Published papers

Many studies have investigated how nanoplastics (NPs) exposure mediates nerve and intestinal toxicity through a dysregulated brain-gut axis interaction, but there are few studies aimed at alleviating those effects. To determine whether and how vitamin D can impact that toxicity, fish were supplemented with a vitamin D-low diet and vitamin D-high diet. Transmission electron microscopy (TEM) showed that polystyrene nanoplastics (PS-NPs) accumulated in zebrafish brain and intestine, resulting in brain blood–brain barrier basement membrane damage and the vacuolization of intestinal goblet cells and mitochondria. A high concentration of vitamin D reduced the accumulation of PS-NPs in zebrafish brain tissues by 20% and intestinal tissues by 58.8% and 52.2%, respectively, and alleviated the pathological damage induced by PS-NPs. Adequate vitamin D significantly increased the content of serotonin (5-HT) and reduced the anxiety-like behavior of zebrafish caused by PS-NPs exposure. Virus metagenome showed that PS-NPs exposure affected the composition and abundance of zebrafish intestinal viruses. Differentially expressed viruses in the vitamin D-low and vitamin D-high group affected the secretion of brain neurotransmitters in zebrafish. Virus AF191073 was negatively correlated with neurotransmitter 5-HT, whereas KT319643 was positively correlated with malondialdehyde (MDA) content and the expression of cytochrome 1a1 (cyp1a1) and cytochrome 1b1 (cyp1b1) in the intestine. This suggests that AF191073 and KT319643 may be key viruses that mediate the vitamin D reduction in neurotoxicity and immunotoxicity induced by PS-NPs. Vitamin D can alleviate neurotoxicity and immunotoxicity induced by PS-NPs exposure by directionally altering the gut virome. These findings highlight the potential of vitamin D to alleviate the brain-gut-virome disorder caused by PS-NPs exposure and suggest potential therapeutic strategies to reduce the risk of NPs toxicity in aquaculture, that is, adding adequate vitamin D to diet.

Inflammatory muscle loss results from excessive inflammatory responses, causing muscle damage and weakness. In the current investigation, we evaluated the protective effects of diphlorethohydroxycarmalol (DPHC) against tumor necrosis factor-alpha (TNF-α)-induced skeletal muscle inflammation and muscle loss and elucidated the underlying mechanisms. Furthermore, the effect of DPHC on swimming performance was confirmed under TNF-α-induced inflammatory muscle loss-conditioned zebrafish by assessing the swimming number, distance moved, time spent swimming, frequency of swimming zebrafishes in an upstream swim track (Zone A). In vivo behavioral endurance test results indicated that TNF-α treatment significantly decreased the number of swimming zebrafish and swimming distance in Zone A compared with the Control. Meanwhile, the DPHC treatment significantly increased the number of swimming zebrafish and swimming distance in Zone A compared to TNF-α-induced zebrafish. These findings indicate that DPHC treatment effectively improved the swimming performance of TNF-α-induced zebrafish. In an additional study, TNF-α significantly induced inflammatory muscle loss by upregulating nuclear factor kappa light chain enhancer of activated B cells (NF-κB) mitogen activated protein kinase (MAPK) associated proteins and MuRF-1 in the skeletal muscle tissues of TNF-α-induced zebrafish. However, DPHC administration significantly counteracted TNF-α-induced inflammation and muscle loss by downregulating NF-Κb and MAPK-associated proteins, as well as the muscle degradation-related proteins MuRF-1 and MAFbx, in the skeletal muscle tissues of TNF-α-induced zebrafish. In summary, our research findings demonstrated that DPHC from Ishige okamurae could be used for the development of nutraceuticals or functional foods targeting inflammatory muscle loss.

The principle of the 3Rs-Reduction, Refinement, and Replacement-encourages minimizing animal use, improving experimental design, and developing alternative models for toxicology testing. Among such models, planaria (aquatic flatworms) have gained increasing attention in pharmacology, regenerative medicine, and toxicology because of their simple anatomy, high environmental sensitivity, exceptional regenerative ability, and ease of laboratory maintenance. In this study, we examined the effects of benzalkonium chloride (BAC)-a commonly used pharmaceutical excipient with antimicrobial and permeability-enhancing properties, as well as a known environmental toxicant-on the locomotor activity of Schmidtea mediterranea using both manual assessment and Lolitrack video-tracking software. Six concentrations of BAC (5-1000 μg/mL) and a negative control were tested. Both approaches showed an overall reduction in locomotor activity over time, though manual analysis indicated a transient stimulation at lower concentrations. The software-based method demonstrated greater reliability, precision, and objectivity, making it preferable for toxicity evaluation in planaria.

Sea trout, Salmo trutta, display a wide range of migratory behaviours, and one aspect of variation comes from freshwater migration distance. The overall aim of this study was to determine if offspring of long‐ and short‐distance migrants exhibited phenotypic differences relating to parental migration distance. For that purpose, we conducted several behavioural tests (dyadic contest, boldness scoring and open field test) and morphological analysis (relative pectoral‐fin length) in multiple freshwater systems across the distribution range of the target species in Europe. It was expected that offspring of long‐distance migrants would be more active, bold and dominant than those of short‐distance migrants and would have longer pectoral fins relative to body length. Additionally, we investigated if boldness varied in relation to latitude. We showed that offspring of long‐distance migrants were more dominant in two cases and more active in one case than those of short‐distance migrants; however, there was no difference in swimming distance or velocity. Boldness and relative pectoral‐fin length were significantly related to site of origin; however, the direction of this relationship differed between systems. Generally, we detected a decrease in boldness with declining latitude. In summary, we have detected variation among juveniles related to location within a stream; however, the drivers and processes behind these are likely more complex than purely parental migratory strategy. Our results can inform suitable management and conservation efforts directed to anadromous Salmo trutta. For example, habitat restoration and removal of migration barriers can increase the possible range of migration distances helping maintain the phenotypic diversity of offspring.

The transfer of information between individuals is fundamental to living systems and requires comprehensive research in various species. Weakly electric fish, Gnathonemus petersii, provides a unique model organism for such investigations due to its advanced electrocommunication via electric organ discharges (EODs). As separating EODs from multiple individuals remains challenging, we developed an unsupervised approach for EOD separation in two free-swimming individuals. Using continuous wavelet transform, t-distributed Stochastic Neighbor Embedding, and hierarchical clustering, we achieved accurate discrimination of EODs without the necessity of any training data. This approach overcomes the supervised algorithms based on previously published methods in accuracy and computational efficiency, simplifies experimental procedures, and supports animal well-being by reducing the number of required measurements. We applied our separation approach in a dyadic fish model, where ketamine was used to induce schizophrenia-like behavior in one fish. We confirmed the ketamine-induced alteration of the intrinsic relationship between locomotion and EOD signaling. Moreover, while ketamine-induced changes in locomotion were socially transferred, correlated changes in EOD signaling were not observed between dyad members, which may be interpreted as a communication deficit. Additionally, we introduced two techniques for EOD sonification, facilitating exploratory analysis of EOD sequences. These advancements lay the groundwork for future studies of EOD-based communication, highlighting the potential of Gnathonemus petersii in neuroethological, psychopharmacological, and translational research.

Marine heatwaves (MHWs), coral bleaching, and chronic local stressors such as eutrophication are accelerating regime shifts from coral‐ to algae‐dominated reefs, increasingly favoring the proliferation of invasive, fast‐growing, and often more grazing‐resistant turf and macroalgae. A central tenet of global reef management strategies is that herbivorous fishes can sustain critical top‐down control of algal proliferation as oceans warm. Here, we challenge this tenet by experimentally evaluating, under controlled laboratory conditions, whether herbivorous coral reef fishes across three key functional groups—browser ( Naso lituratus ), grazer ( Acanthurus triostegus ), and scraper ( Chlorurus spilurus )—can maintain effective algal control across present‐day (24.0°C–27.5°C) temperatures and into projected MHWs (31°C). We assessed (1) whether individuals evacuated thermally stressed conditions, effectively abandoning algal control, and (2) for those that remained, whether they could meet elevated energetic demands by foraging ad libitum on a mixture of Caulerpa spp.—a rapidly spreading and archetypal group of invasive algae in the Indo‐Pacific. All species gained body mass while foraging exclusively on these algae during winter and summer (~0.18%–0.62% per day). However, despite remaining in thermally stressed conditions and maintaining stable foraging rates, all species experienced consistent body mass declines (~0.41%–1.62% per day) under MHW exposure. This precipitous decline in body mass was driven by ~54%–60% increases in basal energetic demands without corresponding increases in food intake. Survival estimates based on body mass loss ranged from ~20–81 days, which is substantially shorter than the projected ~126–152‐day average duration of future MHWs. Our findings reveal that while short‐term algal control may persist during thermal stress, prolonged exposure appears to erode herbivore physiological condition, effectively undermining top‐down control of some algal types. Consequently, as ocean warming intensifies, herbivore protection strategies may become increasingly less effective at staving off algae proliferation and dominance in threatened reef ecosystems.

Groundwater ecosystems play a pivotal role in global biodiversity and ecosystem functioning, yet they face increasing pressures from climate change. The amphipod genus Niphargus, a dominant taxon in European groundwater habitats, has shown evidence of broad thermal adaptability that challenges prevailing theories on narrow thermal niches in groundwater species. This study investigated the locomotory behaviour of Niphargus longicaudatus (Costa, 1851), a stygobitic amphipod, under habitat temperature (9 °C) and preferred temperature (15 °C) using 3D tracking techniques. Individuals at 15 °C displayed significantly higher average swimming speed, increased vertical occupancy, and greater trajectory tortuosity compared to those at 9 °C, despite spending a similar amount of time in movement. These behavioural shifts suggest metabolic adjustments enabling enhanced resource exploration at warmer temperatures. The findings are contextualized within the evolutionary history of the amphipod genus Niphargus, shaped by past climatic, geological and hydrological conditions, which may have selected for eurythermal traits in some lineages. These adaptations highlight potential to exploit habitats across a broad temperature range, not necessarily providing an advantage to N.longicaudatus due to the complex effects of climate change on groundwater ecosystems. This work underscores the importance of integrating behavioural, metabolic, and paleoclimatic perspectives in understanding the impacts of climate change on subterranean biodiversity and distribution.

Invasive bigheaded carps (Hypophthalmichthys molitrix and H. nobilis) have caused substantial ecological and economic damage throughout the Mississippi River Basin and expanded their range threatening the Laurentian Great Lakes. Broadband acoustic deterrents have shown promise in repelling carp and are currently being assessed in navigational lock chambers on the Mississippi River. These nonphysical deterrents permit vessel navigation while reducing carp passage. However, no single deterrent is 100% effective and fish may habituate to the sound after repeated playback. Carp exhibit aversive behaviors to carbon dioxide, which suggests combining these two stimuli into one deterrent system could extend the effective duration of sound and reduce the frequency of carbon dioxide (CO2) application. We conditioned bigheaded carps to associate broadband sound from outboard boat motors (0.06–5 kHz, ~150 dB re. 1 μPa) with CO2 application (~35,000 ppm) in small (80 L) and large (3475 L) two-choice shuttle tanks. We compared negative phonotaxis responses over one to four weeks between fish conditioned with sound and CO2, sound and air, or sound alone. Similar CO2 avoidance thresholds were found across tank sizes and species. Conditioning treatment did not affect time to leave the sound chamber, confirming sound alone remains a deterrent for all fish. Carp conditioned with CO2 took longer to return to the sound chamber than control treatments. Control fish were closer to the speaker during playback than during the pre-sound period, while fish conditioned with CO2 were not significantly closer. Conditioning paradigms may extend the effective duration of nonphysical deterrents for bigheaded carps. Conditioning with CO2 may also increase proactive flight-responses over reactive freeze-responses. Findings could be applied to increase nonphysical barrier effectiveness at locks along the Mississippi River and help protect the Laurentian Great Lakes from invasion.

Deterrent technologies are one component of preventing the spread of invasive fishes to protect aquatic ecosystems from biodiversity loss. Curtains of bubbles can act as a non-physical barrier to deter fish movements, but will not stop all species in all situations. Modifications to bubble curtains that decrease fish movements would help protect aquatic ecosystems. The current study sought to quantify whether adding carbon dioxide gas (CO2) to a bubble curtain would enhance its efficacy to block fish. For this, a choice tank was outfitted with bubble curtains infused with either compressed air alone, or with two different concentrations of CO2 [30 or 100 mg/L]. Passage rates and position of common carp (Cyprinus carpio, an invasive Cyprinid) and black bullhead (Ameiurus melas, a native Ictalurid) exposed to these treatments were compared. Common carp were less likely to pass a bubble curtain when CO2 gas was used relative to the use of compressed air alone, and only 30 mg/L CO2 was needed to reduce passage. Black bullhead passages were not influenced by the bubble curtain, even with the addition of CO2. However, black bullhead, were found 30% further upstream of the curtain when CO2 was used relative to the control and air alone treatments, demonstrating avoidance of CO2. This study shows that CO2 added to a bubble curtain will enhance its ability to block passage of invasive fish.

As more physiologists start to incorporate animal behavior into their experiments, especially in the olfactory behavior research field, some considerations are often overlooked, partly due to the inherited way that physiological experiments are traditionally designed and performed. Here we highlight some of these subtle but important considerations and make a case for why these might affect the results collected from behavioral assays. Our aim is to provide useful suggestions for increased standardization of methods so they can be more easily replicated among different experiments and laboratories. We have focused on areas that are less likely to be mentioned in the materials and methods section of a manuscript such as starvation, preliminary experiments, appropriate sample sizes and considerations when choosing an odorant for an assay. Additionally, we are strongly cautioning against the use of alarm cue to generate behavioral responses due to its highly unstable chemical properties/potency. Instead, we suggest using pure chemicals (made up of one known molecule) such as amino acids, bile acids, or polyamines that are commercially available and easier to make up in known concentrations. Lastly, we strongly suggest using environmentally relevant concentrations of these odorants. We believe these guidelines will help standardize these assays and improve replication of experiments within and between laboratories.

Temperature fluctuations challenge ectothermic species, particularly tropical fish dependent on external temperatures for physiological regulation. However, the molecular mechanisms through which low-temperature stress impacts immune responses in these species, especially in relation to chromatin accessibility and epigenetic regulation, remain poorly understood. In this study, we investigate chromatin and transcriptional changes in the head kidney and thymus tissues of Nile tilapia (Oreochromis niloticus), a tropical fish of significant economic importance, under cold stress. By analyzing cis-regulatory elements in open chromatin regions and their associated transcription factors (TFs), we construct a comprehensive transcriptional regulatory network (TRN) governing immune responses, including DNA damage-induced apoptosis. Our analysis identifies 119 TFs within the TRN, with Stat1 emerging as a central hub exhibiting distinct binding dynamics under cold stress, as revealed by footprint analysis. Overexpression of Stat1 in immune cells leads to apoptosis and increases the expression of apoptosis-related genes, many of which contain Stat1-binding sites in their regulatory regions, emphasizing its critical role in immune cell survival during cold stress. These results provide insights into the transcriptional and epigenetic regulation of immune responses to cold stress in tilapia and highlight Stat1 as a promising target for enhancing cold tolerance in tropical fish species.

Ectotherms, such as fish, are highly dependent on the stability of their environment to regulate body temperature, performance, and metabolism. Increasing temperatures cause behavioural changes in fish which can be observed and used as indices for determining upper thermal limits. The thermal agitation temperature (T ag ) is a recent, and ecologically significant, sublethal index for the upper thermal limit. Previous studies have described thermal agitation as the endpoint, prior to the critical thermal maximum (CT max ), where fish start exhibiting apparent refuge-seeking and thermal avoidance behaviour. It is an assumption that fish are seeking thermal refuge at T ag, but evidence for this is lacking. Therefore, this study aimed to validate this assumption by using zebrafish (Danio rerio) and providing them with thermal refuge while increasing their environment's temperature past T ag. The behavioural responses of D. rerio were observed and their spatial movements were tracked using the animal-tracking software, AnimalTA. The analysis from this study indicated that refuge is sought out prior to T ag and distance between shoal members increases after T ag, indicating D. rerio may trade-off the protective value of a shoal to search for thermal refuge. Our study demonstrates that with refuge available, D. rerio can surpass T ag until refuge itself exceeds T ag, validating that agitation is refuge-seeking behaviour, but a mechanism of last resort. This insight improves our understanding of fish responses to thermal stress and emphasizes the value of using T ag as a sublethal metric alongside CT max in thermal tolerance studies, with potential applications in ecology and conservation contexts.

High phenotypic diversity should provide populations with resilience to environmental change by increasing their capacity to respond to changing conditions. The aim of this study was to identify whether there is consistency in individual behaviours on a reactive-proactive axis in European barbel Barbus barbus ("barbel"), a riverine and aggregatory fish that expresses individual differences in its behaviours in nature. This was tested using three sequential experiments in ex-situ conditions that required individuals to leave a shelter and then explore new habitats (‘open-field test’), respond to social stimuli (‘mirror-image stimulation test’) and forage (‘foraging behaviour test’; assessing exploratory traits). Each suite of experiments was replicated three times per individual (46 hours minimum time between replicates). There was high variability in behaviours both within and among individuals. The most repeatable behaviours were latency to exit the shelter, active time in the shelter, and the number of food items consumed. Principal component scores did, however, indicate a range of consistent behavioural phenotypes across the individuals, distributing them along a reactive-proactive axis in which most of individuals were more reactive phenotypes (shyer, less exploratory, less social). These results suggest that within controlled conditions, there is considerable phenotypic diversity among individuals in their behaviours, suggesting their populations will have some adaptive capacity to environmental change.

The relationship between behavioral thermoregulation and physiological recovery following exhaustive exercise is not well understood. Behavioral thermoregulation could be beneficial for exercise recovery; for example, selection of cooler temperatures could reduce maintenance metabolic cost to preserve aerobic scope for recovery cost, or selection of warmer temperatures could accelerate recovery of exercise metabolites. While post-exercise behavioral thermoregulation has been observed in lizards and frogs, little is known about its importance in fish. We examined the influence of post-exercise recovery temperature on metabolic rate, thermal preference, and metabolite concentrations in juvenile brook char (Salvelinus fontinalis). Fish were acclimated to and exercised at 15 °C, then recovered at either 15 °C or 10 °C while their metabolic rate was measured via respirometry. Metabolite concentrations were measured in fish after exercise at 15 °C and recovery under one of three thermal treatments (to simulate various behavioral thermoregulation scenarios): (i) 6 h recovery at 15 °C, (ii) 6 h recovery at 10 °C, or (iii) 3 h recovery at 10 °C followed by 3 h recovery at 15 °C. Thermal preference was quantified using a static temperature preference system (15 °C vs. 10 °C). Metabolic rates returned to resting faster at 10 °C compared with 15 °C, although at 10 °C there was a tradeoff of delayed metabolite recovery. Specifically, post-exercise plasma osmolality, plasma lactate, and muscle lactate remained elevated for the entire period in fish recovering at 10 °C, whereas these parameters returned to resting levels by 6 h in fish from the other two recovery groups. Regardless, fish did not exhibit clear behavioral thermoregulation (i.e., fish overall did not consistently prefer one temperature) to prioritize either physiological recovery process. The advantage of metabolic rate recovery at cooler temperatures may balance against the advantage of metabolite recovery at warmer temperatures, lessening the usefulness of behavioral thermoregulation as a post-exercise recovery strategy in fish.

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.

Microdosing ketamine is a novel antidepressant for treatment‐resistant depression. Traditional antidepressants, like selective serotonin reuptake inhibitors (SSRIs), inhibit serotonin reuptake, but it is not clear if ketamine shows a similar mechanism. Here, we tested the effects of feeding ketamine and SSRIs to Drosophila melanogaster larvae, which has a similar serotonin system to mammals and is a good model to track depressive behaviors, such as locomotion and feeding. Fast‐scan cyclic voltammetry (FSCV) was used to measure optogenetically stimulated serotonin changes, and locomotion tracking software and blue dye feeding to monitor behavior. We fed larvae various doses (1–100 mM) of antidepressants for 24 h and found that 1 mM ketamine did not affect serotonin, but increased locomotion and feeding. Low doses (≤10 mM) of escitalopram and fluoxetine inhibited dSERT and also increased feeding and locomotion behaviors. At 100 mM, ketamine inhibited dSERT and increased serotonin concentrations, but decreased locomotion and feeding because of its anesthetic properties. Since microdosing ketamine causes behavioral effects, we further investigated behavioral changes with a SERT16 mutant and low doses of other NMDA receptor antagonists and 5‐HT 1A and 2 agonists. Feeding and locomotion changes were similar to ketamine in the mutant, and we found NMDA receptor antagonism increased feeding, while serotonin receptor agonism increased locomotion, which could explain these effects with ketamine. Ultimately, this work shows that Drosophila is a good model to discern antidepressant mechanisms, and that ketamine does not work on dSERT like SSRIs, but effects behavior with other mechanisms that should be investigated further. image

One of the hallmarks of invasive species is their propensity to spread. Removing an invasive species after establishment is virtually impossible, and so considerable effort is invested in preventing the range expansion of invaders. Silver carp were discovered in the Mississippi River in 1981 and have spread throughout the basin. Despite their propensity to expand, the ‘leading edge’ in the Illinois River has stalled south of Chicago, and has remained stable for a decade. Studies have suggested that pollutants in the Chicago Area Waterway System (CAWS) may be contributing to the lack of upstream movement, but this hypothesis has not been tested. This study used a laboratory setting to quantify the role of pollutants in deterring upstream movement of silver carp within the CAWS. For this, water was collected from the CAWS near the upstream edge of the distribution and transported to a fish culture facility. Silver carp and one native species were exposed to CAWS water, and activity, behavior, avoidance and metabolic rates were quantified. Results showed that silver carp experience an elevated metabolic cost in CAWS water, along with reductions in swimming behavior. Together, results suggest a role for components of CAWS water at deterring range expansion.

Exposure of females to stressful conditions during pregnancy or oogenesis has a profound effect on the phenotype of their offspring. For example, offspring behavioural phenotype may show altered patterns in terms of the consistency of behavioural patterns and their average level of performance. Maternal stress can also affect the development of the stress axis in offspring leading to alterations in their physiological stress response. However, the majority of evidence comes from studies utilising acute stressors or exogenous glucocorticoids, and little is known about the effect of chronic maternal stress, particularly in the context of stress lasting throughout entire reproductive lifespan. To bridge this knowledge gap, we exposed female sticklebacks to stressful and unpredictable environmental conditions throughout the breeding season. We quantified the activity, sheltering and anxiety-like behaviour of offspring from three successive clutches of these females, and calculated Intra-class Correlation Coefficients for these behaviours in siblings and half-siblings. We also exposed offspring to an acute stressor and measured their peak cortisol levels. An unpredictable maternal environment had no modifying effect on inter-clutch acute stress responsivity, but resulted in diversification of offspring behaviour, indicated by an increased between-individual variability within families. This may represent a bet-hedging strategy, whereby females produce offspring differing in behavioural phenotype, to increase the chance that some of these offspring will be better at coping with the anticipated conditions.

Titanium dioxide is a type of nanoparticle that is composed of one titanium atom and two oxygen atoms. One of its physicochemical activities is photolysis, which produces different reactive oxygen species (ROS). Atya lanipes shrimp affect detrital processing and illustrate the potential importance of diversity and nutrient availability to the rest of the food web. It is essential in removing sediments, which have an important role in preventing eutrophication. This study aimed to determine the toxic effect of changes in behavior and levels of oxidative stress due to exposure to titanium dioxide nanoparticles in Atya lanipes and to determine the effective concentration (EC50) for behavioral variables. The concentrations of TiO2 NPs tested were 0.0, 0.50, 1.0, 2.0, and 3.0 mg/L with the positive controls given 100 µg/L of titanium and 3.0 mg/L of TiO2 NPs ± 100 µg/L of titanium. After 24 h of exposure, significant hypoactivity was documented. The EC50 was determined to be a concentration of 0.14 mg/L. After the exposure to 10 mg/L of TiO2 NPs, oxidative stress in gastrointestinal and nervous tissues was documented. The toxic effects of this emerging aquatic pollutant in acute exposure conditions were characterized by sublethal effects such as behavior changes and oxidative stress.

Insulin-like growth factor-1 (Igf1) regulates skeletal muscle growth in fishes by increasing protein synthesis and promoting muscle hypertrophy. When fish experience periods of insufficient food intake, they undergo slower muscle growth or even muscle wasting, and those changes emerge in part from nutritional modulation of Igf1 signaling. Here, we examined how food deprivation (fasting) affects Igf1 regulation of liver and skeletal muscle gene expression in gopher rockfish (Sebastes carnatus), a nearshore rockfish of importance for commercial and recreational fisheries in the northeastern Pacific Ocean, to understand how food limitation impacts Igf regulation of muscle growth pathways. Rockfish were either fed or fasted for 14 d, after which a subset of fish from each group was treated with recombinant Igf1 from sea bream (Sparus aurata). Fish that were fasted lost body mass and had lower body condition, reduced hepatosomatic index, and lower plasma Igf1 concentrations, as well as a decreased abundance of igf1 gene transcripts in the liver, increased hepatic mRNAs for Igf binding proteins igfbp1a, igfbp1b, and igfbp3a, and decreased mRNA abundances for igfbp2b and a putative Igf acid labile subunit (igfals) gene. In skeletal muscle, fasted fish showed a reduced abundance of intramuscular igf1 mRNAs but elevated gene transcripts encoding Igf1 receptors A (igf1ra) and B (igf1rb), which also showed downregulation by Igf1. Fasting increased skeletal muscle mRNAs for myogenin and myostatin1, as well as ubiquitin ligase F-box only protein 32 (fbxo32) and muscle RING-finger protein-1 (murf1) genes involved in muscle atrophy, while concurrently downregulating mRNAs for myoblast determination protein 2 (myod2), myostatin2, and myogenic factors 5 (myf5) and 6 (myf6 encoding Mrf4). Treatment with Igf1 downregulated muscle myostatin1 and fbxo32 under both feeding conditions, but showed feeding-dependent effects on murf1, myf5, and myf6/Mrf4 gene expression indicating that Igf1 effects on muscle growth and atrophy pathways is contingent on recent food consumption experience.

Fish adjust rates of somatic growth in the face of changing food consumption. As in other vertebrates, growth in fish is regulated by the growth hormone (Gh)/insulin-like growth factor-1 (Igf1) endocrine axis, and changes in food intake impact growth via alterations to Gh/Igf1 signaling. Understanding the time course by which the Gh/Igf1 axis responds to food consumption is crucial to predict how rapidly changes in food abundance might lead to altered growth dynamics. Here, we looked at the response times of plasma Igf1 and liver Igf1 signaling-associated gene expression to refeeding after food deprivation in juvenile gopher rockfish (Sebastes carnatus), one of several species of northern Pacific Ocean Sebastes rockfishes targeted by fisheries or utilized for aquaculture. Gopher rockfish were fasted for 30 d, after which a subset was fed to satiation for 2 h, while other rockfish continued to be fasted. Refed fish exhibited higher hepatosomatic index (HSI) values and increased Igf1 after food consumption. Gene transcripts for Gh receptor 1 (ghr1), but not ghr2, increased in the liver 2-4 d after eating. Transcripts encoding igf1also increased in the liver of refed fish by 4 d after feeding, only to return to levels similar as continually fasted rockfish by 9 d after feeding. Liver mRNA abundances for Igf binding protein (Igfbp) genes igfbp1a, igfbp1b, and igfbp3a declined within 2 d of feeding. These findings provide evidence that circulating Igf1 in rockfish reflects a fish's feeding experience within the previous few days, and suggest that feeding-induced increases in Igf1 are being mediated in part by altered liver sensitivity to Gh due to upregulated Gh receptor 1 expression.

Food-deprivation state (fed, fasted, starved) affected rock crabs physiological and biochemical responses to hypoxia in Cancer irroratus. Fasted and starved crabs were better adapted to deal with hypoxia than fed animals; however, avoidance behavior is usually considered as the first defense to environmental stressors for decapod crustaceans. We examined the effects of food deprivation on the crab’s behavior to hypoxia using the Loligo® shuttle box system, an automated system with a pair of connected water chambers with regulated flow and oxygen level. Crabs (starved, fasted and fed) that were offered a choice of two different oxygen saturations did not appear to actively avoid the hypoxia regimes tested (50% and 20% oxygen saturation). We used novel algorithms to analyze the data and found that crabs altered rheotaxis (movement towards or away from a current of water) and corresponding moving speed as a function of oxygen saturation. The food-deprivation state did influence thigmotaxis (contact with walls/objects when exploring an open space): starved crabs became bolder and more likely to explore open areas of the apparatus. Technological advancements such as the fully automated shuttle box have improved our ability to collect and analyze behavioral data; however, our study also highlighted some of the potential problems of relying solely on such apparatus to study the behavior of benthic crustaceans.

Laboratory‐based studies examining fish physiological and behavioural responses to temperature can provide important insight into species‐specific habitat preferences and utilisation, and are especially useful in examining vulnerable life stages that are difficult to study in the wild. This study couples shuttle box behavioural experiments with respirometry trials to determine the temperature preferences and metabolic thermal sensitivity of juvenile California horn shark ( Heterodontus francisci ) and leopard shark ( Triakis semifasciata ). As juveniles, these two species often occupy similar estuarine habitats but display contrasting behaviours and activity levels – H. francisci are relatively sedentary, whereas T. semifasciata are more active and mobile. This study shows that juvenile H. francisci and T. semifasciata have comparable thermal preferences and occupy similar temperature ranges, but H. francisci metabolism is more sensitive to acute changes in temperature as expressed through a higher Q 10 ( H. francisci = 2.58; T. semifasciata = 1.97; temperature range: 12–24°C). Underlying chronic temperature acclimation to both warm (21°C) and cool (15°C) representative seasonal temperatures did not appear to significantly affect these parameters. These results are discussed in the context of field studies examining known distributions, habitat and movement patterns of H. francisci and T. semifasciata to better understand the role of temperature in species‐specific behaviour. Juvenile H. francisci likely target thermally stable environments, such as estuaries that are close to their preferred temperature, whereas juvenile T. semifasciata metabolism and behaviour appear less dependent on temperature.

The amphipod Gmelinoides fasciatus has invaded and established in numerous large lakes in Eurasia and, in the process, has displaced the native amphipod, Gammarus lacustris. The mechanism behind its invasion success is unclear and remains an important topic for invasion ecology. Three laboratory experiments were conducted to determine if superior predator avoidance and different types of bottom substrate could be important factors contributing to the invasion success of G. fasciatus. Our results indicate that, on gravel and sand substrates, G. fasciatus exhibited superior digging behaviour to avoid predation by fishes (perch and common roach), contrary to its native counterpart. In addition, G. fasciatus exhibited a more substantial reduction in activity than G. lacustris when in the presence of predatory fish kairomones. However, the presence of kairomones had little effect on digging behaviour. G. fasciatus consistently demonstrated superior predator avoidance abilities over G. lacustris, suggesting that this mechanism might play an important role in the invasion success of G. fasciatus.

Given the threat of climate change to biodiversity, a growing number of studies are investigating the potential for organisms to adapt to rising temperatures. Earlier work has predicted that physiological adaptation to climate change will be accompanied by a shift in temperature preferences, but empirical evidence for this is lacking. Here, we test whether exposure to different thermal environments has led to changes in preferred temperatures in the wild. Our study takes advantage of a “natural experiment” in Iceland, where freshwater populations of threespine sticklebacks ( Gasterosteus aculeatus ) are found in waters warmed by geothermal activity year‐round (warm habitats), adjacent to populations in ambient‐temperature lakes (cold habitats). We used a shuttle‐box approach to measure temperature preferences of wild‐caught sticklebacks from three warm–cold population pairs. Our prediction was that fish from warm habitats would prefer higher water temperatures than those from cold habitats. We found no support for this, as fish from both warm and cold habitats had an average preferred temperature of 13°C. Thus, our results challenge the assumption that there will be a shift in ectotherm temperature preferences in response to climate change. In addition, since warm‐habitat fish can persist at relatively high temperatures despite a lower‐temperature preference, we suggest that preferred temperature alone may be a poor indicator of a population's adaptive potential to a novel thermal environment.

Pseudunio auricularius (Spengler, 1793) is one of the most threatened unionid species worldwide. Translocation is considered one of the ultimate actions that can save this species from extinction in the Iberian Peninsula. Since 2013, massive mortalities have been recorded in the Canal Imperial de Aragón (CIA), an anthropogenic habitat where the highest density of P. auricularius had been recorded in Spain. An adequacy habitat index was calculated assigning scores to different environmental variables to select the most suitable river stretches receiving the translocated specimens. A total of 638 specimens have been translocated: 291 in 2017, 291 in 2018, and 56 in 2019. The first-year survival in the group of individuals translocated in 2017 was 41.6%. The next year, 95% of these specimens were found alive, suggesting a successful initial establishment. Specimens translocated in 2018 and 2019 showed a survival of c. 69% and 49%, respectively. In contrast, the control group left in CIA in 2017 showed a much lower survival rate of 19.7% after one year, which remained equally low during the next two years. Currently, the conditions in the Ebro River seem to allow a higher survival rate for P. auricularius than those in the CIA; nevertheless, future monitoring should confirm their long-term success.

Within Salmonidae, spawning and rearing in brackish water is rare; however, brackish-water resident lake trout (Salvelinus namaycush) have recently been documented in the Arctic. Additionally, early rearing in brackish-water environments increased the fish’s ability to ionoregulate in elevated salinities. Here, we examined the impact of a freshwater (FWR, 0 ppt) or brackish-water (BWR, 5 ppt) rearing environment on salinity preference in lake trout using a dynamic choice experiment. We observed significant differences in salinity preference between our treatments suggesting the importance of early environment in shaping salinity preference. Contrary to our predictions, FWR lake trout selected higher salinity (17.3 ppt) compared to BWR fish (4.8 ppt). Four of the seven FWR fish had preferred salinities near 30 ppt, which is considered physiologically challenging and potentially lethal for lake trout based on direct transfer experiments. Thus, heightened FWR salinity preference might not be a true preference but rather due to a reduced ability to sense differences in salinity, and a result of chance as mean preferred salinity was near half that of the upper and lower thresholds, and variance was larger. Selection of lower salinity by BWR fish suggests that the ability to sense and select different salinities is present in lake trout as a species and appears to be linked to difference in early rearing at elevated salinities.

Temperature and salinity often define the distributions of aquatic organisms. This is at least partially true for Delta Smelt, an imperiled species endemic to the upper San Francisco Estuary. While much is known about the tolerances and distribution of Delta Smelt in relation to these parameters, little is known regarding the temperature and salinity preferences of the species. Therefore, the temperature and salinity preferences of sub-adult Delta Smelt were investigated across a wide range of thermal (8–28 °C) and salinity (0–23 ppt) conditions. Replicates of ten fish were allowed to swim between two circular chambers with different temperature or salinity, and the distribution of fish between the chambers was recorded. We found that Delta Smelt showed no temperature preference below 15 °C, a modest aversion to the warmer tank from 15 to 28 °C, and a strong aversion to the warmer tank with elevated mortality at temperatures above 28 °C. Delta Smelt also preferred lower salinities, and this preference became more pronounced as salinity increased toward 23 ppt. These results indicate that Delta Smelt can tolerate high temperatures and salinities for a short time, and that their preferences for lower temperature and salinity strengthens as these variables increase.

The role of the blood–brain barrier ATP-binding cassette protein transporter P-glycoprotein (P-gp) in protecting zebrafish (Danio rerio) from the central nervous system neurotoxicant ivermectin (IVM, 22,23-dihydroavermectin B1a + 22,23-dihydroavermectin B1b) was examined in the absence and presence of the competitive inhibitor cyclosporin A (CsA). Zebrafish injected intraperitoneally with 1, 2, 5, or 10 µmol/kg IVM exhibited mortality 30 min following administration at the highest dose. At sublethal doses > 1 µmol/kg, IVM altered the swimming performance, exploratory behaviour, motor coordination, escape response and olfactory response in exposed fish. When fish were exposed to IVM in the presence of CsA, alterations in swimming and behaviours increased significantly and at the highest IVM/CsA ratio resulted in a complete lack of exploratory and olfactory behaviours. In separate experiments, fish were either fed or fasted, and the effects of IVM and CsA administration were examined. The effects of IVM administration and the exacerbated effects seen with CsA co-administration were not affected by fasting. This study provides evidence that P-gp provides a protective role in the BBB of fish against environmental neurotoxicants. The results also show that P-gp activity is maintained even under conditions of food deprivation, suggesting that this chemical defence system is prioritized over other energy expenditures during diet limitation.

Wild animals have parasites that can compromise their physiological and/or behavioural performance. Yet, the extent to which parasite load is related to intraspecific variation in performance traits within wild populations remains relatively unexplored. We used pumpkinseed sunfish (Lepomis gibbosus) and their endoparasites as a model system to explore the effects of infection load on host aerobic metabolism and escape performance. Metabolic traits (standard and maximum metabolic rates, aerobic scope) and fast-start escape responses following a simulated aerial attack by a predator (responsiveness, response latency and escape distance) were measured in fish from across a gradient of visible (i.e. trematodes causing black spot disease counted on fish surfaces) and non-visible (i.e. cestodes in fish abdominal cavity counted post-mortem) endoparasite infection. We found that a higher infection load of non-visible endoparasites was related to lower standard and maximum metabolic rates, but not aerobic scope in fish. Non-visible endoparasite infection load was also related to decreased responsiveness of the host to a simulated aerial attack. Visible endoparasites were not related to changes in metabolic traits or fast-start escape responses. Our results suggest that infection with parasites that are inconspicuous to researchers can result in intraspecific variation in physiological and behavioural performance in wild populations, highlighting the need to more explicitly acknowledge and account for the role played by natural infections in studies of wild animal performance.

Nanoparticles are man-made materials defined as materials smaller than 100 nm in at least one dimension. Titanium oxide nanoparticles are of great interest because of their extensive use in self-care products. There is a lack of nanotoxicological studies of TiO2 NPs in benthic organisms to have evidence about the effects of these pollutants in freshwater ecosystems. Atya lanipes is a scraper/filter that can provide a good nanotoxicological model. This study aims to determine how the TiO2 NPs can develop a toxic effect in the larvae of the Atya lanipes shrimp and to document lethal and sublethal effects after acute exposures to TiO2 NP suspensions of: 0.0, 1.0, 10.0, 50.0, 100.0, and 150.0 mg/L. The results show that early exposure to TiO2 NPs in Atya lanipes creates an increase in mortality at 48 and 72 h exposures, hypoactivity in movements, and morphological changes, such as less pigmentation and the presence of edema in exposed larvae. In conclusion, TiO2 NPs are toxic contaminants in the larval stage of the Atya lanipes. It is necessary to regulate these nanoparticles for purposes of the conservation of aquatic biodiversity, especially for freshwater shrimp larvae and likely many other larvae of filter-feeding species.

The cardiovascular performance of salmonids in aquaculture can be impaired by acute climate warming, posing risks for fish survival. Exercise training and functional feeds have been shown to be cardioprotective in mammals but their action on the fish heart and its upper thermal performance has not been studied. To investigate this, rainbow trout were trained at a moderate water velocity of 1 body length per second (bl s -1 ) for 6 h per day, either alone or in combination with one of two functional feed-supplements, allicin and fucoidan. After 6 weeks of exercise training and feeding, maximum heart rate and the temperature coefficient of heart rate were significantly higher in the trained fish as compared to untrained ones. There was a slight increase in hematocrit in trained control fish reared on a normal diet (TC group) compared to untrained fish fed with the same diet (CC). This implies that exercise training enhanced oxygen delivery to trout tissues via an increase of cardiac blood flow in warm water. However, cardiac thermal tolerance was not affected by exercise training or feeding, except from the temperature of peak heart rate which was higher in the trained group fed with fucoidan supplement (TF) as compared to the untrained group fed with same diet (CF). Allicin supplement caused a significant reduction in the maximum heart rate and the temperature coefficient of heart rate, especially in trained fish, while fucoidan supplement did not cause any effect on heart rate. No differences were observed in growth performance among groups. However, fish fed with fucoidan-supplemented diet had a slight reduction in feed conversion efficiency. We suggest further investigations to understand the antagonistic effect of allicin supplemental feeding and exercise training on cardiovascular performance. More studies are also required to investigate if other exercise training intensities could increase cardiac thermal tolerance.

The use of insect meal in aquafeed formulations has recently gained attention. Detailed knowledge about the inclusion levels for pikeperch ( Sander lucioperca ), a promising candidate for intensive aquaculture in Europe remains, however, fragmented. In the present study, 4 isoproteic (45% dry matter) and isoenergetic (21 MJ/kg) diets were formulated, including a control diet (H0) containing 30% fishmeal (FM) on an as-fed basis and the other 3 diets in which FM protein was replaced by defatted black soldier fly ( Hemetia illucens ) meal (HIM) at 25%, 50%, and 100% (diet abbreviation H9, H18 and H36, corresponding to an inclusion level of 9%, 18% and 36%, respectively). The feeding trial was performed in triplicate groups of 50 juvenile pikeperch (mean weight, 68.7 g) fed with experimental diets for 84 d during which the growth performance, nutrient digestibility, fillet quality and economic and environmental sustainability of rearing pikeperch were evaluated. Our findings indicated that pikeperch in H0, H9, and H18 groups displayed better results regarding growth performance indices, except for survival rate where no significant difference among groups was recorded ( P = 0.642). A significantly lower organ-somatic index, including hepatosomatic, viscerosomatic and perivisceral fat index, was found in fish in H18 groups than other groups ( P < 0.05). Inclusion of HIM affected the digestibility of the nutrients and resulted in an almost linear reduction in the apparent digestibility coefficient of dry matter and protein. Concerning the fillet quality, dietary HIM negatively affected the protein and ash contents of the fish fillets, while the crude fat remained unchanged. Dietary HIM did not significantly modify total saturated, monounsaturated and polyunsaturated fatty acids in the fillets of fed pikeperch ( P > 0.05) but did reduce total n-3 fatty acids ( P = 0.001) and increased total n-6 ( P < 0.001). Increasing inclusion levels of HIM reduced the environmental impacts associated with fish in-to-fish out ratio but entailed heavy burdens on energy use and eutrophication. Low and moderate inclusion levels of HIM did not negatively affect land use and water use compared to an HIM-free diet ( P > 0.05). The addition of HIM at a level as low as 9% elicited a similar carbon footprint to that of the control diet. The economic conversion ratio and economic profit index were negatively affected at increased insect meal inclusion levels. This study has shown that the incorporation of HIM in feed formulations for pikeperch is feasible at inclusion levels of 18% without adverse effects on growth performance parameters. The feasibility also highlighted the environmental benefits associated with land use and marine resources required to produce farmed fish.

Oculocutaneous albinism is the result of a combination of homozygous recessive mutations that block the synthesis of the tyrosine and melatonin hormones. This disability is associated with physiological limitations, e.g., visual impairment expressed by lower visual acuity and movement perception, and eventually leads to acrophobia and/or photophobia, suggesting a potentially higher stress level associated with the behavioral responses of individuals with albinism to external stimuli compared to their pigmented conspecifics. However, in fish, differences in behavioral and/or physiological responses and stress levels between these phenotypes have been poorly documented. While acoustic perception of albino individuals is well known, the use of olfactory sensors for social communication, e.g., for the preference for familiar conspecifics, remains poorly understood. We performed two laboratory experiments with albino and pigmented European catfish Silurus glanis to observe: i) their behavioral and physiological responses to short-term stress induced by a combination of air exposure and novel environmental stressors and ii) their ability to use odor keys to recognize of familiar conspecifics and the influence of lateralization on this preference. In response to stress stimuli, albino fish showed higher movement activities and ventilatory frequencies and more often changed their swimming directions compared to their pigmented conspecifics. Blood plasma analysis showed significantly higher values of stress-, deprivation-, and emotional arousal-associated substances, e.g., glucose and lactate, as well as of substances released during intensive muscle activity of hyperventilation and tissue hypoxia, e.g., hemoglobin, mean corpuscular hemoglobin, erythrocytes, and neutrophil granulocytes. A preference test between environments with and without scented water showed the preference by both albino and pigmented catfish for environments with scent of familiar conspecifics, and both groups of fish displayed left-side lateralization associated with the observation of conspecifics and group coordination. The results tended to show higher physiological and behavioral responses of albinos to stress stimuli compared to the responses of their pigmented conspecifics, but the uses of olfactory sensors and lateralization were not differentiated between the two groups.

The photophobia of Conopomorpha sinensis was explored using eight kinds of LED lights, including those with single wavelengths as follows: 400, 460, 520, 600, 660 and 740 nm, and mixed wavelengths: MixW and MixY. The results showed that 460 nm (blue light), 520 nm (green light) and MixW (white light) were the most effective wavelengths for controlling C. sinensis, effectively reducing pest activity by 98.2%, 99.2% and 99.3%, respectively. Egg production decreased by 99.3%, 93.0% and 98.6% under these treatments with a light intensity of 0.5 μmol m−2 s−1, respectively. The C. sinensis damage rate was 72% in the dark control group and 10%, 6%, and 16% under continuous light at night with 460, 520 nm and MixW light, respectively. To further investigate the effect of different wavelengths of light applied to the whole plant at night on Yu-Her-Pao litchi fruit quality, the fruits under 460, 520 nm and MixW light at night were 23.4, 29.8 and 26.6 g in weight and 17.5, 19.3 and 17.8°Brix in total soluble solids, respectively. There was no significant difference in fruit weight or total soluble solids under 520 nm or bagging with no light at night. Thus, 520 nm green light at night had the least effect on fruit quality. HPLC results showed that the sucrose, glucose, fructose, citric acid, L-malic acid, and shikimic acid contents in fruits under 520 nm green light illumination at night were significantly higher than those under other light treatments. Only the fumaric acid content was significantly less than that under the other light treatments. There is no known previous research on the effects of continuous application of light to whole fruit trees and different light wavelengths on fruit quality. The results of this study showed that 520 nm green light at night could prevent and control C. sinensis and maintain fruit quality. These experimental results represent important progress in reducing the use of pesticides in the litchi industry.

Nanoplastics are being detected with increasing frequency in aquatic environments. Although evidence suggests that nanoplastics can cause overt toxicity to biota across different trophic levels, but there is little understanding of how materials such as differently charged polystyrene nanoplastics (PS-NP) impact fish development and behavior. Following exposure to amino-modified (positive charge) PS-NP, fluorescence accumulation was observed in the zebrafish brain and gastrointestinal tract. Positively charged PS-NP induced stronger developmental toxicity (decreased spontaneous movement, heartbeat, hatching rate, and length) and cell apoptosis in the brain and induced greater neurobehavioral impairment as compared to carboxyl-modified (negative charge) PS-NP. These findings correlated well with fluorescence differences indicating PS-NP presence. Targeted neuro-metabolite analysis by UHPLC-MS/MS reveals that positively charged PS-NP decreased levels of glycine, cysteine, glutathione, and glutamic acid, while the increased levels of spermine, spermidine, and tyramine were induced by negatively charged PS-NP. Positively charged PS-NP interacted with the neurotransmitter receptor N-methyl-D-aspartate receptor 2B (NMDA2B), whereas negatively charged PS-NP impacted the G-protein-coupled receptor 1 (GPR1), each with different binding energies that led to behavioral differences. These findings reveal the charge-specific toxicity of nanoplastics to fish and provide new perspective for understanding PS-NP neurotoxicity that is needed to accurately assess potential environmental and health risks of these emerging contaminants.

Ectotherms can respond to climate change via evolutionary adaptation, usually resulting in an increase of their upper thermal tolerance. But whether such adaptation influences the phenotypic plasticity of thermal tolerance when encountering further environmental stressors is not clear yet. This is crucial to understand because organisms experience multiple stressors, besides warming climate, in their natural environment and pollution is one of those. Here, we studied the phenotypic plasticity of thermal tolerance in three-spined stickleback populations inhabiting spatially replicated thermally polluted and pristine areas before and after exposing them to a sublethal concentration of copper for one week. We found that the upper thermal tolerance and its phenotypic plasticity after copper exposure did not depend on the thermal history of fish, suggesting that five decades of thermal pollution did not result in evolutionary adaptation to thermal tolerance. The upper thermal tolerance of fish was, on the other hand, increased by ∼ 1.5 °C after 1-week copper exposure in a sex-specific manner, with males having higher plasticity. To our knowledge this is the first study that shows an improvement of the upper thermal tolerance as a result of metal exposure. The results suggest that three-spined sticklebacks are having high plasticity and they are capable of surviving in a multiple-stressor scenario in the wild and that male sticklebacks seem more resilient to fluctuating environmental conditions than female.

Despite well-known systemic immune reactions in peripheral trauma, little is known about their roles in posttraumatic neurological disorders, such as anxiety, sickness, and cognitive impairment. Leukocyte invasion of the brain, a common denominator of systemic inflammation, is involved in neurological disorders that occur in peripheral inflammatory diseases, whereas the influences of peripheral leukocytes on the brain after peripheral trauma remain largely unclear. In this study, we found that leukocytes, largely macrophages, transiently invaded the brain of zebrafish larvae after peripheral trauma through vasculature-independent migration, which was a part of the systemic inflammation and was mediated by interleukin-1b (il1b). Notably, myeloid cells in the brain that consist of microglia and invading macrophages were implicated in posttraumatic anxiety-like behaviors, such as hyperactivity (restlessness) and thigmotaxis (avoidance), while a reduction in systemic inflammation or myeloid cells can rescue these behaviors. In addition, invading leukocytes together with microglia were found to be responsible for the clearance of apoptotic cells in the brain; however, they also removed the nonapoptotic cells, which suggested that phagocytes have dual roles in the brain after peripheral trauma. More importantly, a category of conserved proteins between zebrafish and humans or rodents that has been featured in systemic inflammation and neurological disorders was determined in the zebrafish brain after peripheral trauma, which supported that zebrafish is a translational model of posttraumatic neurological disorders. These findings depicted leukocyte invasion of the brain during systemic inflammation after peripheral trauma and its influences on the brain through il1b-dependent mechanisms. Invasion of the brain by white blood cells followed tail amputation in zebrafish, the resulting systemic inflammation producing anxiety-like behaviors. Scientists have long recognised an association between systemic inflammation following peripheral traumatic injury such as limb loss and post-traumatic neurological disorders such as anxiety and depression. Raymond Chuen-Chung Chang at the University of Hong Kong, Alvin Chun-Hang Ma at Hong Kong Polytechnic University, China, and co-workers found that following trauma, white cells, mainly macrophages, flowed from neighboring tissues into the hindbrain, before spreading throughout the brain. This influx of white cells, mediated by the small signaling protein interleukin-1b, triggered anxiety-like behaviors such as hyperactivity and avoidance in the zebrafish. The researchers emphasize that the links between systemic inflammation following peripheral trauma and neurological responses require extensive further research.

At early life stages invasive fishes may have no innate or learned behavioral responses to native predators. However, social cues expressed by shoal mates is one strategy species use to assess risk. By shoaling and using social cues, fishes may identify and mimic others with anti-predator behaviors to increase their own survival. Ability of non-native fishes, such as bighead carp (Hypophthalmichthys nobilis), to mimic native species that have experienced predatory threats is not known. In this experimental study, we varied the number of experienced individuals and the species composition to contrast the responses of naïve juvenile bighead carp exposed to predatory kairomones when grouped with differing numbers of either experienced conspecific or experienced heterospecific (golden shiner, Notemigonus crysoleucas) shoal mates. We found fully naïve groups of bighead carp did not respond to largemouth bass kairomones, but that naïve individuals could mimic anti-predatory behaviors of experienced individuals, even when those experienced individuals were heterospecifics. Diverse alarm responses of bighead carp to composition and experience suggest that responses of this species are plastic. Through changing responses based on shoal experience level and composition, plastic social learning highlights how naïve individuals may adapt to novel predator threats, which could inform predictions of non-native persistence in novel waterways.

Interference competition over food and territory can shape population structure and habitat use within and between species. The introduction of invasive species often leads to novel competitive interactions over shared resources and invaders can eventually exclude the native species from preferred habitats. Invasive brook trout ( Salvelinus fontinalis ) introduced to northern Europe have excluded native brown trout ( Salmo trutta ) from numerous headwater streams. The fact that invasive brook trout can displace the more aggressive brown trout is puzzling. However, the earlier spawning and hatching of brook trout, compared to brown trout, may lead to unequal competition due to size advantage and prior resident status of brook trout at the fry stage. In this study, we examine the effect of competition between brown trout and brook trout using the natural size distribution of the two species. In two consecutive experiments, we first measured space use and feeding of a fry (age 0+) in the presence of a juvenile (age 1+). In experiment 2, we assessed territorial interactions between the species at the fry stage (age 0+) and if smaller brown trout could compensate the disadvantage by manipulating residence duration. Fry of brook trout feed sooner and spend more time close to the larger individual than brown trout fry. We also found that brook trout fry won most territorial contests against brown trout, and that increased residence duration led to longer and more aggressive interactions. The results suggest that smaller brown trout are displaced to suboptimal habitats in the presence of a larger brook trout. Therefore, the later emergence from gravel beds resulting in the naturally occurring size disadvantage of brown trout at the fry stage may lead to unequal territorial interactions that could explain why brown trout are displaced from preferred habitats in sympatry with brook trout.

Heat waves constitute a challenge for aquatic ectotherms. However, the thermal tolerance of animals and their individual phenotypic plasticity to respond to heat waves may be influenced by thermal history. We tested these hypotheses by comparing the upper thermal tolerance and the individual capacities of three‐spined sticklebacks from populations with different thermal histories to respond to heat waves. Two populations originated from thermally polluted nuclear power plant (NPP) habitats, while four locations represented geographically adjacent control areas. To disentangle the genetic adaptation from the phenotypic plastic response, we measured the individual upper thermal tolerance and the responses at molecular level in common‐garden conditions before and after a laboratory‐mimicked heat wave. We found that the sticklebacks exhibit considerable phenotypic plasticity in thermal tolerance since the heat wave increased fish upper thermal tolerance significantly. The individual plasticity to respond to the heat wave was also negatively correlated with initial thermal tolerance. On the other hand, neither the thermal tolerance nor the plastic responses differed between NPP and control sites despite detection of significant but low genome‐wide divergence in 10 out of 15 pairwise comparisons. Our results suggest that five decades of NPP activity with warmer water have not resulted in a detectable evolutionary change in either the upper thermal tolerance or its plasticity in three‐spined sticklebacks potentially rendering them sensitive to frequent heat waves.

Flows in river habitats are characterized by unsteady turbulence due to the existence of woody debris, boulders, and vegetation. As a representative aquatic species, fish is important for the riverine ecosystems with its complex behavioral responses to turbulent flows. Previous studies investigated the fish-vortices interaction with vortex streets by placing objects with simplified geometries centred at the flow. However, complex river morphology in natural rivers reuslts in much more spatially heterogeneous flows due to randomly distributed obstructions. Thus, a semi-circular cylinder patch located on one side of flume is used to mimic a vegetation patch at the riverbank. The patch varies in diameter (D0 = 16, 20, 24 cm) and density (φ = 0.04, 0.1), while the flow velocity is fixed at 25 cm/s. Fish are observed to swim in three typical patterns, which are "swim around (Pattern 1)", "spill (Pattern 2)", and "swim through (Pattern 3)". For flow with a dense patch, all three patterns are recorded, but only patterns 1 and 2 are seen in sparse patches. We notice that in patterns 1 and 2, fish prefer to hold place in zones of low velocity and low turbulence. Moreover, variations in patch diameter have little influence on pattern selection. Results showed that tail beat amplitude (TBA*) in each zone displayed more variations compared with tail beat frequency (TBF). In addition, spearman rank tests revealed that TBA* is affected by none of the four hydrodynamic variables ( U, u std, τ xy, Ω z ) whereas flow velocity imposes the most influence on TBF. Both diameter and density of the patch displayed no significant influence on TBA* and TBF. This article is protected by copyright. All rights reserved.

Predator-prey interactions exert significant influence over the survival of juvenile fish cohorts. Therefore, susceptibility of a habitat to invasion is influenced by the capacity of native predators to regulate invasive species through consumption. Closely related predators often share similar characteristics (e.g., odors or body morphometry), and prey species capable of expressing generalized behavioral responses to predators with similar characteristics may increase their chances for survival. Here, we examined how naïve juvenile silver carp (Hypophthalmichthys molitrix), an invasive Asian carp, respond to three predator odors from predators commonly found in Midwestern lakes and rivers of the USA. We tested two congeneric species of bass (largemouth Micropterus salmoides and smallmouth M. dolomieu bass) and one outgroup, longnose gar (Lepisosteus osseus). Additionally, we tested how silver carp conditioned to recognize the odor of one group, largemouth bass, responded to the predator odors of the congeneric species and the outgroup. We found that juvenile silver carp showed no innate response to any of the three predator odors. Additionally, although they could be conditioned to recognize predator odors from largemouth bass, they were unable to generalize predator odors to smallmouth bass or longnose gar odor. These results suggest that invasive species could be less likely to persist in environments with diverse predator communities than environments of equal densities with uniform predator communities and that future studies should continue to explore this area as well as focus on understanding dynamics in predator-prey interactions of invasive species.

Increasing ocean temperatures and the resulting poleward range shifts of species has highlighted the importance of a species preferred temperature and thermal range in shaping ecological communities. Understanding the temperatures preferred and avoided by individual species, and how these are influenced by species interactions is critical in predicting the future trajectories of populations, assemblages, and ecosystems. Using an automated shuttlebox system, we established the preferred temperature and upper and lower threshold temperatures (i.e., avoided temperatures) of a common coral reef fish, the black-axil chromis, Chromis atripectoralis. We then investigated how the presence of conspecifics, heterospecifics (Neopomacentrus bankieri), or a predator (Cephalopholis spiloparaea) influenced the selection of these temperatures. Control C. atripectoralis preferred 27.5 ± 1.0 °C, with individuals avoiding temperatures below 23.5 ± 0.9 °C and above 29.7 ± 0.7 °C. When associating with either conspecifics or heterospecifics, C. atripectoralis selected significantly lower temperatures (conspecifics: preferred = 21.2 ± 1.4 °C, lower threshold = 18.1 ± 0.8 °C; heterospecifics: preferred = 21.1 ± 1.1 °C, lower threshold = 19.2 ± 0.9 °C), but not higher temperatures (conspecifics: preferred = 28.9 ± 1.2 °C, upper threshold = 30.8 ± 0.9 °C; heterospecifics: preferred = 29.7 ± 1.1 °C, upper threshold = 31.4 ± 0.8 °C). The presence of the predator, however, had a significant effect on both lower and upper thresholds. Individual C. atripectoralis exposed themselves to temperatures ~ 5.5 °C cooler or warmer (lower threshold: 18.6 ± 0.5 °C, upper threshold: 35.2 ± 0.5 °C) than control fish before moving into the chamber containing the predator. These findings demonstrate how behavioural responses due to species interactions influence the thermal ecology of a tropical reef fish; however, there appears to be limited scope for individuals to tolerate higher temperatures unless faced with the risk of predation.

Fishes exposed to crude oil have shown reduced sociability and poor habitat selection, which corresponded with increased predation risk. However, the contribution of oil-induced cardiorespiratory impairments to these findings is uncertain. This study explores the effect of oil exposure on predation risk in a model fish species, Sciaenops ocellatus, across a suite of physiological and behavioral end points to elucidate the mechanisms through which any observed effects are manifested. Using mesocosms to assess group predator avoidance, oil exposure to 36.3 μg l -1 ΣPAH reduced the time to 50% mortality from a mean time of 80.0 (74.1-86.0 95% confidence interval [CI]) min to 39.2 (35.6-42.8 95% CI) min. The influence of oil impaired cardiorespiratory and behavioral pathways on predation risk was assessed based on respiratory performance, swim performance, sociability, and routine activity. Swim trials demonstrated that cardiorespiratory and swim performance were unaffected by exposures to 26.6 or 100.8 μg l -1 ΣPAH. Interestingly, behavioral tests revealed that exposure to 26.6 μg l -1 ΣPAH increased distance moved, speed, acceleration, and burst activity. These data indicate that behavioral impairment is more sensitive than cardiorespiratory injury and may be a more important driver of downstream ecological risk following oil exposure in marine species.

Examination of the trade-offs between total assay length and variation in temperature preference in thermal preference experimental design reveals no significant effect of length of acclimation time and trial length on thermal preference of juvenile lake whitefish.

Loss of myofibers during muscle atrophy affects functional capacity and quality of life. Dexamethasone, an inducer of rapid atrophy of skeletal myofibers, has been studied as a glucocorticoid receptor in muscle atrophy or motor neurodegeneration. In this study, we examined dexamethasone-induced muscle atrophy using zebrafish (Danio rerio), a vertebrate model, and assessed whether administration of Lepidium meyenii (maca) as a dietary supplement can prevent muscle atrophy. Changes in skeletal myofibers in zebrafish were evaluated after exposure to dexamethasone for different periods and at different concentrations. Under optimized conditions, zebrafish pre-fed with maca for 3 days were exposed to 0.01% dexamethasone for 1 h/day for 7 days. Thereafter, myofiber loss, damaged muscle contractile proteins, and abnormal exploratory behavior due to the structural and functional impairment of skeletal muscle associated with muscle atrophy were investigated using hematoxylin–eosin, immunofluorescence staining, and behavioral analyses. Our findings suggest that dexamethasone induces muscle atrophy in zebrafish, inhibiting exploratory behavior by inducing myofiber loss, inhibiting muscle contraction, and causing changes in endurance and velocity. Thus, the zebrafish model can be used to screen pharmaceutical agents and to study muscle atrophy. Furthermore, maca is a potential dietary supplement to prevent muscle atrophy, as it protects muscle fibers.

G-protein coupled receptor 139 (GPR139) is an evolutionarily conserved orphan receptor, predominantly expressing in the habenula of vertebrate species. The habenula has recently been implicated in aversive response and its associated learning. Here, we tested the hypothesis that GPR139 signalling in the habenula may play a role in fear learning in the zebrafish. We examined the effect of intraperitoneal injections of a human GPR139-selective agonist (JNJ-63533054) on alarm substance-induced fear learning using conditioned place avoidance paradigm, where an aversive stimulus is paired with one compartment, while its absence is associated with the other compartment of the apparatus. The results indicate that fish treated with 1 µg/g body weight of GPR139 agonist displayed no difference in locomotor activity and alarm substance-induced fear response. However, avoidance to fear-conditioned compartment was diminished, which suggests that the agonist blocks the consolidation of contextual fear memory. On the other hand, fish treated with 0.1 µg/g body weight of GPR139 agonist spent a significantly longer time in the unconditioned neutral compartment as compared to the conditioned (punished and unpunished) compartments. These results suggest that activation of GPR139 signalling in the habenula may be involved in fear learning and the decision-making process in the zebrafish.

Predation is an important factor influencing the strength and success of a year class in many prey fishes. Many adaptations, including chemical cues, benefit shoaling groups by informing members of a possible impending predatory threat. Bighead (Hypophthalmichthys nobilis) and silver carps (Hypophthalmichthys molitrix) are widely dispersed invasive fishes that spend much of their first year at a size vulnerable to predation. The objective of this study was to evaluate the fright response of juvenile bighead and silver carps when exposed to alarm cues from conspecific and heterospecific fishes. Groups of carps were exposed to 5 mL of alarm cue from either bighead carp, silver carp, golden shiner (Notemigonus crysoleucas), or fathead minnow (Pimephales promelas). Behavior was recorded before and after the application of the treatment. Our results indicate that both species were unable to recognize chemical alarm cues from North American cypriniforms, yet both were able to recognize conspecific alarm cues. However, the two species of bigheaded carps respond to conspecific chemicals differently. Silver carp increase distance among individuals, whereas bighead carp reduce distance between individuals. Both show a reduction in activity after chemical application. Though both species recognize conspecific alarm cues, bighead carp exhibit similar fright patterns when exposed to alarm chemicals of silver carp, yet no behavioral responses were detected in silver carp when exposed to alarm chemicals of bighead carp. An understanding of fright response in bighead and silver carps aids understanding of predator interactions and ecological roles of these fishes, and can be utilized in population management applications.

Invasive species exert negative impacts on biodiversity and ecosystems on a global scale, which may be enhanced in the future by climate change. Knowledge of how invasive species respond physiologically and behaviorally to novel and changing environments can improve our understanding of which traits enable the ecological success of these species, and potentially facilitate mitigation efforts. We examined the effects of acclimation to temperatures ranging from 5 to 28°C on aerobic metabolic rates, upper temperature tolerance (critical thermal maximum, CTmax), as well as temperature preference (Tpref) and avoidance (Tavoid) of the round goby (Neogobius melanostomus), one of the most impactful invasive species in the world. We show that round goby maintained a high aerobic scope from 15 to 28°C; that is, the capacity to increase its aerobic metabolic rate above that of its maintenance metabolism remained high across a broad thermal range. Although CTmax increased relatively little with acclimation temperature compared with other species, Tpref and Tavoid were not affected by acclimation temperature at all, meaning that round goby maintained a large thermal safety margin (CTmax−Tavoid) across acclimation temperatures, indicating a high level of thermal resilience in this species. The unperturbed physiological performance and high thermal resilience were probably facilitated by high levels of phenotypic buffering, which can make species readily adaptable and ecologically competitive in novel and changing environments. We suggest that these physiological and behavioral traits could be common for invasive species, which would only increase their success under continued climate change.

Anthropogenic environmental degradation has led to an increase in the frequency and prevalence of aquatic hypoxia (low dissolved oxygen concentration, DO), which may affect habitat quality for water-breathing fishes. The weakly electric black ghost knifefish, Apteronotus albifrons, is typically found in well-oxygenated freshwater habitats in South America. Using a shuttle-box design, we exposed juvenile A. albifrons to a stepwise decline in DO from normoxia (> 95% air saturation) to extreme hypoxia (10% air saturation) in one compartment and chronic normoxia in the other. On average, A. albifrons actively avoided the hypoxic compartment below 22% air saturation. Hypoxia avoidance was correlated with upregulated swimming activity. Following avoidance, fish regularly ventured back briefly into deep hypoxia. Hypoxia did not affect the frequency of their electric organ discharges. Our results show that A. albifrons is able to sense hypoxia at non-lethal levels and uses active avoidance to mitigate its adverse effects.

Global climate change continues to impact fish habitat quality and biodiversity, especially in regard to the dynamics of invasive non-native species. Using individual aquaria and an open channel flume, this study evaluated the effects of water temperature, flow velocity and turbulence interactions on swimming performance of two lentic, invasive non-native fish in the UK, pumpkinseed ( Lepomis gibbosus ) and topmouth gudgeon ( Pseudorasbora parva ). Burst and sustained swimming tests were conducted at 15, 20 and 25°C. Acoustic Doppler velocimetry was used to measure the flume hydrodynamic flow characteristics. Both L. gibbosus and P. parva occupied the near-bed regions of the flume, conserving energy and seeking refuge in the low mean velocities flow areas despite the relatively elevated turbulent fluctuations, a behaviour which depended on temperature. Burst swimming performance and sustained swimming increased by up to 53% as temperature increased from 15 to 20°C and 71% between 15 and 25°C. Furthermore, fish test area occupancy was dependent on thermal conditions, as well as on time-averaged velocities and turbulent fluctuations. This study suggests that invasive species can benefit from the raised temperatures predicted under climate change forecasts by improving swimming performance in flowing water potentially facilitating their further dispersal and subsequent establishment in lotic environments.

Anti-regenerative scarring obstructs spinal cord repair in mammals and presents a major hurdle for regenerative medicine. In contrast, adult zebrafish possess specialized glial cells that spontaneously repair spinal cord injuries by forming a pro-regenerative bridge across the severed tissue. To identify the mechanisms that regulate differential regenerative capacity between mammals and zebrafish, we first defined the molecular identity of zebrafish bridging glia and then performed cross-species comparisons with mammalian glia. Our transcriptomics show that pro-regenerative zebrafish glia activate an epithelial-to-mesenchymal transition (EMT) gene program and that EMT gene expression is a major factor distinguishing mammalian and zebrafish glia. Functionally, we found that localized niches of glial progenitors undergo EMT after spinal cord injury in zebrafish and, using large-scale CRISPR-Cas9 mutagenesis, we identified the gene regulatory network that activates EMT and drives functional regeneration. Thus, non-regenerative mammalian glia lack an essential EMT-driving gene regulatory network that reprograms pro-regenerative zebrafish glia after injury.

Without insights into the threats affecting species across their distributions and throughout their annual cycles, effective conservation management cannot be applied. The Whenua Hou diving petrel Pelecanoides whenuahouensis (WHDP) is a Critically Endangered small seabird whose offshore habits and threats are poorly understood. We tracked WHDPs year-round in 2015/16, 2017/18, and 2018/19 using global location-sensing immersion loggers to identify offshore distribution, movements, behaviour, and overlap with commercial fishing effort. During the breeding period, WHDPs ranged from southern Aotearoa (New Zealand) to Maukahuka (Auckland Islands). After breeding, WHDPs migrated southwest towards the Polar Front south of Australia, exhibited clockwise movements, and returned to their breeding grounds via the Subantarctic Front. During the non-breeding period, WHDPs exhibited extreme aquatic behaviour and spent >95% of their time on, or under, water. The core areas used consistently during breeding and non-breeding periods warrant listing as Important Bird and Biodiversity Areas. Spatiotemporal overlap of commercial fishing effort with breeding distributions was considerable, in contrast with non-breeding distributions. Spatiotemporal management of anthropogenic activity around the breeding colony during the breeding period could help protect WHDPs, but such measures should be subjected to a structured decision-making framework. Our results illustrate the importance of year-round studies to inform conservation of marine species.

Tramadol is a widely used analgesic with additional antidepressant and anxiolytic effects. This compound has been reported in continental waters reaching concentrations of µg/L as a consequence of its inefficient removal in sewage treatment plants and increasing use over time. In this study, European chubs (Squalius cephalus) were exposed to 1 µg/L of tramadol in water for 42 days with a subsequent 14 days of depuration. Our results revealed that chubs exposed to this analgesic underwent changes in their behaviour as compared to the control group. The behavioural outcome was also influenced by the individual concentration of tramadol in brain tissue. In particular, experimental fish presented anxiolytic-like effects, characterized by less bold and less social individuals. Exposed animals were less frequently out of the shelter and moved a shorter distance, indicating that they explored the new environment less during the boldness test. In the novel object recognition experiment, although they distinguished the new item, they examined it less and displayed a reduced activity. Shoal cohesion was disrupted as observed in an increased distance between individuals. After the depuration phase, this alteration remained whereas the boldness effect disappeared. Moreover, the degree of behavioural changes was correlated with the concentration of the substance in brain. According to our findings, chronic presence of tramadol in the environment can impact the fitness of exposed aquatic fauna by altering evolutionary crucial behaviours.

The habenula is an evolutionarily conserved brain structure, which has recently been implicated in fear memory. In the zebrafish, kisspeptin (Kiss1) is predominantly expressed in the habenula, which has been implicated as a modulator of fear response. Hence, in the present study, we questioned whether Kiss1 has a role in fear memory and morphine-induced fear memory impairment using an odorant cue (alarm substances, AS)-induced fear avoidance paradigm in adult zebrafish, whereby the fear-conditioned memory can be assessed by a change of basal place preference (= avoidance) of fish due to AS-induced fear experience. Subsequently, to examine the possible role of Kiss1 neurons-serotonergic pathway, kiss1 mRNA and serotonin levels were measured. AS exposure triggered fear episodes and fear-conditioned place avoidance. Morphine treatment followed by AS exposure, significantly impaired fear memory with increased time-spent in AS-paired compartment. However, fish administered with Kiss1 (10–21 mol/fish) after morphine treatment had significantly lower kiss1 mRNA levels but retained fear memory. In addition, the total brain serotonin levels were significantly increased in AS- and Kiss1-treated groups as compared to control and morphine treated group. These results suggest that habenular Kiss1 might be involved in consolidation or retrieval of fear memory through the serotonin system.

Infectious diseases are key drivers of wildlife populations and agriculture production, but whether and how climate change will influence disease impacts remains controversial. One of the critical knowledge gaps that prevents resolution of this controversy is a lack of high-quality experimental data, especially in marine systems of significant ecological and economic consequence. Here, we performed a manipulative experiment in which we tested the temperature-dependent effects on Atlantic salmon (Salmo salar) of sea lice (Lepeophtheirus salmonis)—a parasite that can depress the productivity of wild-salmon populations and the profits of the salmon-farming industry. We explored sea-louse impacts on their hosts across a range of temperatures (10, 13, 16, 19, and 22 °C) and infestation levels (zero, ‘low’ (mean abundance ± SE = 1.6 ± 0.1 lice per fish), and ‘high’ infestation (6.8 ± 0.4 lice per fish)). We found that the effects of sea lice on the growth rate, condition, and survival of juvenile Atlantic salmon all worsen with increasing temperature. Our results provide a rare empirical example of how climate change may influence the impacts of marine disease in a key social-ecological system. These findings underscore the importance of considering climate-driven changes to disease impacts in wildlife conservation and agriculture.

Climate change and pollution are some of the greatest anthropogenic threats to wild animals. Transgenerational plasticity—when parental exposure to environmental stress leads to changes in offspring phenotype—has been highlighted as a potential mechanism to respond to various environmental and anthropogenic changes across taxa. Transgenerational effects may be mediated via multiple mechanisms, such as transfer of maternal hormones to eggs/foetus. However, sources of variation in hormone transfer are poorly understood in fish, and thus the first step is to characterise whether environmental challenges alter transfer of maternal hormones to eggs. To this end, we explored the population variation and environmental variation (in response to temperature and endocrine disrupting copper) in maternal thyroid hormone (TH), transfer to offspring in a common fish model species, the three-spined stickleback (Gasterosteus aculeatus) using multiple approaches: (i) We compared ovarian TH levels among six populations across a wide geographical range in the Baltic Sea, including two populations at high water temperature areas (discharge water areas of nuclear power plants) and we experimentally exposed fish to (ii) environmentally relevant heat stress and (iii) copper for 7 days. We found that populations did not differ in intraovarian TH levels, and short-term heat stress did not influence intraovarian TH levels. However, copper exposure increased both T4 and T3 levels in ovaries. The next step would be to evaluate if such alterations would lead to changes in offspring phenotype.

Reducing the abundances of invasive species by removals aims to minimize their ecological impacts and enable ecosystem recovery. Removal methods are usually selective, modifying phenotypic traits in the managed populations. However, there is little empirical evidence of how removal‐driven changes in multiple phenotypic traits of surviving individuals of invasive species can affect ecosystem functioning and recovery. Overcoming this knowledge gap is highly relevant because individuals are the elemental units of ecological processes and so integrating individual‐level responses into the management of biological invasions could improve their efficiency. Here we provide novel demonstration that removals by trapping, angling and biocontrol from lakes of the globally invasive crayfish Procambarus clarkii induced substantial changes in multiple phenotypic traits. A mesocosm experiment then revealed that these changes in phenotypic traits constrain recovery of basic ecosystem functions (decomposition of organic matter, benthic primary production) by acting in the opposite direction than the effects of reduced invader abundance. However, only minor ecological impacts of invader abundance and phenotypic traits variation remained a year after its complete eradication. Our study provides quantitative evidence to an original idea that removal‐driven trait changes can dampen recovery of invaded ecosystems even when the abundance of invasive species is substantially reduced. We suggest that the phenotypic responses of invaders to the removal programme have strong effects on ecosystem recovery and should be considered within the management of biological invasions, particularly when complete eradication is not achievable.

Background: Increases in seawater temperatures and in the frequency and severity of hypoxic events are expected with climate change, and may become a challenge for cultured Atlantic salmon and negatively affect their growth, immunology and welfare. Thus, we examined how an incremental temperature increase alone (Warm & Normoxic-WN: 12→20°C; 1°C week -1 ), and in combination with moderate hypoxia (Warm & Hypoxic-WH: ~70% air saturation), impacted salmon’s hepatic transcriptome expression compared to control fish (CT: 12°C, normoxic) using 44K microarrays and qPCR. Results: Overall, we identified 2,894 differentially expressed probes (DEPs, FDR < 5%), that included 1,111 shared DEPs, while 789 and 994 DEPs were specific to WN and WH fish, respectively. Pathway analysis suggested that the cellular mechanisms affected by the two experimental conditions were quite similar, with up-regulated genes functionally associated with heat shock response, ER-stress, apoptosis and immune defence, while genes connected with general metabolic processes, proteolysis and oxidation-reduction were largely suppressed. The qPCR assessment of 41 microarray-identified genes validated that the heat shock response ( hsp90aa1, serpinh1 ), apoptosis ( casp8, jund, jak2 ) and immune responses ( apod, c1ql2, epx ) were up-regulated in WN and WH fish, while oxidative stress and hypoxia sensitive genes were down-regulated ( cirbp, cyp1a1, egln2, gstt1, hif1α, prdx6, rraga, ucp2 ). However, the additional challenge of hypoxia resulted in more pronounced effects on heat shock and immune-related processes, including a stronger influence on the expression of 14 immune-related genes. Finally, robust correlations between the transcription of 19 genes and several phenotypic traits in WH fish suggest that changes in gene expression were related to an impaired physiological and growth performance. Conclusion: Increasing temperature to 20°C alone, and in combination with hypoxia, resulted in the up- and down-regulation of genes involved in similar important pathways in Atlantic salmon. However, the heat shock and immune responses of fish exposed to 20°C and hypoxia were more affected, and their transcriptional dysregulation was related to reduced performance. This study provides valuable information on how these two environmental challenges affect the expression of stress-, metabolic- and immune-related genes and pathways and identifies potential biomarker genes for improving our understanding of fish health and welfare.

Marcot et al. (2019) recently described the risk analysis process by which the U.S. Fish and Wildlife Service (USFWS) chooses species for listing as injurious wildlife. They further compared the Fish Invasiveness Screening Kit (FISK) and the Aquatic Species Invasiveness Screening Kit (AS-ISK), generally unfavorably, with their process/components. We assert that FISK and AS-ISK were largely misrepresented. The USFWS process is a risk analysis framework, whereas FISK and AS-ISK are hazard identification/risk screening tools, addressing only the initial step of a risk analysis scheme. Thus to avoid an apples-to-oranges comparison as done in that paper, FISK/AS-ISK should be compared to the equivalent USFWS tool, the Ecological Risk Screening Summaries (ERSS). The remaining issues that we address concerning FISK/AS-ISK include: (1) need for expert opinion, (2) subjective climate matching, (3) need for regional calibration, and (4) narrower range of information inputs; and concerning the ERSS process: (5) peer-review. Both systems clearly use expert opinion, the FISK/AS-ISK arguably in a more structured and transparent manner. Guidance for FISK/AS-ISK recommends a climate-matching program, but allows for use of Köppen-Geiger climate types or physiological tolerances, potentially increasing subjectivity in some cases but improving climate-match quality in others. Calibration of FISK/AS-ISK follows from the regional nature of invasiveness risk and the use of questions unrelated to climate that are tailored to the risk assessment (RA) area. Climate match is the only element that the ERSS applies specifically to the RA area. The FISK and AS-ISK actually use a much wider range of information than does the ERSS, a system based on invasion history and climate match only. The peer review of ERSS consisted of a five-member expert panel that evaluated the method, whereas the ERSS reports posted online are not peer reviewed. Conversely, FISK applications have resulted in 37 peer-reviewed journal articles, including assessments from over 70 experts in 45 countries. We welcome constructive criticism and improvement of these decision-support tools, but we are concerned that managers may dismiss well-established approaches in favor of a framework that possesses its own method-specific limitations. We recommend that managers evaluate risk-based approaches and adopt systems to support decisions and improve invasive species management.

Animals can gain significant advantages from abilities to detect cues from predators, assess risks, and respond adaptively to reduce the likelihood of injurious interactions. In contrast, predator cue-induced changes in behaviour may interfere with fitness-associated activities such as exploration, foraging and reproduction. Despite the ecological importance of predator-prey interactions in insects, remarkably little is known about the abilities of insects to detect and respond to olfactory cues from predators, or the potential costs of such responses. We here demonstrate that a tephritid fruit fly, the Queensland fruit fly Bactrocera tryoni, is able to detect and respond differentially to volatile olfactory cues from four potential predators (three spiders and an ant) that vary in prevalence and diurnal activity. Male and female flies increased or decreased motility (velocity, active time, distance moved), or exhibited no change in motility, depending on which predator volatiles they encountered. Further, flies significantly reduced foraging, oviposition and mating propensity in the presence of volatiles from any of the predators. This study is the first report of predator-specific responses to olfactory cues in a tephritid fruit fly, and highlights that such anti-predator responses can impose costs on general activity and reproductive behaviour.

Coral reef species, like most tropical species, are sensitive to increasing environmental temperatures, with many species already living close to their thermal maxima. Ocean warming and the increasing frequency and intensity of marine heatwaves are challenging the persistence of reef-associated species through both direct physiological effects of elevated water temperatures and the degradation and loss of habitat structure following disturbance. Understanding the relative importance of habitat degradation and ocean warming in shaping species distributions is critical in predicting the likely biological effects of global warming. Using an automated shuttle box system, we investigated how habitat complexity influences the selection of thermal environments for a common coral reef damselfish, Chromis atripectoralis. In the absence of any habitat (i.e. control), C. atripectoralis avoided temperatures below 22.9 ± 0.8°C and above 31.9 ± 0.6°C, with a preferred temperature (Tpref) of 28.1 ± 0.9°C. When complex habitat was available, individual C. atripectoralis occupied temperatures down to 4.3°C lower (mean ± SE; threshold: 18.6 ± 0.7°C; Tpref: 18.9 ± 1.0°C) than control fish. Conversely, C. atripectoralis in complex habitats occupied similar upper temperatures as control fish (threshold: 31.7 ± 0.4°C; preference: 28.3 ± 0.7°C). Our results show that the availability of complex habitat can influence the selection of thermal environment by a coral reef fish, but only at temperatures below their thermal preference. The limited scope of C. atripectoralis to occupy warmer environments, even when associated with complex habitat, suggests that habitat restoration efforts in areas that continue to warm may not be effective in retaining populations of C. atripectoralis and similar species. This species may have to move to cooler (e.g. deeper or higher latitude) habitats under predicted future warming. The integration of habitat quality and thermal environment into conservation efforts will be essential to conserve of coral reef fish populations under future ocean warming scenarios.

Behavioural changes that occur during the parr–smolt transformation were investigated in juvenile coho salmon Oncorhynchus kisutch. Fish from two populations were examined from the Fraser River catchment in British Columbia, Canada; a short and a long‐distance migrating population. Fish showed a significant decrease in condition factor and significant increase in gill Na + K + ‐ATPase activity during the spring indicating that they became competent smolts, but no difference between populations. Temperature preference trials were conducted using a shuttlebox system throughout the spring. Mean temperature preference did not differ between the two populations, but preferred temperature decreased with development from 16.5 ± 0.3°C for parr to 15.5 ± 0.4°C for smolts. Mean swimming velocity was also greater in smolts than parr, but there was no difference between the two populations. The preference for warmer water temperature observed for parr in early spring may be advantageous for stimulating smolt development. Preference for slightly cooler temperatures observed for smolts would sustain elevated seawater tolerance during the smolt window by a short time and may ensure successful transition to the marine environment.

Occupational, accidental, or suicidal exposure to acrylamide (ACR) may result in a neurotoxic syndrome. Development of animal models of acrylamide neurotoxicity is necessary for increasing our mechanistic understanding of this syndrome and developing more effective therapies. A new model for acute ACR neurotoxicity has been recently developed in adult zebrafish. Whereas the results of the initial characterization were really promising, a further characterization is needed for testing the construct validity of the model. In this study, the presence of gait abnormalities has been investigated by using ZebraGait, software specifically designed to analyze the kinematics of fish swimming in a water tunnel. The results of the kinematic analyses demonstrated that the model exhibits mild-to-moderate gait abnormalities. Moreover, the model exhibited negative scototaxis, a result confirming a phenotype of anxiety comorbid with depression phenotype. Interestingly, depletion of the reduced glutathione levels was found in the brain without a concomitant increase in oxidative stress. Finally, hypolocomotion and positive geotaxis exhibited by this model were fully recovered 5 days after transferring the fish to clean fish-water. All this data support the validity of the ACR acute neurotoxicity model developed in adult zebrafish.

Elevated seawater CO2 can cause a range of behavioural impairments in marine fishes. However, most studies to date have been conducted on small benthic species and very little is known about how higher oceanic CO2 levels could affect the behaviour of large pelagic species. Here, we tested the effects of elevated CO2, and where possible the interacting effects of high temperature, on a range of ecologically important behaviours (anxiety, routine activity, behavioural lateralization and visual acuity) in juvenile yellowtail kingfish, Seriola lalandi. Kingfish were reared from the egg stage to 25 days post-hatch in a full factorial design of ambient and elevated CO2 (∼500 and ∼1000 μatm pCO2) and temperature (21 °C and 25 °C). The effects of elevated CO2 were trait-specific with anxiety the only behaviour significantly affected. Juvenile S. lalandi reared at elevated CO2 spent more time in the dark zone during a standard black-white test, which is indicative of increased anxiety. Exposure to high temperature had no significant effect on any of the behaviours tested. Overall, our results suggest that juvenile S. lalandi are largely behaviourally tolerant to future ocean acidification and warming. Given the ecological and economic importance of large pelagic fish species more studies investigating the effect of future climate change are urgently needed.

Myelin is a specialized membrane that wraps around nerve fibers and is essential for normal axonal conduction in neurons. In the central nervous system, oligodendrocytes are responsible for myelin formation. Recent studies have reported pathological abnormalities in oligodendrocytes in human patients with amyotrophic lateral sclerosis (ALS) and a mouse model of ALS expressing the G93A mutation of the human superoxide dismutase 1 (mtSOD1). However, it is unclear whether oligodendrocyte pathology in ALS represents the primary dysfunction induced by mtSOD1 and how mtSOD1 contributes to oligodendrocyte degeneration and ALS pathogenesis. We analyzed GAL4‐VP16‐UAS transgenic zebrafish selectively expressing mtSOD1 in mature oligodendrocytes. We observed that mtSOD1 directly induced oligodendrocyte degeneration by disrupting the myelin sheath and downregulating monocarboxylate transporter 1 (MCT1), thereby causing spinal motor neuron degeneration. Pathological changes observed in this transgenic zebrafish were similar to the pathology observed in the SOD1 G93A mouse model of ALS, which is characterized by expression of mtSOD1 in all cells. In addition, oligodendrocyte dysfunction induced by mtSOD1 was associated with anxiety‐related behavioral abnormalities, learning impairments, and motor defects in the early symptomatic stage. We also found that treatment with potassium channel inhibitors rescued behavioral abnormalities without rescuing MCT1 expression, suggesting that myelin disruption induces behavioral abnormalities independently of MCT1. These results indicate that mtSOD1‐induced dysfunction of mature oligodendrocytes is sufficient to induce motor neuron degeneration, thus informing future therapeutic strategies targeted at oligodendrocytes in ALS.

Challenging environmental conditions can induce voluntary behavioral avoidance in animals. Dissolved carbon dioxide ( CO 2 ) is an environmental stressor that was previously shown to upregulate the stress axis in fish and also causes voluntary avoidance. Variation in individual state or context, such as whether an animal is fasted or fed, can alter animal behavior, including the response to environmental challenges. In the current study, we sought to define the influence of nutritional status on the response of Largemouth Bass Micropterus salmoides to elevated CO 2. Two groups of Largemouth Bass—one fed group and one fasted group—were first subjected to a CO 2 shuttling protocol to define avoidance thresholds, followed by a CO 2 tolerance protocol to define the time required to lose equilibrium and recover. Data showed that although feeding and fasting had no influence on the avoidance of CO 2, fasted fish required 17% longer to lose equilibrium in elevated CO 2. Avoidance of elevated CO 2 is therefore independent of animal state, but fish in poor nutritional condition from fasting are more tolerant. Thus, managers considering elevated CO 2 as a nonphysical barrier to deter fish movements should be cognizant of food availability, as fasted animals may require increased partial pressures of CO 2 to ensure successful deterrence.

Fish are used in a variety of experimental contexts often in high numbers. To maintain their welfare and ensure valid results during invasive procedures it is vital that we can detect subtle changes in behaviour that may allow us to intervene to provide pain-relief. Therefore, an automated method, the Fish Behaviour Index (FBI), was devised and used for testing the impact of laboratory procedures and efficacy of analgesic drugs in the model species, the zebrafish. Cameras with tracking software were used to visually track and quantify female zebrafish behaviour in real time after a number of laboratory procedures including fin clipping, PIT tagging, and nociceptor excitation via injection of acetic acid subcutaneously. The FBI was derived from activity and distance swum measured before and after these procedures compared with control and sham groups. Further, the efficacy of a range of drugs with analgesic properties to identify efficacy of these agents was explored. Lidocaine (5 mg/L), flunixin (8 mg/L) and morphine (48 mg/L) prevented the associated reduction in activity and distance swum after fin clipping. From an ethical perspective, the FBI represents a significant refinement in the use of zebrafish and could be adopted across a wide range of biological disciplines.

Chemobehavioural phenotypic analysis using small aquatic model organisms is becoming an important toolbox in aquatic ecotoxicology and neuroactive drug discovery. The analysis of the organisms’ behavior is usually performed by combining digital video recording with animal tracking software. This software detects the organisms in the video frames, and reconstructs their movement trajectory using image processing algorithms. In this work we investigated the impact of video file characteristics, video optimization techniques and differences in animal tracking algorithms on the accuracy of quantitative neurobehavioural endpoints. We employed larval stages of a free-swimming euryhaline crustacean Artemia franciscana,commonly used for marine ecotoxicity testing, as a proxy modelto assess the effects of video analytics on quantitative behavioural parameters. We evaluated parameters such as data processing speed, tracking precision, capability to perform high-throughput batch processing of video files. Using a model toxicant the software algorithms were also finally benchmarked against one another. Our data indicates that variability in video file parameters; such as resolution, frame rate, file containers types, codecs and compression levels, can be a source of experimental biases in behavioural analysis. Similarly, the variability in data outputs between different tracking algorithms should be taken into account when designing standardized behavioral experiments and conducting chemobehavioural phenotyping.

Propiconazole is a triazole fungicide that has been widely used in agriculture and has been detected in the aquatic environment. This study aimed to investigate the effects of propiconazole exposure on lipid metabolism in the early life stages of zebrafish for 120 h postfertilization (hpf). Using the early life stages of zebrafish to address scientific questions is lower in cost, more efficient, and suitable to meeting current legislation than those in other traditional fish species. Exposure to propiconazole significantly inhibited the development of zebrafish embryos and larvae. This exposure also caused reduced locomotor activities in zebrafish. Furthermore, total cholesterol levels, lipoprotein lipase, and fatty acid synthase activities were significantly decreased. The expression levels of genes involved in lipid metabolism were significantly up-regulated in response to propiconazole exposure. GC-MS/MS analysis revealed that fatty acids were significantly decreased. Together, the findings indicate the potential environmental risks of propiconazole exposure in the aquatic ecosystem.

The ability to assess welfare is an important refinement that will ensure the good condition of animals used in experimentation. The present study investigated the impact of invasive procedures on the patterns of movement of zebrafish (Danio rerio). Recordings were made before and after fin clipping, PIT tagging and a standard pain test and these were compared with control and sham handled zebrafish. The fractal dimension (FD) from the 3D trajectories was calculated to determine the effect of these treatments on the complexity of movement patterns. While the FD of zebrafish trajectories did not differ over time in either the control or sham group, the FDs of the treatment groups reduced in complexity. The FD of fish injected with different strengths of acetic acid declined in a dose-dependent manner allowing us to develop an arbitrary scale of severity of the treatments. The 3D trajectory plots from some groups indicated the presence of repetitive swimming patterns akin to stereotypical movements. When administered with lidocaine, which has analgesic properties, the movement complexity of fin clipped fish reverted to a pattern that resembled that of control fish. Fractal analysis of zebrafish locomotion could potentially be adopted as a tool for fish welfare assessment.

In expanding populations, the allocation of resources to life-history traits is expected to change rapidly after the colonization of a new area. Understanding these changes is of crucial importance to predict the future changes in distribution ranges, and the possible impacts of expanding species on the colonized environments. Both theoretical and empirical studies have provided some evidence for an increased dispersal at the range edge of expanding populations. Because this phenomenon has been documented on large spatial scales, it seems difficult to determine what proportion of the phenotypic variation is due to the expansion process (spatial sorting) versus spatial variation in the environment (local adaptation). The aim of our study is to determine whether, in a small and recent expanding population, the dispersal capacity is increased at the range edge compared to the range core despite a minimal role of local adaptation. We studied the modification of dispersal capacity at the range edge of an expanding population of the African clawed frog, Xenopuslaevis recently settled in Western France. To do so, we used experiments to compare movements of individuals depending on their location in the population, and capture–mark–recapture field surveys to estimate variation in traits related to dispersal across the invasive range. Both methods consistently showed significantly higher dispersal rates and distances, as well as in the relative length of the hind limb, at the range edge, for both sexes. This result is the first report suggesting that changes in allocation to dispersal, and not only traits associated with dispersal, can occur at a small geographical and temporal scale, where the role of local adaptation is likely minimal. This study fills a gap in an issue where it is currently crucial, for conservation purposes, to understand the mechanisms involved in range expansions.

To meet future global demand for fish protein, more fish will need to be farmed using fewer resources, and this will require the selection of nonaggressive individuals that perform well at high densities. Yet, the genetic changes underlying loss of aggression and adaptation to crowding during aquaculture intensification are largely unknown. We examined the transcriptomic response to aggression and crowding in Nile tilapia, one of the oldest and most widespread farmed fish, whose social structure shifts from social hierarchies to shoaling with increasing density. A mirror test was used to quantify aggression and skin darkening (a proxy for stress) of fish reared at low and high densities, and gene expression in the hypothalamus was analysed among the most and least aggressive fish at each density. Fish reared at high density were darker, had larger brains, were less active and less aggressive than those reared at low density and had differentially expressed genes consistent with a reactive stress‐coping style and activation of the hypothalamus–pituitary–interrenal (HPI) axis. Differences in gene expression among aggressive fish were accounted for by density and the interaction between density and aggression levels, whereas for nonaggressive fish differences in gene expression were associated with individual variation in skin brightness and social stress. Thus, the response to crowding in Nile tilapia is context dependent and involves different neuroendocrine pathways, depending on social status. Knowledge of genes associated with the response to crowding may pave the way for more efficient fish domestication, based on the selection of nonaggressive individuals with increasing tolerance to chronic stress necessary for aquaculture intensification.

The pace-of-life syndrome (POLS) concept predicts that individuals with high baseline metabolic rates demonstrate high boldness, aggressiveness, and activity, especially in food acquisition, with associated relatively greater energy requirements. In fishes, these behaviors may increase individual vulnerability to angling. To test the predictions of the POLS concept, we quantified individual standard metabolic rate (SMR) and boldness in both wild-caught and hatchery-reared Eurasian perch (Perca fluviatilis). We found both SMR and boldness to be repeatable traits but detected no correlation between them. Individual vulnerability to angling was assessed in the hatchery-reared perch, but we found no difference in boldness or SMR between vulnerable and nonvulnerable perch. Wild-caught perch were ice fished using either natural or artificial bait, and we observed no differences in boldness or SMR with respect to bait type or capture order. Our findings do not support the predictions of the POLS concept and, consistent with earlier studies in perch, suggest that angling may not drive selection against boldness in this species.

Inorganic arsenic (As) is one of the most ubiquitous and toxic substances with widespread health effects on human populations and biodiversity. Although arsenic is a frequent surface water pollutant, there is scant evidence about neurotoxicity in aquatic species in different stages of development. In the present study, we investigated the neurobehavioral effects of chronic exposure to environmentally relevant doses of arsenic. We exposed zebrafish to 50 and 500 ppb during the larval, juvenile, and adult stage (from 4 h to 150 days postfertilization). We then used broad behavioral screening to evaluate motor function, social behavior, learning and memory, and anxiety-like behaviors. Our results show that arsenic exposure to 500 ppb alters motor function from the embryo to the adult stage. Furthermore, during the adult phase, associative learning and the sensorimotor response are affected with both high and low doses of As, respectively. Notably, exposure to 500 ppb of As induces behaviors associated with anxiety, during the juvenile and adult phase but not the larval stage, without changes in whole-body cortisol levels. These results indicate that chronic exposure to arsenic during their lifespan is capable of producing alterations in different behavioral markers in aquatic vertebrates.

Broiler chickens are increasingly at the forefront of global meat production but the consequences of fast growth and selection for an increase in body mass on bird health are an ongoing concern for industry and consumers. To better understand the implications of selection we evaluated energetics and behaviour over the 6-week hatch-to-slaughter developmental period in a commercial broiler. The effect of posture on resting metabolic rate becomes increasingly significant as broilers grow, as standing became more energetically expensive than sitting. The proportion of overall metabolic rate accounted for by locomotor behaviour decreased over development, corresponding to declining activity levels, mean and peak walking speeds. These data are consistent with the inference that broilers allocate energy to activity within a constrained metabolic budget and that there is a reducing metabolic scope for exercise throughout their development. Comparison with similarly sized galliforms reveals that locomotion is relatively energetically expensive in broilers.

Animals are bombarded with information about their environment and must select and interpret the relevant cues to make behavioral adjustments critical to survival. How animals integrate and balance the many signals they receive about their environment is rarely assessed. We investigated how signals from the social and physical environment interact to influence environmental preferences in the endemic Tanganyikan cichlid Neolamprologus pulcher. Specifically, we explored how fish respond to the physiological challenge of declining O 2 levels in light of embedded social preferences using a modified shuttle box apparatus to test O 2 preferences. In the presence of a conspecific, the average (preferred) partial pressure of oxygen (Po 2 ) and minimum Po 2 experienced were significantly lower ([Formula: see text] and [Formula: see text] kPa, respectively) than in trials without a conspecific ([Formula: see text] and [Formula: see text] kPa, respectively). Fish with conspecifics also spent more time in the low Po 2 zone of the shuttle box and moved between the high and low Po 2 zones less frequently. Hence, O 2 preferences were modified, and fish willingly remained in an area of continuously declining O 2 availability to associate with a conspecific. The O 2 preferences of an individual during social trials correlated with its excess postexercise O 2 consumption following an exhaustive chase but not with its aerobic scope, routine O 2 consumption rate, or body mass. These results suggest that some aspects of respiratory and metabolic physiology (such as the propensity to use anaerobic metabolism) but not others (such as O 2 transport capacity) underpin some variation in social behavior under environmental stress.

Phenotypic scoring of wild animals under standardized laboratory conditions is important as it allows field ecologists and evolutionary biologists to understand the development and maintenance of interindividual differences in plastic traits (e.g. behaviour and physiology). However, captivity is associated with a shift from a natural familiar environment to an unfamiliar and artificial environment, which may affect estimates of plastic phenotypic traits. In this study, we tested how previous experience with laboratory environments and time spent in captivity affects behavioural (i.e. activity) and metabolic (i.e. standard and maximum metabolic rates) scoring of our model species, wild brown trout Salmo trutta. We found that individuals with previous experience of laboratory captivity (10.5 months earlier) showed higher activity in an open field test than individuals with no prior experience of laboratory captivity. Previous experience with captivity had no significant effect on metabolic rates. However, metabolic rates seemed to increase with increasing time spent in captivity prior to the collection of measurements. Although there are benefits of keeping wild animals in captivity prior to scoring, our results suggest that while allowing for sufficient acclimatization researchers should aim at minimizing time in captivity of wild animals to increase accuracy and ecological relevance of the scoring of plastic phenotypic traits.

Recently, the use of Daphnia magna has been proposed in on-line and real-time biomonitoring programmes as an early warning system for evaluating the effluent quality of sewage treatment plants (STPs). These systems are based on recording behavioural changes in the test organism resulting from the stress caused by the effluents. Indeed, altered behavioural signals could be induced at sublethal concentrations that are significantly lower than the corresponding EC50. However, at present, it is unknown whether the sensitivity of D. magna can be representative of that of other aquatic organisms, particularly benthic macroinvertebrates. An experiment was designed to verify whether D. magna can be employed in biomonitoring programmes for STPs located in alpine areas as a surrogate for cold freshwater best-adapted species. The responses of survival and behaviour alteration to exposure to the effluent of the Tonale Pass plant (Trentino, Italian Alps, 46°N, 10°E; 1799 m a.s.l.) were compared in a laboratory population of D. magna and a wild population of the chironomid Diamesa cinerella. These larvae were collected from the Vermigliana stream 50 metres upstream of the effluent input. Both organisms were exposed for 24 and 48 hrs to the effluent as it is and to three dilutions (/10, /100, /1000). The mortality rate and behavioural responses (using video tracking systems) were recorded. No significant mortality or change in behaviour was observed in the two species when exposed to the undiluted effluent. Exposure to serial dilutions of the treated effluent did not affect the survival of either species but notably altered their behaviour at both exposure times (e.g., the time spent in activity in D. magna and the average speed of movement and the cumulative distance travelled in both), especially when exposed to the ten-times-diluted effluent. Overall, the findings of this study emphasize that even though D. magna and D. cinerella use different behavioural strategies to cope with adverse environmental conditions, their overall sensitivity to treated effluents is similar. Accordingly, the use of D. magna in biological early warning systems protocols seems to also be sufficiently protective for local, cold-adapted species of alpine freshwater ecosystems.

Nickel (Ni) concentrations in aquatic ecosystems can be amplified by anthropogenic activities including resource extraction. Compared with fish and invertebrates, knowledge of Ni toxicity in amphibians is limited, especially for northern species. We examined the effect of Ni on wood frog (Lithobates sylvaticus) tadpoles, the species with the widest and most northern distribution of any anuran in North America. Wood frog tadpoles were exposed to a Ni concentration gradient (0.02–5.5 mg/L of Ni at 164 mg/L as CaCO3 water hardness) for 8 d and examined for lethality, Ni bioaccumulation, and several sublethal endpoints including body condition, food consumption, activity, and chemosensory function. Nickel induced a sublethal effect on body condition (8-d 10 and 20% effect concentrations [EC10 and EC20] of 1.07 ± 0.38 and 2.44 ± 0.51 mg/L of Ni ± standard error [SE], respectively) but not on food consumption, activity, or chemosensory function. Nickel accumulation in tadpole tissues was positively related to an increase in aqueous Ni concentration but was not lethal. Both the acute and chronic US Environmental Protection Agency water quality guideline concentrations for Ni (0.71 and 0.08 mg/L at 164 mg/L as CaCO3 water hardness, respectively) were protective against lethal and sublethal effects in wood frog tadpoles. In the present study, wood frog tadpoles were protected by current water quality guidelines for Ni and are likely not as useful as other taxa for environmental effects monitoring for this particular metal. Environ Toxicol Chem 2018;37:2458–2466. © 2018 SETAC The body condition of wood frog tadpoles decreases with an increase in the accumulation of nickel (Ni) in the whole body.

The main objective of the present study was to investigate possible links between sub-individual and supra-individual levels (i.e. population level) biomarkers in D. magna exposed to sublethal concentrations of the insecticide chlorpyrifos (CPF). To achieve the aim, 8-day old individuals were exposed for 96 h to two environmentally relevant concentrations of CPF (50 and 250 ng/L). Sub-individual level effects were investigated by measuring the activity of antioxidant (SOD, CAT, and GPx) and detoxifying (GST) enzymes, as well as by measuring the acetylcholinesterase (AChE) inhibition. In addition, the effects at supra-individual level were assessed by using a video-tracking system and analyzing changes in swimming capabilities (i.e. percentage of activity time, distance moved, and velocity). Our data have shown that daphnids exposed to both CPF concentrations were in a condition of stress which was highlighted by changes in both sub- and supra-individual biomarkers. Moreover, our results highlighted that the lowest tested CPF concentration did not modulate the antioxidant and detoxifying enzymes, whereas, an inhibition of AChE and a decrease of some parameters related to swimming behaviour (distance moved and velocity) were noted. On the contrary, significant changes in all the sub-individual biomarkers were measured at the highest tested concentration. In addition, organisms recovered the movement capability (distance moved) and also activate a mechanism of avoidance (increased swimming velocity). On the other hand, a reduction in the percent of active time was measured and this was attributed to the energy spent by organisms to activate antioxidant and detoxifying enzymes and the mechanism of avoidance. Based on these results, our study suggests the existence of a link between sub- and supra-individual levels, as the activation or non-activation in the antioxidant and detoxifying enzymes activities can led to different modifications of the swimming behaviour in D. magna. Effects of chlorpyrifos on biomarker responses and swimming behaviour.

Organismal responses to stressors can be influenced by several internal and external factors including physiological condition and inherent behavioral type. Carbon dioxide (CO2), a known stressor for fish, is naturally increasing in fresh water, and has been proposed as a non-physical barrier to prevent invasive fish movement. Intraspecific differences in how fish respond to CO2 challenges have been noted, with some individuals responding at low partial pressures of CO2 (pCO2), and others responding at higher pCO2. Sensitivity to pCO2 may play a role in avoidance behaviors with respect to CO2 barriers and may predict how fish respond to naturally occurring CO2 challenges. We sought to determine the role that both physiological condition (i.e., elevated cortisol) and personality (i.e., boldness) play in influencing behavioral responses. To accomplish this goal, a shuttle box assay was used to determine the pCO2 that elicited avoidance in cortisol-injected or non-injected largemouth bass (Micropterus salmoides), as well as bold or shy bluegill (Lepomis macrochirus). Cortisol-injected largemouth bass shuttled at 45% higher pCO2 than control fish, but personality of bluegill had no effect on shuttling. It appears that an individual’s cortisol level can affect CO2 avoidance, likely mediated through the effects of cortisol on acid–base balance at the gill, or through the effects of cortisol on coping styles. Our finding has important implications for how fish respond to either natural or anthropogenically-driven changes in CO2, as stressed fish with high cortisol would appear to be more tolerant of elevated CO2, independent of personality type.

Effective behavioral deterrents are needed to prevent aquatic invasive species from entering novel ecosystems. One deterrent strategy that shows promise is elevated carbon dioxide (CO2) concentrations in water which can alter the behavior of freshwater fishes, including invasive bigheaded carps (Hypophthalmichthys spp.). However, few studies have evaluated behavioral responses to elevated CO2 concentrations at different water temperatures. The objective of this study was to quantify CO2 concentrations needed to achieve avoidance (voluntary response) and narcosis (involuntary response observed by loss of equilibrium) behaviors in silver carp (H. molitrix) and bighead carp (H. nobilis) at 5, 15, and 25 °C. Overall, silver carp and bighead carp displayed avoidance and narcosis behaviors to CO2 at each water temperature, however bighead carp responded at higher CO2 concentrations than silver carp. Behavioral avoidance and narcosis were observed at approximately 40% lower CO2 concentrations in 5 °C water relative to 25 °C suggesting considerable influence of water temperature on a CO2 stimulus for both species. Results indicate that fluctuating water temperature (e.g., spatial and temporal variation across management sites) can influence how fish respond to elevated CO2, and may usefully be considered when applying CO2 as a behavioral deterrent.

Preventing the spread of invasive fishes is an important aspect of management programs, but is challenging due to the behavior of fish and the nature of aquatic environments. The use of dissolved carbon dioxide (CO2) has recently gained traction as a non-physical barrier for invasive fishes due to its ability to elicit avoidance behaviors in fish. Research to date has focused on the development of CO2 barriers using static water environments. Because CO2 barriers have been proposed for flowing water (i.e., in rivers or shipping canals), understanding the dynamics between fish and elevated CO2 in flowing water is essential. Our study aims to define threshold levels required to alter behavior of bluegill (Lepomis macrochirus) and largemouth bass (Micropterus salmoides) in flowing water, and to quantify behavioral metrics of fish exposed to < 200 [ambient], 25,000, 50,000, and 100,000 µatm pCO2. We also sought to quantify the impacts of repeated CO2 exposure on fish behavior. Bluegill showed increased activity at 25,000 µatm, while largemouth bass showed increased activity at 100,000 µatm. When repeatedly exposed to cycles of 50,000 µatm pCO2, bluegill exhibited increased activity followed by a diminished response after the second exposure. Results from this study define threshold levels required to elicit behavioral responses, and show that the effects that multiple exposures of elevated pCO2 can decline, possibly due to habituation. Results will help shape the development and deployment of a CO2 barrier to control the movements of invasive fishes.

Homozygous SMN1 loss causes spinal muscular atrophy (SMA), the most common lethal genetic childhood motor neuron disease. SMN1 encodes SMN, a ubiquitous housekeeping protein, which makes the primarily motor neuron-specific phenotype rather unexpected. SMA-affected individuals harbor low SMN expression from one to six SMN2 copies, which is insufficient to functionally compensate for SMN1 loss. However, rarely individuals with homozygous absence of SMN1 and only three to four SMN2 copies are fully asymptomatic, suggesting protection through genetic modifier(s). Previously, we identified plastin 3 (PLS3) overexpression as an SMA protective modifier in humans and showed that SMN deficit impairs endocytosis, which is rescued by elevated PLS3 levels. Here, we identify reduction of the neuronal calcium sensor Neurocalcin delta (NCALD) as a protective SMA modifier in five asymptomatic SMN1-deleted individuals carrying only four SMN2 copies. We demonstrate that NCALD is a Ca 2+ -dependent negative regulator of endocytosis, as NCALD knockdown improves endocytosis in SMA models and ameliorates pharmacologically induced endocytosis defects in zebrafish. Importantly, NCALD knockdown effectively ameliorates SMA-associated pathological defects across species, including worm, zebrafish, and mouse. In conclusion, our study identifies a previously unknown protective SMA modifier in humans, demonstrates modifier impact in three different SMA animal models, and suggests a potential combinatorial therapeutic strategy to efficiently treat SMA. Since both protective modifiers restore endocytosis, our results confirm that endocytosis is a major cellular mechanism perturbed in SMA and emphasize the power of protective modifiers for understanding disease mechanism and developing therapies.

Transport of coral reef fish larvae is driven by advection in ocean currents and larval swimming. However, for swimming to be advantageous, larvae must use external stimuli as guides. One potential stimulus is “odor” emanating from settlement sites (e.g., coral reefs), signaling the upstream location of desirable settlement habitat. However, specific chemicals used by fish larvae have not been identified. Dimethyl sulfide (DMS) is produced in large quantities at coral reefs and may be important in larval orientation. In this study, a choice-chamber (shuttle box) was used to assess preference of 28 pre-settlement stage larvae from reef fish species for seawater with DMS. Swimming behavior was examined by video-tracking of larval swimming patterns in control and DMS seawater. We found common responses to DMS across reef fish taxa - a preference for water with DMS and change in swimming behavior - reflecting a switch to “exploratory behavior”. An open water species displayed no response to DMS. Affinity for and swimming response to DMS would allow a fish larva to locate its source and enhance its ability to find settlement habitat. Moreover, it may help them locate prey accumulating in fronts, eddies, and thin layers, where DMS is also produced.

Aquatic toxicity testing in environmental monitoring and chemical risk assessment is critical to assess water quality for human use as well as predict impact of pollutants on ecosystems. In recent years, studies have increasingly focused on the relevance of sub-lethal effects of environmental contaminants. Sub-lethal toxicity endpoints such as behavioural responses are highly integrative and have distinct benefits for assessing water quality because they occur rapidly and thus can be used to sense the presence of toxicants. Our work describes a Lab-on-a-Chip system for the automated analysis of freshwater cladoceran Daphnia magna locomotory responses to water-born toxicants. The design combines a Lab-on-a-Chip system for Daphnia sp. culture under perfusion with time-resolved videomicroscopy and software tracking locomotory activity of multiple specimens. The application of the system to analyse the swimming behaviour of water fleas exposed to different concentrations of water-born toxicants demonstrated that Lab-on-a-Chip devices can become important research tools for behavioural ecotoxicology and water quality biomonitoring.

Both adult and larval zebrafish have been demonstrated to show behavioural responses to noxious stimulation but also to potentially stress- and fear or anxiety- eliciting situations. The pain or nociceptive response can be altered and modulated by these situations in adult fish through a mechanism called stress-induced analgesia. However, this phenomenon has not been described in larval fish yet. Therefore, this study explores the behavioural changes in larval zebrafish after noxious stimulation and exposure to challenges that can trigger a stress, fear or anxiety reaction. Five-day post fertilization zebrafish were exposed to either a stressor (air emersion), a predatory fear cue (alarm substance) or an anxiogenic (caffeine) alone or prior to immersion in acetic acid 0.1%. Pre- and post-stimulation behaviour (swimming velocity and time spent active) was recorded using a novel tracking software in 25 fish at once. Results show that larvae reduced both velocity and activity after exposure to the air emersion and alarm substance challenges and that these changes were attenuated using etomidate and diazepam, respectively. Exposure to acetic acid decreased velocity and activity as well, whereas air emersion and alarm substance inhibited these responses, showing no differences between pre- and post-stimulation. Therefore, we hypothesize that an antinociceptive mechanism, activated by stress and/or fear, occur in 5dpf zebrafish, which could have prevented the larvae to display the characteristic responses to pain.

Management of invasive species is addressed in both national and international regulations regarding the protection of marine habitats and biodiversity and in regulations of aquaculture. The geographical range of the invasive Pacific oyster, Crassostrea gigas, is expanding, both through human mediated vectors and by natural dispersal. The species is now spreading in Scandinavia. In order to optimize the management of the oyster, including targeted monitoring and mitigation activities, knowledge on the present and future distribution and impact on the ecosystem is important. The development of the population and the potential impacts on native ecosystems were analyzed, based on the present scientific knowledge on the distribution in Scandinavia, data on new settlements and existing literature. Data was first evaluated by 14 experts (including the authors) during a workshop, relating the current status of habitats where Pacific oysters are found in Scandinavia (Low energy rock, Littoral sand and mudflats, Littoral biogenic reefs, Sublittoral sand and Sublittoral biogenic reefs) to a predicted development, thereafter assessed in relation to impact on the habitats. The assessment was done as a function of climate change in a long-term IPCC climate scenario (A1B). We conclude that Littoral biogenic reefs are at risk to obtain the highest expected increase, while all other habitats are at risk of low to moderate development of the oyster populations. Accordingly, Littoral Biogenic reefs was assessed as the habitat type at risk of the largest ecosystem effects as high densities of oysters already exist in these areas, and the densities are expected to increase rapidly until reaching a threshold density. Low energy rock and Littoral sand and mud were assessed as being subjected to moderate to high ecosystem effects. Sub-littoral sand and Sub-littoral biogenic reefs were assessed as currently being at risk of moderate ecosystem effects as there are low densities of oysters in these habitats, although densities in sublittoral biogenic reefs has the potential to increase. We discuss management and mitigation strategies based on the forecasted development and effects of the Pacific oyster populations.

The pyrethroid prallethrin, an AI in DUET™ (Clarke Mosquito Control, St. Charles, IL), is widely marketed ultra-low volume (ULV) mosquito adulticide. Volatilized prallethrin is intended to stimulate mosquito flight, increasing its adulticide effectiveness. However, field tests using volatilized prallethrin have not produced significant differences in mosquito trap catches, leading to questions regarding prallethrin's behavioral impact efficacy. Thus, we conducted laboratory tests of prallethrin's effect on flight behavior of adult female Asian tiger mosquitoes, Aedes albopictus. Mosquitoes were divided into 3 groups: untreated control, exposed to volatilized prallethrin, and exposed to a liquid spray calibrated to simulate a ULV application at label rates. After exposure, mosquito behavior in an airstream of 0.5 m/sec was recorded and analyzed using motion-tracking software. No significant differences in flight behavior were found between the control and treated mosquitoes exposed to volatilized prallethrin. The ULV-sprayed mosquitoes exhibited a significant increase in the number of flight events, the turning frequency, overall movement speed, and flight speed compared to the control-a significant difference in locomotor stimulation response that would increase exposure to a ULV spray cloud. However, our results showed that volatilization alone was insufficient to increase ULV efficacy in the field and suggested that incorporating a more volatile flight stimulant into ULV adulticides would provide a measurable improvement in mosquito control.

Research has recently demonstrated that larval zebrafish show similar molecular responses to nociception to those of adults. Our study explored whether unprotected larval zebrafish exhibited altered behaviour after exposure to noxious chemicals and screened a range of analgesic drugs to determine their efficacy to reduce these responses. This approach aimed to validate larval zebrafish as a reliable replacement for adults as well as providing a high-throughput means of analysing behavioural responses. Zebrafish at 5 days post-fertilization were exposed to known noxious stimuli: acetic acid (0.01%, 0.1% and 0.25%) and citric acid (0.1%, 1% and 5%). The behavioural response of each was recorded and analysed using novel tracking software that measures time spent active in 25 larvae at one time. Subsequently, the efficacy of aspirin, lidocaine, morphine and flunixin as analgesics after exposure to 0.1% acetic acid was tested. Larvae exposed to 0.1% and 0.25% acetic acid spent less time active, whereas those exposed to 0.01% acetic acid and 0.1–5% citric acid showed an increase in swimming activity. Administration of 2.5 mg l−1 aspirin, 5 mg l−1 lidocaine and 48 mg l−1 morphine prevented the behavioural changes induced by acetic acid. These results suggest that larvae respond to a noxious challenge in a similar way to adult zebrafish and other vertebrates and that the effect of nociception on activity can be ameliorated by using analgesics. Therefore, adopting larval zebrafish could represent a direct replacement of a protected adult fish with a non-protected form in pain- and nociception-related research.

A phenotypic syndrome refers to complex patterns of integration among functionally related traits in an organism that defines how the organism interacts with its environment and sustains itself. Human‐induced biological invasions have become important sources of environmental modifications. However, the extent to which invasive species affect the phenotypic syndromes of individuals in a native is currently unknown. Such knowledge has important implications for understanding ecological interactions and the management of biological invasions. Here, field monitoring in a natural stream were combined with standardized estimates of behavioral, physiological and morphological traits to address the hypothesis that coexistence with a non‐native invader induces a novel environmental pressure that disrupts the adaptive integration among phenotypic traits of the native species. We compared the strength of integration among key phenotypic traits (i.e. aerobic scope, standard metabolic rate, body growth, activity, and body shape) and ecological niche traits (i.e. spring and summer diet, home range size, daily movements) of an allopatric group of native brown trout ( Salmo trutta ) with a group of brown trout living in sympatry with non‐native brook trout ( Salvelinus fontinalis ). We found that the integration of phenotypic traits was substantially reduced in the sympatric brown trout and that allopatric and sympatric brown trout differed in key phenotypic and ecological niche traits. Brown trout living in sympatry with non‐native brook trout consumed more terrestrial prey, had smaller home ranges, and a stouter body shape. Sympatric brown trout also had lower specific growth rate, suggesting a lower fitness. The results are generally in line with our hypothesis suggesting that the reduction in fitness observed in sympatric brown trout is caused by the breakdown of their adaptive phenotypic syndrome. This may be caused by differences in the plasticity of the response of phenotypic traits to the novel selection pressure induced by the non‐native species. Our results may help explaining deleterious effects of non‐native species reported in the absence of direct competition with the native species. A lay summary is available for this article.

Permethrin, a type 1 pyrethroid insecticide, was previously reported to promote adipogenesis in 3T3-L1 adipocytes and insulin resistance in C2C12 muscle cells; however, the effects of permethrin exposure on glucose and lipid metabolisms in vivo remain unknown. The purpose of this study was to investigate the effects of permethrin exposure on glucose and lipid homeostasis as well as voluntary movement in female mice in response to dietary fat. We tested three doses of permethrin (50, 500, & 5000 μg/kg body weight/day) in low fat diet-fed (4% w/w of diet) and high fat diet-fed (20% w/w of diet) female C57BL/6J mice for twelve weeks. Our results demonstrated that permethrin treatment potentiated high fat diet-induced insulin resistance as indicated by insulin tolerance tests, glucose tolerance tests, and homeostasis model assessment - insulin resistance (HOMA-IR) without altering weight or fat mass. Permethrin treatment significantly decreased voluntary movement and elevated blood glucose and insulin levels. Western blot results further showed that permethrin impaired insulin signaling via the Akt signaling pathway in the gastrocnemius muscle. Taken together, these results suggest that oral administration of permethrin potentiated high fat diet-induced insulin resistance, possibly increasing the risk of type 2 diabetes without altering weight gain in female C57BL/6J mice. Keywords: Permethrin, insecticide, glucose metabolism, voluntary activities

Many species are migratory, resulting in a life cycle divided into periodic stages occurring in different habitats occupied for a limited amount of time. Estimating the time spent in each habitat is crucial to understanding how individuals modulate their activities and thus to evolutionary ecology and conservation biology. Several methods, including some recent promising advances, can be used to estimate stopover duration as well as arrival and departure probabilities at sites where individuals are monitored using capture–recapture sampling. Our objectives in this study were to (1) describe the available models to estimate stopover duration, (2) illustrate with an original data set what kinds of questions can be addressed using the most recent methods, and (3) to provide in a detailed appendix a practical guide for implementing these methods in E‐ SURGE software. To illustrate the potential of these models for testing biological hypotheses, we used a capture–recapture data set on marbled newts ( Triturus marmoratus ). We used time‐dependent and time‐elapsed‐since‐arrival effects (using both Markovian and semi‐Markov processes for the latter) to model stopover duration and the probability of arriving in and departing from a breeding pond for this species and compared the relative performance of the resulting models. Our findings showed a strong sex effect on stopover duration: females stayed on average 5.63 weeks in a breeding pond whereas males stayed only 3.03 weeks. In both sexes, the retention probability was mainly influenced by the time already spent there. Consequently, individuals of the same sex stayed a similar amount of time in a pond, although they did not arrive simultaneously but successively. The selected data set demonstrated the flexibility of these methods and their potential relevance for applications in evolutionary ecology and conservation.

Temperature influences the geographic range, physiology, and behavior of many ectothermic species, including the invasive lionfish Pterois sp. Thermal parameters were experimentally determined for wild-caught lionfish at different acclimation temperatures (13, 20, 25 and 32 °C). Preferences and avoidance were evaluated using a videographic shuttlebox system, while critical thermal methodology evaluated tolerance. The lionfish thermal niche was compared experimentally to two co-occurring reef fishes (graysby Cephalopholis cruentata and schoolmaster Lutjanus apodus) also acclimated to 25 °C. The physiologically optimal temperature for lionfish is likely 28.7 ± 1 °C. Lionfish behavioral thermoregulation was generally linked to acclimation history; tolerance and avoidance increased significantly at higher acclimation temperatures, but final preference did not. The tolerance polygon of lionfish shows a strong correlation between thermal limits and acclimation temperature, with the highest CTmax at 39.5 °C and the lowest CTmin at 9.5 °C. The tolerance range of invasive lionfish (24.61 °C) is narrower than those of native graysby (25.25 °C) and schoolmaster (26.87 °C), mostly because of lower thermal maxima in the former. Results show that lionfish display “acquired” thermal tolerance at higher and lower acclimation temperatures, but are no more eurythermal than other tropical fishes. Collectively, these results suggest that while lionfish range expansion in the western Atlantic is likely over the next century from rising winter sea temperatures due to climate change, the magnitude of poleward radiation of this invasive species is limited and will likely be equivalent to native tropical and subtropical fishes with similar thermal minima.

Freshwater fish behaviors have the potential to be impacted by acidification due to increases in dissolved carbon dioxide (CO2). Recent work in the marine environment suggests that increased CO2 levels due to climate change can negatively affect fishes homing to natal environments, while also hindering their ability to detect predators and perform aerobically. The potential for elevated CO2 to have similar negative impacts on freshwater communities remains understudied. The objective of our study was to quantify the effects of elevated CO2 on the behaviors of fathead minnows (Pimephales promelas) and silver carp (Hypophthalmichthys molitrix) following exposure to conspecific skin extracts (alarm cues). In fathead minnows, their response to conspecific skin extracts was significantly impaired following exposure to elevated CO2 levels for at least 96 h, while silver carp behaviors were unaltered. However, fathead minnow behaviors did return to pre-CO2 exposure in high-CO2-exposed fish following 14 days of holding at ambient CO2 levels. Overall, this study suggests there may be potential impacts to freshwater fishes alarm cue behaviors following CO2 exposure, but these responses may be species-specific and will likely be abated should the CO2 stressor be removed.

Aquatic hypercarbia, either naturally occurring or anthropogenically induced, can have extensive impacts on aquatic environments and resident organisms. While the impact of acute hypercarbia exposure on the behavior and physiology of fishes has been well studied, relatively little work has examined the physiological impact and acclimation capacity of fishes to chronic hypercarbia. To better understand the impacts of prolonged hypercarbia exposure, largemouth bass were held at ambient CO2 (13 mg L−1) and elevated CO2 (31 mg L−1; ≈21,000 µatm) for 58 days. Following this acclimation period, fish were subjected to three separate, yet complementary, experiments: (1) acute hypercarbia challenge of 120 mg L−1 CO2 for 1 h to quantify physiological and molecular responses; (2) hypercarbia avoidance challenge to compare CO2 agitation and avoidance responses; and (3) swim performance challenge to quantify burst swimming performance. Acclimation to 31 mg L−1 CO2 resulted in a significant constitutive upregulation of c-fos expression in erythrocytes, combined with significant constitutive expression of hsp70 in both gill and erythrocytes, relative to controls. Largemouth bass acclimated to elevated CO2 also had a reduced glucose response (relative to controls) following an acute CO2 exposure, indicating a reduced stress response to CO2 stressors. In addition, largemouth bass acclimated to elevated CO2 conditions required 50 % higher CO2 concentrations to illicit agitation behaviors and displayed prolonged burst swimming abilities in high CO2 environments relative to controls. Together, results demonstrate that largemouth bass exposed to chronic hypercarbia may possess a physiological advantage during periods of elevated CO2 relative to naïve fish, which may permit increased performance in hypercarbia.

Ongoing climate change is affecting animal physiology in many parts of the world. Using metabolism, the oxygen- and capacity-limitation of thermal tolerance (OCLTT) hypothesis provides a tool to predict the responses of ectothermic animals to variation in temperature, oxygen availability and pH in the aquatic environment. The hypothesis remains controversial, however, and has been questioned in several studies. A positive relationship between aerobic metabolic scope and animal activity would be consistent with the OCLTT but has rarely been tested. Moreover, the performance model and the allocation model predict positive and negative relationships, respectively, between standard metabolic rate and activity. Finally, animal activity could be affected by individual morphology because of covariation with cost of transport. Therefore, we hypothesized that individual variation in activity is correlated with variation in metabolism and morphology. To test this prediction, we captured 23 wild European perch (Perca fluviatilis) in a lake, tagged them with telemetry transmitters, measured standard and maximal metabolic rates, aerobic metabolic scope and fineness ratio and returned the fish to the lake to quantify individual in situ activity levels. Metabolic rates were measured using intermittent flow respirometry, whereas the activity assay involved high-resolution telemetry providing positions every 30 s over 12 days. We found no correlation between individual metabolic traits and activity, whereas individual fineness ratio correlated with activity. Independent of body length, and consistent with physics theory, slender fish maintained faster mean and maximal swimming speeds, but this variation did not result in a larger area (in square metres) explored per 24 h. Testing assumptions and predictions of recent conceptual models, our study indicates that individual metabolism is not a strong determinant of animal activity, in contrast to individual morphology, which is correlated with in situ activity patterns.

Global warming results in increasing water temperature, which may represent a threat to aquatic ectotherms. The rising temperature affects ecology through physiology, by exerting a direct limiting effect on the individual. The mechanism controlling individual thermal tolerance is still elusive, but some evidence shows that the heart plays a central role, and that insufficient transport of oxygen to the respiring tissues may determine the thermal tolerance of animals. In this study, the influence of the heart in thermal limitation was investigated by measurements of aerobic scope in the European eel ( Anguilla anguilla ) together with measurements of cardiac output during rest and activity. Aerobic capacity was not limited by an acutely increased temperature in the European eel. Oxygen demand was met by an increase in heart rate and arteriovenous extraction. These findings suggest that thermal tolerance during exposure to acute temperature changes is not defined by oxygen transport capacity in the eel, and other mechanisms may play a central role in limiting thermal tolerance in these fish.

Aquatic chemical ecology is an important and growing field of research that involves understanding how organisms perceive and respond to chemical cues in their environment. Research assessing the preference or avoidance of a water source containing specific chemical cues has increased in popularity in recent years, and a variety of methods have been described in the scientific literature. Two‐current choice flumes have seen the greatest increase in popularity, perhaps because of their potential to address the broadest range of research questions. Here, we review the literature on two‐current choice flumes and show that there is a clear absence of standardized methodologies that make comparisons across studies difficult. Some of the main issues include turbulent flows that cause mixing of cues, inappropriate size of choice arenas for the animals, short experiments with stressed animals, failure to report how experiment and researcher biases were eliminated, general underreporting of methodological details, underutilization of collected data and inappropriate data analyses. In this review, we present best practice guidelines on how to build, test and use two‐current choice flumes to measure the behavioural responses of aquatic animals to chemical cues, and provide blueprints for flume construction. The guidelines include steps that can be taken to avoid problems commonly encountered when using two‐current choice flumes and analysing the resulting data. This review provides a set of standards that should be followed to ensure data quality, transparency and replicability in future studies in this field.

Salinity preference and responses to predatory chemical cues were examined both separately and simultaneously in freshwater ( FW ) and saltwater ( SW )‐acclimated sailfin mollies Poecilia latipinna, a euryhaline species. It was hypothesized that P. latipinna would prefer FW over SW, move away from chemical cues from a crayfish predator, and favour predator avoidance over osmoregulation when presented with both demands. Both FW and SW ‐acclimated P. latipinna preferred FW and actively avoided predator cues. When presented with FW plus predator cues v. SW with no cues, P. latipinna were more often found in FW plus predator cues. These results raise questions pertaining to the potential osmoregulatory stress of salinity transitions in euryhaline fishes relative to the potential fitness benefits and whether euryhalinity is utilized for predator avoidance. This study sheds light on the potential benefits and consequences of being salt tolerant or intolerant and complicates the understanding of the selection pressures that have favoured the different osmoregulatory mechanisms among fishes.

Previous studies hailed thermal tolerance and the capacity for organisms to acclimate and adapt as the primary pathways for species survival under climate change. Here we challenge this theory. Over the past decade, more than 365 tropical stenothermal fish species have been documented moving poleward, away from ocean warming hotspots where temperatures 2–3 °C above long‐term annual means can compromise critical physiological processes. We examined the capacity of a model species – a thermally sensitive coral reef fish, Chromis viridis (Pomacentridae) – to use preference behaviour to regulate its body temperature. Movement could potentially circumvent the physiological stress response associated with elevated temperatures and may be a strategy relied upon before genetic adaptation can be effectuated. Individuals were maintained at one of six temperatures (23, 25, 27, 29, 31 and 33 °C) for at least 6 weeks. We compared the relative importance of acclimation temperature to changes in upper critical thermal limits, aerobic metabolic scope and thermal preference. While acclimation temperature positively affected the upper critical thermal limit, neither aerobic metabolic scope nor thermal preference exhibited such plasticity. Importantly, when given the choice to stay in a habitat reflecting their acclimation temperatures or relocate, fish acclimated to end‐of‐century predicted temperatures (i.e. 31 or 33 °C) preferentially sought out cooler temperatures, those equivalent to long‐term summer averages in their natural habitats (~29 °C). This was also the temperature providing the greatest aerobic metabolic scope and body condition across all treatments. Consequently, acclimation can confer plasticity in some performance traits, but may be an unreliable indicator of the ultimate survival and distribution of mobile stenothermal species under global warming. Conversely, thermal preference can arise long before, and remain long after, the harmful effects of elevated ocean temperatures take hold and may be the primary driver of the escalating poleward migration of species.

Cubomedusae all have a similar set of six eyes on each of their four rhopalia. Still, there is a great variation in activity patterns with some species being strictly day active while others are strictly night active. Here we have examined the visual ecology of the medusa of the night active Copula sivickisi from Okinawa using optics, morphology, electrophysiology, and behavioral experiments. We found the lenses of both the upper and the lower lens eyes to be image forming but under-focused, resulting in low spatial resolution in the order of 10–15°. The photoreceptor physiology is similar in the two lens eyes and they have a single opsin peaking around 460 nm and low temporal resolution with a flicker fusion frequency (fff) of 2.5 Hz indicating adaptions to vision in low light intensities. Further, the outer segments have fluid filled swellings, which may concentrate the light in the photoreceptor membrane by total internal reflections, and thus enhance the signal to noise ratio in the eyes. Finally our behavioral experiments confirmed that the animals use vision when hunting. When they are active at night they seek out high prey-concentration by visual attraction to areas with abundant bioluminescent flashes triggered by their prey.

The hypothesis that interindividual differences in the activity of brown trout alter the exposure to parasitic freshwater pearl mussel glochidia was tested in a Swedish stream. Wild yearling brown trout (N = 103) were caught, individually tagged for identification and scored for open‐field activity during standardized laboratory tests in June. Fifty gravid freshwater pearl mussels were relocated to the stream, where after the trout were released back into the stream. The fish were recaptured in October (N = 35), checked for glochidia encystment (infested individuals: n = 6) and re‐scored for open‐field activity traits. Swimming velocity during the test was higher in fish infected with glochidia, suggesting that high activity could increase their exposure to glochidia. Potentially, as metabolism rate and ventilation rate typically increase with activity, elevated activity may lead to an increased likelihood of glochidia passing over the gills. This novel finding suggests that glochidia infestation is non‐random and that the behaviour of the host fish can influence the likelihood of glochidia infestation.

The lampricide, 3-trifluoromethyl-4-nitrophenol (TFM), is a primary component to sea lamprey control in the Laurentian Great Lakes. Though the lethal effects of TFM are well-known, the sublethal effects on fishes are virtually unknown. Here we studied the effects of TFM on the olfactory capabilities and behavior of young-of-the-year (YOY) lake sturgeon (Acipenser fulvescens). At ecologically relevant concentrations of TFM there was reduced olfactory response to all three cues (l-alanine, taurocholic acid, food cue) tested, suggesting that TFM inhibits both olfactory sensory neurons tested. Sturgeon exposed to TFM also showed a reduced attraction to the scent of food and reduced consumption of food relative to unexposed fish. Exposed fish were more active than control fish, but with slower acceleration. Fish were able to detect the scent of TFM, but failed to avoid it in behavioral trials. The connection between neurophysiological and behavioral changes, and the commonality of habitats between sturgeon and lamprey ammocoetes, suggests that there may be effects at the ecosystem level in streams that undergo lamprey control treatments.

Habitat heterogeneity is a key driver of biodiversity in many ecosystems. Wildlife inhabiting the native prairies of North America evolved in a heterogeneous mosaic of habitat conditions created by fire and grazing by native ungulates. Current rangeland management practices in the tallgrass prairie ecosystem evenly distribute fire and grazing across management units and promote homogeneous habitat conditions. Patch‐burn grazing is a rangeland management strategy that seeks to restore heterogeneity to rangelands via fire‐grazing interactions. Our 3.5‐year study tested the effects of patch‐burn grazing on habitat heterogeneity and small mammal community dynamics in the Flint Hills ecoregion of eastern Kansas. To study the ecological effects of patch‐burn grazing, we sampled habitat conditions and the small mammal community. We assessed habitat conditions once each growing season in a negative control that was annually burned and grazed, a positive control that was burned every four years and ungrazed, and within each of three units of a patch‐burn grazing experiment (PBG) managed with rotational fire. Habitat conditions were significantly different among treatments, and a principal components analysis showed that the patch‐burn grazing treatment had higher canopy cover of forbs and habitat heterogeneity than our two control units. To sample the small mammal community, we conducted monthly live trapping of small mammals on two randomly located trap grids in each of our two controls and three units of our PBG treatment. Small mammal diversity was significantly higher in the patch‐burn grazing treatment and in the positive control, vs. the negative control. Moreover, a canonical correspondence analysis showed that a fire‐grazing interaction was the major driver structuring small mammal communities. Patch‐burn grazing is an effective strategy for restoring heterogeneity to vegetative structure and composition, and can increase biodiversity of small mammals in managed rangelands in the tallgrass prairie ecosystem.

Theory suggests that high activity levels in animals increase growth at the cost of increased mortality. This growth‐mortality tradeoff has recently been incorporated into the wider framework of the pace‐of‐life syndrome ( POLS ) hypothesis. However, activity is often quantified only in the laboratory and on a diurnal basis, leaving open the possibility that animals manage predation risk and feeding efficiency in the wild by modulating their circadian activity rhythms. Here we investigate how laboratory activity in wild brown trout parr ( Salmo trutta L.) associates with circadian activity, growth, and mortality in their natal stream. We found that individuals with high activity in the laboratory displayed high dispersal and cathemeral activity in their natal stream. In contrast, trout with low laboratory activity showed variation of activity in the wild, which was negatively related to the light intensity. Our results do not support the growth‐mortality trade‐off of the POLS hypothesis as highly active, fast‐growing individuals showed higher survival than inactive conspecifics. These novel results show for the first time that active and inactive individuals, as scored in the lab, can show different circadian patterns of behavior in the wild driven by light intensity. This implies that studies conducted under a narrow range of light conditions can bias our understanding of individual behavioral variation and its fitness consequences in the wild.

Water temperature influences the behavior and distribution patterns of both larval and adult American lobster Homarus americanus. However, very little is known about the responses of juvenile lobsters. The juvenile life stage is a critical period; high levels of mortality, combined with specific behavioral responses, can disconnect larval settlement from patterns of abundance of adults. We assessed behavioral thermoregulation in juvenile lobsters, and determined how thermal preferences can be altered by the presence of shelter and food. Juvenile lobsters avoided temperatures higher than 20 °C and lower than 8 °C, and had a mean temperature preference of 16.2 ± 1 °C. This preference was unaffected by prior acclimation, origin (laboratory-raised or wild), or size. When the animals were subjected to a temperature change (5-20 °C), activity rates peaked at 15 °C, and remained stable thereafter. Activity rates did not change when a shelter was added. The addition of food resulted in an increase in activity associated with food handling. When juvenile lobsters were offered a choice between temperature, shelter, and food, they always chose the environment with a shelter, even when it was in a thermally unfavorable temperature. Juveniles also spent more time in a thermally unfavorable environment when food was present; however, acquisition of a shelter was prioritized over food. Although juveniles had a similar thermal preference to adults, they are more vulnerable to predation; the innate shelter-seeking behavior of juveniles overrode their thermal preference. While temperature is an important environmental factor affecting the physiology, distribution, and growth of aquatic ectotherms, our findings suggest that trade-off behaviors occur in order to maintain optimal fitness and survival of the individual.

The lemon shark is a large coastal species that is currently classified as Near Threatened by the International Union for the Conservation of Nature. As juveniles of the species utilize shallow, near shore habitat, they are particularly susceptible to anthropogenic impacts. The abundance, size characteristics, growth, condition factor and movement patterns of juvenile lemon sharks within the Bell Sound Nature Reserve were compared with those outside the Reserve. Sharks were more abundant within the Reserve, which also accommodated a greater proportion of smaller size classes. Although there was no difference in linear growth‐rate between sharks inside and outside the Reserve, those outside exhibited higher condition factor values. It is suggested that these results are due mainly to habitat preference on the part of the sharks, with the predominantly shallower near shore waters of the Reserve offering a greater level of protection from predation. Regardless of the causative factor, the area occupied by the Bell Sound Nature Reserve appears to be highly important to the local juvenile lemon shark population. Copyright © 2016 John Wiley & Sons, Ltd.

Bigheaded carps are non-native invasive fishes that have quickly become the most abundant fishes in many portions of the Midwestern United States. While the spread of bigheaded carps into the Great Lakes is currently impeded by three electrified barriers, these fish have the potential to negatively impact the Great Lakes ecosystem if this barrier is breached, and these barriers may be particularly vulnerable to the passage of small fishes. As such, novel barrier technologies would provide an additional mechanism to prevent bigheaded carps from invading the Great Lakes, and provide much needed redundancy to the current electric barrier. The current study used a combination of molecular and behavioral experiments to determine the effectiveness of carbon dioxide as a chemical deterrent for larval and juvenile fishes, with an emphasis on bigheaded carps. Juvenile silver carp (Hypophthalmichthys molitrix), bighead carp (H. nobilis), bluegill (Lepomis macrochirus) and largemouth bass (Micropterus salmoides) showed avoidance of elevated CO2 environments at approximately 200 mg/L. Additionally, exposure to 120 mg/L CO2 resulted in the induction of hsp70 mRNA in 8 days old silver carp fry, while gill c-fos transcripts increased following hypercarbia exposure in all juvenile species examined. Together, our results show that CO2 has potential to deter the movement of larval and juvenile fishes.

Biologging technology has enhanced our understanding of the ecology of marine animals and has been central to identifying how oceanographic conditions drive patterns in their distribution and behavior. Among these environmental influences, there is increasing recognition of the impact of dissolved oxygen on the distribution of marine animals. Understanding of the impact of oxygen on vertical and horizontal movements would be advanced by contemporaneous in situ measurements of dissolved oxygen from animal-borne sensors instead of relying on environmental data that may not have appropriate spatial or temporal resolution. Here, we demonstrate the capabilities of dissolved oxygen pop-up satellite archival tags (DO-PATs) by presenting the results from calibration experiments and trial deployments of two prototype tags on bluntnose sixgill sharks (Hexanchus griseus). The DO-PATs provided fast, accurate, and stable measurements in calibration trials and demonstrated high correlation with vertical profiles obtained via traditional ship-borne oceanographic instruments. Deployments on bluntnose sixgill sharks recorded oxygen saturations as low as 9.4 % and effectively captured the oceanography of the region when compared with World Ocean Atlas 2013 values. This is the first study to use an animal-borne device to autonomously measure and record in situ dissolved oxygen saturation from non-air-breathing marine animals. The DO-PATs maintained consistency over time and yielded measurements equivalent to industry standards for environmental sampling. Acquiring contemporaneous in situ measurements of dissolved oxygen saturation alongside temperature and depth data will greatly improve our ability to investigate the spatial ecology of marine animals and make informed predictions of the impacts of global climate change. The information returned from DO-PATs is relevant not only to the study of the ecology of marine animals but will also become a useful new tool for investigating the physical structure of the oceans.

Intraspecific variation and trade-off in aerobic and anaerobic traits remain poorly understood in aquatic locomotion. Using gilthead sea bream (Sparus aurata) and Trinidadian guppy (Poecilia reticulata), both axial swimmers, this study tested four hypotheses: (1) gait transition from steady to unsteady (i.e., burst-assisted) swimming is associated with anaerobic metabolism evidenced as excess post exercise oxygen consumption (EPOC); (2) variation in swimming performance (critical swimming speed; U crit) correlates with metabolic scope (MS) or anaerobic capacity (i.e., maximum EPOC); (3) there is a trade-off between maximum sustained swimming speed (U sus) and minimum cost of transport (COTmin); and (4) variation in U sus correlates positively with optimum swimming speed (U opt; i.e., the speed that minimizes energy expenditure per unit of distance traveled). Data collection involved swimming respirometry and video analysis. Results showed that anaerobic swimming costs (i.e., EPOC) increase linearly with the number of bursts in S. aurata, with each burst corresponding to 0.53 mg O2 kg(-1). Data are consistent with a previous study on striped surfperch (Embiotoca lateralis), a labriform swimmer, suggesting that the metabolic cost of burst swimming is similar across various types of locomotion. There was no correlation between U crit and MS or anaerobic capacity in S. aurata indicating that other factors, including morphological or biomechanical traits, influenced U crit. We found no evidence of a trade-off between U sus and COTmin. In fact, data revealed significant negative correlations between U sus and COTmin, suggesting that individuals with high U sus also exhibit low COTmin. Finally, there were positive correlations between U sus and U opt. Our study demonstrates the energetic importance of anaerobic metabolism during unsteady swimming, and provides intraspecific evidence that superior maximum sustained swimming speed is associated with superior swimming economy and optimum speed.

Changes in behavioral traits exhibited by small aquatic invertebrates are increasingly postulated as ethically acceptable and more sensitive endpoints for detection of water-born ecotoxicity than conventional mortality assays. Despite importance of such behavioral biotests, their implementation is profoundly limited by the lack of appropriate biocompatible automation, integrated optoelectronic sensors, and the associated electronics and analysis algorithms. This work outlines development of a proof-of-concept miniaturized Lab-on-a-Chip (LOC) platform for rapid water toxicity tests based on changes in swimming patterns exhibited by Artemia franciscana (Artoxkit MTM) nauplii. In contrast to conventionally performed end-point analysis based on counting numbers of dead/immobile specimens we performed a time-resolved video data analysis to dynamically assess impact of a reference toxicant on swimming pattern of A. franciscana. Our system design combined: (i) innovative microfluidic device keeping free swimming Artemia sp. nauplii under continuous microperfusion as a mean of toxin delivery; (ii) mechatronic interface for user-friendly fluidic actuation of the chip; and (iii) miniaturized video acquisition for movement analysis of test specimens. The system was capable of performing fully programmable time-lapse and video-microscopy of multiple samples for rapid ecotoxicity analysis. It enabled development of a user-friendly and inexpensive test protocol to dynamically detect sub-lethal behavioral end-points such as changes in speed of movement or distance traveled by each animal.

Whirling disease, caused by the myxozoan parasite Myxobolus cerebralis, has high economical and ecological importance worldwide. Susceptibility to the disease varies considerably among salmonid species. In brown trout (Salmo trutta) the infection is usually subclinical with low mortality, which increases the risk of parasite dissemination, especially when farm fish are used for stocking natural habitats. The influence of intraspecific genetic differences (especially the level of homozygosity) on susceptibility is unknown. Therefore, we examined the possible correlations between parental genetic diversity and offspring susceptibility of brown trout stocks to whirling disease. Two brown trout brood stocks from a German and a Hungarian fish farm were genetically characterized using microsatellite and lineage-specific genetic markers. The individual inbreeding coefficient f and pairwise relatedness factor r were estimated based on eight microsatellite markers. Brood stock populations were divided into groups according to low and high f and r value estimates and subjected to selective fertilization. The offspring from these separate groups were exposed to M. cerebralis actinospores, and the infection prevalence and intensity was measured and statistically analysed. The analysis of phylogeographic lineage heritage revealed high heterogeneity in the Hungarian brood stock since > 50% of individuals were Atlantic-Danubian hybrids, while only pure Atlantic-descending specimens were detected in the German population. Based on f msat and r msat estimations, classified non-inbred (NIB), inbred (IB) and a group of closely related fish (REL) were created. The susceptibility of their offspring varied considerably. Although there was no significant difference in the prevalence of M. cerebralis infection, the mean intensity of infection differed significantly between NIB and IB groups. In REL and IB groups, a high variability was observed in infection intensity. No external clinical signs were observed in the exposed brown trout groups. Our findings indicate that the allelic diversity of brown trout brood stock may constitute a significant factor in disease susceptibility, i.e. the intensity of parasite infection in the subsequent generation.

There are controversies whether learning of conditioned eyeblink responses primarily takes place within the cerebellar cortex, the interposed nuclei, or both. It has also been suggested that the cerebellar cortex may be important during early stages of learning, and that there is a shift to the cerebellar nuclei during later stages. As yet, human studies have provided little to resolve this question. In the present study, we established a setup that allows ultra-high-field 7T functional magnetic resonance imaging (fMRI) of the cerebellar cortex and interposed cerebellar nuclei simultaneously during delay eyeblink conditioning in humans. Event-related fMRI signals increased concomitantly in the cerebellar cortex and nuclei during early acquisition of conditioned eyeblink responses in 20 healthy human subjects. ANOVAs with repeated-measures showed significant effects of time across five blocks of 20 conditioning trials in the cortex and nuclei ( p

This study investigated the relationships between behavioural responses of Atlantic salmon Salmo salar smolts to saltwater ( SW ) exposure and physiological characteristics of smolts in laboratory experiments. It concurrently described the behaviour of acoustically tagged smolts with respect to SW and tidal cycles during estuary migration. Salmo salar smolts increased their use of SW relative to fresh water ( FW ) from April to June in laboratory experiments. Mean preference for SW never exceeded 50% of time in any group. Preference for SW increased throughout the course of smolt development. Maximum continuous time spent in SW was positively related to gill Na +, K + ‐ ATPase ( NKA ) activity and osmoregulatory performance in full‐strength SW (measured as change in gill NKA activity and plasma osmolality). Smolts decreased depth upon reaching areas of the Penobscot Estuary where SW was present, and all fish became more surface oriented during passage from head of tide to the ocean. Acoustically tagged, migrating smolts with low gill NKA activity moved faster in FW reaches of the estuary than those with higher gill NKA activity. There was no difference in movement rate through SW reaches of the estuary based on gill NKA activity. Migrating fish moved with tidal flow during the passage of the lower estuary based on the observed patterns in both vertical and horizontal movements. The results indicate that smolts select low‐salinity water during estuary migration and use tidal currents to minimize energetic investment in seaward migration. Seasonal changes in osmoregulatory ability highlight the importance of the timing of stocking and estuary arrival.

Conjugated linoleic acid (CLA) has been reported to prevent body weight gain and fat accumulation in part by improving physical activity in mice. However, the effects of postweaning administration of CLA on the development of obesity later in life have not yet been demonstrated. The current study investigated the role of postweaning CLA treatment on skeletal muscle energy metabolism in genetically induced inactive adult-onset obese model, nescient basic helix-loop-helix 2 knockout (N2KO) mice. Four-week-old male N2KO and wild type mice were fed either control or a CLA-containing diet (0.5%) for 4 weeks, and then CLA was withdrawn and control diet provided to all mice for the following 8 weeks. Postweaning CLA supplementation in wild type animals, but not N2KO mice, may activate AMP-activated protein kinase (AMPK) and peroxisome proliferator-activated receptor-δ (PPARδ) as well as promote desensitization of phosphatase and tensin homologue (PTEN) and sensitization of protein kinase B (AKT) at threonine 308 in gastrocnemius skeletal muscle, improving voluntary activity and glucose homeostasis. We suggest that postweaning administration of CLA may in part stimulate the underlying molecular targets involved in muscle energy metabolism to reduce weight gain in normal animals, but not in the genetically induced inactive adult-onset animal model.

Most studies on the impact of near-future levels of carbon dioxide on fish behaviour report behavioural alterations, wherefore abnormal behaviour has been suggested to be a potential consequence of future ocean acidification and therefore a threat to ocean ecosystems. However, an increasing number of studies show tolerance of fish to increased levels of carbon dioxide. This variation among studies in susceptibility highlights the importance of continued investigation of the possible effects of elevated pCO2. Here, we investigated the impacts of increased levels of carbon dioxide on behaviour using the goldsinny wrasse (Ctenolabrus rupestris), which is a common species in European coastal waters and widely used as cleaner fish to control sea lice infestation in commercial fish farming in Europe. The wrasses were exposed to control water conditions (370 μatm) or elevated pCO2 (995 μatm) for 1 month, during which time behavioural trials were performed. We investigated the possible effects of CO2 on behavioural lateralization, swimming activity, and prey and predator olfactory preferences, all behaviours where disturbances have previously been reported in other fish species after exposure to elevated CO2. Interestingly, we failed to detect effects of carbon dioxide for most behaviours investigated, excluding predator olfactory cue avoidance, where control fish initially avoided predator cue while the high CO2 group was indifferent. The present study therefore shows behavioural tolerance to increased levels of carbon dioxide in the goldsinny wrasse. We also highlight that individual fish can show disturbance in specific behaviours while being apparently unaffected by elevated pCO2 in other behavioural tests. However, using experiments with exposure times measured in weeks to predict possible effects of long-term drivers, such as ocean acidification, has limitations, and the behavioural effects from elevated pCO2 in this experiment cannot be viewed as proof that these fish would show the same reaction after decades of evolution.

Social isolation in early life deregulates the serotonergic system of the brain, compromising reproductive function. Gonadotropin-inhibitory hormone (GnIH) neurons in the dorsomedial hypothalamic nucleus are critical to the inhibitory regulation of gonadotropin-releasing hormone neuronal activity in the brain and release of luteinizing hormone by the pituitary gland. Although GnIH responds to stress, the role of GnIH in social isolation-induced deregulation of the serotonin system and reproductive function remains unclear. We investigated the effect of social isolation in early life on the serotonergic–GnIH neuronal system using enhanced green fluorescent protein (EGFP)-tagged GnIH transgenic rats. Socially isolated rats were observed for anxious and depressive behaviors. Using immunohistochemistry, we examined c-Fos protein expression in EGFP–GnIH neurons in 9-week-old adult male rats after 6 weeks post-weaning isolation or group housing. We also inspected serotonergic fiber juxtapositions in EGFP–GnIH neurons in control and socially isolated male rats. Socially isolated rats exhibited anxious and depressive behaviors. The total number of EGFP–GnIH neurons was the same in control and socially isolated rats, but c-Fos expression in GnIH neurons was significantly reduced in socially isolated rats. Serotonin fiber juxtapositions on EGFP–GnIH neurons were also lower in socially isolated rats. In addition, levels of tryptophan hydroxylase mRNA expression in the dorsal raphe nucleus were significantly attenuated in these rats. These results suggest that social isolation in early-life results in lower serotonin levels, which reduce GnIH neuronal activity and may lead to reproductive failure.

Recently, conjugated linoleic acid (CLA) has been reported to prevent body weight gain and fat accumulation in part by improving physical activity in mice. The current study was conducted to determine the role of CLA on skeletal muscle metabolism in nescient basic helix‐loop‐helix 2 knock‐out (N2KO) mice, an adult‐onset inactivity induced obese model. Five‐week‐old female N2KO and wild type mice were fed either control or CLA containing diet (0.5%) for 10 wk. Voluntary activity was determined biweekly and markers for muscle metabolisms were determined from the gastrocnemius muscle. CLA fed N2KO animals showed significant increased voluntary movement and gastrocnemius muscle mass compared to control group, whereas in wild type animals, no differences were observed. CLA treatment up‐regulated AMP‐activated protein kinase (AMPK), mitochondria biogenic markers, peroxisome proliferator‐activated receptor‐δ (PPARδ), and mitochondrial transcription factor A (Tfam) compared to control animals. These observations indicate that CLA supplementation activates AMPKα‐PPARδ and/or ‐Tfam signaling cascade for stimulating mitochondria biogenesis. Taken together, these results suggest that CLA may in part activate the underlying biomarkers involved in muscle metabolism via stimulation of mitochondrial biogenesis, resulting in increased voluntary activity and muscle mass, potentially contributing to regulating weight gain. Practical applications: Lack of physical activity is a global public health problem, which induces obesity and its associated pathologies. Approximately, 3.2 million deaths per year are attributable to lack of physical activity. CLA has previously been reported to increase voluntary and endurance activities in mice. However, the exact mode of action is not completely understood. Thus, the purpose of the study was to shed light on positive effects of CLA on physical activity through modulation of molecular targets in skeletal muscle of Nhlh2 knockout mice, which are an adult‐onset inactivity induced obesity model. The current results suggest that CLA acts as a potential exercise‐mimetic, resulting in increased voluntary activity and muscle mass, potentially contributing to regulating weight gain. Conjugated linoleic acid partially improves muscle metabolism via stimulation of mitochondrial biogenesis, resulting in increased voluntary activity.

As global temperatures increase, fish populations at low latitudes are thought to be at risk as they are adapted to narrow temperature ranges and live at temperatures close to their thermal tolerance limits. Behavioural movements, based on a preference for a specific temperature (T pref), may provide a strategy to cope with changing conditions. A temperature-sensitive coral reef cardinalfish (Cheilodipterus quinquelineatus) was exposed to 28 °C (average at collection site) or 32 °C (predicted end-of-century) for 6 weeks. T pref was determined using a shuttlebox system, which allowed fish to behaviourally manipulate their thermal environment. Regardless of treatment temperature, fish preferred 29.5 ± 0.25 °C, approximating summer average temperatures in the wild. However, 32 °C fish moved more frequently to correct their thermal environment than 28 °C fish, and daytime movements were more frequent than night-time movements. Understanding temperature-mediated movements is imperative for predicting how ocean warming will influence coral reef species and distribution patterns.

Using rainbow trout Oncorhynchus mykiss, the present study demonstrated that: (1) calcium (Ca) increased the range of copper (Cu) concentrations that O. mykiss avoided; (2) Ca conserved the maintenance of pre‐exposure swimming activity during inescapable acute (10 min) Cu exposure. Data showed that when presented with a choice of Cu‐contaminated water (ranging from 0 to 454 µg Cu l −1 ) and uncontaminated water in a choice tank, O. mykiss acclimated and tested at low Ca concentration (3 mg Ca l −1 ) avoided the 10 µg Cu l −1 only. By contrast, O. mykiss acclimated and tested at high Ca concentration (158 mg Ca l −1 ) avoided all the Cu concentrations ≥37 µg l −1. The Cu avoidance was connected with increased spontaneous swimming speed in the Cu‐contaminated water. When subjected to inescapable Cu exposure (35 µg Cu l −1 ), O. mykiss acclimated and tested at low Ca concentration reduced their spontaneous swimming speed, whereas no response was observed in O. mykiss acclimated and tested at high Ca concentration. Collectively, the data support the conclusion that in O. mykiss the behavioural responses to acute Cu exposure are Ca‐dependent.

Most animals experience temperature variations as they move through the environment. For ectotherms, in particular, temperature has a strong influence on habitat choice. While well studied at the species level, less is known about factors affecting the preferred temperature of individuals; especially lacking is information on how physiological traits are linked to thermal preference and whether such relationships are affected by factors such feeding history and growth trajectory. This study examined these issues in the common minnow Phoxinus phoxinus, to determine the extent to which feeding history, standard metabolic rate ( SMR ) and aerobic scope ( AS ), interact to affect temperature preference. Individuals were either: 1) food deprived (FD) for 21 days, then fed ad libitum for the next 74 days; or 2) fed ad libitum throughout the entire period. All animals were then allowed to select preferred temperatures using a shuttle‐box, and then measured for SMR and AS at 10 °C, estimated by rates of oxygen uptake. Activity within the shuttle‐box under a constant temperature regime was also measured. In both FD and control fish, SMR was negatively correlated with preferred temperature. The SMR of the FD fish was increased compared with the controls, probably due to the effects of compensatory growth, and so these growth‐compensated fish preferred temperatures that were on average 2·85 °C cooler than controls fed a maintenance ration throughout the study. Fish experiencing compensatory growth also displayed a large reduction in activity. In growth‐compensated fish and controls, activity measured at 10 °C was positively correlated with preferred temperature. Individual fish prefer temperatures that vary predictably with SMR and activity level, which are both plastic in response to feeding history and growth trajectories. Cooler temperatures probably allow individuals to reduce maintenance costs and divert more energy towards growth. A reduction in SMR at cooler temperatures, coupled with a decrease in spontaneous activity, could also allow individuals to increase surplus AS for coping with environmental stressors. In warming climates, however, aquatic ectotherms could experience frequent fluctuations in food supply with long‐lasting effects on metabolic rate due to compensatory growth, while simultaneously having limited access to preferred cooler habitats.

The response to tissue damage is a complex process, which involves the coordinated regulation of multiple proteins to ensure tissue repair. In order to investigate the effect of tissue damage in a lower vertebrate, samples were taken from rainbow trout ( Oncorhynchus mykiss ) at day 7 after damage and proteins were separated using 2DE. The experimental design included two groups of rainbow trout, which were fed organic feed either with or without astaxanthin. In total, 96 proteins were found to be affected by tissue damage, clearly demonstrating in this lower vertebrate the complexity and magnitude of the cellular response, in the context of a regenerative process. Using a bioinformatics approach, the main biological function of these proteins were assigned, showing the regulation of proteins involved in processes such as apoptosis, iron homeostasis, and regulation of muscular structure. Interestingly, it was established that exclusively within the astaxanthin feed group, three members of the annexin protein family (annexin IV, V, and VI) were regulated in response to tissue damage.

Dietary restriction (DR) extends lifespan and promotes metabolic health in evolutionary distinct species. DR is widely believed to promote longevity by causing an energy deficit leading to increased mitochondrial respiration. We here show that inhibitors of mitochondrial complex I promote physical activity, stress resistance as well as lifespan of Caenorhabditis elegans despite normal food uptake, i.e. in the absence of DR. However, complex I inhibition does not further extend lifespan in dietarily restricted nematodes, indicating that impaired complex I activity mimics DR. Promotion of longevity due to complex I inhibition occurs independently of known energy sensors, including DAF-16/FoxO, as well as AAK-2/AMPK and SIR-2.1/sirtuins, or both. Consistent with the concept of mitohormesis, complex I inhibition transiently increases mitochondrial formation of reactive oxygen species (ROS) that activate PMK-1/p38 MAP kinase and SKN-1/NRF-2. Interference with this retrograde redox signal as well as ablation of two redox-sensitive neurons in the head of the worm similarly prevents extension of lifespan. These findings unexpectedly indicate that DR extends organismal lifespan through transient neuronal ROS signaling rather than sensing of energy depletion, providing unexpected pharmacological options to promote exercise capacity and healthspan despite unaltered eating habits.

Injection of the seaweed toxin kainic acid (KA) in rats induces a severe status epilepticus initiating complex neuropathological changes in limbic brain areas and subsequently spontaneous recurrent seizures. Although neuropathological changes have been intensively investigated in the hippocampus proper and the dentate gyrus in various seizure models, much less is known about changes in parahippocampal areas. We now established telemetric EEG recordings combined with continuous video monitoring to characterize the development of spontaneous seizures after KA-induced status epilepticus, and investigated associated neurodegenerative changes, astrocyte and microglia proliferation in the subiculum and other parahippocampal brain areas. The onset of spontaneous seizures was heterogeneous, with an average latency of 15 ± 1.4 days (range 3-36 days) to the initial status epilepticus. The frequency of late spontaneous seizures was higher in rats in which the initial status epilepticus was recurrent after its interruption with diazepam compared to rats in which this treatment was more efficient. Seizure-induced neuropathological changes were assessed in the subiculum by losses in NeuN-positive neurons and by Fluoro-Jade C staining of degenerating neurons. Neuronal loss was already prominent 24 h after KA injection and only modestly progressed at the later intervals. It was most severe in the proximal subiculum and in layer III of the medial entorhinal cortex and distinct Fluoro-Jade C labeling was observed there in 75% of rats even after 3 months. Glutamatergic neurons, labeled by in situ hybridization for the vesicular glutamate transporter 1 followed a similar pattern of cell losses, except for the medial entorhinal cortex and the proximal subiculum that appeared more vulnerable. Glutamate decarboxylase65 (GAD65) mRNA expressing neurons were generally less vulnerable than glutamate neurons. Reactive astrocytes and microglia were present after 24 h, however, became prominent only after 8 days and remained high after 30 days. In the proximal subiculum, parasubiculum and entorhinal cortex the number of microglia cells was highest after 30 days. Although numbers of reactive astrocytes and microglia were reduced again after 3 months, they were still present in most rats. The time course of astrocyte and microglia proliferation parallels that of epileptogenesis.

The rising atmospheric CO2 level is continuously driving the dissolution of more CO2 into the oceans, and some emission scenarios project that the surface waters may reach 1000 μatm by the end of the century. It is not known if fish can detect moderately elevated CO2 levels, and if they avoid areas with high CO2. If so, avoidance behaviour to water with high CO2 could affect movement patterns and migrations of fish in the future. It is also being increasingly recognized that fish behaviour can be altered by exposure to CO2. Therefore this study investigated how long-term exposure to elevated pCO2 affects predator avoidance and CO2 avoidance in juvenile Atlantic cod (Gadus morhua). The fish were exposed to control water or CO2-enriched water (1000 μatm) for six weeks before being subjected to tests of behaviour. Despite long term exposure to elevated pCO2 the cod still strongly avoided the smell of a predator. These data are surprising because several coral reef fish have demonstrated reversal of olfactory responses after CO2 exposure, turning avoidance of predator cues into preference for predator cues. Fish from both treatment groups also demonstrated strong avoidance of CO2 when presented with the choice of control or CO2-acidified water, indicating that habituation to the CO2 sensory stimuli is negligible. As Atlantic cod maintained normal behavioural responses to olfactory cues, they may be tolerant to CO2-induced behavioural changes. The results also suggest that despite the long-term exposure to CO2-acidified water, the fish still preferred the control water over CO2-acidified water. Therefore, in the future, fish may alter their movements and migrations in search of waters with a lower CO2 content.

Molecular markers (PCR–RFLP and microsatellite) were used to investigate the genetic background of the only brown trout (Salmo trutta m fario) broodstock in Hungary which due to the hydrogeography of the country should theoretically belong to the Danubian lineage. PCR–RFLP (mitochondrial DNA control region and lactate dehydrogenase and somatolactin genes) as well as microsatellite (BFRO002, OMM1064, Ssa408uos, SsoSL417, SsoSL438) markers were used to distinguish between Danubian and Atlantic lineages of brown trout in the Lillafüred broodstock. Altogether 435 fish were tagged during the experimental period. Due to mortalities, fin clips were collected from 401 individuals. According to the genetic analysis of the mitochondrial DNA, the Danubian haplotype is present in only one individual (0.2 %) of the broodstock. Analysis of the nuclear markers revealed that alleles characteristic of both the Atlantic and the Danubian lineages are found in the population. However, Atlantic alleles dominate throughout the broodstock which is in Hardy–Weinberg equilibrium according to the investigated markers. Results indicate that the original broodstock that was introduced to the farm following its construction in 1933 was of the Atlantic lineage. Although later fish from a local stream were collected and added to the broodstock, the number of these was limited and they were almost exclusively males. Fish from this farm that are stocked by anglers can have a significant genetic impact on trout populations of natural streams.

Arsenite is one of the most toxic chemical substances known and is assumed to exert detrimental effects on viability even at lowest concentrations. By contrast and unlike higher concentrations, we here find that exposure to low‐dose arsenite promotes growth of cultured mammalian cells. In the nematode C. elegans, low‐dose arsenite promotes resistance against thermal and chemical stressors and extends lifespan of this metazoan, whereas higher concentrations reduce longevity. While arsenite causes a transient increase in reactive oxygen species ( ROS ) levels in C. elegans, co‐exposure to ROS scavengers prevents the lifespan‐extending capabilities of arsenite, indicating that transiently increased ROS levels act as transducers of arsenite effects on lifespan, a process known as mitohormesis. This requires two transcription factors, namely DAF ‐16 and SKN ‐1, which employ the metallothionein MTL ‐2 as well as the mitochondrial transporter TIN ‐9.1 to extend lifespan. Taken together, low‐dose arsenite extends lifespan, providing evidence for nonlinear dose‐response characteristics of toxin‐mediated stress resistance and longevity in a multicellular organism.

Understanding the interplay among the external environment, physiology and adaptive behaviour is crucial for understanding how animals survive in their natural environments. The external environment can have wide ranging effects on the physiology of animals, while behaviour determines which environments are encountered. Here, we identified changes in the behavioural selection of external salinity in Fundulus heteroclitus, an estuarine teleost, as a consequence of digesting a meal. Fish that consumed high levels of dietary calcium exhibited a higher preferred salinity compared with unfed fish, an effect that was exaggerated by elevated dietary sodium chloride. The mean swimming speed (calculated as a proxy of activity level) was not affected by consuming a diet of any type. Constraining fish to water of 22 p.p.t. salinity during the digestion of a meal did not alter the amount of calcium that was absorbed across the intestine. However, when denied the capacity to increase their surrounding salinity, the compromised ability to excrete calcium to the water resulted in significantly elevated plasma calcium levels, a potentially hazardous physiological consequence. This study is the first to show that fish behaviourally exploit their surroundings to enhance their ionoregulation during digestion, and to pinpoint the novel role of dietary calcium and sodium in shaping this behaviour. We conclude that in order to resolve physiological disturbances in ion balance created by digestion, fish actively sense and select the environment they inhabit. Ultimately, this may result in transient diet-dependent alteration of the ecological niches occupied by fishes, with broad implications for both physiology and ecology.

Passive integrated transponder tags are commonly used in fisheries science to individually identify fish in studies that assume high fish survival and tag retention rates, and have recently been used in studies on burbot Lota lota, although information on retention and survival rates is unavailable. Burbot (310–676-mm TL) surgically implanted with 23-mm PIT tags had 93% survival and 100% retention over 60 d; the survival of these burbot was not significantly different from that of a control group. This suggests that this marking method is a viable means of individually identifying burbot for short-term studies. Received January 21, 2012; accepted July 4, 2012

Aquatic low O2 (hypoxia) is not uncommon but mobile organisms do not have to suffer low O2 stress if they engage in a safe avoidance strategy by moving away early and quickly. Within a behavioural choice chamber system at 20°C, the Cape silverside (Atherina breviceps), a common species from South Africa, was found to seek out and select high O2 saturated water when water O2 saturations declined to 23% saturation. Atherina breviceps also avoided low O2 with a modest increase in swimming speed but no correlation between hypoxic swimming speed and avoidance could be found implying that heightened swimming speed does not necessarily lead to faster avoidance on an individual basis. The overall low O2 response of A. breviceps was seen to reflect an effective avoidance strategy that is compatible with the survival of this species facing low O2 challenge in the wild.

The supplementation of conjugated linoleic acid (CLA) has been shown to improve endurance by enhancing fat oxidation during exercise in rodents and humans. This study was designed to investigate the isomer-specific effects of CLA on endurance capacity and energy metabolism in mice during exercise. Male 129Sv/J mice were divided into three dietary groups and fed treatment diet for 6 weeks; control, 0.5 % cis-9,trans-11 (c9,t11) CLA, or 0.5 % trans-10,cis-12 (t10,c12) CLA. Dietary t10,c12 CLA induced a significant increase in maximum running time and distance until exhaustion with a dramatic reduction of total adipose depots compared to a control group, but there were no significant changes in endurance with the c9,t11 CLA treatment. Serum triacylglycerol and non-esterified fatty acid concentrations were significantly lower in the t10,c12 fed mice after exercise compared to control and the c9,t11 CLA fed-animals. Glycogen contents in livers of the t10,c12 fed-mice were higher than those in control mice, concomitant with reduction of serum l-lactate level. There were no differences in non-exercise physical activity among all treatment groups. In addition, the mRNA expression levels of carnitine palmitoyl transferase 1β, uncoupling protein 2 and peroxisome proliferator-activated receptor δ (PPARδ) in skeletal muscle during exercise were significantly up-regulated by the t10,c12 CLA but not the c9,t11 CLA. These results suggest that the t10,c12 CLA is responsible for improving endurance exercise capacity by promoting fat oxidation with a reduction of the consumption of stored liver glycogen, potentially mediated via PPARδ dependent mechanisms.

It is well established that the lack of physical activity can lead to weight gain or obesity. However, there is limited information on influences of diet components on physical activity. Thus the purpose of this study was to investigate the role of major dietary components on energy expenditure by affecting nonexercise physical activity in C57BL/6J mice. All mice were assigned to 1 of the following 4 dietary groups based on their body weight and baseline physical activity; low fat/normal protein, high fat/normal protein, low fat/low protein, or low fat/high protein. After 3 mo, the highest weight gain was observed in animals fed with high‐fat/normal‐protein diet, and the caloric intake was significantly lower in low‐fat/high‐protein diet‐fed mice compared to other groups. However, there were no significant changes in nonexercise physical activity during experimental periods in all groups. The respiratory quotient and energy expenditure were not significantly different among the dietary groups. These findings suggest that diet‐induced obesity is not explainable by levels of physical activity and energy expenditure. Practical Application: The understanding the link between diet and nonexercise physical activity would provide important knowledge that will potentially assist appropriate food choices to control obesity and its related health problems.

Sensory quality of fish meat is influenced by many parameters prior to slaughter. In the present study, it was examined if previous infections or damages in the muscle tissue influence product quality parameters in fish. Fillets from rainbow trout ( Oncorhynchus mykiss ) reared in seawater at a commercial fish farm were sensory evaluated for more than a year after recovery following physical tissue damage or infection by the bacterial pathogens Yersinia ruckeri and Vibrio anguillarum. The effect of vaccination was also included as some fish were vaccinated before bacterial challenge. The fish fillets were sensory examined as heat‐treated and cold‐smoked. Heat‐treated fillets from nonvaccinated fish previously infected by V. anguillarum had changed textural characteristics and were less flaky, had a lower oiliness and a higher toughness and fibrousness in comparison with control fish. This article was the first to describe a correlation between previous infections in fish and changes in sensory‐quality parameters. PRACTICAL APPLICATIONS This work contributes with knowledge about sensory‐quality parameters of fish meat after recovery from infections and physical‐tissue damage. Because the results demonstrate an influence on the texture from previous disease, the practical potentials of the results are valuable for the aquaculture industry. In order to minimize the effects of previous diseases on the sensory quality regarding the texture, these fishes should be processed as cold‐smoked instead of being sold as raw meat. The established correlation between disease history stresses the importance of disease prevention in aquaculture production, e.g., vaccination of the fish.

The purpose of this study was to evaluate whether conjugated linoleic acid (CLA) exposure during the developmental period increases voluntary activity, which would influence obesity outcome later in life. The effects of dietary supplementation of 0.5% CLA in a high fat diet were evaluated in nescient basic helix-loop-helix 2 (Nhlh2) knock-out (N2KO) mice, which is a unique animal model representing inactivity-induced obesity in a pre-obese condition. Male wild type and N2KO mice were fed either control or CLA (0.5%) diet for 8 weeks. As expected, control diet fed N2KO animals showed greater body weight with decreased physical activity in the late stage of the experimental period compared with wild type control. Dietary CLA significantly decreased body weight and adipose depots in both wild type and N2KO mice, and the body weights of both genotypes fed CLA were similar during the experimental period. CLA exposure during the developmental period significantly improved the impairment of physical activity in N2KO mice, but the wild type did not show any effect of CLA. In both genotypes, CLA significantly reduced serum triglycerides levels and down-regulated the mRNA expressions of CCAAT/enhancer binding protein α (C/EBPα) and leptin in white adipose tissue. These findings suggest that early CLA exposure could prevent obesity with improved voluntary physical activity in N2KO mice.

Individuals often show consistent behavioural differences where behaviours can form integrated units across functionally different contexts. However, the factors causing and maintaining behavioural syndromes in natural populations remain poorly understood. In this study, we provide evidence for the emergence of a behavioural syndrome during the first months of life in wild brown trout ( Salmo trutta ). Behavioural traits of trout were scored before and after a 2‐month interval covering a major survival bottleneck, whereupon the consistency and covariance of behaviours were analysed. We found that selection favoured individuals with high activity levels in an open‐field context, a personality trait consistent throughout the duration of the experiment. In addition, a behavioural syndrome emerged over the 2 months in the wild, linking activity to aggressiveness and exploration tendency. These novel results suggest that behavioural syndromes can emerge rapidly in nature from interaction between natural selection and behavioural plasticity affecting single behaviours.

Cubozoan medusae have a stereotypic set of 24 eyes, some of which are structurally similar to vertebrate and cephalopod eyes. Across the approximately 25 described species, this set of eyes varies surprisingly little, suggesting that they are involved in an equally stereotypic set of visual tasks. During the day Tripedalia cystophora is found at the edge of mangrove lagoons where it accumulates close to the surface in sun-lit patches between the prop roots. Copula sivickisi (formerly named Carybdea sivickisi) is associated with coral reefs and has been observed to be active at night. At least superficially, the eyes of the two species are close to identical. We studied the diurnal activity pattern of these two species both in the wild and under controlled conditions in laboratory experiments. Despite the very similar visual systems, we found that they display opposite patterns of diurnal activity. T. cystophora is active exclusively during the day, whereas C. sivickisi is actively swimming at night, when it forages and mates. At night T. cystophora is found on the muddy bottom of the mangrove lagoon. C. sivickisi spends the day attached to structures such as the underside of stones and coral skeletons. This species difference seems to have evolved to optimize foraging, since the patterns of activity follow those of the available prey items in their respective habitats.

The frequency of low O2 (hypoxia) has increased in coastal marine areas but how fish avoid deleterious water masses is not yet clear. To assess whether the presence and oxygen pressure (PO2) level of an O2 refuge affects the hypoxia avoidance behaviour of fish, individual Atlantic cod (Gadus morhua L.) were exposed to a range of O2 choices in a 2-way choice chamber at 11.4°C over two different experiments. Cod in the first experiment were allowed access to a fixed O2 refuge (fully air-saturated seawater) whilst oxygen pressure (PO2) on the other side was reduced in steps to a critically low level, i.e. 4.3 kPa—a point where cod can no longer regulate O2 consumption. Under these conditions, cod did not avoid any level of hypoxia and fish swimming speed also remained unchanged. In contrast, strong avoidance reactions were exhibited in a second experiment when fish were again exposed to 4.3 kPa but the safety, i.e. PO2, of the refuge was reduced. Fish not only spent less time at 4.3 kPa as a result of fewer sampling visits but they also swam at considerably slower speeds. The presence of an avoidance response was thus strongly related to refuge PO2 and it is unlikely that cod, and possibly other fish species, would enter low O2 to feed in the wild if a sufficiently safe O2 refuge was not available. It is therefore hypothesized that the feeding range of fish may be heavily compressed if hypoxia expands and intensifies in future years.

The objective of this study was to identify behavioural adjustments leading to avoidance of hypoxia. Using the oxygen-sensitive species rainbow trout Oncorhynchus mykiss as a model, individual fish were recorded while moving freely between two sides of a test arena: one with normoxia and one with stepwise progressive hypoxia [80-30% dissolved oxygen (DO) air saturation]. The results demonstrated a gradual decrease in the total time spent in hypoxia starting at 80% DO air saturation. At this DO level, the avoidance of hypoxia could not be attributed to changes in spontaneous swimming speed, neither in normoxia nor in hypoxia. Reducing the DO level to 60% air saturation resulted in decreased spontaneous swimming speed in normoxia, yet the number of trips to the hypoxic side of the test arena remained unchanged. Moreover, data revealed increased average residence time per trip in normoxia at DO levels ≤60% air saturation and decreased average residence time per trip in hypoxia at DO levels ≤50% air saturation. Finally, the spontaneous swimming speed in hypoxia increased at DO levels ≤40% air saturation and the number of trips to hypoxia decreased at the 30% DO air saturation level. Thus, avoidance of the deepest hypoxia was connected with a reduced number of trips to hypoxia as well as decreased and increased spontaneous swimming speed in normoxia and hypoxia, respectively. Collectively, the data support the conclusions that the mechanistic basis for avoidance of hypoxia may (1) not involve changes in swimming speed during mild hypoxia and (2) depend on the severity of hypoxia.

Field observations were supplemented with laboratory experiments to reveal patterns of salinity selection and preference for grey snapper Lutjanus griseus ( c. 21 cm total length, L T ), an ecologically and economically important species in the south‐eastern U.S.A. Fish abundance data were examined from a long‐term field survey conducted in the mangrove habitats of Biscayne Bay, Florida, where salinities ranged from <1 to 40. First, regression analyses indicated significant, positive linear relationships with salinity for both L. griseus frequency of occurrence and concentration (density when present). These patterns are inconsistent with physiological expectations of minimizing energetic osmoregulatory costs. Next, the salinity preference and swimming activity of 11 L. griseus (ranging from 18 to 23 cm L T ) were investigated using a newly developed electronic shuttlebox system. In the laboratory, fish preferred intermediate salinities in the range of 9–23. Swimming activity (measured in terms of spontaneous swimming speed) followed a parabolic relationship with salinity, with reduced activity at salinity extremes perhaps reflecting compensation for higher osmoregulatory costs. It is suspected that the basis of the discrepancy between laboratory and field observations for size classes at or near maturity ultimately relates to the reproductive imperative to move towards offshore (high‐salinity) coral‐reef habitats, a necessity that probably overrides the strategy of minimizing osmoregulatory energetic costs.

Oxygen consumption (MO2) was measured for gilthead seabream (Sparus aurata) during spontaneous and forced activities. During spontaneous activity, the swimming pattern was analysed for the effect on MO2 on the average speed (U), turning rate ( ) and change in speed (DU). All swimming characteristics contributed significantly to the source of spontaneous swimming costs, and the models explained up to 58 % of the variation in

The objective of this study was to identify kinematic variables correlated with oxygen consumption during spontaneous labriform swimming. Kinematic variables (swimming speed, change of speed, turning angle, turning rate, turning radius and pectoral fin beat frequency) and oxygen consumption (MO2) of spontaneous swimming in Embiotoca lateralis were measured in a circular arena using video tracking and respirometry, respectively. The main variable influencing MO2 was pectoral fin beat frequency (r 2 = 0.71). No significant relationship was found between swimming speed and pectoral fin beat frequency. Complementary to other methods within biotelemetry such as EMG it is suggested that such correlations of pectoral fin beat frequency may be used to measure the energy requirements of labriform swimming fish such as E. lateralis in the field, but need to be taken with great caution since movement and oxygen consumption patterns are likely to be quite different in field situation compared to a small lab tank. In addition, our methods could be useful to measure metabolic costs of growth and development, or bioassays for possible toxicological effects on fish.

The short‐lived annual fish Nothobranchius furzeri shows extremely short captive life span and accelerated expression of age markers, making it an interesting model system to investigate the effects of experimental manipulations on longevity and age‐related pathologies. Here, we tested the effects of dietary restriction (DR) on mortality and age‐related markers in N. furzeri. DR was induced by every other day feeding and the treatment was performed both in an inbred laboratory line and a longer‐lived wild‐derived line. In the inbred laboratory line, DR reduced age‐related risk and prolonged maximum life span. In the wild‐derived line, DR induced early mortality, did not reduce general age‐related risk and caused a small but significant extension of maximum life span. Analysis of age‐dependent mortality revealed that DR reduced demographic rate of aging, but increased baseline mortality in the wild‐derived strain. In both inbred‐ and wild‐derived lines, DR prevented the expression of the age markers lipofuscin in the liver and Fluoro‐Jade B (neurodegeneration) in the brain. DR also improved performance in a learning test based on conditioning (active avoidance in a shuttle box). Finally, DR induced a paradoxical up‐regulation of glial fibrillary acidic protein in the brain.

Animal social networks can be extremely complex and are characterized by highly non-random interactions between group members. However, very little is known about the underlying factors affecting interaction preferences, and hence network structure. One possibility is that behavioural differences between individuals, such as how bold or shy they are, can affect the frequency and distribution of their interactions within a network. We tested this using individually marked three-spined sticklebacks ( Gasterosteus aculeatus ), and found that bold individuals had fewer overall interactions than shy fish, but tended to distribute their interactions more evenly across all group members. Shy fish, on the other hand, tended to associate preferentially with a small number of other group members, leading to a highly skewed distribution of interactions. This was mediated by the reduced tendency of shy fish to move to a new location within the tank when they were interacting with another individual; bold fish showed no such tendency and were equally likely to move irrespective of whether they were interacting or not. The results show that animal social network structure can be affected by the behavioural composition of group members and have important implications for understanding the spread of information and disease in social groups.

Pikeperch (Sander lucioperca) are non-native in the United Kingdom. It is important to understand how environmental factors, such as salinity, influence the behaviour and activity of introduced fish species to identify their dispersal potential. Previous studies have shown that pikeperch, traditionally recognised as a freshwater fish, can tolerate brackish waters and demonstrate physiological acclimation. However, their behavioural responses to brackish waters are unknown. The aim of the present study was therefore to investigate the activity and swimming behaviour of pikeperch obtained from freshwater canals in southern England. In the laboratory, fish were exposed to a 12-h simulated tidal cycle and a 12-day stepped salinity challenge where salinity was increased by 4 every 2 days, up to a salinity of 20. In both regimes, fish showed increased swimming activity in response to increasing salinity, which may represent an avoidance response. The most dramatic changes, including vertical movements, occurred at salinities above ~16. At these higher salinities, head shaking and coughing behaviours were also observed, suggesting significant stress and respiratory impairment. However, during the simulated tidal cycle, normal behaviour was rapidly restored once salinity was reduced. The results of this study may have implications in understanding the dispersal of non-native fish in the wild.

Remaining populations of westslope cutthroat trout (Oncorhynchus clarkii iewisi) in western North America are primarily confined to cold headwaters whereas nonnative rainbow trout (Oncorhynchus mykiss) predominate in warmer, lower elevation stream sections historically occupied by westslope cutthroat trout. We tested whether differing thermal preferences could account for the spatial segregation observed in the field. Thermal preferences of age-1 westslope cutthroat trout and rainbow trout (125 to 150 mm total length) were assessed in the laboratory using a modified annular preference chamber at acclimation temperatures of 10, 12, 14, and 16°C. Final preferred temperature of westslope cutthroat trout (14.9°C) was similar to that of rainbow trout (143°C) when tested in a thermal gradient of 11–17°C. The high degree of overlap in thermal preference indicates the two species have similar thermal niches and a high potential for competition. We suggest several modifications to the annular preference chamber to improve performance in future studies.

A previous study has reported that Atlantic cod can be conditioned to detect ultrasonic sound pulses of high intensity. This capability has been proposed as a mean for detection and avoidance of echolocating toothed whales that emit intense ultrasonic clicks. In this study, we use acoustic playbacks to test the hypotheses that unconditioned cod can detect and respond to intense ultrasound from toothed whales and from echosounders. Intense ultrasound exposure of 210 dB re. 1μPa (pp) did not cause a short-term stress response in the form of bradycardia in unconditioned cod. Free-swimming cod exposed to ultrasonic clicks and echosounder pulses with received levels of more than 204 dB re. 1 μPa (pp) did not elicit flight responses as seen in ultrasound detecting Alosinae. Furthermore, we tested the debilitating effects of high intensity ultrasound on swimming cod with no detected changes in swimming ability when exposed to more than 213 dB re. 1 μPa (pp). It is concluded that intense ultrasound exposure induces neither an antipredator nor a stress response in Atlantic cod, and that echosounder pulses and biosonar clicks therefore most probably play no ecophysiological role in wild cod populations.

Lesser sandeel (Ammodytes tobianus) is abundant in near-shore areas where it is a key prey. It exhibits the behaviour of alternating between swimming in schools and lying buried in the sediment. We first determined the species’ standard metabolic rate (SMR), critical partial pressure of oxygen $$(P_{{\rm O}_{2{\rm crit}}})$$ and maximal oxygen uptake $$(M_{{\rm O}_{2{\rm max}}}).$$ The sandeel were then exposed to an acute stepwise decline in water oxygen pressure (18.4, 13.8, 9.8, 7.5, 5.8, 4.0, and 3.1 kPa $$P_{{\rm O}_{2}}$$ ). Swimming speed and routine- and post-experimental blood lactate levels were measured, in addition to levels associated with strenuous exercise. The SMR was 69.0 ± 8.4 mg O2 kg−1 h−1 and the $$M_{{\rm O}_{2{\rm max}}}$$ about seven times as high. The $$P_{{\rm O}_{2{\rm crit}}}$$ was found to be 4.1 kPa. A rapid decrease (within 1 h) in $$P_{{\rm O}_{2}}$$ from 18.4 to 3.1 kPa had no significant effect on routine swimming speed (0.9 ± 0.06 bl s−1), but steady levels at the lowest $$P_{{\rm O}_{2}}$$ (3.1 kPa) gradually reduced the swimming speed by 95% after 40 min. The routine blood lactate levels were 2.2 ± 0.6 mmol l−1, while the levels in the strenuously exercised groups were significantly higher with 5.4 ± 1.6 and 5.8 ± 1.3 mmol l−1. The highest levels were observed in post-experimental fish with 7.5 ± 2.7 mmol l−1. We argue that, as sandeel showed no decrease in swimming speed (to offset stress) nor an increased speed to escape the hypoxia, the fish either rely on a low SMR and being a reasonable strong oxygen regulator $$(\hbox{low}\;P_{{\rm O}_{2{\rm crit}}})$$ as a mean to cope when exposed to acute hypoxia, or that the hypoxia simply developed too fast for the fish to decide on an appropriate strategy. Not showing a behavioural response may in the present case be maladaptive, as the consequence was major physiological stress which the fish however appears tolerant towards. The high routine blood lactate levels suggest that anaerobic metabolism is associated with swimming in sandeel, which may be related to the specific lifestyle of the fish where they regularly bury in the sediment.

In order to determine the potential for the invasive fishes sunbleak Leucaspius delineatus and topmouth gudgeon Pseudorasbora parva to disperse through saline waters their behaviour and physiology were investigated during exposure to salinities of 10·0 and 12·5. Increased salinity caused an increase in whole body cortisol in both species, but sunbleak and topmouth gudgeon showed very different metabolic and behavioural responses to the salinity stress. Sunbleak displayed increased swimming activity in brackish water, which may be important for dispersal through saline waters in the wild, although there were increased metabolic costs associated with this behaviour. Conversely, topmouth gudgeon showed a reduction in both swimming activity and metabolic rate in brackish waters. A pronounced depression in food intake (70–80%) was shown by both species during the salinity exposures. Both sunbleak and topmouth gudgeon, however, showed a full recovery of food intake within 24 h following return to fresh water. Despite the fact that exposure to saline waters is stressful, and affects both physiology and behaviour, rapid recovery of appetite after return to fresh water suggests that short‐term use of brackish waters is a feasible dispersal route for sunbleak and topmouth gudgeon in the wild.

We examined the effects of turbidity on habitat preference of juvenile Atlantic cod in the laboratory, using a shuttle box where fish could select between two different habitats. In the first experiment, we compared three turbidity levels of kaolin (3, 8 and 21 beam attenuation m−1). In the second experiment, we looked at the effect of turbidity media (kaolin versus algae), after controlling for spectral differences between turbidity media. Although cod preferred an intermediate turbidity of kaolin over low turbidity water, comparisons between low and high turbidity, and intermediate and high turbidity did not significantly influence habitat preference. Algae did not influence habitat preference by cod. Although other studies have found that turbidity affects both foraging and antipredator behaviour of juvenile cod, this study has shown that gradients in turbidity per se do not have a strong effect on their habitat preference.

The multiple responses of fishes to changes in dissolved oxygen saturations have been studied widely in the laboratory. In contrast, only a few studies have included field observa- tions. The objective of the present study was to evaluate the performance of a novel acoustic dissolved oxygen transmitter for field biotelemetry. The results demonstrated that the output of the transmitter was unaffected by three different temperatures (10 to 30°C) and described the dissolved oxygen saturation with high accuracy (r 2 > 0.99) over the entire range of 0 to 191% saturation. The response time (≥ 90% of end value) of the transmitter was 12 s both in terms of decreasing (100 to 0%) and increasing (0 to 100%) oxygen saturations. When externally attached to fishes, the present findings support the use of the transmitter for reliable dissolved oxygen measurements on individuals living in environments that may change both temporally and spatially with regard to ambient temperature and dissolved oxygen saturation.