after 18 days of feeding (One-Way ANOVA, p < 0.05). Treatment's comparisons were also performed between the 18- and 85-days feeding trials through Two-Way ANOVA (p < 0.05). MS/MS raw data are available via ProteomeXChange with identifiers PXD019610 and 10.6019/PXD019610 (18-days dataset); and PXD019622 and 10.6019/PXD019622 (85-days dataset). This dataset corresponds to fish sampled after 18-days of experimental trial and is made available to support the study conducted in the afore-mentioned article, by performing the analysis during a short-term period of feeding. The data presented may be further used in other nutritional studies e.g., addressing hepatic changes mediated by Met.
- Development and application of a mechanistic nutrient-based model for precision fish farmingPublication . Soares, Filipe M. R. C.; Nobre, Ana M. D.; Raposo, Andreia I. G.; Mendes, Rodrigo; Engrola, Sofia; Rema, Paulo J. A. P.; Conceição, Luís E. C.; Silva, Tomé S.This manuscript describes and evaluates the FEEDNETICS model, a detailed mechanistic nutrient-based model that has been developed to be used as a data interpretation and decisionsupport tool by fish farmers, aquafeed producers, aquaculture consultants and researchers. The modelling framework comprises two main components: (i) fish model, that simulates at the individual level the fish growth, composition, and nutrient utilization, following basic physical principles and prior information on the organization and control of biochemical/metabolic processes; and (ii) farm model, that upscales all information to the population level. The model was calibrated and validated for five commercially relevant farmed fish species, i.e., gilthead seabream (Sparus aurata), European seabass (Dicentrarchus labrax), Atlantic salmon (Salmo salar), rainbow trout (Oncorhynchus mykiss), and Nile tilapia (Oreochromis niloticus), using data sets covering a wide range of rearing and feeding conditions. The results of the validation of the model for fish growth are consistent between species, presenting a mean absolute percentage error (MAPE) between 11.7 and 13.8%. Several uses cases are presented, illustrating how this tool can be used to complement experimental trial design and interpretation, and to evaluate nutritional and environmental effects at the farm level. FEEDNETICS provides a means of transforming data into useful information, thus contributing to more efficient fish farming
- Dietary antioxidant supplementation promotes growth in senegalese sole postlarvaePublication . Xavier, Maria J; Engrola, Sofia; Conceição, Luis E C; Manchado, Manuel; Carballo, Carlos; Gonçalves, Renata; Colen, R.; Figueiredo, Vera; Valente, Luisa M PSomatic growth is a balance between protein synthesis and degradation, and it is largely influenced by nutritional clues. Antioxidants levels play a key role in protein turnover by reducing the oxidative damage in the skeletal muscle, and hence promoting growth performance in the long-term. In the present study, Senegalese sole postlarvae (45 days after hatching, DAH) were fed with three experimental diets, a control (CTRL) and two supplemented with natural antioxidants: curcumin (CC) and grape seed (GS). Trial spanned for 25 days and growth performance, muscle cellularity and the expression of muscle growth related genes were assessed at the end of the experiment (70 DAH). The diets CC and GS significantly improved growth performance of fish compared to the CTRL diet. This enhanced growth was associated with larger muscle cross sectional area, with fish fed CC being significantly different from those fed the CTRL. Sole fed the CC diet had the highest number of muscle fibers, indicating that this diet promoted muscle hyperplastic growth. Although the mean fiber diameter did not differ significantly amongst treatments, the proportion of large-sized fibers (>25 μm) was also higher in fish fed the CC diet suggesting increased hypertrophic growth. Such differences in the phenotype were associated with a significant up-regulation of the myogenic differentiation 2 (myod2) and the myomaker (mymk) transcripts involved in myocyte differentiation and fusion, respectively, during larval development. The inclusion of grape seed extract (GS diet) resulted in a significant increase in the expression of myostatin1. These results demonstrate that both diets (CC and GS) can positively modulate muscle development and promote growth in sole postlarvae. This effect is more prominent in CC fed fish, where increased hyperplastic and hypertrophic growth of the muscle was associated with an upregulation of myod2 and mymk genes.
- Dietary curcumin promotes gilthead seabream larvae digestive capacity and modulates oxidative statusPublication . Xavier, Maria João; Dardengo, Gian Marco; Navarro-Guillén, Carmen; Lopes, Andre D.; Colen, R.; Valente, Luisa M. P.; Conceição, Luís E. C.; Engrola, SofiaThe larval stage is highly prone to stress due to the ontogenetic and metabolic alterations occurring in fish. Curcumin inclusion in diets has been shown to improve growth by modulating oxidative status, immune response, and/or feed digestibility in several fish species. The aim of the present work was to assess if dietary curcumin could promote marine fish larvae digestive maturation and improve robustness. Gilthead seabream larvae were fed a diet supplemented with curcumin at dose of 0 (CTRL), 1.5 (LOW), or 3.0 g/Kg feed for 27 days. From 4 to 24 days after hatching (DAH), no differences were observed in growth performance. At the end of the experiment (31 DAH) LOW larvae had a better condition factor than CTRL fish. Moreover, HIGH larvae showed higher trypsin and chymotrypsin activity when compared to CTRL fish. LOW and HIGH larvae were able to maintain the mitochondrial reactive oxygen species production during development, in contrast to CTRL larvae. In conclusion, curcumin supplementation seems to promote larvae digestive capacity and modulate the oxidative status during ontogeny. Furthermore, the present results provide new insights on the impacts of dietary antioxidants on marine larvae development and a possible improvement of robustness in the short and long term.
- Dietary methionine supplementation improves the European seabass (Dicentrarchus labrax) immune status following long-term feeding on fishmeal-free dietsPublication . Machado, Marina; Engrola, Sofia; Colen, R.; Conceicao, Luis E. C.; Dias, Jorge; Costas, BenjaminMethionine is a limiting amino acid (AA) in fish diets, particularly in those containing high levels of plant protein (PP), and is key in the immune system. Accordingly, outcome on the fish immune mechanisms of methionine-deficient and methionine-supplemented diets within the context of 0 % fishmeal formulation, after a short and prolonged feeding period, was studied in European seabass (Dicentrarchus labrax). For this, seabass juveniles were fed a (i) fishmeal-free diet, meeting AA requirements, but deficient in methionine (MET0·65); (ii) as control, the MET0·65 supplemented with L-methionine at 0·22 % of feed weight (CTRL); (iii) two diets, identical to MET0·65 but supplemented at 0·63 and 0·88 % of feed weight of L-methionine (MET1·25 and MET1·5, respectively); and (iv) a fishmeal-based diet (FM), as positive control. After 2 and 12 weeks of feeding, blood and plasma were sampled for leucocyte counting and humoral parameter assays and head-kidney collected for gene expression. After 2 weeks of feeding, a fishmeal-free diet supplemented with methionine led to changes in the expression of methionine- and leucocyte-related genes. A methionine immune-enhancer role was more evident after 12 weeks with an increased neutrophil percentage and a decreased expression of apoptotic genes, possibly indicating an enhancement of fish immunity by methionine dietary supplementation. Furthermore, even though CTRL and FM present similar methionine content, CTRL presented a reduced expression of several immune-related genes indicating that in a practical PP-based diet scenario, the requirement level of methionine for an optimal immune status could be higher.