Browsing by Author "Costas, B."
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- Changes in liver proteome expression of Senegalese sole (Solea senegalensis) in response to repeated handling stressPublication . Cordeiro, O.; Silva, Tomé S.; Alves, Ricardo N.; Costas, B.; Wulff, Tune; Richard, Nadège; Vareilles, Mahaut; Conceição, L. E. C.; Rodrigues, PedroThe Senegalese sole, a high-value flatfish, is a good candidate for aquaculture production. Nevertheless, there are still issues regarding this species’ sensitivity to stress in captivity. We aimed to characterize the hepatic proteome expression for this species in response to repeated handling and identify potential molecular markers that indicate a physiological response to chronic stress. Two groups of fish were reared in duplicate for 28 days, one of them weekly exposed to handling stress (including hypoxia) for 3 min, and the other left undisturbed. Two-dimensional electrophoresis enabled the detection of 287 spots significantly affected by repeated handling stress (Wilcoxon–Mann–Whitney U test, p<0.05), 33 of which could be reliably identified by peptide mass spectrometry. Chronic exposure to stress seems to have affected protein synthesis, folding and turnover (40S ribosomal protein S12, cathepsin B, disulfide-isomerase A3 precursor, cell-division cycle 48, and five distinct heat shock proteins), amino acid metabolism, urea cycle and methylation/folate pathways (methionine adenosyltransferase I α, phenylalanine hydroxylase, mitochondrial agmatinase, serine hydroxymethyltransferase, 3-hydroxyanthranilate 3,4-dioxygenase, and betaine homocysteine methyltransferase), cytoskeletal (40S ribosomal protein SA, α-actin, β-actin, α-tubulin, and cytokeratin K18), aldehyde detoxification (aldehyde dehydrogenase 4A1 family and aldehyde dehydrogenase 7A1 family), carbohydrate metabolism and energy homeostasis (fatty acid-binding protein, enolase 3, enolase 1, phosphoglycerate kinase, glyceraldehyde-3-phosphate dehydrogenase, aconitase 1, mitochondrial ATP synthase α-subunit, and electron-transfer flavoprotein α polypeptide), iron and selenium homeostasis (transferrin and selenium binding protein 1), steroid hormone metabolism (3-oxo-5-β-steroid 4-dehydrogenase), and purine salvage (hypoxanthine phosphoribosyltransferase). Further characterization is required to fully assess the potential of these markers for the monitoring of fish stress response to chronic stressors of aquaculture environment.
- Changes in plasma amino acid levels in a euryhaline fish exposed to different environmental salinitiesPublication . Aragão, C.; Costas, B.; Vargas-Chacoff, L.; Ruiz-Jarabo, I.; Dinis, Maria Teresa; Mancera, J. M.; Conceição, L. E. C.Previous studies have shown that Senegalese sole is partially euryhaline in the juvenile phase, being able to adapt to a wide range of salinities in a short-time period, due to changes at the osmoregulatory and metabolic level. This study aimed to assess the effects of acclimation of sole to a wide range of salinities, with a special emphasis on the role of plasma amino acids during this process. Sole juveniles were acclimated for 2 weeks to different salinities: 5, 15, 25, 38, and 55 g L−1. Plasma levels of cortisol, glucose, osmolality, and free amino acids were assessed at the end. Changes in plasma levels of cortisol, glucose, and amino acids indicate that fish reared at 5 and 55 g L−1 were facing extra energy costs. Amino acids seem to play an important role during salinity acclimation, either as energy sources or as important osmolytes for cell volume regulation.
- Dietary nitrogen and fish welfarePublication . Conceição, L. E. C.; Aragão, C.; Dias, J.; Costas, B.; Terova, G.; Martins, C.; Tort, L.Little research has been done in optimizing the nitrogenous fraction of the fish diets in order to minimize welfare problems. The purpose of this review is to give an overview on how amino acid (AA) metabolism may be affected when fish are under stress and the possible effects on fish welfare when sub-optimal dietary nitrogen formulations are used to feed fish. In addition, it intends to evaluate the current possibilities, and future prospects, of using improved dietary nitrogen formulations to help fish coping with predictable stressful periods. Both metabolomic and genomic evidence show that stressful husbandry conditions affect AA metabolism in fish and may bring an increase in the requirement of indispensable AA. Supplementation in arginine and leucine, but also eventually in lysine, methionine, threonine and glutamine, may have an important role in enhancing the innate immune system. Tryptophan, as precursor for serotonin, modulates aggressive behaviour and feed intake in fish. Bioactive peptides may bring important advances in immunocompetence, disease control and other aspects of welfare of cultured fish. Fishmeal replacement may reduce immune competence, and the full nutritional potential of plant-protein ingredients is attained only after the removal or inactivation of some antinutritional factors. This review shows that AA metabolism is affected when fish are under stress, and this together with sub-optimal dietary nitrogen formulations may affect fish welfare. Furthermore, improved dietary nitrogen formulations may help fish coping with predictable stressful events.
- Different environmental temperatures affect amino acid metabolism in the eurytherm teleost Senegalese sole (Solea senegalensis Kaup, 1858) as indicated by changes in plasma metabolitesPublication . Costas, B.; Aragão, C.; Ruiz-Jarabo, I.; Vargas-Chacoff, L.; Arjona, F. J.; Mancera, J. M.; Dinis, Maria Teresa; Conceição, L. E. C.Senegalese sole (Solea senegalensis) is a eurytherm teleost that under natural conditions can be exposed to annual water temperature fluctuations between 12 and 26C. This study assessed the effects of temperature on sole metabolic status, in particular in what concerns plasma free amino acid changes during thermal acclimation.
- Effects of dietary amino acids and repeated handling on stress response and brain monoaminergic neurotransmitters in Senegalese sole (Solea senegalensis) juvenilesPublication . Costas, B.; Aragão, C.; Soengas, J. L.; Míguez, J. M.; Rema, P.; Dias, J.; Afonso, A.; Conceição, L. E. C.The present study aimed to assess the effects of increased availability of dietary amino acids (AA) on brain monoamine neurotransmitters and the metabolic processes resulting from stressful situations in fish. Senegalese sole (Solea senegalensis) juveniles (24.2 ± 0.4 g wet mass) were weekly subjected to an acute handling stressor (HDLG) or remained undisturbed (CTL). Additionally, both treatments were fed a control or a high protein (HP) diet (CTL, CTL HP, HDLG and HDLG HP). The HP diet slightly increased the levels of digestible indispensable AA, together with tyrosine and cysteine. Repeated handling induced a stress response after 14 and 28 days in fish held at both HDLG and HDLG HP treatments. While dietary treatment and handling stress activated the serotonergic system at 14 days, these effects were not observed after 28 days. In addition, the HP diet minimized the decrease in plasma indispensable AA due to repeated handling stress after 28 days. It was concluded that HP diet decreased post-stress plasma glucose and lactate levels in HDLG HP specimens only at 14 days of treatment. Moreover, dietary treatment was also effective in stimulating DA synthesis and release, thus dietary phenylalanine supplementation can increase DA biosynthesis in fish.
- Feed deprivation in Senegalese sole (Solea senegalensis Kaup, 1858) juveniles: Effects on blood plasma metabolites and free amino acid levelsPublication . Costas, B.; Aragão, C.; Ruiz-Jarabo, I.; Vargas-Chacoff, L.; Arjona, F. J.; Dinis, Maria Teresa; Mancera, J. M.; Conceição, L. E. C.The effects of prolonged feed deprivation were assessed on blood plasma metabolites and free amino acid levels in Solea senegalensis. Juvenile specimens were maintained at two experimental conditions (24 h fasted and 21 days feed-deprived). In feed-deprived fish, relative growth rate and hepatosomatic index as well as plasma triglycerides and proteins levels were significantly lower.
- Health status in gilthead seabream (Sparus aurata) juveniles fed diets devoid of fishmeal and supplemented with Phaeodactylum tricornutumPublication . Reis, B.; Ramos-Pinto, L.; Martos-Sitcha, J. A.; Machado, M.; Azeredo, R.; Fernandez-Boo, S.; Engrola, S.; Unamunzaga, C.; Calduch-Giner, J.; Conceicao, L. E. C.; Silva, T.; Dias, J.; Costas, B.; Perez-Sanchez, J.To enhance fish general health, feeds can be supplemented with health-promoting additives, reducing the need to use chemotherapeutics. Incorporation of marine algae biomasses in aquafeeds has been shown to improve fish immune status by enhancing innate immune response. This study evaluated the effects of Phaeodactylum tricornutum incorporation in feed by two different processes, either as freeze-dried biomass or broken cell wall biomass, on fish health status and performance. Triplicate groups of gilthead seabream juveniles (13.3 +/- 0.3 g) were either fed a control diet (CTRL) with an extreme (i.e., 0% fishmeal), nutritionally balanced, formulation, or two experimental diets formulated as the CTRL with 1% inclusion of the microalga P. tricornutum at the expense of wheat meal: BC diet contains P. tricornutum broken cells and WC diet microalgae whole cells. After 2 and 12 weeks of feeding, blood was collected for hematological procedures, whereas plasma and mucus were sampled for immune parameters. Head-kidney, liver, and white skeletal muscle were also collected for gene expression measurements. No major differences were observed in hematological nor plasma humoral parameters after 12 weeks irrespective of dietary treatment. Arrays of 29-31 genes were analyzed in the different tissues, revealing an early dietary effect (2 weeks) in a tissue-specific pattern. In the liver, the major effect was found in the GH/IGF axis and in muscle there was a late downregulation of myostatin (mstn) gene, mainly due to WC diet, even though all fish had similar growth performance. Regarding the head-kidney, BC diet led to alpha-2-macroglobulin (a2m) gene upregulation. Also, the same treatment showed increased mucus alternative complement pathway and bactericidal activity at 2 and 12 weeks, respectively. Hence, it seems that BC diet has a potential stimulatory effect that might be relevant as a prophylactic measure before a predictable stressful event.
- High stocking density induces crowding stress and affects amino acid metabolism in Senegalese sole Solea senegalensis (Kaup 1858) juvenilesPublication . Costas, B.; Aragão, C.; Mancera, J. M.; Dinis, Maria Teresa; Conceição, L. E. C.; Refojos, B. C.Fish held at high stocking densities are generally exposed to chronic stress situations that impose severe energy demands and may predispose the ¢sh to infection. Senegalese sole Solea senegalensis (Kaup) juveniles (78.8 18.9 g body weight) were maintained at low (LSD; 4 kg m2 at the end of the experiment), medium (MSD; 9 kg m2 ) and high (HSD;14 kg m2 ) stocking densities during a period of 63 days. Although disease outbreaks were observed in ¢sh reared at HSD, growth and food consumption did not vary among di¡erent treatments. Results from plasma cortisol and free amino acids (FAAs) showed signi¢- cant di¡erences among di¡erent rearing densities pointing to HSD as stressful rearing condition.
- Individual differences in metabolism predict coping styles in fishPublication . Martins, C. I. M.; Castanheira, Maria Filipa; Engrola, S.; Costas, B.; Conceição, L. E. C.Studies on metabolism usually rely on measurements of oxygen consumption obtained in respirometry chambers. Despite rigorous standardization there is still considerable interindividual variation in metabolic rates which is often ignored. Furthermore, housing in respirometry chambers implies handling and exposure to a new environment, conditions known to be a source of stress in animals. The extent of stress response is known to be dependent on an individual’s coping style. Therefore, the extent of individual variation in respirometry data may be related to the expression of coping styles. The goal of this study is to determine whether the variation in oxygen consumption after a stressor is the resultant of an individual’s coping style. Our approach was to determine oxygen consumption in fish immediately after transfer to respiratory chambers and during the subsequent 22 h. In addition, the same individuals were subjected to 3 respirometry runs (22 h each) to address the question whether habituation occurs when fish are used in multiple runs and whether habituation potential is linked to coping styles. The same individuals were then subjected to an acute stressor and the escape behaviour and plasma cortisol were determined to assess coping style. Taken as a whole, the findings of this study suggest that individual variation in oxygen consumption in respirometry chambers are related to the expression of coping styles: individuals with higher oxygen consumption at the start and throughout the measurement period were shown to react passively (longer latency to escape) in a confinement stress. In addition, successive respirometry runs should not be assumed to result in habituation, as depending on an individual’s coping style either habituation (in the case of passive copers) or sensitization (in the case of active copers) can occur.
- Interactive effects of a high-quality protein diet and high stocking density on the stress response and some innate immune parameters of Senegalese sole Solea senegalensisPublication . Costas, B.; Aragão, C.; Dias, J.; Afonso, A.; Conceição, L. E. C.Amino acids (AA) regulate key metabolic pathways, including some immune responses. Therefore, this study aimed to assess whether an increased availability of dietary AA can mitigate the expected increase in plasma cortisol and metabolites levels due to high stocking density and its subsequent immunosuppression. Senegalese sole (Solea senegalensis) were maintained at low stocking density (LSD; 3.5 kg m−2) or high stocking density (HSD; 12 kg m−2) for 18 days. Additionally, both treatments were fed a control or a high protein (HP) diet (LSD, LSD HP, HSD and HSD HP). The HP diet slightly increased the levels of digestible indispensable AA, together with tyrosine and cysteine. HSD was effective in inducing a chronic stress response after 18 days of treatment since fish held at HSD presented higher plasma cortisol, glucose and lactate levels. Moreover, this increase in stress indicators translated in a decrease in plasma lysozyme, alternative complement pathway (ACP) and peroxidase activities, suggesting some degree of immunosuppression. Interestingly, while plasma glucose and lactate levels in HSD HP specimens decreased to similar values than LSD fish, plasma lysozyme, ACP and peroxidase activities increased, with even higher values than LSD groups for ACP activity. It is suggested that the HP diet may be used as functional feed since it may represent a metabolic advantage during stressful events and may counteract immunosuppression in sole.