Increased intestinal carbonate precipitate abundance in the sea bream (Sparus aurata L.) in response to ocean acidification

Marine fish contribute to the carbon cycle by producing mineralized intestinal precipitates generated as by-products of their osmoregulation. Here we aimed at characterizing the control of epithelial bicarbonate secretion and intestinal precipitate presence in the gilthead sea bream in response to predicted near future increases of environmental CO2. Our results demonstrate that hypercapnia (950 and 1800 μatm CO2) elicits higher intestine epithelial HCO3- secretion ex vivo and a subsequent parallel increase of intestinal precipitate presence in vivo when compared to present values (440 μatm CO2). Intestinal gene expression analysis in response to environmental hypercapnia revealed the up-regulation of transporters involved in the intestinal bicarbonate secretion cascade such as the basolateral sodium bicarbonate co-transporter slc4a4, and the apical anion transporters slc26a3 and slc26a6 of sea bream. In addition, other genes involved in intestinal ion uptake linked to water absorption such as the apical nkcc2 and aquaporin 1b expression, indicating that hypercapnia influences different levels of intestinal physiology. Taken together the current results are consistent with an intestinal physiological response leading to higher bicarbonate secretion in the intestine of the sea bream paralleled by increased luminal carbonate precipitate abundance and the main related transporters in response to ocean acidification.Agência financiadora Fundacao para a Ciencia e a Tecnologia (FCT) SFRH/BD/113363/2015 PTDC/MAR-BIO/3034/2014 Fundacao para a Ciencia e a Tecnologia (FCT) UID/Multi/04326/2019 Ministry of Science and Higher Education, Polandinfo:eu-repo/semantics/publishedVersio

Physiology: an important tool to assess the welfare of aquatic animals

The assessment of welfare in aquatic animals is currently under debate, especially concerning those kept by humans. The classic concept of animal welfare includes three elements: The emotional state of the organism (including the absence of negative experiences), the possibility of expressing normal behaviors, and the proper functioning of the organism. While methods for evaluating their emotions (such as fear, pain, and anguish) are currently being developed for aquatic species and understanding the natural behavior of all aquatic taxa that interact with humans is a task that requires more time, the evaluation of internal responses in the organisms can be carried out using analytical tools. This review aims to show the potential of the physiology of crustaceans, cephalopods, elasmobranchs, teleosts, and dipnoans to serve as indicators of their wellbeing. Since the classical methods of assessing welfare are laborious and time-consuming by evaluation of fear, pain, and anguish, the assessment may be complemented by physiological approaches. This involves the study of stress responses, including the release of hormones and their effects. Therefore, physiology may be of help in improving animal welfare

Acute-Stress Biomarkers in Three Octopodidae Species After Bottom Trawling

Several Octopodidae species have a great potential for the diversification of worldwide aquaculture. Unfortunately, the lack of stress-related biomarkers in this taxon results an obstacle for its maintenance in conditions where animal welfare is of paramount relevance. In this study, we made a first approach to uncover physiological responses related to fishing capture in Eledone moschata, Eledone cirrhosa, and Octopus vulgaris. Captured octopus from all three species were individually maintained in an aquaculture system onboard of oceanographic vessel in south-western waters of Europe. Haemolymph plasma and muscle were collected in animals at the moment of capture, and recovery was evaluated along a time-course of 48 h in Eledone spp., and 24 h for O. vulgaris. Survival rates of these species captured in spring and autumn were evaluated. Physiological parameters such as plasma pH, total CO2, peroxidase activity, lysozyme, hemocyanin, proteases, pro-phenoloxidase, anti-proteases, free amino acids, lactate and glucose levels, as well as muscle water percentage, free amino acids, lactate, glycogen and glucose values were analyzed. The immune system appears to be compromised in these species due to capture processes, while energy metabolites were mobilized to face the acute-stress situation, but recovery of all described parameters occurs within the first 24 h after capture. Moreover, this situation exerts hydric balance changes, as observed in the muscle water, being these responses depending on the species assessed. In conclusion, three Octopodidae species from south-western waters of Europe have been evaluated for stress-related biomarkers resulting in differentiated mechanisms between species. This study may pave the way to further study the physiology of stress in adult octopuses and develop new methodologies for their growth in aquaculture conditions

Cortisol and Dexamethasone Mediate Glucocorticoid Actions in the Lesser Spotted Catshark (Scyliorhinus canicula)

Corticosteroids are hormones produced in vertebrates exerting gluco- and mineralocorticoid actions (GC and MC) mediated by specific receptors (GR and MR, respectively). In elasmobranchs, the major circulating corticosteroid is the 1 -hydroxycorticosterone (1 -OHB). This hormone acts as a MC, but to date its role as a GC has not been established. As there is no 1 -OHB standard available, here we employed a set of in vivo and ex vivo approaches to test GC actions of other corticosteroids in the lesser spotted catshark (Scyliorhinus canicula). Dexamethasone (DEX, a synthetic corticosteroid) slow-release implants decreased plasma 1 -OHB levels after 7 days, and modified carbohydrates metabolism in liver and white muscle (energy stores and metabolic enzymes). In addition, ex vivo culture of liver and white muscle explants confirmed GC actions of corticosteroids not naturally present in sharks (cortisol and DEX) by increasing glucose secretion from these tissues. Dose–response curves induced by cortisol and DEX, altogether with the use of specific GR inhibitor mifepristone, confirmed the involvement of GR mediating glucose secretion. This study highlights the influence of corticosteroids in the glucose balance of S. canicula, though the role of 1 -OHB as a GC hormone in sharks should be further confirmed

Regulation of Stanniocalcin Secretion by Calcium and PTHrP in Gilthead Seabream (Sparus aurata)

Calcium balance is of paramount importance for vertebrates. In fish, the endocrine modulators of calcium homeostasis include the stanniocalcin (STC), and some members of the parathyroid hormone (PTH) family, such as the PTH-related protein (PTHrP), acting as antagonists. STC is ubiquitously expressed in higher vertebrates. In turn, bony fish exhibit specific STC-producing glands named the corpuscles of Stannius (CS). Previous studies pointed to a calcium-sensing receptor (CaSR) involvement in the secretion of STC, but little is known of the involvement of other putative regulators. The CS provides a unique model to deepen the study of STC secretion. We developed an ex vivo assay to culture CS of fish and a competitive ELISA method to measure STC concentrations. As expected, STC released from the CS responds to CaSR stimulation by calcium, calcimimetics, and calcilytic drugs. Moreover, we uncover the presence (by PCR) of two PTHrP receptors in the CS, e.g., PTH1R and PTH3R. Thus, ex vivo incubations revealed a dose-response inhibition of STC secretion in response to PTHrP at basal Ca2+ concentrations. This inhibition is achieved through specific and reversible second messenger pathways (transmembrane adenylyl cyclases and phospholipase C), as the use of specific inhibitors highlights. Together, these results provide evidence for endocrine modulation between two antagonist hormones, STC and PTHrP

Crecimiento y cambios metabólicos anuales en corvina Argyrosomus regius cultivada en esteros

Metabolic modifications associated with natural environmental conditions were assessed in the meagre Argyrosomus regius cultured in earthen ponds under natural photoperiod and temperature. Juvenile specimens (90-100 g initial weight) were sampled (plasma, liver and muscle) every two months for 18 months (between December 2004 and May 2006). Specimens showed seasonal variations in growth rate, with the highest values in spring and summer. Plasmatic, hepatic and muscular metabolite levels and hepatic and muscular metabolic enzymes also showed significant variations throughout the year. Enzymatic activity related to carbohydrate metabolism in the liver (HK, FBPase and G6PDH) showed great modifications in summer, increasing glycogenogenic pathways, while amino acid metabolism (GDH and GOT activity) was enhanced in spring and summer. However lipid-related (G3PDH activity) metabolic enzymes did not show a clear seasonal pattern. In muscle, enzymatic activity related to amino acid, lipid and lactate metabolism (LDH-O activity), but not carbohydrate metabolism, showed seasonal changes in parallel with changes in growth rate. Thus A. regius specimens showed a trend to grow in summer months and mobilize their energy reserves in winter. Differences in the hepatic level were observed between the first and the second year of the study, suggesting the possible existence of metabolic changes related to specimen age or size. Our results indicate that growth and metabolic responses in A. regius are environmentally dependent and that this species is a very good candidate for diversification in aquaculture.Modificaciones metabólicas asociadas a condiciones ambientales temporales fueron evaluadas en la corvina Argyrosomus regius, cultivadas en esteros con fotoperiodo y temperatura natural. Ejemplares juveniles (90-100 g de peso inicial) fueron muestreados (plasma, hígado y músculo) cada dos meses durante 18 meses (entre diciembre de 2004 y mayo de 2006). Las muestras mostraron variaciones estacionales en la tasa de crecimiento, con valores más altos durante la primavera y el verano. Niveles de metabolitos plasmáticos, hepáticos y musculares, así como las actividades de enzimas metabólicas hepáticas y musculares también presentaron variaciones significativas a lo largo del año. La actividad enzimática relacionada con el metabolismo de carbohidratos en el hígado (HK, FBPasa y G6PDH) mostró altas modificaciones en el verano, el aumento de las vías glucogenogénicas, mientras el metabolismo de aminoácidos (actividades de GDH y GOT) se incrementó en temporadas de primavera y verano. Sin embargo la actividad de G3PDH (enzima metabólica relacionada con los lípidos) no mostró un claro patrón estacional. En el músculo, la actividad enzimática respecto a los aminoácidos, lípidos y el metabolismo del lactato (LDH-O) presentó cambios estacionales en paralelo con los cambios en la tasa de crecimiento, pero el metabolismo de los carbohidratos no presentó cambios estacionales. Así especímenes de A. regius mostraron una tendencia a crecer en los meses de verano y movilizar sus reservas de energía en invierno. Se observaron diferencias en el nivel hepático entre el primer y el segundo año de estudio, lo que sugiere la posible existencia de cambios metabólicos relacionados con la edad o el tamaño del espécimen. Nuestros resultados indican que el crecimiento y las respuestas metabólicas en A. regius dependen del medio ambiente, pero también indican que esta especie es una muy buena candidata para la diversificación de la acuicultura

Yearly growth and metabolic changes in earthen pond-cultured meagre <em>Argyrosomus regius</em>

Metabolic modifications associated with natural environmental conditions were assessed in the meagre Argyrosomus regius cultured in earthen ponds under natural photoperiod and temperature. Juvenile specimens (90-100 g initial weight) were sampled (plasma, liver and muscle) every two months for 18 months (between December 2004 and May 2006). Specimens showed seasonal variations in growth rate, with the highest values in spring and summer. Plasmatic, hepatic and muscular metabolite levels and hepatic and muscular metabolic enzymes also showed significant variations throughout the year. Enzymatic activity related to carbohydrate metabolism in the liver (HK, FBPase and G6PDH) showed great modifications in summer, increasing glycogenogenic pathways, while amino acid metabolism (GDH and GOT activity) was enhanced in spring and summer. However lipid-related (G3PDH activity) metabolic enzymes did not show a clear seasonal pattern. In muscle, enzymatic activity related to amino acid, lipid and lactate metabolism (LDH-O activity), but not carbohydrate metabolism, showed seasonal changes in parallel with changes in growth rate. Thus A. regius specimens showed a trend to grow in summer months and mobilize their energy reserves in winter. Differences in the hepatic level were observed between the first and the second year of the study, suggesting the possible existence of metabolic changes related to specimen age or size. Our results indicate that growth and metabolic responses in A. regius are environmentally dependent and that this species is a very good candidate for diversification in aquaculture

Bienestar Animal en la Acuicultura de Peces : Atenuación del Estrés a través de la Dieta y mediante el Empleo de Anestésicos durante el Transporte

La acuicultura de peces genera situaciones de estrés que disminuyen las condiciones de bienestar animal y el rendimiento económico del cultivo. En este artículo exponemos dos posibilidades para la atenuación del estrés en la dorada (Sparus aurata L.), una de las especies emblemáticas de la acuicultura nacional y europea.Fish aquaculture generates stressful situations that reduce animal welfare conditions and the economic benefits. In this article, we expose two possibilities for stress attenuation in the gilthead seabream (Sparus aurata L.), an emblematic species for the national and European aquaculture

Acute Stress in Lesser-Spotted Catshark (Scyliorhinus canicula Linnaeus, 1758) Promotes Amino Acid Catabolism and Osmoregulatory Imbalances

Simple Summary In catsharks (Scyliorhinus canicula), air exposure induces amino acid catabolism altogether with osmoregulatory imbalances. This study describes a novel NHE isoform being expressed in gills that may be involved in ammonium excretion. Acute-stress situations in vertebrates induce a series of physiological responses to cope with the event. While common secondary stress responses include increased catabolism and osmoregulatory imbalances, specific processes depend on the taxa. In this sense, these processes are still largely unknown in ancient vertebrates such as marine elasmobranchs. Thus, we challenged the lesser spotted catshark (Scyliorhinus canicula) to 18 min of air exposure, and monitored their recovery after 0, 5, and 24 h. This study describes amino acid turnover in the liver, white muscle, gills, and rectal gland, and plasma parameters related to energy metabolism and osmoregulatory imbalances. Catsharks rely on white muscle amino acid catabolism to face the energy demand imposed by the stressor, producing NH4+. While some plasma ions (K+, Cl- and Ca2+) increased in concentration after 18 min of air exposure, returning to basal values after 5 h of recovery, Na+ increased after just 5 h of recovery, coinciding with a decrease in plasma NH4+. These changes were accompanied by increased activity of a branchial amiloride-sensitive ATPase. Therefore, we hypothesize that this enzyme may be a Na+/H+ exchanger (NHE) related to NH4+ excretion. The action of an omeprazole-sensitive ATPase, putatively associated to a H+/K+-ATPase (HKA), is also affected by these allostatic processes. Some complementary experiments were carried out to delve a little deeper into the possible branchial enzymes sensitive to amiloride, including in vivo and ex vivo approaches, and partial sequencing of a nhe1 in the gills. This study describes the possible presence of an HKA enzyme in the rectal gland, as well as a NHE in the gills, highlighting the importance of understanding the relationship between acute stress and osmoregulation in elasmobranchs.This studywas partially supported by grant PID2020-117557RB-C22 (funded byMCIN/AEI/10.13039/501100011033 and by the European Union) and Project DISCARDLIFE (Programa Pleamar, Ministerio de Agricultura y Pesca, Alimentacion y Medio Ambiente, Spain) to JMM. I. Ruiz-Jarabo and J.M. Mancera belong to the Fish Welfare and Stress Network (AGL2016-81808-REDT), supported by the Agencia Estatal de Investigacion (MINECO, Spanish Government)

Microbiota composition and intestinal integrity remain unaltered after the inclusion of hydrolysed Nannochloropsis gaditana in Sparus aurata diet

The use of lysed microalgae in the diet of carnivorous fish can increase the bioavailability of proteins and bioactive compounds, such as unsaturated fatty acids or vitamins in the digestive tract. These are essential molecules for the proper physiological development of fish in aquaculture. However, some antinutritional components and other undesirable molecules can be released from an excess of microalgae supplied, compromising the integrity of the intestine. The inclusion of small amounts of hydrolized microalgae in the fish diet can be a good strategy to avoid negative effects, improving the availability of beneficial compounds. Nannochloropsis gaditana is an interesting microalgae as it contains nutraceuticals. Previous studies reported beneficial effects after its inclusion in the diet of Sparus aurata, a widely cultured species in Europe and in all Mediterranean countries. However, administration of raw microalgae can produce intestinal inflammation, increased intestinal permeability, bacterial translocation and disturbance of digestion and absorption processes. The aim of this study was to evaluate changes in the intestinal microbiota and barrier stability of S. aurata fed with low inclusion (5%) hydrolysed N. gaditana. Intestinal microbiota was analyzed using Illumina MiSeq technology and libraries were constructed using variable regions V3-V4 of 16S rDNA molecules. Analysis were based in the identification, quantification and comparison of sequences. The predictive intestinal microbial functionality was analyzed with PICRUSt software. The results determined that the intestinal microbiota bacterial composition and the predictive intestinal microbiota functionality did not change statistically after the inclusion of N. gaditana on the diet. The study of gene expression showed that genes involved in intestinal permeability and integrity were not altered in fish treated with the experimental diet. The potential functionality and bacterial taxonomic composition of the intestinal microbiota, and the expression of integrity and permeability genes in the intestine of the carnivorous fish S. aurata were not affected by the inclusion of hydrolysed 5% N. gaditana microalgae