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Repeated fish handling may cause stress, which biases experiments and so affects the results. In order to reduce this, the present study investigates the benefit of using bioimpedance analysis to estimate morphological parameters. Bioimpedance analysis is a non-lethal and integrative electrical measurement that can be used to estimate several kinds of physiological parameters and is used in medicine and ecological studies. In particular, bioimpedance can monitor the individual growth of fish, which is a prerequisite for most biological and ecological studies, as body size is one of the best predictors for numerous life history traits and ecological parameters. After a short review on the use of bioimpedance measurement in medicine and ecology, we illustrate the potential of bioimpedance spectroscopy, as opposed to single frequency measurement, for experimental studies on fish. Using a monolithic four-contact electrode and a cost-effective portable system, we conducted bioimpedance spectroscopy on 159 sardines. The association between the length, weight, and electrical parameters obtained at different frequencies from the bioimpedance spectroscopy was investigated. Our results show that accounting for more than one frequency substantially improves the prediction of length and weight. We conclude that bioimpedance could potentially be a powerful tool for monitoring fish growth in ecological studies.

Aquatic ecosystems can exhibit seasonal variation in resource availability and animals have evolved to cope with the associated caloric restriction. During winter in the NW Mediterranean Sea, the European sardine Sardina pilchardus naturally experiences caloric restriction due to a decrease in diversity and quantity of plankton. Ongoing global warming has, however, had deleterious effects on plankton communities such that food shortages may occur throughout the year, especially under warm conditions in the summer. We investigated the interactive effects of temperature and food availability on sardine metabolism, by continuously monitoring whole-animal respiration of groups of control (fed) and food-deprived sardines over a 60-day experiment in winter (12°C) or summer (20°C) conditions under natural photoperiod. In addition, we measured mitochondrial respiration of red muscle fibres, biometric variables and energy reserves, of individuals sampled at 30 and 60 days. This revealed that winter food deprivation elicits energy saving mechanisms at whole animal and cellular levels by maintaining a low metabolism to preserve energy reserves, allowing high survival. By contrast, despite energy saving mechanisms at the mitochondrial level, whole animal metabolic rate was high during food deprivation in summer, causing increased consumption of energy reserves at the muscular level and high mortality after 60 days. Furthermore, a 5-day refeeding did not improve survival and mortalities actually continued, suggesting that long-term food deprivation at high temperatures caused profound stress in sardines that potentially impaired nutrient absorption.

By evaluating genetic variation across the entire genome, one can address existing questions in a novel way while raising new ones. The latter include how different local environments influence adaptive and neutral genomic variation within and among populations, providing insights into local adaptation of natural populations and their responses to global change. Here, under a seascape genomic approach, ddRAD data of 4609 SNPs from 398 sardines (Sardina pilchardus) collected in 11 Mediterranean and one Atlantic site were generated. These were used along with oceanographic and ecological information to detect signals of adaptive divergence with gene flow across environmental gradients. The studied sardines constitute two clusters (FST=0.07), a pattern attributed to outlier loci, highlighting putative local adaptation. The trend in the number of days with sea surface temperature above 19oC, critical threshold for successful sardine spawning, was crucial at all levels of population structuring with implications on species’ key biological processes. Outliers link candidate SNPs to the region's environmental heterogeneity. Our findings provide evidence for a dynamic equilibrium where population structure is maintained by physical and ecological factors under the opposing influences of migration and selection. This dynamic in nature system, warrants continuous monitoring under a seascape genomic approach that might benefit from a temporal and more detailed spatial dimension. Our results may contribute to complementary studies aimed at providing deeper insights into the mechanistic processes underlying population structuring. Those are key for understanding and predicting future changes and responses of this highly exploited species in the face of climate change.

By evaluating genetic variation across the entire genome, one can address existing questions in a novel way while raising new ones. The latter includes how different local environments influence adaptive and neutral genomic variation within and among populations, providing insights into local adaptation of natural populations and their responses to global change. Here, under a seascape genomic approach, ddRAD data of 4609 single nucleotide polymorphisms (SNPs) from 398 sardines (Sardina pilchardus) collected in 11 Mediterranean and one Atlantic site were generated. These were used along with oceanographic and ecological information to detect signals of adaptive divergence with gene flow across environmental gradients. The studied sardines constitute two clusters (FST = 0.07), a pattern attributed to outlier loci, highlighting putative local adaptation. The trend in the number of days with sea surface temperature above 19°C, a critical threshold for successful sardine spawning, was crucial at all levels of population structuring with implications on the species' key biological processes. Outliers link candidate SNPs to the region's environmental heterogeneity. Our findings provide evidence for a dynamic equilibrium in which population structure is maintained by physical and ecological factors under the opposing influences of migration and selection. This dynamic in a natural system warrants continuous monitoring under a seascape genomic approach that might benefit from a temporal and more detailed spatial dimension. Our results may contribute to complementary studies aimed at providing deeper insights into the mechanistic processes underlying population structuring. Those are key to understanding and predicting future changes and responses of this highly exploited species in the face of climate change

This research represents the first approach to the study of the shelf zooplankton community at a Mediterranean scale, using acoustic standardized data collected in June-July in 10 different geographical sub-areas (GSAs) established by the General Fisheries Commission for the Mediterranean (GFCM) within the framework of the MEDiterranean International Acoustic Survey (MEDIAS). The analysis of the zooplankton layers based on their acoustic characteristics has revealed the potential of these surveys for the study of zooplankton at a Mediterranean basin scale and, also, the need to collect biological samples to interpret the acoustic records in terms of species. The fish population’s direct assessment is established in the MEDIAS framework, but the integration of zooplankton community data would constitute a qualitative step for the understanding of the fluctuations of fish populations and therefore to achieve the objective of an ecosystem-based management. Results have revealed the ubiquity of the zooplankton layers and its ability to form layers detectable by scientific echosounders throughout the Mediterranean at the common fish assessment frequency (38 kHz). In addition, the use of two frequencies (38 and 120 kHz) has allowed to apply the dB difference method, observing changes in the difference of the Mean Volume Backscattering Strength (∆MVBS) at 38 and 120 kHz frequencies, which would be related to changes in the composition of the zooplankton community.

In the Gulf of Lions, small pelagic fish have shown reduced body size and body condition after 2007 that would result from changes in zooplankton community. We therefore examined zooplankton density, body size, and taxonomic composition at the closest long-term monitoring station (1995–2019): the coastal Point-B. To cover a broader spectrum of zooplankton community, samples obtained from two nets, the WP2 (200 µm mesh size) and the Regent (690 µm), were analysed with the imaging Zooscan method. One important result was the high stability through time of the zooplankton community. No long-term monotonous trends in density, size, and taxonomic composition were detected. Interannual variations in zooplankton size and density were not significantly correlated to any environmental variable, suggesting the possible importance of biotic interactions. Still, an increase in temperature was followed by a sharp decrease of zooplankton density in 2015, after which only gelatinous groups recovered. No change in the zooplankton community was detected around 2007 to support bottom-up control on small pelagic fish. Whether this derives from different local processes between the Gulf of Lions and the Ligurian Sea cannot be excluded, highlighting the need for simultaneous monitoring of different ecosystem compartments to fully understand the impact of climate change.

Telomeres are specialized non-coding DNA sequences located at the end of chromosomes and that protect genetic information. Telomere loss over lifespan is generally viewed as a phenomenon associated with aging in animals. Recently, telomere elongation after hibernation has been described in several mammals. Whether this pattern is an adaptation to repair DNA damage caused during rewarming from torpor or if it coevolved as a mechanism to promote somatic maintenance in preparation for the upcoming reproductive effort remains unclear. In a longitudinal study measuring telomere length using buccal swabs, we tested if telomere elongation was related to reproductive success in wild adult female Columbian ground squirrels (Urocitellus columbianus) that were monitored from emergence from hibernation to the end of the reproductive season. We found three key results. First, female telomere length increased at the start of the breeding season, both in breeding and non-breeding individuals. Second, post-emergence telomere lengthening was unrelated to female future reproductive output. Third, telomere length decreased in breeding females during lactation, but remained stable in non-breeding females over a similar period. Within breeders, telomeres shortened more in females producing larger and heavier litters. We concluded that telomere lengthening after hibernation did not constrain immediate female reproductive capacities. It was more likely to be part of the body recovery process that takes place after hibernation. Telomere erosion that occurs after birth may constitute a physiological cost of female reproduction.

Since 2008, there has been a major decrease in the numbers of old and large sardines in the Gulf of Lions, which has had a major effect on regional fisheries. A bottom-up process involving a shift in diet towards smaller planktonic prey has been suggested as the main driver of this development. Yet, the reproductive capacities of the sardines have not changed, suggesting potential modifications in energy allocation trade-offs. Whether this could also affect maintenance, in particular at the end of the winter reproductive period, and explain the lower adult survival and the disappearance of older individuals remains unclear. We therefore experimentally investigated the consequences of seasonal food availability (summer vs. winter) on life-history traits and energy allocation trade-offs at the individual and population levels. Our results indicate that food resources during summer had a major effect on energy reserves and growth, limiting the maximum size and body condition reached at the end of reproduction. In addition, food restrictions during growth and/or reproduction periods led to physiological costs mediated by increased oxidative damage. Mediterranean sardines did not show any ability for compensatory growth and did not appear to be capital breeders. Instead, they displayed individual differences in coping with physiological constraints and displayed various life-history strategies regardless of food availability. We highlighted 3 main individual energy allocation strategies: (1) preferential allocation to body condition or (2) to growth, or (3) simultaneous allocation to reproduction and growth. These issues are key, as climate change is expected to favour smaller phytoplankton, which might amplify the deterioration in the condition of pelagic fish.

Forage fish populations often undergo large and rapid fluctuations in abundance. However, most of their predators are buffered against such fluctuations owing to their slower pace of life, which allows them to maintain more stable populations, at least during short periods of food scarcity. In this study, we investigated top‐down processes exerted by seabirds on forage fish stocks in five contrasted marine ecosystems, compiling numerous data sets on seabird counts, diets, energetic needs and prey energy content and abundance. Off Norway, South Africa, Peru, Sweden and Scotland, we found that predation pressure—estimated as the proportion of a fish stock consumed by seabirds—was generally low (median = 1%), but increased sharply at low levels of prey abundance. When prey biomass decreased below 15–18% of its maximum recorded value, predation by seabirds became a source of important additional pressure on prey stocks (~20% of prey biomass is consumed by seabirds). An earlier empirical study advocated for keeping forage stocks from falling below a threshold of 33% of long‐term maximum prey biomass in order to safeguard seabird breeding success, but here we further suggest that a threshold of 18% should be considered as a limit not to be exceeded for the sake of the forage fish themselves, and below which extra cautious management of fisheries may be required. Nevertheless, despite exceptionally high rates of predation on some occasions, predation pressure was not correlated with prey dynamics, suggesting an absence of prey entrapment due to seabirds alone in these five ecosystems.

Independence of science and best available science are fundamental pillars of the UN-FAO code of conduct for responsible fisheries and are also applied to the European Union (EU) Common Fishery Policy (CFP), with the overarching objective being the sustainable exploitation of the fisheries resources. CFP is developed by DG MARE, the department of the European Commission responsible for EU policy on maritime affairs and fisheries, which has the Scientific, Technical and Economic Committee for Fisheries (STECF) as consultant body. In the Mediterranean and Black Sea, the General Fisheries Commission for the Mediterranean (FAO-GFCM), with its own Scientific Advisory Committee on Fisheries (GFCM-SAC), plays a critical role in fisheries governance, having the authority to adopt binding recommendations for fisheries conservation and management. During the last years, advice on the status of the main stocks in the Mediterranean and Black Sea has been provided both by GFCM-SAC and EU-STECF, often without a clear coordination and a lack of shared rules and practices. This has led in the past to: i) duplications of the advice on the status of the stocks thus adding confusion in the management process and, ii) a continuous managers’ interference in the scientific process by DG MARE officials hindering its transparency and independence. Thus, it is imperative that this stalemate is rapidly resolved and that the free role of science in Mediterranean fisheries assessment and management is urgently restored to assure the sustainable exploitation of Mediterranean marine resources in the future.

Animal mortality is difficult to observe in marine systems, preventing a mechanistic understanding of major drivers of fish population dynamics. In particular, starvation is known to be a major cause of mortality at larval stages, but adult mortality is often unknown. In this study, we used a laboratory food-deprivation experiment, on wild caught sardine Sardina pilchardus from the Gulf of Lions. This population is interesting because mean individual phenotype shifted around 2008, becoming dominated by small, young individuals in poor body condition, a phenomenon that may result from declines in energy availability. Continuous monitoring of body mass loss and metabolic rate in 78 captive food-deprived individuals revealed that sardines could survive for up to 57 days on body reserves. Sardines submitted to long-term caloric restriction prior to food-deprivation displayed adaptive phenotypic plasticity, reducing metabolic energy expenditure and enduring starvation for longer than sardines that had not been calorie-restricted. Overall, entry into critical fasting phase 3 occurred at a body condition of 0.72. Such a degree of leanness has rarely been observed over 34 years of wild population monitoring. Still, the proportion of sardines below this threshold has doubled since 2008 and is maximal in January and February (the peak of the reproductive season), now reaching almost 10% of the population at that time. These results indicate that the demographic changes observed in the wild may result in part from starvation-related adult mortality at the end of the winter reproductive period, despite adaptive plastic responses.

Acoustic surveys represent the standard fishery-independent method worldwide for evaluating the biomass and spatial distribution of small pelagic fish populations. Considering the peculiarities of the spatial behaviour of pelagic fishes, the efficiency of the survey design in determining their biomass and spatial distribution is related to the ability to capture the portion of the patches accounting for a larger part of the total biomass. However, the spatial structure of the patches could be strongly influenced by ecosystem characteristics as well as by changes in total biomass related to a density-dependent mechanism. This is of particular interest for anchovies and sardines, which are known for their wide fluctuations and high sensitivity to the environment. In this study, we analysed the efficiency of acoustic surveys targeting European anchovies (Engraulis encrasicolus) and European sardines (Sardina pilchardus) in 10 different areas of the Mediterranean Sea spanning three years of different biomass levels. Using the geostatistical coefficient of variation (CVgeo) of the average occurrence probability of high/medium density values, we showed different patterns in terms of survey design efficiency among areas and species. Anchovies usually showed a lower CVgeo than sardines in the Alboran Sea. In 4 out of 20 cases, CVgeo values showed a consistent decrease with increasing biomass, while in the remaining cases, the CVgeo did not follow any clear pattern, suggesting the presence of important environmental effects. Higher survey design efficiency was found in highly productive sectors influenced by river run-off, allowing us to hypothesize that higher productivity along with the presence of well-localized enrichment mechanisms could favour a spatially consistent distribution and coherent organization of fish populations, leading to higher precision estimates with a given transect design. While most surveys displayed CVgeo close to 10% or less even at low biomass, i

Data on Sardinella aurita (round sardinella) and Sprattus sprattus (European sprat) in the Mediterranean Sea are limited due to their scarce commercial interest, at least in European countries. Yet, these two small pelagic fish, sitting at opposite ends of the thermal range, could be interesting sentinel species to monitor the effects of climate change in the basin. Using the Mediterranean International Acoustic Surveys (MEDIAS) - the most extensive source of information on these species - we analysed their biomass in several geographical subareas of the central and western Mediterranean Sea in relation to satellite-derived environmental parameters. Our findings highlight that the S. aurita biomass responded to temperature, salinity, chlorophyll concentration and sea level anomaly, depending on the GSA examined, whereas the S. sprattus biomass correlated significantly with salinity in GSA 6, with salinity and chlorophyll concentration in GSA 7 and with sea level anomaly in GSA 17. These data widen our knowledge of the factors that contribute to the ecology of these species. Further studies of their spatial distribution and of their interactions with other small pelagic species, predators and prey are needed to depict a more comprehensive scenario.

Size at first sexual maturity (L50) represents an important life-history trait that needs to be considered in the development of management measures as it provides fundamental information for avoiding the exploitation of younger individuals. L50 is known to display variability due to fishing pressure, geographical gradients, and environmental features. In this study, to investigate L50 variability among areas in the Mediterranean and Black Seas, maturity ogives of anchovies (Engraulis encrasicolus) were estimated by considering samples collected during the anchovy spawning period in the framework of the MEDiterranean International Acoustic Survey (MEDIAS) program. Anchovy size and sexual maturity data from several geographical subareas (GSAs), i.e., northern Spain, Gulf of Lion, Tyrrhenian Sea, Strait of Sicily, Adriatic Sea and Black Sea, were gathered according to a standard methodological protocol. Maturity ogives were estimated by a logistic regression considering total length, condition factor, sex and GSA. The obtained results showed a significant effect of the condition factor, in that fish in better condition reached maturity earlier, and the results also indicated differences in L50 values among the areas and between the sexes, with males reaching maturity at lower lengths than females. Even though the obtained L50 estimates are relative to the spawning period only, the variability observed at the Mediterranean basin scale highlights the importance of explicitly considering specific habitat characteristics when providing management advice based on an ecosystem approach for fisheries.

In this study we consider the applicability of Bergmann’s rule to the populations of small pelagic fish species in the Mediterranean Sea. Under Bergmann’s rule, body size increases with decreasing temperature and increasing latitude. Although this macroecological pattern in body sizes is well established for many taxa of endotherms and ectotherms, it does not remain universal, and the proposed mechanisms underlying it are multiple and still lack consensus. Here we explored the occurrence of geographical body size clines using measurements of average body sizes of 10 species collected in pelagic trawl hauls carried out during acoustic surveys in the Mediterranean Sea. Bergmann’s rule was evaluated by correlating body sizes with latitude and temperature for each species while accounting for potential confounding variables and sampling bias. For 5 species, namely anchovy, sardine, Atlantic chub mackerel, bogue and blue jack mackerel, we demonstrate that they follow Bergmann’s rule, with a decline in average body size by about 3.01, 3.43, 3.67, 3.82 and 3.76 % per 1°C of warming, respectively, although this did not translate with an increase in size with latitude. The adherence of these 5 pelagic fish to Bergmann’s rules strongly suggest that temperature is a major determinant of their body sizes and enables them to act as sentinel species for identifying the drivers and consequences of warming in the Mediterranean ecosystems.

Global warming is causing profound modifications of aquatic ecosystems and one major outcome appears to be a decline in adult size of many fish species. Over the last decade, sardine populations in the Gulf of Lions (NW Mediterranean Sea) have shown severe declines in body size and condition as well as disappearance of the oldest individuals, which could not be related to overfishing, predation pressure or epizootic diseases. In this study, we investigated whether this situation reflects a bottom-up phenomenon caused by reduced size and availability of prey that could lead to energetic constraints. We fed captive sardines with food items of two different sizes eliciting a change in feeding mode (filter-feeding on small items and directly capturing larger ones) at two different rations for several months, and then assessed their muscle bioenergetics to test for changes in cellular function. Feeding on smaller items was associated with a decline in body condition, even at high ration, and almost completely inhibited growth by comparison to sardines fed large items at high ration. Sardines fed on small items presented specific mitochondrial adjustments for energy sparing, indicating a major bioenergetic challenge. Moreover, mitochondria from sardines in poor condition had low basal oxidative activity but high efficiency of ATP production. Notably, when body condition was below a threshold value of 1.07, close to the mean observed in the wild, it was directly correlated with basal mitochondrial activity in muscle. The results show a link between whole-animal condition and cellular bioenergetics in the sardine, and reveal physiological consequences of a shift in feeding mode. They demonstrate that filter-feeding on small prey leads to poor growth, even under abundant food and an increase in the efficiency of ATP production. These findings may partially explain the declines in sardine size and condition observed in the wild.

Mate choice is expected to be important for the fitness of both sexes for species in which successful reproduction relies strongly on shared and substantial parental investment by males and females. Reciprocal selection may then favour the evolution of morphological signals providing mutual information on the condition/quality of tentative partners. However, because males and females often have differing physiological constraints, it is unclear which proximate physiological pathways guarantee the honesty of male and female signals in similarly ornamented species. We used the monomorphic king penguin (Aptenodytes patagonicus) as a model to investigate the physiological qualities signalled by colour and morphological ornaments known to be under sexual selection (coloration of the beak spots and size of auricular feather patches). In both sexes of this slow-breeding seabird, we investigated the links between ornaments and multiple indices of individual quality; including body condition, immunity, stress and energy status. In both sexes, individual innate immunity, resting metabolic rate, and the ability to mount a stress response in answer to an acute disturbance (capture) were similarly signalled by various aspects of beak coloration or auricular patch size. However, we also reveal interesting and contrasting relationships between males and females in how ornaments may signal individual quality. Body condition and oxidative stress status were signalled by beak coloration, although in opposite directions for the sexes. Over an exhaustive set of physiological variables, several suggestive patterns indicated the conveyance of honest information about mate quality in this monomorphic species. However, sex-specific patterns suggested that monomorphic ornaments may signal different information concerning body mass and oxidative balance of males and females, at least in king penguins.

Investigating the drivers of fish assemblage trophic structure is a critical question, in order to better understand ecosystem functioning, predict the effects of perturbations and implement integrated management of exploited marine ecosystems. Ecosystemic surveys enabled the determination of the trophic structure of the fish assemblages in three French marine ecosystems, namely the Eastern English Channel, the Bay of Biscay and the Gulf of Lions, through the simultaneous collection of qualitative (stable isotopes and energy content) and quantitative (biomass) data. In the Bay of Biscay and in the Gulf of Lions, pelagic primary production supported at least 80% of the fish biomass production, and explained the dominance of pelagic species, but with differences resulting from the different productivity. The lower productivity in the oligotrophic Gulf of Lions led to a lower total biomass, energy density as well as the predominance of zooplankton feeders. In contrast, fluxes in the Bay of Biscay were sufficient to support a higher biomass of pelagic piscivores, and of species with higher energy content. In the shallow Eastern English Channel, the respective contributions of pelagic and benthic sources were similar. Bentho-demersal species of higher trophic level dominated this assemblage, because of their ability to exploit both pathways. Results of the present study confirmed that fisheries-focused surveys can be used as efficient platforms to address questions about ecosystem functioning. Here it confirmed the expected differences between ecosystems and the importance of primary production and environment as drivers of fish assemblage structure and functioning. Future studies should nevertheless develop new methods to better assess the paramount role of low trophic level consumers.

An important decrease in small pelagic fish condition and size has been observed in the most productive ecosystem of the Mediterranean Sea, the Gulf of Lions, since 2008, leading to an important fishery crisis. Previous studies suggested bottom-up control to be the most probable cause for these changes. Here, we investigate whether an environmental change might have caused such a situation. In the absence of zooplankton time series, this study aims at describing temporal changes in key abiotic factors for the planktonic and fish production of the Gulf of Lions, such as SST, meso-scale fronts, wind-induced coastal upwelling, river discharge, water stratification and deep convection and then at understanding potential link on Chl-a concentration as well as small pelagic fish populations. Our results indicate that the environmental conditions have broadly changed in the Gulf of Lion, with a major change in the mid-2000s, affecting the Chla concentration (which showed a regime shift in 2007), but also the SST, the upwelling and frontal activities, the Rhone river discharge (and particularly the N and P nutrients inputs) as well as the deep winter convection. Those changes could have affected the plankton production and consequently the small pelagic fish community that displayed similar patterns of variations as the environmental conditions.

We investigated links between swimming behavior and muscle bioenergetics in two emblematic Mediterranean fish species that have very different ecologies and activity levels. European sardines Sardina pilchardus are pelagic, they swim aerobically, school constantly and have high muscle fat content. Gilthead seabream Sparus aurata are bentho-pelagic, they show discontinuous spontaneous swimming patterns and store less fat in their muscle. Estimating the proportion of red and white muscle phenotypes, sardine exhibited a larger proportion of red muscle (~10% of the body mass) compared to gilthead seabream (~5% of the body mass). We firstly studied red and white muscle fiber bioenergetics, using high-resolution respirometers, showing a 4-fold higher oxidation capacity for red compared to white muscle. Secondly, we aimed to compare the red muscle ability to oxidize either lipids or carbohydrates. Sardine red muscle had a 3-fold higher oxidative capacity than gilthead seabream and a greater capacity to oxidize lipids. This study provides novel insights into physiological mechanisms underlying the different lifestyles of these highly-prized species.

Various protocols are currently used to study marine lipids, but there is a growing interest in working on dry samples that are easier to transport. However, reference protocols are still lacking for dry samples. In order to make recommendations on this use, lipid classes and fatty acids (FA) obtained from six analytical protocols using two different tissue states (dry vs wet) and three extraction methods (automat vs manual potter vs leaving the solvent to work on tissue) were compared. Three dry storage modes of tissue (freezer vs gas nitrogen vs dry room) during one and three months were also compared. These comparisons were made on seven marine species with different lipid profiles, including fishes, crustaceans and mollusks. Lipid classes and FA obtained from wet and dry tissues were similar, but they were affected by the extraction methods. Regardless of tissue state, “Leave to work” methods obtained the highest lipid quantities, followed by manual potter and automat methods (ca. 90% and 80% of “Leave to work” methods, respectively). Linear relationships allowed correction for lipid classes and FA concentrations obtained from different protocols. The repeatability of all protocols still needs to be improved, especially for fish species. Increasing the replicate number for each sample might be an indirect way to improve lipid quantification. Our results show that storing dry tissues in the freezer for more than one month was associated with a decrease in lipids, which is also observed for other storage methods. For qualitative studies of FA (expressed in %), a three-month storage of dry tissue in freezer did not affect the relative composition of species/tissues with a lipid content below 20% of dry weight.

Around 2008, an ecosystem shift occurred in the Gulf of Lions, highlighted by considerable changes in biomass and fish mean weight of its two main small pelagic fish stocks (European anchovy, Engraulis encrasicolus; European sardine, Sardina pilchardus). Surprisingly these changes did not appear to be mediated by a decrease in fish recruitment rates (which remained high) or by a high fishing pressure (exploitation rates being extremely low). Here, we review the current knowledge on the population's dynamics and its potential causes. We used an integrative ecosystem approach exploring alternative hypotheses, ranging from bottom-up to top-down control, not forgetting epizootic diseases. First, the study of multiple population characteristics highlighted a decrease in body condition for both species as well as an important decrease in size resulting both from a slower growth and a progressive disappearance of older sardines. Interestingly, older sardines were more affected by the decrease in condition than younger ones, another sign of an unbalanced population structure. While top-down control by bluefin tuna or dolphins, emigration and disease were mostly discarded as important drivers, bottom-up control mediated by potential changes in the plankton community appeared to play an important role via a decrease in fish energy income and hence growth, condition and size. Isotopic and stomach content analyses indicated a dietary shift pre- and post-2008 and modeled mesozooplankton abundance was directly linked to fish condition. Despite low energy reserves from 2008 onwards, sardines and anchovies maintained if not increased their reproductive investment, likely altering the life-history trade-off between reproduction and survival and resulting in higher natural mortality. The current worrying situation might thus have resulted from changes in plankton availability/diversity, which remains to be thoroughly investigated together with fish phenotypic plasticity.