Spatial and ontogenetic modulation of fatty acid composition in juvenile European sea bass (Dicentrarchus labrax) from two French estuaries.

This study evaluated how estuary of origin and ontogenetic stage influence the fatty acid (FA) composition in the tissues of wild European sea bass juvenile. We evidenced tissue-specific patterns, with the brain exhibiting a distinct FA composition from the liver and muscle. Ontogenetic stage and estuary influenced the general FA profile, and particularly the essential FA (EFA) like docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and arachidonic acid (ARA) in all tissues. The data also revealed the ability of wild sea bass to modulate, at the molecular level, FA biosynthesis pathways and suggest a potential dietary DHA limitation in the natural environment. The distribution of FA within tissues might reflect shifts in diet, metabolic demands, or adaptations to environmental conditions. This study provides insights about FA dynamics in euryhaline fish during juvenile life stage, improving our understanding of the metabolism need and EFA trophic availability in a changing environment.

Non-lethal sampling method for the analysis of white muscle fatty acid profiles in European sea bass (Dicentrarchus labrax).

This study presents a novel non-lethal sampling method for assessing fatty acid (FA) composition in juvenile European sea bass (Dicentrarchus labrax) using subcutaneous white muscle biopsies. This research aimed to evaluate the suitability of the biopsy for FA analysis using two lipid extraction protocols and comparing them to a lethal routine method. The results showed that a mass of fresh tissue as low as 1.4 mg provided good quality FA chromatograms for both reserve and membrane lipids. Although the biopsy method displayed high variability in terms of FA quantity among intra-individual replicates, it showed good FA profile repeatability in both reserve and membrane lipids. The study highlights the potential of this non-lethal approach for studying FA dynamics in fish, with its application being particularly promising for ecological and experimental studies. However, careful biopsy implementation is recommended to account for potential lipid droplet and lipid distribution variability within the tissue.

Lipid-correction models for δ13C values across small pelagic fishes (Clupeiformes) from the Atlantic Ocean.

The interpretation of δ13C values in trophic ecology requires standardization of the lipid content of organisms estimated through their C:N ratio. To avoid time-consuming lipid extractions, the use of mathematical corrections has been developed for many years, and the conclusions generally point in the direction of species-specific adjustment of the models. This study aimed at defining the maximum taxonomic level required to obtain the best corrected δ13C values in small pelagic fish of the order Clupeiformes. δ13C values of six species were analyzed bulk and lipid-free, and were used to fit and validate linear and mass-balance models at different taxonomic levels. Despite a species effect combined with the C:N ratio effect, the corrected δ13C values produced by a global model for the Clupeiformes were as good as or better when compared to lipid-free samples than those produced by species-specific models, paving the way for possible generalization to other species in this order. At the order level, the linear model outperformed the mass-balance model.

Growth regulation in brook charr Salvelinus fontinalis.

Fish growth can be modulated through genetic selection. However, it is not known whether growth regulatory mechanisms modulated by genetic selection can provide information about phenotypic growth variations among families or populations. Following a five-generation breeding program that selected for the absence of early sexual maturity and increased growth in brook charr we aimed to understand how the genetic selection process modifies the growth regulatory pathway of brook charr at the molecular level. To achieve this, we studied the regulation of growth traits at three different levels: 1) between lines-one under selection, the other not, 2) among-families expressing differences in average growth phenotypes, which we termed family performance, and 3) among individuals within families that expressed extreme growth phenotypes, which we termed slow- and fast-growing. At age 1+, individuals from four of the highest performing and four of the lowest performing families in terms of growth were sampled in both the control and selected lines. The gene expression levels of three reference and ten target genes were analyzed by real-time PCR. Results showed that better growth performance (in terms of weight and length at age) in the selected line was associated with an upregulation in the expression of genes involved in the growth hormone (GH)/insulin growth factor-1 (IGF-1) axis, including the igf-1 receptor in pituitary; the gh-1 receptor and igf-1 in liver; and ghr and igf-1r in white muscle. When looking at gene expression within families, family performance and individual phenotypes were associated with upregulations of the leptin receptor and neuropeptid Y-genes related to appetite regulation-in the slower-growing phenotypes. However, other genes related to appetite (ghrelin, somatostatin) or involved in muscle growth (myosin heavy chain, myogenin) were not differentially expressed. This study highlights how transcriptomics may improve our understanding of the roles of different key endocrine steps that regulate physiological performance. Large variations in growth still exist in the selected line, indicating that the full genetic selection potential has not been reached.

Risk and benefit assessment of seafood consumption harvested from the Pertuis Charentais region of France.

Seafood is well recognized as a major source of Long Chain n-3 Polyunsaturated Fatty Acids (LC n-3 PUFA, especially ecosapentaenoic acid, i.e. EPA and docosaheaxaenoic acid, i.e. DHA) and essential trace elements (As, Cu, Fe, Mn, Se, and Zn). It is also a source of non-essential trace elements (Ag, Cd, Hg, Pb) that can be deleterious for health even at low concentrations. Edible parts of sixteen species (fish, cephalopods, crustaceans and bivalves) of great importance in the Pertuis Charentais region, one of the main shellfish farming and fishing areas along the french coastline, were sampled in winter and analyzed to determine their fatty acid (FA) composition and trace element concentrations. Based on these analyses, a suite of indices was calculated to estimate risk and benefit of seafood consumption: the n-6/n-3 ratio, the atherogenic index, the thrombogenic index, the EPA + DHA daily recommended portion, as well as the maximum safe consumption. The results showed that fish contributed the most to LC n-3 PUFA supply, while bivalves and crustaceans were more beneficial in essential trace elements. Whatever the species, the concentrations of non-essential elements were not limiting for seafood consumption, as important amounts of the analyzed species can be eaten daily or weekly before becoming hazardous to consumers. Yet, concentrations of Hg in dogfish and seabass can become a concern for frequent seafood consumers (>three meals a week), confirming that varying seafood items is a key point for consumers to optimize the benefits of diverse seafood resources. Considering FA composition, whiting and pilchard are the most beneficial fish species for human diet, while surmullet was the least beneficial one. However, using an index integrating the relative risk due to Hg content, the surmullet appears as one of the most beneficial. This study provides a temporal shot of the quality of marine resources consumed in winter period in the studied area and highlights the complexity of a quantitative risk and benefit assessment with respect to the biochemical attributes of selected seafood.

Post-mortem storage conditions and cooking methods affect long-chain omega-3 fatty acid content in Atlantic mackerel (Scomber scombrus).

Long-chain omega-3 fatty acids such as eicosapentaenoic (EPA) and docosahexaenoic acids (DHA) are health beneficial lipids found in high concentration in pelagic fishes, including Atlantic mackerel. While EPA and DHA are sensitive to oxidation during fish storage and processing, post-mortem degradation in the first hours following fish death is poorly documented. Here, we stored fish at two temperatures (2-4 °C and 18-20 °C) and monitored EPA + DHA content in dorsal fillet 6, 12 and 24 h after fish death and after cooking (grill or steam). Storage duration was the only influencing factor, and EPA + DHA loss was faster at 18-20 °C. Six hours after fish death, EPA + DHA content decreased by 1.3 ± 1.3 mg.g-1 dw (9.6 ± 9.5% of the initial content) but it was highly variable among individuals. Handling between fishing and storage should be as short and as cool as possible to preserve EPA + DHA and food safety. Regarding cooking, EPA + DHA and mono-unsaturated fatty acids increased in grilled fillets.

The relationship between membrane fatty acid content and mitochondrial efficiency differs within- and between- omega-3 dietary treatments.

An important, but underappreciated, consequence of climate change is the reduction in crucial nutrient production at the base of the marine food chain: the long-chain omega-3 highly unsaturated fatty acids (n-3 HUFA). This can have dramatic consequences on consumers, such as fish as they have limited capacity to synthesise n-3 HUFA de novo. The n-3 HUFA, such as docosahexaenoic acid (DHA, 22:6n-3) and eicosapentaenoic acid (EPA, 20:5n-3), are critical for the structure and function of all biological membranes. There is increasing evidence that fish will be badly affected by reductions in n-3 HUFA dietary availability, however the underlying mechanisms remain obscure. Hypotheses for how mitochondrial function should change with dietary n-3 HUFA availability have generally ignored ATP production, despite its importance to a cell's total energetics capacity, and in turn, whole-animal performance. Here we (i) quantified individual variation in mitochondrial efficiency (ATP/O ratio) of muscle and (ii) examined its relationship with content in EPA and DHA in muscle membrane of a primary consumer fish, the golden grey mullet Chelon auratus, receiving either a high or low n-3 HUFA diet. Mitochondria of fish fed on the low n-3 HUFA diet had higher ATP/O ratio than those of fish maintained on the high n-3 HUFA diet. Yet, mitochondrial efficiency varied up about 2-fold among individuals on the same dietary treatment, resulting in some fish consuming half the oxygen and energy substrate to produce the similar amount of ATP than conspecific on similar diet. This variation in mitochondrial efficiency among individuals from the same diet treatment was related to individual differences in fatty acid composition of the membranes: a high ATP/O ratio was associated with a high content in EPA and DHA in biological membranes. Our results highlight the existence of interindividual differences in mitochondrial efficiency and its potential importance in explaining intraspecific variation in response to food chain changes.

Maturation of the European sardine Sardina pilchardus under experimental conditions strengthens bioenergetic estimate.

This study aims at (1) experimentally estimating first sexual maturation of the European sardine S. pilchardus, (2) using the results to calibrate existing bioenergetic models. During the 183 days-experiment, fish growth and body condition were assessed by biometry, and gonads were weighed when present. Age, wet weight and total length at first maturity were estimated at 262 days, 10.79 ± 0.75 g, and 11.26 ± 0.21 cm, respectively. Including these traits in biphasic Von Bertalanffy models did not significantly improve simulations for either length or weight data, meaning that energy allocation was not impacted by these traits. The implementation of the results in the Dynamic Energy Budget (DEB) calibration procedure strengthened the parameter set of the existing model, but resulted in significant changes in the energy allocation. Our results are a first step that will allow the design of new experiments to further quantify maturation and reproduction rates in diverse environmental conditions, consolidating DEB model calibration.

Fish facing global change: are early stages the lifeline?

The role of phenotypic plasticity in the acclimation and adaptive potential of an organism to global change is not currently accounted for in prediction models. The high plasticity of marine fishes is mainly attributed to their early stages, during which morphological, structural and behavioural functions are particularly sensitive to environmental constraints. This developmental plasticity can determine later physiological performances and fitness, and may further affect population dynamics and ecosystem functioning. This review asks the essential question of what role early stages play in the ability of fish to later cope with the effects of global change, considering three key environmental factors (temperature, hypoxia and acidification). After having identified the carry-over effects of early exposure reported in the literature, we propose areas that we believe warrant the most urgent attention for further research to better understand the role of developmental plasticity in the responses of marine organisms to global change.

Ocean warming combined with lower omega-3 nutritional availability impairs the cardio-respiratory function of a marine fish.

Highly unsaturated fatty acids of the omega-3 series (HUFA) are major constituents of cell membranes, yet are poorly synthesised de novo by consumers. Their production, mainly supported by aquatic microalgae, has been decreasing with global change. The consequences of such reductions may be profound for ectotherm consumers, as temperature tightly regulates the HUFA content in cell membranes, maintaining their functionality. Integrating individual, tissue and molecular approaches, we examined the consequences of the combined effects of temperature and HUFA depletion on the key cardio-respiratory functions of the golden grey mullet, an ectotherm grazer of high ecological importance. For 4 months, fish were exposed to two contrasting HUFA diets [4.8% eicosapentaenoic acid (EPA)+docosahexaenoic acid (DHA) on dry matter (DM) versus 0.2% EPA+DHA on DM] at 12 and 20°C. Ventricular force development coupled with gene expression profiles measured on cardiac muscle suggest that combining HUFA depletion with warmer temperatures leads to: (1) a proliferation of sarcolemmal and sarcoplasmic reticulum Ca2+ channels and (2) a higher force-generating ability by increasing extracellular Ca2+ influx via sarcolemmal channels when the heart has to sustain excessive effort due to stress and/or exercise. At the individual scale, these responses were associated with a greater aerobic scope, maximum metabolic rate and net cost of locomotion, suggesting the higher energy cost of this strategy. This impaired cardiac performance could have wider consequences for other physiological performance such as growth, reproduction or migration, all of which greatly depend on heart function.

Depletion of Essential Fatty Acids in the Food Source Affects Aerobic Capacities of the Golden Grey Mullet Liza aurata in a Warming Seawater Context.

The objective of this study was to evaluate the combined effects of thermal acclimation and n-3 highly unsaturated fatty acids (n-3 HUFA) content of the food source on the aerobic capacities of fish in a thermal changing environment. The model used was the golden grey mullet Liza aurata, a species of high ecological importance in temperate coastal areas. For four months, fish were exposed to two food sources with contrasting n-3 HUFA contents (4.8% ecosapentaenoic acid EPA + docosahexaenoic acid DHA on the dry matter DM basis vs. 0.2% EPA+DHA on DM) combined with two acclimation temperatures (12°C vs. 20°C). The four experimental conditions were LH12, LH20, HH12 and HH20. Each group was then submitted to a thermal challenge consisting of successive exposures to five temperatures (9°C, 12°C, 16°C, 20°C, 24°C). At each temperature, the maximal and minimal metabolic rates, metabolic scope, and the maximum swimming speed were measured. Results showed that the cost of maintenance of basal metabolic activities was particularly higher when n-3 HUFA food content was low. Moreover, fish exposed to high acclimation temperature combined with a low n-3 HUFA dietary level (LH20) exhibited a higher aerobic scope, as well as a greater expenditure of energy to reach the same maximum swimming speed as other groups. This suggested a reduction of the amount of energy available to perform other physiological functions. This study is the first to show that the impact of lowering n-3 HUFA food content is exacerbated for fish previously acclimated to a warmer environment. It raises the question of the consequences of longer and warmer summers that have already been recorded and are still expected in temperate areas, as well as the pertinence of the lowering n-3 HUFA availability in the food web expected with global change, as a factor affecting marine organisms and communities.

Coding Gene SNP Mapping Reveals QTL Linked to Growth and Stress Response in Brook Charr (Salvelinus fontinalis).

Growth performance and reduced stress response are traits of major interest in fish production. Growth and stress-related quantitative trait loci (QTL) have been already identified in several salmonid species, but little effort has been devoted to charrs (genus Salvelinus). Moreover, most QTL studies to date focused on one or very few traits, and little investigation has been devoted to QTL identification for gene expression. Here, our objective was to identify QTL for 27 phenotypes related to growth and stress responses in brook charr (Salvelinus fontinalis), which is one of the most economically important freshwater aquaculture species in Canada. Phenotypes included 12 growth parameters, six blood and plasma variables, three hepatic variables, and one plasma hormone level as well as the relative expression measurements of five genes of interest linked to growth regulation. QTL analysis relied on a linkage map recently built from S. fontinalis consisting of both single-nucleotide polymorphism (SNP, n = 266) and microsatellite (n =81) markers in an F(2) interstrain hybrid population (n = 171). We identified 63 growth-related QTL and four stress-related QTL across 18 of the 40 linkage groups of the brook charr linkage map. Percent variance explained, confidence interval, and allelic QTL effects also were investigated to provide insight into the genetic architecture of growth- and stress-related QTL. QTL related to growth performance and stress response that were identified could be classified into two groups: (1) a group composed of the numerous, small-effect QTL associated with some traits related to growth (i.e., weight) that may be under the control of a large number of genes or pleiotropic genes, and (2) a group of less numerous QTL associated with growth (i.e., gene expression) and with stress-related QTL that display a larger effect, suggesting that these QTL are under the control of a limited number of genes of major effect. This study represents a first step toward the identification of genes potentially linked to phenotypic variation of growth and stress response in brook charr. The ultimate goal is to provide new tools for developing Molecular Assisted Selection for this species.

Coding Gene Single Nucleotide Polymorphism Mapping and Quantitative Trait Loci Detection for Physiological Reproductive Traits in Brook Charr, Salvelinus fontinalis.

A linkage map of 40 linkage groups (LGs) was developed for brook charr, Salvelinus fontinalis, using an F(2) interstrain hybrid progeny (n = 171) and 256 coding gene SNP developed specifically for brook charr and validated from a large (>1000) subset of putative SNP, as well as 81 microsatellite markers. To identify quantitative trait loci (QTL) related to reproduction functions, these fish were also phenotyped at six physiological traits, including spermatozoid head diameter, sperm concentration, plasma testosterone, plasma 11-keto-testosterone, egg diameter, and plasma 17ß-estradiol. Five significant QTL were detected over four LGs for egg diameter and plasma 17ß-estradiol concentration in females, and sperm concentration as well as spermatozoid head diameter in males. In females, two different QTLs located on LG 11 and LG 34 were associated with the egg number, whereas one QTL was associated with plasma 17ß-estradiol concentration (LG 8). Their total percent variance explained (PVE) was 26.7% and 27.6%, respectively. In males, two QTL were also detected for the sperm concentration, and their PVE were estimated at 18.58% and 14.95%, respectively. The low QTL number, associated with the high PVE, suggests that the variance in these reproductive physiological traits was either under the control of one major gene or a small number of genes. The QTL associated with sperm concentration, plasma 17ß-estradiol, and egg diameter appeared to be under a dominance effect, whereas the two others were under a negative additive effect. These results show that genes underlying the phenotypic variance of these traits are under different modes of action (additive vs. dominance) and may be used to predict an increase or a decrease in their phenotypic values in subsequent generations of selective breeding. Moreover, this newly developed panel of mapped SNP located in coding gene regions will be useful for screening wild populations, especially in the context of investigating the genetic impact of massive stocking of domestic brook charr to support the angling industry throughout eastern North America.

Ontogenic effects of early feeding of sea bass (Dicentrarchus labrax) larvae with a range of dietary n-3 highly unsaturated fatty acid levels on the functioning of polyunsaturated fatty acid desaturation pathways.

Four replicated groups of sea bass (Dicentrarchus labrax) larvae were fed diets containing an extra-high level of highly unsaturated fatty acids (HUFA) (XH; 3.7 % EPA+DHA), a high level of HUFA (HH; 1.7 %), a low level of HUFA (LH; 0.7 %) or an extra-low level of HUFA (XLH; 0.5 %) from day 6 to day 45 (experiment 1; XH1, HH1, LH1, XLH1). After a subsequent 1-month period feeding a commercial diet (2.7 % EPA+DHA), the capacity of the four initial groups to adapt to an n-3 HUFA-restricted diet (0.3 % EPA+DHA; R-groups: XH2R, HH2R, LH2R, XLH2R) was tested for 35 d. Larval dietary treatments had no effect on larval and juvenile survival rates. The wet weight of day 45 larvae was higher in XH1 and HH1 (P < 0.001), but the R-juvenile mass gains were similar in all treatments. Delta-6-desaturase (Delta6D) mRNA level was higher in LH1 and XLH1 at day 45 (P < 0.001), and higher in LH2R and XLH2R, with a significant increase at day 118.Concomitantly, PPARalpha and PPARbeta mRNA levels were higher in XLH1 at day 45, and PPARbeta and gamma mRNA levels were higher in XLH2R at day 118, suggesting possible involvement of PPAR in stimulation of Delta6D expression, when drastic dietary larval conditioning occurred. The low DHA content in the polar lipids (PL) of LH1 and XLH1 revealed an n-3-HUFA deficiency in these groups. Larval conditioning did not affect DHA content in the PL of R-juveniles. The present study showed (i) a persistent Delta6D mRNA enhancement in juveniles pre-conditioned with an n-3 HUFA-deficient larval diet, over the 1-month intermediate period, and (ii) brought new findings suggesting the involvement of PPAR in the Delta6D mRNA level stimulation. However, such nutritional conditioning had no significant effect on juvenile growth and lipid composition.