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Global warming has already caused a loss of almost 50% Arctic sea-ice coverage since the 1980s. Sea-ice loss strengthens summer stratification of the ocean’s water column and, consequently, hypoxic zones in the deep-water layers may form. The present study investigated the response of an Arctic keystone species, the Polar cod,Boreogadus saida, to hypoxia and warming. We measured the respiratory capacity (standard, routine and maximum metabolic rates, SMR, RMR, MMR, aerobic scope, critical oxygen saturation (Pcrit)) and swimming performance of Polar cod under progressive hypoxia at 2.4 °C and after warm acclimation to close to the species’ thermal limit (10.0 °C) via flow-through and swim tunnel respirometry. We observed clear and stable patterns that were similar in both thermal regimes: Polar cod displayed oxygen-regulating behaviour under progressive hypoxia, with SMR never below aerobic baseline metabolism and a very stable AS. Our study revealed that Polar cod can handle exceptionally low oxygen saturations down to aPcritof 5.9 % air saturation at typical habitat temperatures. Closer to critical temperatures (10.0 °C),Pcritrose to 21.6 % air saturation. However, the pertinent question remains whether the observed behaviour can be summarized under classic hypoxia tolerance, as we a) did not observe any metabolic downregulation and b) no anaerobic component of the hypoxia response in Polar cod, which are usually put forward in the definition of hypoxia tolerance. Therefore, we describe the observed metabolic response to hypoxia rather as metabolic hypoxia compensation than hypoxia tolerance as the mechanisms involved here actively seek to improve oxygen supply instead of (anaerobically) tolerating hypoxia through metabolic depression.

Although Atlantic cod has been observed in Svalbard waters since the 1880s, knowledge about the presence in the Arctic shallow water zone is limited. The regular catch of juvenile Atlantic cod in Kongsfjorden since 2008 is in line with an overall northward shift of boreal fish species toward the Arctic. This is the first study showing the age class composition, growth rates, and stomach content of Atlantic cod in the shallow water zone of Kongsfjorden, Svalbard. From 2012 to 2014 a total of 721 specimens were sampled in 3 to 12 m water depth. The primary age classes were identified as 0+, 1+, and 2+ using otolith age analysis. The different cohorts of these specimens show stable growth rates during the polar day and night. By stomach content analysis, we show that these specimens primarily feed on benthic food sources. These observations support the assumption that the shallow water zone of Kongsfjorden is likely to be a nursery ground for Atlantic cod.

Special issue The Weddell Sea and the ocean off Dronning Maud Land: unique oceanographic conditions shape circumpolar and global processes – a multi-disciplinary study (OS/BG/TC inter-journal SI).-- 30 pages, 5 figures.-- Data availability: The data (sets) used for the illustrations in some figures are referenced in the figure captions: bathymetry (water depth) in Fig. 1 (Dorschel et al., 2022); bathymetry in Figs. 2a and 3 (Arndt et al., 2013); and sea ice, chlorophyll, and temperature in Fig. 4 (Lavergne et al., 2019, and http://oceandata.sci.gsfc.nasa.gov/, last access: 23 August 2022). No addition data sets were used in this publication, Systematic long-term studies on ecosystem dynamics are largely lacking from the East Antarctic Southern Ocean, although it is well recognized that they are indispensable to identify the ecological impacts and risks of environmental change. Here, we present a framework for establishing a long-term cross-disciplinary study on decadal timescales. We argue that the eastern Weddell Sea and the adjacent sea to the east, off Dronning Maud Land, is a particularly well suited area for such a study, since it is based on findings from previous expeditions to this region. Moreover, since climate and environmental change have so far been comparatively muted in this area, as in the eastern Antarctic in general, a systematic long-term study of its environmental and ecological state can provide a baseline of the current situation, which will be important for an assessment of future changes from their very onset, with consistent and comparable time series data underpinning and testing models and their projections. By establishing an Integrated East Antarctic Marine Research (IEAMaR) observatory, long-term changes in ocean dynamics, geochemistry, biodiversity, and ecosystem functions and services will be systematically explored and mapped through regular autonomous and ship-based synoptic surveys. An associated long-term ecological research (LTER) programme, including experimental and modelling work, will allow for studying climate-driven ecosystem changes and interactions with impacts arising from other anthropogenic activities. This integrative approach will provide a level of long-term data availability and ecosystem understanding that are imperative to determine, understand, and project the consequences of climate change and support a sound science-informed management of future conservation efforts in the Southern Ocean, Stefanie Arndt, Heike Link and Christoph Held have received financial support from the Deutsche Forschungsgemeinschaft (DFG) in the framework of the priority programme 1158 “Antarctic Research with Comparative Investigations in Arctic Sea Ice Areas”. The article processing charges for this open-access publication were covered by the Alfred Wegener Institute, Helmholtz Centre for Polar and Marine Research (AWI), With the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S)

The current and projected environmental change of the Arctic Ocean contrasts sharply with the limited knowledge of its genetic biodiversity. Polar cod Boreogadus saida (Lepechin, 1774) is an abundant circumpolar marine fish and ecological key species. The central role of polar cod in the Arctic marine food web warrants a better understanding of its population structure and connectivity. In this study, the genetic population structure of 171 juveniles, collected from several fjords off West-Svalbard (Billefjorden, Hornsund and Kongsfjorden), the northern Sophia Basin and the Eurasian Basin of the Arctic Ocean, was analysed using nine DNA microsatellite loci. Genetic analyses indicated moderate to high genetic diversity, but absence of spatial population structure and isolation-by-distance, suggesting ongoing gene flow between the studied sampling regions. High levels of connectivity may be key for polar cod to maintain populations across wide spatial scales. The adaptive capacity of the species will be increasingly important to face challenges such as habitat fragmentation, ocean warming and changes in prey composition. In view of a limited understanding of the population dynamics and evolution of polar cod, a valuable next step to predict future developments should be an integrated biological evaluation, including population genomics, a life-history approach, and habitat and biophysical dispersal modelling. ispartof: JOURNAL OF FISH BIOLOGY vol:99 issue:1 pages:49-60 ispartof: location:England status: published

Mitochondrial function can provide key insights into how fish will respond to climate change, due to its important role in heart performance, energy metabolism and oxidative stress. However, whether warm acclimation can maintain or improve the energetic status of the fish heart when exposed to short-term heat stress is not well understood. We acclimated Atlantic salmon, a highly aerobic eurythermal species, to 12 and 20 °C, then measured cardiac mitochondrial functionality and integrity at 20 °C and at 24, 26 and 28 °C (this species’ critical thermal maximum ± 2 °C). Acclimation to 20 °C vs. 12 °C enhanced many aspects of mitochondrial respiratory capacity and efficiency up to 24 °C, and preserved outer mitochondrial membrane integrity up to 26 °C. Further, reactive oxygen species (ROS) production was dramatically decreased at all temperatures. These data suggest that salmon acclimated to ‘normal’ maximum summer temperatures are capable of surviving all but the most extreme ocean heat waves, and that there is no ‘tradeoff’ in heart mitochondrial function when Atlantic salmon are acclimated to high temperatures (i.e., increased oxidative phosphorylation does not result in heightened ROS production). This study suggests that fish species may show quite different acclimatory responses when exposed to prolonged high temperatures, and thus, susceptibility to climate warming.

When organisms are unable to feed ad libitum they may be more susceptible to negative effects of environmental stressors such as ocean acidification and warming (OAW). We reared sea bass (Dicentrarchus labrax) at 15 or 20 °C and at ambient or high PCO2 (650 versus 1750 µatm PCO2; pH = 8.1 or 7.6) at ad libitum feeding and observed no discernible effect of PCO2 on the size-at-age of juveniles after 277 (20 °C) and 367 (15 °C) days. Feeding trials were then conducted including a restricted ration (25% ad libitum). At 15 °C, growth rate increased with ration but was unaffected by PCO2. At 20 °C, acidification and warming acted antagonistically and low feeding level enhanced PCO2 effects. Differences in growth were not merely a consequence of lower food intake but also linked to changes in digestive efficiency. The specific activity of digestive enzymes (amylase, trypsin, phosphatase alkaline and aminopeptidase N) at 20 °C was lower at the higher PCO2 level. Our study highlights the importance of incorporating restricted feeding into experimental designs examining OAW and suggests that ad libitum feeding used in the majority of the studies to date may not have been suitable to detect impacts of ecological significance.

1.AbstractThe aim of this study was to investigate the effect of ocean acidification (OA) and warming (OW) as well as the transgenerational effect of OA on larval and juvenile growth and metabolism of a large economically important fish species with a long generation time. Therefore we incubated European sea bass from Brittany (France) for two generations (>5 years in total) under current and predicted OA conditions (PCO2: 650 and 1700 µatm). In the F1 generation both OA condition were crossed with OW (temperature: 15-18°C and 20-23°C). We found that OA alone did not affect larval or juvenile growth and OW increased developmental time and growth rates, but OAW decreased larval size at metamorphosis. Larval routine metabolic rate (RMR) and juvenile standard metabolic rate (SMR) were significantly lower in cold compared to warm conditioned fish and also lower in F0 compared to F1 fish. We did not find any effect of OA on RMR or SMR. JuvenilePO2critwas not affected by OA, OW or OAW in both generations.We discuss the potential underlying mechanisms resulting in beneficial effects of OW on F1 larval growth and RMR and in resilience of F0 and F1 larvae and juveniles to OA, but on the other hand resulting in vulnerability of F1, but not F0 larvae to OAW.. With regard to the ecological perspective, we conclude that recruitment of larvae and early juveniles to nursery areas might decrease under OAW conditions but individuals reaching juvenile phase might benefit from increased performance at higher temperatures.Summary statementWe found that OA did not affect developmental time, growth, RMR and SMR, while OW increased these traits. OAW decreased larval size at metamorphosis. We discuss underlying mechanisms and the ecological perspective resulting from these results and conclude that recruitment to nursery areas might decrease under OAW conditions but individuals reaching juvenile phase might benefit from increased performance at higher temperatures in Atlantic waters.

Rapid climate change is placing many marine species at risk of local extinction. Recent studies show that epigenetic mechanisms (e.g. DNA methylation, histone modifications) can facilitate both within and transgenerational plasticity to cope with changing environments. However, epigenetic reprogramming (erasure and re-establishment of epigenetic marks) during gamete and early embryo development may hinder transgenerational epigenetic inheritance. Most of our knowledge about reprogramming stems from mammals and model organisms, whereas the prevalence and extent of reprogramming among non-model species from wild populations is rarely investigated. Moreover, whether reprogramming dynamics are sensitive to changing environmental conditions is not well known, representing a key knowledge gap in the pursuit to identify mechanisms underlying links between parental exposure to changing climate patterns and environmentally-adapted offspring phenotypes. Here, we investigated epigenetic reprogramming (DNA methylation/hydroxymethylation) and gene expression across gametogenesis and embryogenesis of marine stickleback (Gasterosteus aculeatus) under three ocean warming scenarios (ambient, + 1.5 °C and + 4 °C). We found that parental acclimation to ocean warming led to dynamic and temperature-sensitive reprogramming throughout offspring development. Both global methylation/hydroxymethylation and expression of genes involved in epigenetic modifications were strongly and differentially affected by the increased warming scenarios. Comparing transcriptomic profiles from gonads, mature gametes, and early embryonic stages showed sex-specific accumulation and temperature sensitivity of several epigenetic actors. DNA methyltransferase induction was primarily maternally inherited (suggesting maternal control of remethylation), whereas induction of several histone-modifying enzymes was shaped by both parents. Importantly, massive, temperature-specific changes to the epigenetic landscape occurred in blastula, a critical stage for successful embryo development, which could, thus, translate to substantial consequences for offspring phenotype resilience in warming environments. In summary, our study identified key stages during gamete and embryo development with temperature-sensitive reprogramming and epigenetic gene regulation, reflecting potential "windows of opportunity" for adaptive epigenetic responses under future climate change.

Mitochondrial respiration is a multi-step pathway that involves matrix and membrane-associated enzymes and plays a key role in acclimation to variable environmental conditions, but until now it has not been clear which of these steps would be most important in acclimation to changing temperatures and CO2 levels. Considering scenarios of ocean warming and acidification we assessed the role and limitation to phenotypic plasticity in the hearts of two Gadoid species adapted to different thermal ranges: the polar cod (Boreogadus saida), an Arctic stenotherm, and the Northeast Arctic population of Atlantic cod (NEAC, Gadus morhua), a cold eurytherm. We analysed the capacity of single enzymes involved in mitochondrial respiration [citrate synthase (CS), succinate dehydrogenase (SDH), cytochrome c oxidase (CCO)], the capacity of the electron transport system and the lipid class composition of the cellular membranes. Juveniles of the two species were held for four months at four temperatures (0, 3, 6, 8 °C for polar cod and 3, 8, 12, 16 °C for NEAC), at both ambient and elevated $$P_{{{\text{CO}}_{2} }}$$ (400 µatm and 1170 µatm, respectively). Polar cod showed no changes in mitochondrial enzyme capacities and in the relative lipid class composition in response to altered temperature or elevated $$P_{{{\text{CO}}_{2} }} .$$ The lack of cardiac cellular plasticity together with evidence at the whole-animal level coming from other studies is indicative of little or no ability to overcome stenothermy, in particular during acclimation to 8 °C. In contrast, eurythermal NEAC exhibited modifications of membrane composition towards a more rigid structure and altered enzyme capacities to preserve functionality at higher temperatures. Furthermore, in NEAC, the capacities of SDH, CCO and CS were increased by high levels of CO2 if combined with high temperatures (12 and 16 °C), suggesting the compensation of an inhibitory effect. These results indicate that the cold eurythermal species (NEAC) is able to alter its mitochondrial function to a far greater extent than the Arctic stenotherm (polar cod), indicating greater resilience to variable environmental conditions. This difference in plasticity may underpin differences in the resilience to climate change and affect future species distributions and, eventually, survival.

In the last few decades, numerous studies have investigated the impacts of simulated ocean acidification on marine species and communities, particularly those inhabiting dynamic coastal systems. Despite these research efforts, there are many gaps in our understanding, particularly with respect to physiological mechanisms that lead to pathologies. In this review, we trace how carbonate system disturbances propagate from the coastal environment into marine invertebrates and highlight mechanistic links between these disturbances and organism function. We also point toward several processes related to basic invertebrate biology that are severely understudied and prevent an accurate understanding of how carbonate system dynamics influence organismic homeostasis and fitness-related traits. We recommend that significant research effort be directed to studying cellular phenotypes of invertebrates acclimated or adapted to elevated seawater pCO2 using biochemical and physiological methods.

The aryl hydrocarbon receptor (AhR) pathway mediates many, if not all, responses of fish to dioxin-like compounds. The Southern Ocean is progressively exposed to increasing concentrations of anthropogenic pollutants. Antarctic fish are known to accumulate those pollutants, yet nothing is known about their capability to induce chemical biotransformation via the AhR pathway. The objective of the present study was to investigate whether Antarctic eelpout, Pachycara brachycephalum, respond to anthropogenic pollutants by activation of the AhR and its target gene cytochrome P4501A (CYP1A), and of superoxide dismutase (SOD), which served as a representative for oxidative stress. We exposed P. brachycephalum to 10 and 100 mg benzo[a]pyrene (BaP)/kg body weight for 10 d and measured the expression of AhR, CYP1A, and SOD in liver tissue via quantitative polymerase chain reaction. We identified two distinct AhR isoforms in the liver of P. brachycephalum. Antarctic eelpout responded to both BaP exposures by an up-regulation of AhR and SOD, and by a particularly strong induction of CYP1A expression, which remained high until day 10 of the exposure time. Our data suggest that P. brachycephalum possesses the potential to up-regulate xenobiotic biotransformation pathways, at least at the gene expression level. The time course of the AhR and CYP1A response points to an efficient but slow xenobiotics metabolism. Moreover, BaP exposure could include adverse effects such as oxidative stress. Environ Toxicol Chem 2018;37:1487-1495. © 2018 SETAC.

International audience; Since sensory system allows organisms to perceive and interact with their external environment, any disruption in their functioning may have detrimental consequences on their survival. Ocean acidification has been shown to potentially impair olfactory system in fish and it is therefore essential to develop biological tools contributing to better characterize such effects. The olfactory marker protein (omp) gene is involved in the maturation and the activity of olfactory sensory neurons in vertebrates. In teleosts, two omp genes (ompa and ompb) originating from whole genome duplication have been identified. In this study, bioinformatic analysis allowed characterization of the ompa and ompb genes from the European seabass (Dicentrarchus labrax) genome. The European seabass ompa and ompb genes differ in deduced amino acid sequences and in their expression pattern throughout the tissues. While both ompa and ompb mRNA are strongly expressed in the olfactory epithelium, ompb expression was further observable in different brain areas while ompa expression was also detected in the eyes and in other peripheral tissues. Expression levels of ompa and ompb mRNA were investigated in adult seabass (4 years-old, F0) and in their offspring (F1) exposed to pH of 8 (control) or 7.6 (ocean acidification, OA). Under OA ompb mRNA was down-regulated while ompa mRNA was up-regulated in the olfactory epithelium of F0 adults, suggesting a long-term intragenerational OA-induced regulation of the olfactory sensory system. A shift in the expression profiles of both ompa and ompb mRNA was observed at early larval stages in F1 under OA, suggesting a disruption in the developmental process. Contrary to the F0, the expression of ompa and ompb mRNA was not anymore significantly regulated under OA in the olfactory epithelium of juvenile F1 fish. This work provides evidence for long-term impact of OA on sensorial system of European seabass as well as potential intergenerational acclimation of omp genes expression to OA in European seabass.

Observations of climate impacts on ecosystems highlight the need for an understanding of organismal thermal ranges and their implications at the ecosystem level. Where changes in aquatic animal populations have been observed, the integrative concept of oxygen- and capacity-limited thermal tolerance (OCLTT) has successfully characterised the onset of thermal limits to performance and field abundance. The OCLTT concept addresses the molecular to whole-animal mechanisms that define thermal constraints on the capacity for oxygen supply to the organism in relation to oxygen demand. The resulting ‘total excess aerobic power budget’ supports an animal's performance (e.g. comprising motor activity, reproduction and growth) within an individual's thermal range. The aerobic power budget is often approximated through measurements of aerobic scope for activity (i.e. the maximum difference between resting and the highest exercise-induced rate of oxygen consumption), whereas most animals in the field rely on lower (i.e. routine) modes of activity. At thermal limits, OCLTT also integrates protective mechanisms that extend time-limited tolerance to temperature extremes – mechanisms such as chaperones, anaerobic metabolism and antioxidative defence. Here, we briefly summarise the OCLTT concept and update it by addressing the role of routine metabolism. We highlight potential pitfalls in applying the concept and discuss the variables measured that led to the development of OCLTT. We propose that OCLTT explains why thermal vulnerability is highest at the whole-animal level and lowest at the molecular level. We also discuss how OCLTT captures the thermal constraints on the evolution of aquatic animal life and supports an understanding of the benefits of transitioning from water to land.

Antarctic fish belonging to Notothenioidei represent an extraordinary example of radiation in the cold. In addition to the absence of hemoglobin, icefish show a number of other striking peculiarities including large-diameter blood vessels, high vascular densities, mitochondria-rich muscle cells, and unusual mitochondrial architecture. In order to investigate the bases of icefish adaptation to the extreme Southern Ocean conditions we sequenced the complete genome of the icefish Chionodraco myersi. Comparative analyses of the icefish genome with those of other teleost species, including two additional white-blooded and five red-blooded notothenioids, provided a new perspective on the evolutionary loss of globin genes. Muscle transcriptome comparative analyses against red-blooded notothenioids as well as temperate fish revealed the peculiar regulation of genes involved in mitochondrial function in icefish. Gene duplication and promoter sequence divergence were identified as genome-wide patterns that likely contributed to the broad transcriptional program underlying the unique features of icefish mitochondria.

The parasitic marine dinoflagellate Amoebophrya is the first known eukaryote with aerobic mitochondria without a genome., Dinoflagellates are microbial eukaryotes that have exceptionally large nuclear genomes; however, their organelle genomes are small and fragmented and contain fewer genes than those of other eukaryotes. The genus Amoebophrya (Syndiniales) comprises endoparasites with high genetic diversity that can infect other dinoflagellates, such as those forming harmful algal blooms (e.g., Alexandrium). We sequenced the genome (~100 Mb) of Amoebophrya ceratii to investigate the early evolution of genomic characters in dinoflagellates. The A. ceratii genome encodes almost all essential biosynthetic pathways for self-sustaining cellular metabolism, suggesting a limited dependency on its host. Although dinoflagellates are thought to have descended from a photosynthetic ancestor, A. ceratii appears to have completely lost its plastid and nearly all genes of plastid origin. Functional mitochondria persist in all life stages of A. ceratii, but we found no evidence for the presence of a mitochondrial genome. Instead, all mitochondrial proteins appear to be lost or encoded in the A. ceratii nucleus.

Antarctic fish belonging to Notothenioidei represent an extraordinary example of radiation in the cold. In addition to the absence of hemoglobin, icefish show a number of other striking peculiarities including large-diameter blood vessels, high vascular densities, mitochondria-rich muscle cells, and unusual mitochondrial architecture. In order to investigate the bases of icefish adaptation to the extreme Southern Ocean conditions we sequenced the complete genome of the icefish Chionodraco myersi. Comparative analyses of the icefish genome with those of other teleost species, including two additional white-blooded and five red-blooded notothenioids, provided a new perspective on the evolutionary loss of globin genes. Muscle transcriptome comparative analyses against red-blooded notothenioids as well as temperate fish revealed the peculiar regulation of genes involved in mitochondrial function in icefish. Gene duplication and promoter sequence divergence were identified as genome-wide patterns that likely contributed to the broad transcriptional program underlying the unique features of icefish mitochondria., Bargelloni et al. assemble the genome of the icefish Chionodraco myersi and use comparative analysis to examine the basis of adaptation to extreme Southern Ocean conditions. They show evidence for the evolutionary loss of globin genes and the peculiar regulation of genes involved in mitochondrial function.

1. Abstract The world’s oceans are acidifying and warming due to increasing amounts of atmospheric CO2. Thermal tolerance of fish much depends on the cardiovascular ability to supply the tissues with oxygen. The heart itself is highly dependent on oxygen and heart mitochondria thus might play a key role in shaping an organism’s tolerance to temperature. The present study aimed to investigate the effects of acute and chronic warming on respiratory capacities of European sea bass (Dicentrarchus labrax L.) heart mitochondria. We hypothesized that acute warming would impair mitochondrial respiratory capacities, but be compensated after long-term. Increasing PCO2 may cause intracellular changes, likely further constricting cellular energy metabolism. We found increased leak respiration rates in acutely warmed heart mitochondria of cold-conditioned fish in comparison to measurements at their rearing temperature, suggesting a lower aerobic capacity to synthesize ATP. However, thermal acclimation led to increased mitochondrial functionality, e.g. higher RCRo in heart mitochondria of warm-conditioned compared to cold-conditioned fish. Exposure to high PCO2 synergistically amplified the effects of acute and long-term warming, but did not result in changes by itself. We explained the high ability to maintain mitochondrial function under OA with the fact that seabass are moving between various environmental conditions. Improved mitochondrial capacities after warm conditioning could be due to the origin of this species in the warm waters of the Mediterranean. Our results also indicate that seabass are not yet fully adapted to the colder temperatures in their northern distribution range and might benefit from warmer temperatures. Summary statement Mitochondria of juvenile European sea bass hearts are impaired by acute warming, but seem to benefit from acclimation to warmer temperatures, they are only marginally impacted by ocean acidification. 2. List of abbreviations Δ500: Acidification condition (ambient PCO2 + 500 µatm) Δ1000: Acidification condition (ambient PCO2 + 1000 µatm) A: Ambient PCO2 condition C: Cold life conditioned group CI: Complex I of the electron transport system CII: Complex II of the electron transport system CIV: Complex IV of the electron transport system dph: Days post hatch IPCC: Intergovernmental Panel on Climate Change Leak-CI: State 4 respiration of complex I MS-222: Tricaine methane sulfonate OA: Ocean acidification OAW: Ocean acidification and warming OW: Ocean warming OXPHOS: Full state 3 respiration of the electron transport system OXPHOS-CI: State 3 respiration of complex I OXPHOS-CII: State 3 respiration of complex II OXPHOS-cytC: State 3 respiration after addition of cytochrome c PCO2: Partial pressure of CO2 RCP: Representative concentration pathway RCRo: Respiratory control ratio State 4o: State 4 respiration after addition of oligomycin W: Warm life conditioned group

Transgenerational effects can buffer populations against environmental change, yet little is known about underlying mechanisms, their persistence or the influence of environmental cue timing. We investigated mitochondrial respiratory capacity (MRC) and gene expression of marine sticklebacks that experienced acute or developmental acclimation to simulated ocean warming (21°C) across three generations. Previous work showed that acute acclimation of grandmothers to 21°C led to lower (optimized) offspring MRCs. Here, developmental acclimation of mothers to 21°C led to higher, but more efficient offspring MRCs. Offspring with a 21°C × 17°C grandmother‐mother environment mismatch showed metabolic compensation: their MRCs were as low as offspring with a 17°C thermal history across generations. Transcriptional analyses showed primarily maternal but also grandmaternal environment effects: genes involved in metabolism and mitochondrial protein biosynthesis were differentially expressed when mothers developed at 21°C, whereas 21°C grandmothers influenced genes involved in hemostasis and apoptosis. Genes involved in mitochondrial respiration all showed higher expression when mothers developed at 21° and lower expression in the 21°C × 17°C group, matching the phenotypic pattern for MRCs. Our study links transcriptomics to physiology under climate change, and demonstrates that mechanisms underlying transgenerational effects persist across multiple generations with specific outcomes depending on acclimation type and environmental mismatch between generations.

Polar cod (Boreogadus saida) is an important prey species in the Arctic ecosystem, yet its habitat is changing rapidly: climate change, through rising seawater temperatures and CO2 concentrations, is projected to be most pronounced in Arctic waters. This study aimed to investigate the influence of ocean acidification and warming on maximum performance parameters of B. saida as indicators for the species' acclimation capacities under environmental conditions projected for the end of this century. After 4 months at four acclimation temperatures (0, 3, 6, 8°C) each combined with two PCO2 levels (390 and 1170 µatm), aerobic capacities and swimming performance of B. saida were recorded following a Ucrit protocol. At both CO2 levels, standard metabolic rate (SMR) was elevated at the highest acclimation temperature indicating thermal limitations. Maximum metabolic rate (MMR) increased continuously with temperature, suggesting an optimum temperature for aerobic scope for exercise (ASex) at 6°C. Aerobic swimming performance (Ugait) increased with acclimation temperature irrespective of CO2 levels, while critical swimming speed (Ucrit) did not reveal any clear trend with temperature. Hypercapnia evoked an increase in MMR (and thereby ASex). However, swimming performance (both Ugait and Ucrit) was impaired under elevated near-future PCO2 conditions, indicating reduced efficiencies of oxygen turnover. The contribution of anaerobic metabolism to swimming performance was very low overall, and further reduced under hypercapnia. Our results revealed high sensitivities of maximum performance parameters (MMR, Ugait, Ucrit) of B. saida to ocean acidification. Impaired swimming capacity under ocean acidification may reflect reduced future competitive strength of B. saida., Summary: Polar cod is a key species in the Arctic ecosystem; its swimming capacity decreases under future water conditions, likely reducing its survival.

Science is built on skepticism. We therefore appreciate the high interest in our paper (Portner et al., 2017) and welcome a debate that has been going on for some time. Our commentary started as a draft correspondence with specific criticism of a paper, and was then invited by the journal editor to

Atlantic herring (Clupea harengus) is a benthic spawner, therefore its eggs are prone to encounter different water conditions during embryonic development, with bottom waters often depleted of oxygen and enriched in CO2. Some Atlantic herring spawning grounds are predicted to be highly affected by ongoing Ocean Acidification and Warming with water temperature increasing by up to +3°C and CO2 levels reaching ca. 1000 μatm (RCP 8.5). Although many studies investigated the effects of high levels of CO2 on the embryonic development of Atlantic herring, little is known about the combination of temperature and ecologically relevant levels of CO2. In this study, we investigated the effects of Ocean Acidification and Warming on embryonic metabolic and developmental performance such as mitochondrial function, respiration, hatching success (HS) and growth in Atlantic herring from the Oslo Fjord, one of the spawning grounds predicted to be greatly affected by climate change. Fertilized eggs were incubated under combinations of two PCO2 conditions (400 μatm and 1100 μatm) and three temperatures (6, 10 and 14°C), which correspond to current and end-of-the-century conditions. We analysed HS, oxygen consumption (MO2) and mitochondrial function of embryos as well as larval length at hatch. The capacity of the electron transport system (ETS) increased with temperature, reaching a plateau at 14°C, where the contribution of Complex I to the ETS declined in favour of Complex II. This relative shift was coupled with a dramatic increase in MO2 at 14°C. HS was high under ambient spawning conditions (6–10°C), but decreased at 14°C and hatched larvae at this temperature were smaller. Elevated PCO2 increased larval malformations, indicating sub-lethal effects. These results indicate that energetic limitations due to thermally affected mitochondria and higher energy demand for maintenance occur at the expense of embryonic development and growth.

Background Ocean acidification and warming are happening fast in the Arctic but little is known about the effects of ocean acidification and warming on the physiological performance and survival of Arctic fish. Results In this study we investigated the metabolic background of performance through analyses of cardiac mitochondrial function in response to control and elevated water temperatures and PCO2 of two gadoid fish species, Polar cod (Boreogadus saida), an endemic Arctic species, and Atlantic cod (Gadus morhua), which is a temperate to cold eurytherm and currently expanding into Arctic waters in the wake of ocean warming. We studied their responses to the above-mentioned drivers and their acclimation potential through analysing the cardiac mitochondrial function in permeabilised cardiac muscle fibres after 4 months of incubation at different temperatures (Polar cod: 0, 3, 6, 8 °C and Atlantic cod: 3, 8, 12, 16 °C), combined with exposure to present (400μatm) and year 2100 (1170μatm) levels of CO2. OXPHOS, proton leak and ATP production efficiency in Polar cod were similar in the groups acclimated at 400μatm and 1170μatm of CO2, while incubation at 8 °C evoked increased proton leak resulting in decreased ATP production efficiency and decreased Complex IV capacity. In contrast, OXPHOS of Atlantic cod increased with temperature without compromising the ATP production efficiency, whereas the combination of high temperature and high PCO2 depressed OXPHOS and ATP production efficiency. Conclusions Polar cod mitochondrial efficiency decreased at 8 °C while Atlantic cod mitochondria were more resilient to elevated temperature; however, this resilience was constrained by high PCO2. In line with its lower habitat temperature and higher degree of stenothermy, Polar cod has a lower acclimation potential to warming than Atlantic cod.

1) Our study addresses the role of non-genetic and genetic inheritance in shaping the adaptive potential of populations under a warming ocean scenario. We used a combined experimental approach (transgenerational plasticity and quantitative genetics) to partition the relative contribution of maternal vs. paternal (additive genetic) effects to offspring body size (a key component of fitness), and investigated a potential physiological mechanism (mitochondrial respiration capacities) underlying whole organism growth/size responses. 2) In very early stages of growth (up to 30 days), offspring body size of marine sticklebacks benefited from maternal transgenerational plasticity (TGP): offspring of mothers acclimated to17°C were larger when reared at 17°C, and offspring of mothers acclimated to 21°C were larger when reared at 21°C. The benefits of maternal TGP on body size were stronger and persisted longer (up to 60 days) for offspring reared in the warmer (21°C) environment, suggesting that maternal effects will be highly relevant for climate change scenarios in this system. 3) Mitochondrial respiration capacities measured on mature offspring (F1 adults) matched the pattern of TGP for juvenile body size, providing an intuitive mechanistic basis for the maternal acclimation persisting into adulthood. Size differences between temperatures seen at early growth stages remained in the F1 adults, linking offspring body size to maternal inheritance of mitochondria. 4) Lower maternal variance components in the warmer environment were mostly driven by mothers acclimated to ambient (colder) conditions, further supporting our tenet that maternal effects were stronger at elevated temperature. Importantly, all parent-offspring temperature combination groups showed genotype x environment (GxE) interactions, suggesting that reaction norms have the potential to evolve. 5) To summarise, transgenerational plasticity and genotype x environment interactions work in concert to mediate impacts of ocean warming on metabolic capacity and early growth of marine sticklebacks. TGP can buffer short-term detrimental effects of climate warming and may buy time for genetic adaptation to catch up, therefore markedly contributing to the evolutionary potential and persistence of populations under climate change.

The identification of loci under selection (out- liers) is a major challenge in evolutionary biology, being critical to comprehend evolutionary processes leading to population differentiation and speciation, and for conser- vation purposes, also in light of recent climate change. However, detection of selected loci can be difficult when populations are weakly differentiated. This is the case of marine fish populations, often characterized by high levels of gene flow and connectivity, and particularly of fish living in the Antarctic marine environment, characterized by a complex and strong circulating system promoting individual dispersal all around the continent. With the final aim of identifying outlier loci putatively under selection in the Chionodraco genus, we used 21 microsatellites, including both genomic (Type II) and EST-linked loci (Type I), to investigate the genetic differentiation among the three recently derived Chionodraco species that are endemic to the freezing Antarctic waters. Neutrality tests were applied in interspecific comparisons in order to identify candidate loci showing high levels of genetic dif- ferentiation, which might reveal imprints of past selection. Three outlier loci were identified, detecting a higher dif- ferentiation between species than did neutral loci. Outliers showed sequence similarity to a calmodulin gene, to an antifreeze glycoprotein/trypsinogen-like protease gene and to nonannotated fish mRNAs. Selective pressures acting on outlier loci identified in this study might reflect past evo- lutionary processes, which led to species divergence and local adaptation in the Chionodraco genus. Used loci will provide a valuable tool for future population genetic studies in Antarctic notothenioids.

Thermal tolerance windows serve as a powerful tool for estimating the vulnerability of marine species and their life stages to increasing temperature means and extremes. However, it remains uncertain to which extent additional drivers, such as ocean acidification, modify organismal responses to temperature. This study investigated the effects of CO2 -driven ocean acidification on embryonic thermal sensitivity and performance in Atlantic cod, Gadus morhua, from the Kattegat. Fertilized eggs were exposed to factorial combinations of two PCO2 conditions (400 μatm vs. 1100 μatm) and five temperature treatments (0, 3, 6, 9 and 12 °C), which allow identifying both lower and upper thermal tolerance thresholds. We quantified hatching success, oxygen consumption (MO2 ) and mitochondrial functioning of embryos as well as larval morphometrics at hatch and the abundance of acid-base-relevant ionocytes on the yolk sac epithelium of newly hatched larvae. Hatching success was high under ambient spawning conditions (3-6 °C), but decreased towards both cold and warm temperature extremes. Elevated PCO2 caused a significant decrease in hatching success, particularly at cold (3 and 0 °C) and warm (12 °C) temperatures. Warming imposed limitations to MO2 and mitochondrial capacities. Elevated PCO2 stimulated MO2 at cold and intermediate temperatures, but exacerbated warming-induced constraints on MO2 , indicating a synergistic interaction with temperature. Mitochondrial functioning was not affected by PCO2 . Increased MO2 in response to elevated PCO2 was paralleled by reduced larval size at hatch. Finally, ionocyte abundance decreased with increasing temperature, but did not differ between PCO2 treatments. Our results demonstrate increased thermal sensitivity of cod embryos under future PCO2 conditions and suggest that acclimation to elevated PCO2 requires reallocation of limited resources at the expense of embryonic growth. We conclude that ocean acidification constrains the thermal performance window of embryos, which has important implication for the susceptibility of cod to projected climate change.

Oceans are experiencing increasing acidification in parallel to a distinct warming trend in consequence of ongoing climate change. Rising seawater temperatures are mediating a northward shift in distribution of Atlantic cod (Gadus morhua), into the habitat of polar cod (Boreogadus saida), that is associated with retreating cold water masses. This study investigates the competitive strength of the co-occurring gadoids under ocean acidification and warming (OAW) scenarios. Therefore, we incubated specimens of both species in individual tanks for 4 months, under different control and projected temperatures (polar cod: 0, 3, 6, 8 °C, Atlantic cod: 3, 8, 12, 16 °C) and PCO2 conditions (390 and 1170 µatm) and monitored growth, feed consumption and standard metabolic rate. Our results revealed distinct temperature effects on both species. While hypercapnia by itself had no effect, combined drivers caused nonsignificant trends. The feed conversion efficiency of normocapnic polar cod was highest at 0 °C, while optimum growth performance was attained at 6 °C; the long-term upper thermal tolerance limit was reached at 8 °C. OAW caused only slight impairments in growth performance. Under normocapnic conditions, Atlantic cod consumed progressively increasing amounts of feed than individuals under hypercapnia despite maintaining similar growth rates during warming. The low feed conversion efficiency at 3 °C may relate to the lower thermal limit of Atlantic cod. In conclusion, Atlantic cod displayed increased performance in the warming Arctic such that the competitive strength of polar cod is expected to decrease under future OAW conditions.

Global climate change affects marine fish through drivers such as ocean warming, acidification and oxygen depletion, causing changes in marine ecosystems and socioeconomic impacts. While experimental and observational results can inform about anticipated effects of different drivers, linking between these results and ecosystem-level changes requires quantitative integration of physiological and ecological processes into models to advance research and inform management. We give an overview of important physiological and ecological processes affected by environmental drivers. We then provide a review of available modelling approaches for marine fish, analysing their capacities for process-based integration of environmental drivers. Building on this, we propose approaches to advance important research questions. Examples of integration of environmental drivers exist for each model class. Recent extensions of modelling frameworks increase the potential for including detailed mechanisms and improving model projections. Experimental results on energy allocation, behaviour and physiological limitations will advance the understanding of organism-level trade-offs and thresholds in response to multiple drivers. More explicit representation of life cycles and biological traits can improve description of population dynamics and adaptation, and data on food web topology and feeding interactions help to detail the conditions for possible regime shifts. Identification of relevant processes will also benefit the coupling of different models to investigate spatial–temporal changes in stock productivity and integrated responses of social–ecological systems. Thus, a more process-informed foundation for models will promote the integration of experimental and observational results and increase the potential for model-based extrapolations into a future under changing environmental conditions.

The cuttlefish Sepia officinalis expresses several hemocyanin isoforms with potentially different pH optima, indicating their reliance on efficient pH regulation in the blood. Ongoing ocean warming and acidification could influence the oxygen-binding properties of respiratory pigments in ectothermic marine invertebrates. This study examined whether S. officinalis differentially expresses individual hemocyanin isoforms to maintain optimal oxygen transport during development and acclimation to elevated seawater pCO2 and temperature. Using quantitative PCR, we measured relative mRNA expression levels of three different hemocyanin isoforms in several ontogenetic stages (embryos, hatchlings, juveniles, and adults), under different temperatures and elevated seawater pCO2. Our results indicate moderately altered hemocyanin expression in all embryonic stages acclimated to higher pCO2, while hemocyanin expression in hatchlings and juveniles remained unaffected. During the course of development, total hemocyanin expression increased independently of pCO2 or thermal acclimation status. Expression of isoform 3 is reported for the first time in a cephalopod in this study and was found to be generally low but highest in the embryonic stages (0.2% of total expression). Despite variable hemocyanin expression, hemolymph total protein concentrations remained constant in the experimental groups. Our data provide first evidence that ontogeny has a stronger influence on hemocyanin isoform expression than the environmental conditions chosen, and they suggest that hemocyanin protein abundance in response to thermal acclimation is regulated by post-transcriptional/translational rather than by transcriptional modifications. J. Exp. Zool. 317A:511523, 2012. (C) 2012 Wiley Periodicals, Inc.

90 pages, This paper is the result of an international initiative and is a first attempt to develop guidelines for the care and welfare of cephalopods (i.e. nautilus, cuttlefish, squid and octopus) following the inclusion of this Class of ∼700 known living invertebrate species in Directive 2010/63/EU. It aims to provide information for investigators, animal care committees, facility managers and animal care staff which will assist in improving both the care given to cephalopods, and the manner in which experimental procedures are carried out. Topics covered include: implications of the Directive for cephalopod research; project application requirements and the authorisation process; the application of the 3Rs principles; the need for harm-benefit assessment and severity classification. Guidelines and species-specific requirements are provided on: i. supply, capture and transport; ii. environmental characteristics and design of facilities (e.g. water quality control, lighting requirements, vibration/noise sensitivity); iii. accommodation and care (including tank design), animal handling, feeding and environmental enrichment; iv. assessment of health and welfare (e.g. monitoring biomarkers, physical and behavioural signs); v. approaches to severity assessment; vi. disease (causes, prevention and treatment); vii. scientific procedures, general anaesthesia and analgesia, methods of humane killing and confirmation of death. Sections covering risk assessment for operators and education and training requirements for carers, researchers and veterinarians are also included. Detailed aspects of care and welfare requirements for the main laboratory species currently used are summarised in Appendices. Knowledge gaps are highlighted to prompt research to enhance the evidence base for future revision of these guidelines., CephRes, AISAL and FELASA support this work. This publication is also supported by COST (European Cooperation in Science and Technology).

Background Octopods have successfully colonised the world’s oceans from the tropics to the poles. Yet, successful persistence in these habitats has required adaptations of their advanced physiological apparatus to compensate impaired oxygen supply. Their oxygen transporter haemocyanin plays a major role in cold tolerance and accordingly has undergone functional modifications to sustain oxygen release at sub-zero temperatures. However, it remains unknown how molecular properties evolved to explain the observed functional adaptations. We thus aimed to assess whether natural selection affected molecular and structural properties of haemocyanin that explains temperature adaptation in octopods. Results Analysis of 239 partial sequences of the haemocyanin functional units (FU) f and g of 28 octopod species of polar, temperate, subtropical and tropical origin revealed natural selection was acting primarily on charge properties of surface residues. Polar octopods contained haemocyanins with higher net surface charge due to decreased glutamic acid content and higher numbers of basic amino acids. Within the analysed partial sequences, positive selection was present at site 2545, positioned between the active copper binding centre and the FU g surface. At this site, methionine was the dominant amino acid in polar octopods and leucine was dominant in tropical octopods. Sites directly involved in oxygen binding or quaternary interactions were highly conserved within the analysed sequence. Conclusions This study has provided the first insight into molecular and structural mechanisms that have enabled octopods to sustain oxygen supply from polar to tropical conditions. Our findings imply modulation of oxygen binding via charge-charge interaction at the protein surface, which stabilize quaternary interactions among functional units to reduce detrimental effects of high pH on venous oxygen release. Of the observed partial haemocyanin sequence, residue 2545 formed a close link between the FU g surface and the active centre, suggesting a role as allosteric binding site. The prevalence of methionine at this site in polar octopods, implies regulation of oxygen affinity via increased sensitivity to allosteric metal binding. High sequence conservation of sites directly involved in oxygen binding indicates that functional modifications of octopod haemocyanin rather occur via more subtle mechanisms, as observed in this study. Electronic supplementary material The online version of this article (doi:10.1186/s12862-015-0411-4) contains supplementary material, which is available to authorized users.

Population substructure of the cuttlefish Sepia officinalis (Mollusca: Cephalopoda), as investigated by genetic variation of microsatellite loci, has been reported to be significantly extensive around the Iberian Peninsula with F(ST)=0.061 [Pérez-Losada, M., Guerra, A., Carvalho, G.R., Sanjuan, A., Shaw, P.W., 2002. Extensive population subdivision of the cuttlefish Sepia officinalis (Mollusca: Cephalopoda) around the Iberian Peninsula indicated by microsatellite DNA variation. Heredity 89, 417-424] and panmictic in the semi-enclosed Adriatic Sea with F(ST)=0.011 [Garoia, F., Guarniero, I., Ramsak, A., Ungaro, N., Landi, M., Piccinetti, C., Mannini, P., Tinti, F., 2004. Microsatellite DNA variation reveals high gene flow and panmictic populations in the Adriatic shared stocks of the European squid and cuttlefish (Cephalopoda). Heredity 93, 166-174]. Yet, no verified genetic information on population substructure existed for the northern distribution range of this species in the English Channel and the Bay of Biscay. So far, reproductive and migration behaviour and in vitro oxygen binding properties of haemocyanin have suggested the existence of separate populations in the English Channel and the Bay of Biscay. Examination of genetic variation at seven microsatellite loci within samples from the Bay of Biscay, the English Channel and the southern North Sea indicated low levels of genetic differentiation in this area but also a breakdown of free gene flow at highly significant average F(ST)=0.018. Although there is a considerable genetic exchange between populations of S. officinalis in the English Channel and the Bay of Biscay, they cannot be regarded as a single, freely interbreeding population. Earlier reported biological differences might thus be due to genetic variability between the populations.

Uncoupling proteins (UCP), capable of increasing proton leakage across the inner mitochondrial membrane, may play a role in the temperature-dependent setting of energy turnover in animals (and their mitochondria). Therefore, the genes and expression of fish UCP were investigated in the Antarctic eelpout Pachycara brachycephalum and a temperate confamilial species, the common eelpout Zoarces viviparus. UCP full-length cDNA was amplified from liver and muscle using RT-PCR and rapid amplification of cDNA ends (RACE). The fish UCP mRNA consists of 1906 bp in P. brachycephalum and of 1876 bp in Z. viviparus. Both zoarcid sequences contain open reading frames of 939 bp, encoding 313 amino acids, with 98% and 99% identity, respectively. Protein sequences of zoarcid UCP are closest related to fish and mammalian UCP2. For analysis of temperature-dependent expression common eelpouts were cold-acclimated from 10 degrees C to 2 degrees C and Antarctic eelpouts were warm-acclimated from 0 degrees C to 5 degrees C. Identical cDNA probes for both species were developed to investigate fish UCP mRNA expression, and protein expression levels were detected by Western Blot in the enriched membrane fraction. During cold-acclimation in Z. viviparus, mRNA levels increased by a factor up to 2.0, protein levels increased up to 1.5, in line with mitochondrial proliferation during cold-acclimation. Despite decreased mitochondrial protein content, in Antarctic eelpout UCP levels rose upon warm acclimation by a factor up to 2.0 (mRNA) and 1.6 (protein), respectively. Besides the ongoing discussion of UCP function in vertebrates, the data are indicative of a significant role of fish UCP in thermal adaptation of fish mitochondria.

Introduction Exposure to elevated seawater PCO2 limits the thermal tolerance of crustaceans but the underlying mechanisms have not been comprehensively explored. Larval stages of crustaceans are even more sensitive to environmental hypercapnia and possess narrower thermal windows than adults. Results In a mechanistic approach, we analysed the impact of high seawater CO2 on parameters at different levels of biological organization, from the molecular to the whole animal level. At the whole animal level we measured oxygen consumption, heart rate and activity during acute warming in zoea and megalopa larvae of the spider crab Hyas araneus exposed to different levels of seawater PCO2. Furthermore, the expression of genes responsible for acid–base regulation and mitochondrial energy metabolism, and cellular responses to thermal stress (e.g. the heat shock response) was analysed before and after larvae were heat shocked by rapidly raising the seawater temperature from 10°C rearing temperature to 20°C. Zoea larvae showed a high heat tolerance, which decreased at elevated seawater PCO2, while the already low heat tolerance of megalopa larvae was not limited further by hypercapnic exposure. There was a combined effect of elevated seawater CO2 and heat shock in zoea larvae causing elevated transcript levels of heat shock proteins. In all three larval stages, hypercapnic exposure elicited an up-regulation of genes involved in oxidative phosphorylation, which was, however, not accompanied by increased energetic demands. Conclusion The combined effect of seawater CO2 and heat shock on the gene expression of heat shock proteins reflects the downward shift in thermal limits seen on the whole animal level and indicates an associated capacity to elicit passive thermal tolerance. The up-regulation of genes involved in oxidative phosphorylation might compensate for enzyme activities being lowered through bicarbonate inhibition and maintain larval standard metabolic rates at high seawater CO2 levels. The present study underlines the necessity to align transcriptomic data with physiological responses when addressing mechanisms affected by an interaction of elevated seawater PCO2 and temperature extremes. Electronic supplementary material The online version of this article (doi:10.1186/s12983-014-0087-4) contains supplementary material, which is available to authorized users.

Oxygen demand elicited by the main cellular energy consumers was examined in isolated hepatocytes of sub-Antarctic (Lepidonotothen larseni) and high-Antarctic notothenioids (Trematomus eulepidotus, Trematomus pennellii, Trematomus lepidorhinus, Trematomus bernacchii, Artedidraco orianae) and in a zoarcid (Pachycara brachycephalum) fish with respect to the role of cellular metabolism in co-defining thermal tolerance. The relative proportions of energy allocated to protein and RNA/DNA synthesis, ion regulation and ATP synthesis were quantified between 0°C and 15°C by analysis of inhibitor sensitive cellular respiration. In all the investigated species, protein synthesis constituted 25–37%, RNA synthesis 24–35%, Na+/K+-ATPase 40–45% and mitochondrial ATP synthesis 57–65% of total respiration. The sub-Antarctic nototheniid L. larseni displayed lower cellular protein synthesis rates but somewhat higher active ion regulation activities than its high-Antarctic confamilials, as is typical for more eurythermal species. Assumed thermal optima were mirrored in minimized overall cellular energy demand. In the sub-Antarctic L. larseni and P. brachycephalum, minima of oxygen consumption were located between 3°C and 6°C, indicating elevated energy turnover below and above these temperatures. In contrast, the high-Antarctic species displayed progressively rising respiration rates during warming with a cellular energetic minimum at 0°C. The sub-Antarctic nototheniid and the zoarcid showed signs of cold-eurythermy and appear to live close to their lower limit of thermal tolerance, while high-Antarctic notothenioids show high degrees of energetic efficiency at 0°C. All cellular preparations maintained energy budgets over a wide thermal range, supporting the recent concept that thermal limits are set by oxygen and associated energy limitations at the whole organism level.

In various phyla of marine invertebrates limited capacities of both ventilatory and circulatory performance were found to set the borders of the thermal tolerance window with limitations in aerobic scope and onset of hypoxia as a first line of sensitivity to both cold and warm temperature extremes. The hypothesis of oxygen limited thermal tolerance has recently been investigated in fish using a combination of non-invasive nuclear magnetic resonance (NMR) methodology with invasive techniques. In contrast to observations in marine invertebrates arterial oxygen tensions in fish were independent of temperature, while venous oxygen tensions displayed a thermal optimum. As the fish heart relies on venous oxygen supply, limited cardio-circulatory capacity is concluded to set the first level of thermal intolerance in fish. Nonetheless, maximized ventilatory capacity is seen to support circulation in maintaining the width of thermal tolerance windows. The interdependent setting of low and high tolerance limits is interpreted to result from trade-offs between optimized tissue functional capacity and baseline oxygen demand and energy turnover co-determined by the adjustment of mitochondrial densities and functional properties to a species-specific temperature range. At temperature extremes, systemic hypoxia will elicit metabolic depression, thereby widening the thermal window transiently sustained especially in those species preadapted to hypoxic environments.

The Atlantic cod (Gadus morhua) is an economically important marine fish species exploited by both fishery and aquaculture, especially in the North Atlantic and Arctic oceans. Ongoing climate changes are happening faster in the high latitude oceans with a higher increase of temperature and a steeper decrease in water pH due to anthropogenic CO2 than in the temperate regions threatening the existence of the Atlantic cod in the areas of its maximum exploitation. In this study, we investigated the mitochondrial physiology of two life-stages of cod under the sea water temperatures and pCO2 conditions forecasted for the year 2100 in the North Atlantic (+ 5 °C, 1000 μatm CO2). In embryos, the metabolism during development showed to be sensitive to rising temperatures with a general increase in respiratory activity until 9 °C (5 °C over the natural range) and a drop in activity at 12 °C mainly caused by a dramatic decrease in Complex I activity, which was not compensated by Complex II. In the adults, already well known for their metabolic plasticity, mitochondria from liver and heart are not affected by either increasing temperature or pCO2. However, in heart mitochondria of animals that were reared under warm hypercapnia (10 °C + 1000 μatm CO2), we found OXPHOS to exploit already 100% of the ETS capacity. This suggests that a further increase in temperature or pCO2 might lead to a mismatch in the ATP demand/production and consequently decrease heart performances. The different mitochondrial plasticities of the two life-stages reflect the sensitivity range at population level and thus can provide a more realistic reading frame of the potential survival of the North Atlantic cod population under climate change.

Summary Oxygen equilibrium curves have been widely used to understand oxygen transport in numerous organisms. A major challenge has been to monitor oxygen binding characteristics and concomitant pH changes as they occur in vivo, in limited sample volumes. Here we report a technique allowing highly resolved and simultaneous monitoring of pH and blood pigment saturation in minute blood volumes. We equipped a gas diffusion chamber with a broad range fibre optic spectrophotometer and a micro-pH optode and recorded changes of pigment oxygenation along PO2 and pH gradients to test the setup. Oxygen binding parameters derived from measurements in only 15 μl of haemolymph from the cephalopod Octopus vulgaris showed low instrumental error (0.93%) and good agreement with published data. Broad range spectra, each resolving 2048 data points, provided detailed insight into the complex absorbance characteristics of diverse blood types. After consideration of photobleaching and intrinsic fluorescence, pH optodes yielded accurate recordings and resolved a sigmoidal shift of 0.03 pH units in response to changing PO2 from 0-21 kPa. Highly resolved continuous recordings along pH gradients conformed to stepwise measurements at low rates of pH changes. In this study we showed that a diffusion chamber upgraded with a broad range spectrophotometer and an optical pH sensor accurately characterizes oxygen binding with minimal sample consumption and manipulation. We conclude that the modified diffusion chamber is highly suitable for experimental biologists who demand high flexibility, detailed insight into oxygen binding as well as experimental and biological accuracy combined in a single set up.

24 páginas, 4 tablas.-- Graziano Fiorito ... et al.-- This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited., Cephalopods have been utilised in neuroscience research for more than 100 years particularly because of their phenotypic plasticity, complex and centralised nervous system, tractability for studies of learning and cellular mechanisms of memory (e.g. long-term potentiation) and anatomical features facilitating physiological studies (e.g. squid giant axon and synapse). On 1 January 2013, research using any of the about 700 extant species of “live cephalopods” became regulated within the European Union by Directive 2010/63/EU on the “Protection of Animals used for Scientific Purposes”, giving cephalopods the same EU legal protection as previously afforded only to vertebrates. The Directive has a number of implications, particularly for neuroscience research. These include: (1) projects will need justification, authorisation from local competent authorities, and be subject to review including a harm-benefit assessment and adherence to the 3Rs principles (Replacement, Refinement and Reduction). (2) To support project evaluation and compliance with the new EU law, guidelines specific to cephalopods will need to be developed, covering capture, transport, handling, housing, care, maintenance, health monitoring, humane anaesthesia, analgesia and euthanasia. (3) Objective criteria need to be developed to identify signs of pain, suffering, distress and lasting harm particularly in the context of their induction by an experimental procedure. Despite diversity of views existing on some of these topics, this paper reviews the above topics and describes the approaches being taken by the cephalopod research community (represented by the authorship) to produce “guidelines” and the potential contribution of neuroscience research to cephalopod welfare, D.M. has been supported by POR Campania FSE 2007–2013, Project MODO-Model Organism.

Sensitivity of marine crustaceans to anthropogenic CO2 emissions and the associated acidification of the oceans may be less than that of other, especially lower, invertebrates. However, effects on critical transition phases or carry-over effects between life stages have not comprehensively been explored. Here we report the impact of elevated seawater PCO2 values (3100 µatm) on Hyas araneus during the last 2 weeks of their embryonic development (pre-hatching phase) and during development while in the consecutive zoea I and zoea II larval stages (post-hatching phase). We measured oxygen consumption, dry weight, developmental time and mortality in zoea I to assess changes in performance. Feeding rates and survival under starvation were investigated at different temperatures to detect differences in thermal sensitivities of zoea I and zoea II larvae depending on pre-hatch history. When embryos were pre-exposed to elevated PCO2 during maternal care, mortality increased about 60% under continued CO2 exposure during the zoea I phase. The larvae that moulted into zoea II, displayed a developmental delay by about 20 days compared to larvae exposed to control PCO2 during embryonic and zoeal phases. Elevated PCO2 caused a reduction in zoea I dry weight and feeding rates, while survival of the starved larvae was not affected by the seawater CO2 concentration. In conclusion, CO2 effects on egg masses under maternal care carried over to the first larval stages of crustaceans and reduced their survival and development to levels below those previously reported in studies exclusively focussing on acute PCO2 effects on the larval stages.

Mitochondrial plasticity plays a central role in setting the capacity for acclimation of aerobic metabolism in ectotherms in response to environmental changes. We still lack a clear picture if and to what extent the energy metabolism and mitochondrial enzymes of Antarctic fish can compensate for changing temperatures or PCO2 and whether capacities for compensation differ between tissues. We therefore measured activities of key mitochondrial enzymes (citrate synthase (CS), cytochrome c oxidase (COX)) from heart, red muscle, white muscle and liver in the Antarctic fish Notothenia rossii after warm- (7 degrees C) and hypercapnia- (0.2kPa CO2) acclimation vs. control conditions (1 degrees C, 0.04kPa CO2). In heart, enzymes showed elevated activities after cold-hypercapnia acclimation, and a warm-acclimation-induced upward shift in thermal optima. The strongest increase in enzyme activities in response to hypercapnia occurred in red muscle. In white muscle, enzyme activities were temperature-compensated. CS activity in liver decreased after warm-normocapnia acclimation (temperature-compensation), while COX activities were lower after cold- and warm-hypercapnia exposure, but increased after warm-normocapnia acclimation. In conclusion, warm-acclimated N. rossii display low thermal compensation in response to rising energy demand in highly aerobic tissues, such as heart and red muscle. Chronic environmental hypercapnia elicits increased enzyme activities in these tissues, possibly to compensate for an elevated energy demand for acid-base regulation or a compromised mitochondrial metabolism, that is predicted to occur in response to hypercapnia exposure. This might be supported by enhanced metabolisation of liver energy stores. These patterns reflect a limited capacity of N. rossii to reorganise energy metabolism in response to rising temperature and PCO2.

Early life stages of marine crustaceans respond sensitively to elevated seawater P CO 2 . However, the underlying physiological mechanisms have not been studied well. We therefore investigated the effects of elevated seawater P CO 2 on oxygen consumption, dry weight, elemental composition, median developmental time (MDT) and mortality in zoea I larvae of the spider crab Hyas araneus (Svalbard 79°N/11°E; collection, May 2009; hatch, December 2009). At the time of moulting, oxygen consumption rate had reached a steady state level under control conditions. In contrast, elevated seawater P CO 2 caused the metabolic rate to rise continuously leading to a maximum 1.5-fold increase beyond control level a few days before moulting into the second stage (zoea II), followed by a pronounced decrease. Dry weight of larvae reared under high CO 2 conditions was lower than in control larvae at the beginning of the moult cycle, yet this difference had disappeared at the time of moulting. MDT of zoea I varied between 45 ± 1 days under control conditions and 42 ± 2 days under the highest seawater CO 2 concentration. The present study indicates that larval development under elevated seawater P CO 2 levels results in higher metabolic costs during premoulting events in zoea I. However, H. araneus zoea I larvae seem to be able to compensate for higher metabolic costs as larval MDT and survival was not affected by elevated P CO 2 level

Research investigating the genetic basis of physiological responses has significantly broadened our understanding of the mechanisms underlying organismic response to environmental change. However, genomic data are currently available for few taxa only, thus excluding physiological model species from this approach. In this study we report the transcriptome of the model organism Hyas araneus from Spitsbergen (Arctic). We generated 20,479 transcripts, using the 454 {GS} {FLX} sequencing technology in combination with an Illumina HiSeq sequencing approach. Annotation by Blastx revealed 7159 blast hits in the {NCBI} non-redundant protein database. The comparison between the spider crab H. araneus transcriptome and {EST} libraries of the European lobster Homarus americanus and the porcelain crab Petrolisthes cinctipes yielded 3229/2581 sequences with a significant hit, respectively. The clustering by the Markov Clustering Algorithm (MCL) revealed a common core of 1710 clusters present in all three species and 5903 unique clusters for H. araneus. The combined sequencing approaches generated transcripts that will greatly expand the limited genomic data available for crustaceans. We introduce the {MCL} clustering for transcriptome comparisons as a simple approach to estimate similarities between transcriptomic libraries of different size and quality and to analyze homologies within the selected group of species. In particular, we identified a large variety of reverse transcriptase (RT) sequences not only in the H. araneus transcriptome and other decapod crustaceans, but also sea urchin, supporting the hypothesis of a heritable, anti-viral immunity and the proposed viral fragment integration by host-derived {RTs} in marine invertebrates.

Antarctic notothenioid fish are characterized by their evolutionary adaptation to the cold, thermostable Southern Ocean, which is associated with unique physiological adaptations to withstand the cold and reduce energetic requirements but also entails limited compensation capacities to environmental change. This study compares the capacities of mitochondrial acclimation to ocean warming and acidification between the Antarctic nototheniid Notothenia rossii and the sub-Antarctic Lepidonotothen squamifrons, which share a similar ecology, but different habitat temperatures. After acclimation of L. squamifrons to 9°C and N. rossii to 7°C (normocapnic/hypercapnic, 0.2 kPa CO2/2000 ppm CO2) for 4–6 weeks, we compared the capacities of their mitochondrial respiratory complexes I (CI) and II (CII), their P/O ratios (phosphorylation efficiency), proton leak capacities and mitochondrial membrane fatty acid compositions. Our results reveal reduced CII respiration rates in warm-acclimated L. squamifrons and cold hypercapnia-acclimated N. rossii. Generally, L. squamifrons displayed a greater ability to increase CI contribution during acute warming and after warm-acclimation than N. rossii. Membrane unsaturation was not altered by warm or hypercapnia-acclimation in both species, but membrane fatty acids of warm-acclimated L. squamifrons were less saturated than in warm normocapnia−/hypercapnia-acclimated N. rossii. Proton leak capacities were not affected by warm or hypercapnia-acclimation of N. rossii. We conclude that an acclimatory response of mitochondrial capacities may include higher thermal plasticity of CI supported by enhanced utilization of anaplerotic substrates (via oxidative decarboxylation reactions) feeding into the citrate cycle. L. squamifrons possesses higher relative CI plasticities than N. rossii, which may facilitate the usage of energy efficient NADH-related substrates under conditions of elevated energy demand, possibly induced by ocean warming and acidification. The observed adjustments of electron transport system complexes with a higher flux through CI under warming and acidification suggest a metabolic acclimation potential of the sub-Antarctic L. squamifrons, but only limited acclimation capacities for N. rossii.

In the eurythermal cuttlefish Sepia officinalis, performance depends on hearts that ensure systemic oxygen supply over a broad range of temperatures. We therefore aimed to identify adjustments in energetic cardiac capacity and underlying mitochondrial function supportin g therma l acclimati on th at cou ld be criti cal for the cuttlef ish’s comp etitive success in variable environments. Two genetically distinct cuttlefish populations were acclimated to 11°C, 16°C and 21°C, respectively. Subsequently, skinned and permeabilised heart fibres were used to assess mitochondrial functioning by means of high-resolution respirometry and a substrate-inhibitor protocol, followed by measurements of cardiac citrate synthase and cytosolic enzyme activities. In cuttlefish hearts, thermal sensitivity of mitochondrial substrate oxidation was high for proline and pyruvate but low for succinate. Oxygen efficiency of catabolism rose from 11°C to 21°C via shifts to oxygen-conserving oxidation of proline and pyruvate as well as via reduced relative proton leak. Acclimation to 21°C decreased mitochondrial complex I activity in Adriatic cuttlefish and increased complex IV activity in English Channel cuttlefish. However, compensation of mitochondrial capacities did not occur during cold acclimation to 11°C. Moreover, temperate English Channel cuttlefish had larger hearts with lower mitochondrial capacities than subtropical Adriatic cuttlefish. The changes observed for substrate oxidation, mitochondrial complexes, relative proton leak or heart weights improve energetic efficiency and essentially seem to extend tolerance to high temperatures and reduce associated tissue hypoxia. We conclude that cuttlefish sustain cardiac performance and thus, systemic oxygen delivery over short and long-term changes of temperature and environmental conditions by multiple adjustments in cellular and mitochondrial energetics.

Antarctic marine ecosystems are increasingly threatened by climate change and are considered to be particularly sensitive because of the adaptation of most organisms to cold and stable environmental conditions. Fishes play a central role in the Antarctic marine food web and might be affected by climate change in different ways: (i) directly by increasing water temperatures, decreasing seawater salinity and/or increasing concentrations of CO2; (ii) indirectly by alterations in the food web, in particular by changes in prey composition, and (iii) by alterations and loss of habitat due to sea ice retreat and increased ice scouring on the sea floor. Based on new data and data collected from the literature, we analyzed the vulnerability of the fish community to these threats. The potential vulnerability and acting mechanisms differ among species, developmental stages and habitats. The icefishes (family Channichthyidae) are one group that are especially vulnerable to a changing South Polar Sea, as are the pelagic shoal fish species Pleuragramma antarcticum. Both will almost certainly be negatively affected by abiotic alterations and changes in food web structure associated with climate change, the latter additionally by habitat loss. The major bottleneck for the persistence of the majority of populations appears to be the survival of early developmental stages, which are apparently highly sensitive to many types of alterations. In the long term, if climate projections are realized, species loss seems inevitable: within the demersal fish community, the loss or decline of one species might be compensated by others, whereas the pelagic fish community in contrast is extremely poor in species and dominated by P. antarcticum. The loss of this key species could therefore have especially severe consequences for food web structure and the functioning of the entire ecosystem.

There is an increasing need to predict the effects of global climate change on ecologically important marine organisms and a demand for proactive solutions to reduce CO2 emissions. CO2 sequestration is one such method. While this offers a practical solution, recognition should be given to the potential for considerable localised effects on marine organisms in the event of leakage. This laboratory study quantifies the impact of exposure to elevated pCO2 conditions on the physiological and behavioural responses of a relatively tolerant marine organism. Burrowing shrimps Upogebia deltaura were exposed to CO2-enriched seawater for 35 d to treatments of 1396 μatm (pH 7.64), 2707 μatm (pH 7.35) and 14 110 μatm (pH 6.71). CO2 levels represented scenarios which included coastal ocean acidification and extremely elevated CO2 associated with geological CO2 sequestration leaks. Results were compared with those from shrimps maintained in a control treatment (pH 7.99). U. deltaura appeared to be tolerant to elevated pCO2 predicted to occur in the year 2100 (1396 μatm, pH 7.64). However, at 2707 μatm (pH 7.35) shrimps experienced extracellular acidosis, but no difference in haemolymph bicarbonate concentration, suggesting they have little or no buffering capacity, although there was no evidence of other physiological costs in terms of metabolism, osmotic regulation, shell mineralogy, growth and overall activity. At pH 6.71, before 100% mortality occurred, significant differences in activity were observed compared with shrimps in other pH treatments. Results suggest deleterious consequences for benthic ecosystems in the event of a CO2 sequestration leakage.

Mechanisms that affect thermal tolerance of ectothermic organisms have recently received much interest, mainly due to global warming and climate-change debates in both the public and in the scientific community. In physiological terms, thermal tolerance of several marine ectothermic taxa can be linked to oxygen availability, with capacity limitations in ventilatory and circulatory systems contributing to oxygen limitation at extreme temperatures. The present review briefly summarizes the processes that define thermal tolerance in a model cephalopod organism, the cuttlefish Sepia officinalis, with a focus on the contribution of the cephalopod oxygen-carrying blood pigment, hemocyanin. When acutely exposed to either extremely high or low temperatures, cuttlefish display a gradual transition to an anaerobic mode of energy production in key muscle tissues once critical temperatures (T crit ) are reached. At high temperatures, stagnating metabolic rates and a developing hypoxemia can be correlated with a progressive failure of the circulatory system, well before T crit is reached. However, at low temperatures, declining metabolic rates cannot be related to ventilatory or circulatory failure. Rather, we propose a role for hemocyanin functional characteristics as a major limiting factor preventing proper tissue oxygenation. Using information on the oxygen binding characteristics of cephalopod hemocyanins, we argue that high oxygen affinities (= low P 50 values), as found at low temperatures, allow efficient oxygen shuttling only at very low venous oxygen partial pressures. Low venous PO 2 s limit rates of oxygen diffusion into cells, thus eventually causing the observed transition to anaerobic metabolism. On the basis of existing blood physiological, molecular, and crystallographical data, the potential to resolve the role of hemocyanin isoforms in thermal adaptation by an integrated molecular physiological approach is discussed.