Heat shocks during egg incubation led to developmental, morphological, and behavioral differences in Arctic charr (Salvelinus alpinus).

Temperature variation is affecting fish biodiversity worldwide, causing changes in geographic distribution, phenotypic structure, and even species extinction. Incubation is a critical stage for stenothermic species, which are vulnerable to large temperature fluctuations, and its effects on the phenotype at later developmental stages are understudied, despite the fact that the phenotype being essential for organism ecology and evolution. In this study, we tested the effects of heat shocks during the embryonic period on the phenotype of Arctic charr (Salvelinus alpinus). We repeatedly quantified multiple phenotypic traits, including morphology, development, and behavior, over a period of 4 months, from hatching to juvenile stage in individuals that had experienced heat shocks (+ 5°C on 24 h, seven times) during their embryonic stage and those that had not. We found that heat shocks led to smaller body size at hatching and a lower sociability. Interestingly, these effects weakened throughout the development of individuals and even reversed in the case of body size. We also found an accelerated growth rate and a higher body condition in the presence of heat shocks. Our study provides evidence that heat shocks experienced during incubation can have long-lasting effects on an individual's phenotype. This highlights the importance of the incubation phase for the development of ectothermic organisms and suggests that temperature fluctuations may have significant ecological and evolutionary implications for Arctic charr. Given the predicted increase in extreme events and the unpredictability of temperature fluctuations, it is critical to further investigate their effects on development by examining fluctuations that vary in frequency and intensity.

Changes in vertical and horizontal diversities mediated by the size structure of introduced fish collectively shape food-web stability.

Species introductions can alter local food-web structure by changing the vertical or horizontal diversity within communities, largely driven by their body size distributions. Increasing vertical and horizontal diversities is predicted to have opposing effects on stability. However, their interactive effects remain largely overlooked. We investigated the independent and collective effects of vertical and horizontal diversities on food-web stability in alpine lakes stocked with variable body size distributions of introduced fish species. Introduced predators destabilize food-webs by increasing vertical diversity through food chain lengthening. Alternatively, increasing horizontal diversity results in more stable food-web topologies. A non-linear interaction between vertical and horizontal diversities suggests that increasing vertical diversity is most destabilizing when horizontal diversity is low. Our findings suggest that the size structure of introduced predators drives their impacts on stability by modifying the structure of food-webs, and highlights the interactive effects of vertical and horizontal diversities on stability.

Automatic detection, identification and counting of anguilliform fish using in situ acoustic camera data: Development of a cross-camera morphological analysis approach.

Acoustic cameras are increasingly used in monitoring studies of diadromous fish populations, even though analyzing them is time-consuming. In complex in situ contexts, anguilliform fish may be especially difficult to identify automatically using acoustic camera data because the undulation of their body frequently results in fragmented targets. Our study aimed to develop a method based on a succession of computer vision techniques, in order to automatically detect, identify and count anguilliform fish using data from multiple models of acoustic cameras. Indeed, several models of cameras, owning specific technical characteristics, are used to monitor fish populations, causing major differences in the recorded data shapes and resolutions. The method was applied to two large datasets recorded at two distinct monitoring sites with populations of European eels with different length distributions. The method yielded promising results for large eels, with more than 75% of eels automatically identified successfully using datasets from ARIS and BlueView cameras. However, only 42% of eels shorter than 60 cm were detected, with the best model performances observed for detection ranges of 4-9 m. Although improvements are required to compensate for fish-length limitations, our cross-camera method is promising for automatically detecting and counting large eels in long-term monitoring studies in complex environments.

Evolution of pace-of-life syndrome under conditions of maternal PCB contamination and global warming in early life stages of cold stenothermic fish (Arctic char).

The end of the 20th century was characterised by rapid modifications of ecosystem functioning under different pressures (such as eutrophication and toxic pollution). Increasing temperatures in the context of global warming could have indirect consequences, such as increased bioavailability of hydrophobic organic pollutants amongst aquatic species. According to the "pace-of-life syndrome" (POLS) theory, these stressors could lead to covariations in many life traits. Lake Bourget is the largest natural lake in France and has been highly polluted from the fifties to the eighties both with a high load of nutrients (wastewater discharge) and polychlorinated biphenyls (PCBs) (industrial effluent discharge). Despite improvements in water quality since the 21st century, PCB levels are still higher than the United States Environmental Protection Agency cut-off for wildlife protection. The population of Arctic char, a cold stenothermic salmonid, has remained low in Lake Bourget for the last ten years despite restocking efforts and complete re-oligotrophication. We hypothesised that PCB pollution can affect the Arctic char population and that the increase in water temperature could magnify the effects of PCB. Thus, this study aimed to investigate the effects of maternal PCB contamination on offspring using a multiparametric and multiscale approach. Female Arctic char were contaminated with PCB before spawning, and each fertilised spawn was incubated at two temperatures (4 and 8.5 °C). The results showed that co-exposure to increased temperature and maternal PCB contamination influenced biodemographic, physiological, and behavioural parameters. The effects were highly dependant on the developmental stage. Based on the POLS theory, a continuum of life traits that may reflect potential physiological and behavioural modifications in response to these concurrent stressors is highlighted.

Ultra slow acoustic energy transport in dense fish aggregates.

A dramatic slowing down of acoustic wave transport in dense fish shoals is observed in open-sea fish cages. By employing a multi-beam ultrasonic antenna, we observe the coherent backscattering phenomenon. We extract key parameters of wave transport such as the transport mean free path and the energy transport velocity of diffusive waves from diffusion theory fits to the experimental data. The energy transport velocity is found to be about 10 times smaller than the speed of sound in water, a value that is exceptionally low compared with most observations in acoustics. By studying different models of the fish body, we explain the basic mechanism responsible for the observed very slow transport of ultrasonic waves in dense fish shoals. Our results show that, while the fish swim bladder plays an important role in wave scattering, other organs have to be considered to explain ultra-low energy transport velocities.

Mitigation of ecological impacts on fish of large reservoir sediment management through controlled flushing - The case of the Verbois dam (Rhône River, Switzerland).

Sediment trapping within reservoirs is a worldwide phenomenon which impairs the ecological functioning of upstream and downstream ecosystems. It also reduces reservoir water storage volume, which lessens the services dams provide such as hydropower production or flood control and questions their sustainability. Hydraulic flushing is a widely used operation to recover the reservoir volume, but ecological impacts are massive. Recently, environmental awareness led dam operators to modify their management practices: 'Controlled Sediment Flushing Operations' (CSFOs) include environmental objectives in their implementation and are designed to be less harmful for aquatic ecosystems by controlling the flow and Suspended Sediment Concentration (SSC) downstream. However, CSFOs are not yet widespread, their ecological impacts are poorly documented, and comparisons with 'classical' flushing operations are unreported. Here, we analysed impacts on fish of the first CSFO of the Verbois reservoir in 2016, both upstream and downstream of the dam, and compared these with those from the empty flushing of 2012 using the same methodology (Grimardias et al., 2017). Time-series of hydroacoustics surveys enabled us to estimate the fish abundance in the reservoir, while radiotelemetry measured movements and apparent survival below the dam for four representative species. The 2016 CSFO lasted 10 days, and released a mean Suspended Sediment Concentration (SSC) of 3.47 g·L-1 (peak = 11.98 g·L-1). The fish density as assessed by the mean acoustic scattering strength SA in the reservoir did not change significantly pre- and post-CSFO, and SA seasonal estimates of year 2016 did not differ from those of 2015 and 2017. The apparent survival estimated from capture-recapture survey of marked fish (N = 118) decreased significantly during the CSFO for all species and differed across species, while the distances moved downstream increased. By comparison with the 2012 empty flushing, the 2016 CSFO allowed fish to remain in the reservoir, while impacts below the dam were mostly behavioural rather than lethal. Overall, despite significant impacts, the CSFO advantageously replaced 'classical' flushing from an ecological viewpoint. Provided that an acceptable balance between economical, ecological and technical aspects is found, CSFO can be considered for many reservoirs while accounting for their biological and physical site-specificity.

Acoustic density estimation of dense fish shoals.

Multiple scattering of acoustic waves offers a noninvasive method for density estimation of a dense shoal of fish where traditional techniques such as echo-counting or echo-integration fail. Through acoustic experiments with a multi-beam sonar system in open sea cages, multiple scattering of sound in a fish shoal, and, in particular, the coherent backscattering effect, can be observed and interpreted quantitatively. Furthermore, a volumetric scan of the fish shoal allows isolation of a few individual fish from which target strength estimations are possible. The combination of those two methods allows for fish density estimation in the challenging case of dense shoals.

Manual fish length measurement accuracy for adult river fish using an acoustic camera (DIDSON).

Total lengths (LT ) of 50 free-swimming fish in a tank, silver carp Hypophthalmichthys molitrix and rainbow trout Oncorhynchus mykiss, were measured using a DIDSON (Dual-frequency IDentification SONar) camera. Using Sound Metrics software, multiple measurements of each fish (LT , side aspect angle and distance from the camera) at different times were analysed by two experienced operators while a subset of data was analysed by two inexperienced operators. The main result showed high variability in intra-fish LT measurements. The number of measurements required to minimise errors and to obtain robust fish measurements (true LT ± 3 cm) was estimated by a bootstrap method. Three to five measurements per fish were recommended for fish surveys in rivers. In this experimental study, aiming to reproduce river conditions, no evidence of fish position (side aspect angle and distance from the camera) effect was detected, but an operator effect (partially explained by training) was observed. General linear mixed models also showed that lengths of the smallest fish (LT < 57 cm) were overestimated and lengths of the largest fish (LT > 57 cm) were underestimated in comparison with their true lengths. In conclusion, we highlight that this technology, like any monitoring methods, returns imperfect observations. We advise DIDSON users to ensure that measurements are carried out correctly in order to draw accurate conclusion from this new technology.

Parasitic versus nutritional regulation of natural fish populations.

Although parasites are expected to affect their host's fitness, quantitative proof for impacts of parasitism on wild populations is hampered by confounding environmental factors, including dietary resource. Herein, we evaluate whether the physiological conditions of European perch (Perca fluviatilis) in three large peri-alpine lakes (Geneva, Annecy, and Bourget) depend on (a) the nutritional status of the juvenile fish, as revealed by stable isotope and fatty acid compositions, (b) the prevalence of the tapeworm Triaenophorus nodulosus, a parasite transmitted to perch through copepod preys, or (c) interactive effects of both factors. At the scale of lake populations, the deficit in growth and fat storage of juvenile perch during their first summer coincides with a high parasite prevalence and also a low quality of dietary resource. Yet, at the individual level, parasites had no evident effect on the growth of the juvenile perch, while impacts on fat storage appeared only at the highest prevalence of the most infected lake. Fatty acid and stable isotope analyses of fish tissue do not reveal any impact of T. nodulosus on diet, physiology, and feeding behaviour of fish within lakes. Overall, we found a low impact of parasitism on the physiological condition and trophic status of juvenile perch at the end of their first summer. We find instead that juvenile perch growth and fat storage, both factors tied to their winter survival, are under strong nutritional constraints. However, the coinciding nutritional constraints and parasite prevalence of perch juveniles in these three lakes may result from the indirect effect of lake nutrient concentrations, which, as a major control of zooplankton communities, simultaneously regulate both the dietary quality of fish prey and the host-parasite encounter rates.

Drawdown flushing of a hydroelectric reservoir on the Rhône River: Impacts on the fish community and implications for the sediment management.

Sediment flushings of hydropower reservoirs are commonly performed to maintain water resource uses and ecosystem services, but may have strong impacts on fish communities. Despite the worldwide scope of this issue, very few studies report quantitative in situ evaluations of these impacts. In June 2012, the drawdown flushing of the Verbois reservoir (Rhône River) was performed and subsequent impacts on the fish community were assessed, both inside the reservoir (fish densities by hydroacoustic surveys) and downstream (short-term movement and survival of radio tracked adult fish). Results showed that after the flushing fish acoustic density decreased by 57% in the reservoir, and no recolonization process was observed over the following 16 months. Downstream of the dam, the global apparent survival of fish to the flushing was estimated at 74%, but differed between species. The nine-year delay from the previous flushing and thus the amount of sediments to remove were too stressful for the low-resilience fish community of the Rhône River. Alternative flushing schedules are discussed to reduce these impacts.

Trophic transfer of microcystins through the lake pelagic food web: evidence for the role of zooplankton as a vector in fish contamination.

An in situ study was performed to investigate the role of zooplankton as a vector of microcystins (MCs) from Planktothrix rubescens filaments to fish during a metalimnic bloom of P. rubescens in Lake Hallwil (Switzerland). The concentrations of MCs in P. rubescens and various zooplanktonic taxa (filter-feeders and predators) were assessed in different water strata (epi-, meta- and hypolimnion) using replicated sampling over a 24-hour survey. The presence of P. rubescens in the gut content of various zooplanktonic taxa (Daphnia, Bosmina and Chaoborus) was verified by targeting the cyanobacterial nucleic acids (DNA). These results highlighted that cyanobacterial cells constitute a part of food resource for herbivorous zooplanktonic taxa during metalimnic bloom periods. Furthermore, presence of MCs in Chaoborus larvae highlighted the trophic transfer of MCs between herbivorous zooplankton and their invertebrate predators. Our results suggest that zooplanktonic herbivores by diel vertical migration (DVM) act as vectors of MCs by encapsulating grazed cyanobacteria. As a consequence, they largely contribute to the contamination of zooplanktonic predators, and in fine of zooplanktivorous whitefish. Indeed, we estimated the relative contribution of three preys of the whitefish (i.e. Daphnia, Bosmina and Chaoborus) to diet contamination. We showed that Chaoborus and Daphnia were the highest contributor as MC vectors in the whitefish diet (74.6 and 20.5% of MC-LR equivalent concentrations, respectively). The transfer of MCs across the different trophic compartments follows complex trophic pathways involving various trophic levels whose relative importance in fish contamination might vary at daily and seasonal scale.

Are cyanobacterial blooms trophic dead ends?

Cyanobacterial blooms induce significant costs that are expected to increase in the near future. Cyanobacterial resistance to zooplankton grazing is one factor thought to promote bloom events. Yet, numerous studies on zooplankton ability to graze upon cyanobacteria have been producing contradictory results and such a puzzle might arise from the lack of direct observations in situ. Our objective was to track, using fatty acid (FA) and fatty acid stable isotope analyses (FA-SIA), the fate of cyanobacterial organic matter in the food web of a lake subjected to summer blooms of Planktothrix rubescens. A metalimnetic bloom of P. rubescens occurred in Lake Bourget (France) during the study period (May-November 2009). The bloom was especially rich in α-linolenic acid, 18:3(n-3), but none of the considered zooplankton taxa exhibited spiking content in this particular FA. FA-SIA revealed, however, that over a quarter of 18:3(n-3) in small zooplankton (<500 µm) was provided by P. rubescens while large cladocerans (>500 µm) did not benefit from it. P. rubescens 18:3(n-3) could be tracked up to perch (Perca fluviatilis) young of the year (YOY) to which it contributed to ~15 % of total 18:3(n-3). Although transferred with a much lower efficiency than micro-algal organic matter, the P. rubescens bloom supported a significant share of the pelagic secondary production and did not constitute, sensu stricto, a 'trophic dead end'. The cyanobacterial bloom also provided perch YOY with components of high nutritional values at a season when these are critical for their recruitment. This cyanobacterial bloom might thus be regarded as a significant dietary bonus for juvenile fish.

Impact of toxic cyanobacterial blooms on Eurasian perch (Perca fluviatilis): experimental study and in situ observations in a peri-alpine lake.

Due to the importance of young-of-the-year (YOY) perch in the peri-alpine regions where they are consumed, the microcystin (MC) contamination of YOY perch was analysed both in field (Lake Bourget, France) and experimentally using force-feeding protocols with pure MCs. In-situ, schools of YOY perch present in the epilimnion of the lake were never found in direct contact with the P. rubescens blooms that were present in the metalimnion. However, MCs were detected in the muscles and liver of the fish and were thus assumed to reach YOY perch through dietary routes, particularly via the consumption of MC-containing Daphnia. Force-feeding experiment demonstrates the existence of MC detoxification/excretion processes and suggests that in situ, YOY perch could partly detoxify and excrete ingested MCs, thereby limiting the potential negative effects on perch populations under bloom conditions. However, because of chronic exposure these processes could not allow for the complete elimination of MCs. In both experimental and in situ studies, no histological change was observed in YOY perch, indicating that MC concentrations that occurred in Lake Bourget in 2009 were too low to cause histological damage prone to induce mortality. However, Deoxyribonucleic acid (DNA) damages were observed for both the high and low experimental MC doses, suggesting that similar effects could occur in situ and potentially result in perch population disturbance during cyanobacterial blooms. Our results indicate the presence of MCs in wild perch, the consumption of this species coming from Lake Bourget is not contested but more analyses are needed to quantify the risk.

Short-term uptake of microcystin-LR by Coregonus lavaretus: GST activity and genotoxicity.

In the present study, juvenile whitefish weighing 2 g were exposed by force-feeding to two ecologically relevant doses (0.05 and 0.5 µg per fish) of microcystin-LR (MC-LR). Then over 96 h the MC uptake in fish liver and muscle was measured, as the activity of the detoxification enzyme glutathione S-transferase (GST) in the liver, and the genotoxicity impact on red blood cells. Results show that (1) the MC-LR equivalent concentrations increased for both doses and in both organs of whitefish with approximately threefold lower concentrations for the low dose compared to the high dose in both organs and threefold lower concentrations in the muscle compared to the liver for each dose (2) the liver GST activity increased during the first 48 h of exposure with fivefold higher GST activity for the highest dose at 48 h compared to control and (3) MC-LR leads to deoxyribonucleic acid strand breaks that were detected by the comet assay and shown to be partially repaired. This work demonstrates that European whitefish could be impacted by cyanobacteria toxins due to rapid microcystin uptake, especially in the context of chronic contamination, which can occur during long bloom episodes.

Gaucher's disease and fatal hepatic fibrosis despite prolonged enzyme replacement therapy.

We report on the case of a girl with type 1 Gaucher's disease, treated from age 9 to 15 with high-dose enzyme replacement therapy. This treatment did not avert the development of an extensive mutilating hepatic fibrosis warranting a liver transplantation, which was followed by death. In some cases of Gaucher's disease, alternative strategies such as fractionated or further increased ERT, gene therapy, or glucosyltransferase inhibitor should be explored.