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The identification of biological pattern is often complicated by the lack of methodologically consistent data with broad geographic coverage, especially when considering functional characteristics of organisms that differ greatly in body size and morphology. In our study (Dahlke et al. 2020), we addressed the problem of data scarcity by using different types of observational and experimental data together with statistical (phylogenetic) data imputation, and by placing our analysis into the context of a physiological concept, which provides a mechanism-based explanation for the observed pattern (ontogenetic shift in thermal tolerance of fish) and with respect to transition from sublethal to lethal thresholds. Here, we show with comparative examples that our results were not affected by the use of methodologically inconsistent data.

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)

Atlantic cod (Gadus morhua) is one of the most commercially important fish species in the North Atlantic. Environmental factors, such as water temperatures, influence growth of individuals over time, thus forming population-specific growth patterns across climatic regions. Here we develop an integrative approach to investigate the role of temperature in shaping geographic differences of cod growth in the Celtic Sea, North Sea, Iceland, and Barents Sea. We combine a physiology-based growth model and 50-years observational temperature data of 0.5 × 0.5° spatial resolution to simulate continuous growth of cod. The model generated weight-at-age data for the period 1959–2007 which we compared to observational data from fishery-independent scientific surveys. In the Celtic and the northern North Sea, simulated growth matches well observational data. We also show that relatively warm temperatures in the Celtic Sea facilitate maximum growth rates; future warming is likely to have a negative impact on growth of these cod stocks. Growth simulations in Icelandic waters and the Barents Sea are less consistent with local observational data. More complex growth patterns in these regions are probably shaped by ontogenetic shifts in temperature regimes, feeding conditions and physiological adaptations. These findings should stimulate further research on critical processes to be considered in population-specific projections of growth of cod and productivity.

Over the past decades, three major challenges to marine life have emerged as a consequence of anthropogenic emissions: ocean warming, acidification and oxygen loss. While most experimental research has targeted the first two stressors, the last remains comparatively neglected. Here, we implemented sequential hierarchical mixed-model meta-analyses (721 control–treatment comparisons) to compare the impacts of oxygen conditions associated with the current and continuously intensifying hypoxic events (1–3.5 O2 mg l−1) with those experimentally yielded by ocean warming (+4 °C) and acidification (−0.4 units) conditions on the basis of IPCC projections (RCP 8.5) for 2100. In contrast to warming and acidification, hypoxic events elicited consistent negative effects relative to control biological performance—survival (–33%), abundance (–65%), development (–51%), metabolism (–33%), growth (–24%) and reproduction (–39%)—across the taxonomic groups (mollusks, crustaceans and fish), ontogenetic stages and climate regions studied. Our findings call for a refocus of global change experimental studies, integrating oxygen concentration drivers as a key factor of ocean change. Given potential combined effects, multistressor designs including gradual and extreme changes are further warranted to fully disclose the future impacts of ocean oxygen loss, warming and acidification. A meta-analysis of experimental effects of stressors on marine organisms shows that hypoxia could harm crustaceans, mollusks and fish to a larger extent than warming and acidification.

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.

Ocean acidification induces strong behavioural alterations in marine fish as a conse- quence of acid−base regulatory processes in response to increasing environmental CO2 partial pressure. While these changes have been investigated in tropical and temperate fish species, nothing is known about behavioural effects on polar species. In particular, fishes of the Arctic Ocean will experience much greater acidification and warming than temperate or tropical species. Also, possible interactions of ocean warming and acidification are still understudied. Here we analysed the combined effects of warming and acidification on behavioural patterns of 2 fish species co-occurring around Svalbard, viz. polar cod Boreogadus saida and Atlantic cod Gadus morhua. We found a significant temperature effect on the spontaneous activity of B. saida, but not of G. morhua. Environmental CO2 did not significantly influence activity of either species. In con- trast, behavioural laterality of B. saida was affected by CO2 but not by temperature. Behavioural laterality of G. morhua was not affected by temperature or CO2; however, in this species, a possi- ble temperature dependency of CO2 effects on relative laterality may have been missed due to sample size restrictions. This study indicates that fish in polar ecosystems may undergo some, albeit less intense, behavioural disturbances under ocean acidification and in combination with ocean warming than observed in tropical species. It further accentuates species-specific differ- ences in vulnerability.

The vulnerability of fish embryos and larvae to environmental factors is often attributed to a lack of adult-like organ systems (gills) and thus insufficient homeostatic capacity. However, experimental data supporting this hypothesis are scarce. Here, by using Atlantic cod (Gadus morhua) as a model, the relationship between embryo vulnerability (to projected ocean acidification and warming) and homeostatic capacity was explored through parallel analyses of stage-specific mortality and in vitro activity and expression of major ion pumps (ATP-Synthase, Na+/K+-ATPase, H+-ATPase) and co-transporters (NBC1, NKCC1). Immunolocalization of these transporters was used to study ionocyte morphology in newly-hatched larvae. Treatment-related embryo mortality until hatch (+20% due to acidification and warming) occurred primarily during an early period (gastrulation) characterized by extremely low ion transport capacities. Thereafter, embryo mortality decreased in parallel with an exponential increase in activity and expression of all investigated ion transporters. Significant changes in transporter activity and expression in response to acidification (+15% activity) and warming (-30% expression) indicate some potential for short-term acclimatization, although likely associated with energetic trade-offs. Interestingly, whole-larvae enzyme capacities (supported by abundant epidermal ionocytes) reached levels similar to those previously measured in gill tissue of adult cod, suggesting that early-life stages without functional gills are better equipped in terms of ion homeostasis than previously thought. This study implies that the gastrulation period represents a critical transition from inherited (maternal) defenses to active homeostatic regulation, which facilitates enhanced resilience of later stages to environmental factors.

BackgroundBrachyuran crabs can effectively modulate cardiac stroke volume independently of heart rate in response to abiotic drivers. Non-invasive techniques can help to improve the understanding of cardiac performance parameters of these animals. This study demonstrates the in vivo quantification of cardiac performance parameters through magnetic resonance imaging (MRI) on the edible crabCancer pagurus. Furthermore, the suitability of signal integrals of infra-red photoplethysmographs as a qualitative tool is assessed under severe hypoxia.ResultsMulti-slice self-gated cardiac cinematic (CINE) MRI revealed the structure and motion of the ventricle to quantify heart rates, end-diastolic volume, end-systolic volume, stroke volume and ejection fraction. CINE MRI showed that stroke volumes increased under hypoxia because of a reduction of end-systolic volumes at constant end-diastolic volumes. Plethysmograph recordings allowed for automated heart rate measurements but determination of a qualitative stroke volume proxy strongly depended on the position of the sensor on the animal. Both techniques revealed a doubling in stroke volumes after 6 h under severe hypoxia (waterPO2 = 15% air saturation).ConclusionsMRI has allowed for detailed descriptions of cardiac performance in intact animals under hypoxia. The temporal resolution of quantitative non-invasive CINE MRI is limited but should encourage further refining. The stroke volume proxy based on plethysmograph recordings is feasible to complement other cardiac measurements over time. The presented methods allow for non-destructive in vivo determinations of multiple cardiac performance parameters, with the possibility to study neuro-hormonal or environmental effects on decapod cardio physiology.

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.

The maintenance of ion and pH homeostasis despite changes in ambient temperature is crucial for ectothermic organisms. Thermal sensitivity of Na(+)/K(+) ATPase mRNA expression, protein expression and activity was determined in gills of North Sea cod (NC) and Northeastern Arctic cod (NEAC), acclimated for 6 weeks at 4 and 10 °C and compared to field samples of North Sea cod (sNC), acclimatized to early spring (4 °C) and summer (18 °C) conditions. The same analyses were conducted in gills of the confamiliar whiting, acclimated at 4 and 10 °C. Branchial Na(+)/K(+) ATPase capacities remained uncompensated at functional and protein levels in NC and NEAC at both acclimation temperatures. Na(+)/K(+) ATPase mRNA expression in NEAC acclimated at 10 °C was about twofold higher compared to NC, indicating some population-specific differentiation at this level. Lower Na(+)/K(+) ATPase capacities in gills of warm-acclimatized sNC at common assay temperatures indicate thermal compensation between seasonal extremes, and post-translational modifications contributed to this mitigation at high assay temperature. Together, cod compensates Na(+)/K(+) ATPase capacities on the warm edge of the thermal window and below 4 °C, respectively. In contrast, whiting Na(+)/K(+) ATPase capacities were cold compensated at 4 °C, supported by 1.5-fold higher mRNA and protein expression. Besides, capacities were lower in whiting compared to NC and NEAC at optimum temperature, which may be advantageous in terms of reduced maintenance cost, but at temperatures ≤4 °C, compensation may represent an energy trade-off to maintain homeostasis. The species-specific response of gadid Na(+)/K(+) ATPase indicates certain threshold temperatures beyond which compensation of the pump is elicited, possibly related to the different biogeography of these species.

Anthropogenic climate change confronts marine organisms with rapid trends of concomitant warming and CO2 induced ocean acidification. The survival and distribution of species partly depend on their ability to exploit their physiological plasticity during acclimatization. Therefore, in laboratory studies the effects of simulated future ocean acidification on thermal tolerance, energy metabolism and acid–base regulation capacity of the North Sea population of the blue mussel Mytilus edulis were examined. Following one month of pre-acclimation to 10 °C and control CO2 levels, mussels were exposed for two weeks to control and projected oceanic CO2 levels (390, 750 and 1120 μatm) before being subjected to a stepwise warming protocol between 10 °C and 31 °C (+ 3 °C each night). Oxygen consumption and heart rates, anaerobic metabolite levels and haemolymph acid–base status were determined at each temperature. CO2 exposure left oxygen consumption rate unchanged at acclimation temperature but caused a somewhat stronger increase during acute warming and thus mildly higher Q10-values than seen in controls. Interestingly, the thermally induced limitation of oxygen consumption rate set in earlier in normocapnic than in hypercapnic (1120 μatm CO2) mussels (25.2 °C vs. 28.8 °C), likely due to an onset of metabolic depression in the control group following warming. However, the temperature induced increase in heart rate became limited above 25 °C in both groups indicating an unchanged pejus temperature regardless of CO2 treatment. An upper critical temperature was reached above 28 °C in both treatments indicated by the accumulation of anaerobic metabolites in the mantle tissue, paralleled by a strong increase in haemolymph PCO2 at 31 °C. Ocean acidification caused a decrease in haemolymph pH. The extracellular acidosis remained largely uncompensated despite some bicarbonate accumulation. In all treatments animals developed a progressive warming-induced extracellular acidosis. A stronger pH drop at around 25 °C was followed by stagnating heart rates. However, normocapnic mussels enhanced bicarbonate accumulation at the critical limit, a strategy no longer available to hypercapnic mussels. In conclusion, CO2 has small effects on the response patterns of mussels to warming, leaving thermal thresholds largely unaffected. High resilience of adult North Sea mussels to future ocean acidification indicates that sensitivity to thermal stress is more relevant in shaping the response to future climate change.

During development crustaceans experience a range of oxygen tensions (PO2) in their various developmental environments. Therefore, it is likely that they developed varying strategies to respond to hypoxic events during their ontogeny. We investigated short-term responses to various oxygen levels in early life stages of two anomuran decapods Petrolisthes laevigatus and Lithodes santolla that develop in different habitats. Embryos and juveniles of P. laevigatus inhabit the upper rocky intertidal and are regularly exposed to hypoxia during low tide, while larvae of this species inhabit well-oxygenated surface waters. L. santolla is a subtidal species that inhabits fjords and is frequently exposed to hypoxia due to stratification in this environment. Oxygen consumption (MO2), lactate content, RNA/DNA ratio, lactate dehydrogenase and superoxide dismutase (SOD) activities were measured in embryos, larvae and juveniles after exposure to PO2s of 3–21 kPa for 24 h. Results indicated considerable variation in hypoxia tolerances among the ontogenetic stages of both species. Embryos of both species relied on anaerobiosis at all tested PO2 yet were able to regulate MO2 until 15 kPa. In contrast, larvae of L. santolla were oxyconformers, while those of P. laevigatus continued to oxyregulate in conjunction with a decrease in SOD activities in response to low PO2. Juveniles of P. laevigatus were more sensitive to hypoxia than those of L. santolla as all tested parameters were affected at higher PO2 in the former. This study shows that aerobic and anaerobic metabolism, antioxidant defence and protein synthesis of early life stages are diverse and reflect the variety of inhabited environments.

Ocean acidification causes an accumulation of CO2 in marine organisms and leads to shifts in acid–base parameters. Acid–base regulation in gill breathers involves a net increase of internal bicarbonate levels through transmembrane ion exchange with the surrounding water. Successful maintenance of body fluid pH depends on the functional capacity of ion-exchange mechanisms and associated energy budget. For a detailed understanding of the dependence of acid–base regulation on water parameters, we investigated the physiological responses of the shore crab Carcinus maenas to 4 weeks of ocean acidification [OA, P(CO2)w = 1800 µatm], at variable water bicarbonate levels, paralleled by changes in water pH. Cardiovascular performance was determined together with extra-(pHe) and intracellular pH (pHi), oxygen consumption, haemolymph CO2 parameters, and ion composition. High water P(CO2) caused haemolymph P(CO2) to rise, but pHe and pHi remained constant due to increased haemolymph and cellular [HCO3−]. This process was effective even under reduced seawater pH and bicarbonate concentrations. While extracellular cation concentrations increased throughout, anion levels remained constant or decreased. Despite similar levels of haemolymph pH and ion concentrations under OA, metabolic rates, and haemolymph flow were significantly depressed by 40 and 30%, respectively, when OA was combined with reduced seawater [HCO3−] and pH. Our findings suggest an influence of water bicarbonate levels on metabolic rates as well as on correlations between blood flow and pHe. This previously unknown phenomenon should direct attention to pathways of acid–base regulation and their potential feedback on whole-animal energy demand, in relation with changing seawater carbonate parameters.

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

In this article, we pointed out that understanding the physiology of differential climate change effects on organisms is one of the many urgent challenges faced in ecology and evolutionary biology. We explore how physiological ecology can contribute to a holistic view of climate change impacts on organisms and ecosystems and their evolutionary responses. We suggest that theoretical and experimental efforts not only need to improve our understanding of thermal limits to organisms, but also to consider multiple stressors both on land and in the oceans. As an example, we discuss recent efforts to understand the effects of various global change drivers on aquatic ectotherms in the field that led to the development of the concept of oxygen and capacity limited thermal tolerance (OCLTT) as a framework integrating various drivers and linking organisational levels from ecosystem to organism, tissue, cell, and molecules. We suggest seven core objectives of a comprehensive research program comprising the interplay among physiological, ecological, and evolutionary approaches for both aquatic and terrestrial organisms. While studies of individual aspects are already underway in many laboratories worldwide, integration of these findings into conceptual frameworks is needed not only within one organism group such as animals but also across organism domains such as Archaea, Bacteria, and Eukarya. Indeed, development of unifying concepts is relevant for interpreting existing and future findings in a coherent way and for projecting the future ecological and evolutionary effects of climate change on functional biodiversity. We also suggest that OCLTT may in the end and from an evolutionary point of view, be able to explain the limited thermal tolerance of metazoans when compared to other organisms.

Background Exposure to future ocean acidification scenarios may alter the behaviour of marine teleosts through interference with neuroreceptor functioning. So far, most studies investigated effects of ocean acidification on the behaviour of fish, either isolated or in combination with environmental temperature. However, only few physiological studies on this issue were conducted despite the putative neurophysiological origin of the CO2-induced behavioural changes. Here, we present the metabolic consequences of long-term exposure to projected ocean acidification (396–548 μatm PCO2 under control and 915–1272 μatm under treatment conditions) and parallel warming in the brain of two related fish species, polar cod (Boreogadus saida, exposed to 0 °C, 3 °C, 6 °C and 8 °C) and Atlantic cod (Gadus morhua, exposed to 3 °C, 8 °C, 12 °C and 16 °C). It has been shown that B. saida is behaviourally vulnerable to future ocean acidification scenarios, while G. morhua demonstrates behavioural resilience. Results We found that temperature alters brain osmolyte, amino acid, choline and neurotransmitter concentrations in both species indicating thermal responses particularly in osmoregulation and membrane structure. In B. saida, changes in amino acid and osmolyte metabolism at the highest temperature tested were also affected by CO2, possibly emphasizing energetic limitations. We did not observe changes in neurotransmitters, energy metabolites, membrane components or osmolytes that might serve as a compensatory mechanism against CO2 induced behavioural impairments. In contrast to B. saida, such temperature limitation was not detected in G. morhua; however, at 8 °C, CO2 induced an increase in the levels of metabolites of the glutamate/GABA-glutamine cycle potentially indicating greater GABAergic activity in G.morhua. Further, increased availability of energy-rich substrates was detected under these conditions. Conclusions Our results indicate a change of GABAergic metabolism in the nervous system of Gadus morhua close to the optimum of the temperature range. Since a former study showed that juvenile G. morhua might be slightly more behaviourally resilient to CO2 at this respective temperature, we conclude that the observed change of GABAergic metabolism could be involved in counteracting OA induced behavioural changes. This may serve as a fitness advantage of this respective species compared to B. saida in a future warmer, more acidified polar ocean. Electronic supplementary material The online version of this article (10.1186/s12983-017-0238-5) contains supplementary material, which is available to authorized users.

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.

Eulimnogammarus verrucosusis an amphipod endemic to the unique ecosystem of Lake Baikal and serves in particular as an emerging model in ecotoxicological studies. We report here on a survey sequencing of its genome as a first step to establish sequence resources for this species. From a sin gle lane of paired-end sequencing data we estimated the genome size as nearly 10 Gb and we obtained an overview of the repeat content. At least two thirds of the genome are non-unique DNA, and a third of the genomic DNA is composed of just five families of repetitive elements, including low-complexity sequences. Attempts to use off-the-shelf assembly tools failed on the available low-coverage data both before and after removal of highly repetitive components. Using a seed-based approach we nevertheless assembled short contigs covering 33 pre-microRNAs and the homeodomain-containing exon of nine Hox genes. The absence of clear evidence for paralogs implies that a genome duplication did not contribute to the large genome size. We furthermore report the assembly of the mitochondrial genome using a new, guided “crystallization” procedure. The initial results presented here set the stage for a more compl ete sequencing and analysis of this large genome.

According to the Membrane Pacemaker Theory of metabolism (MPT) allometric scaling of metabolic rate in animals is determined by the composition of cellular and mitochondrial membranes that changes with body size in a predictable manner. MPT has been elaborated from interspecific comparisons in mammals. It projects that the degree of unsaturation of membrane phospholipids decreases in larger organisms, thereby lowering ion permeability of the membranes and making cellular and thus whole animal metabolism more efficient. Here we tested the applicability of the MPT to a marine ectotherm, the mussel Mytilus edulis at the intraspecific level. We determined effects of body mass on whole organism, tissue and cellular oxygen consumption rates, on heart rate, metabolic enzyme activities and on the lipid composition of membranes. In line with allometric patterns the organismal functions and processes such as heart rate, whole animal respiration rate and phospholipid contents showed a mass-dependent decline. However, the allometry of tissue and cellular respiration and activity of metabolic enzymes was poor; fatty acid unsaturation of membrane phospholipids of gill tissue was independent of animal size. It is thus conceivable that most of the metabolic allometry observed at the organismal level is determined by systemic functions. These whole organism patterns may be supported by energy savings associated with growing cell size but not by structural changes in membranes. Overall, the set of processes contributing to metabolic allometry in ectotherms may differ from that operative in mammals and birds, with a reduced involvement of the mechanisms proposed by the MPT.

Increased maintenance costs at cellular, and consequently organism level, are thought to be involved in shaping the sensitivity of marine calcifiers to ocean acidification (OA). Yet, knowledge of the capacity of marine calcifiers to undergo metabolic adaptation is sparse. In Kiel Fjord, blue mussels thrive despite periodically high seawater PCO2, making this population interesting for studying metabolic adaptation under OA. Consequently, we conducted a multi-generation experiment and compared physiological responses of F1 mussels from ‘tolerant’ and ‘sensitive’ families exposed to OA for 1 year. Family classifications were based on larval survival; tolerant families settled at all PCO2 levels (700, 1120, 2400 µatm) while sensitive families did not settle at the highest PCO2 (≥99.8% mortality). We found similar filtration rates between family types at the control and intermediate PCO2 level. However, at 2400 µatm, filtration and metabolic scope of gill tissue decreased in tolerant families, indicating functional limitations at the tissue level. Routine metabolic rates (RMR) and summed tissue respiration (gill and outer mantle tissue) of tolerant families were increased at intermediate PCO2, indicating elevated cellular homeostatic costs in various tissues. By contrast, OA did not affect tissue and routine metabolism of sensitive families. However, tolerant mussels were characterised by lower RMR at control PCO2 than sensitive families, which had variable RMR. This might provide the energetic scope to cover increased energetic demands under OA, highlighting the importance of analysing intra-population variability. The mechanisms shaping such difference in RMR and scope, and thus species’ adaptation potential, remain to be identified.

With global climate change, ocean warming and acidification occur concomitantly. In this study, we tested the hypothesis that increasing CO2 levels affect the acid–base balance and reduce the activity capacity of the Arctic spider crab Hyas araneus, especially at the limits of thermal tolerance. Crabs were acclimated to projected oceanic CO2 levels for 12 days (today: 380, towards the year 2100: 750 and 1,120 and beyond: 3,000 μatm) and at two temperatures (1 and 4 °C). Effects of these treatments on the righting response (RR) were determined (1) at acclimation temperatures followed by (2) righting when exposed to an additional acute (15 min) heat stress at 12 °C. Prior to (resting) and after the consecutive stresses of combined righting activity and heat exposure, acid–base status and lactate contents were measured in the haemolymph. Under resting conditions, CO2 caused a decrease in haemolymph pH and an increase in oxygen partial pressure. Despite some buffering via an accumulation of bicarbonate, the extracellular acidosis remained uncompensated at 1 °C, a trend exacerbated when animals were acclimated to 4 °C. The additional combined exposure to activity and heat had only a slight effect on blood gas and acid–base status. Righting activity in all crabs incubated at 1 and 4 °C was unaffected by elevated CO2 levels or acute heat stress but was significantly reduced when both stressors acted synergistically. This impact was much stronger in the group acclimated at 1 °C where some individuals acclimated to high CO2 levels stopped responding. Lactate only accumulated in the haemolymph after combined righting and heat stress. In the group acclimated to 1 °C, lactate content was highest under normocapnia and lowest at the highest CO2 level in line with the finding that RR was largely reduced. In crabs acclimated to 4 °C, the RR was less affected by CO2 such that activity caused lactate to increase with rising CO2 levels. In line with the concept of oxygen and capacity limited thermal tolerance, all animals exposed to temperature extremes displayed a reduction in scope for performance, a trend exacerbated by increasing CO2 levels. Additionally, the differences seen between cold- and warm-acclimated H. araneus after heat stress indicate that a small shift to higher acclimation temperatures also alleviates the response to temperature extremes, indicating a shift in the thermal tolerance window which reduces susceptibility to additional CO2 exposure.

The ongoing process of ocean acidification already affects marine life, and according to the concept of oxygen and capacity limitation of thermal tolerance, these effects may be intensified at the borders of the thermal tolerance window. We studied the effects of elevated CO2 concentrations on clapping performance and energy metabolism of the commercially important scallop Pecten maximus. Individuals were exposed for at least 30 days to 4 °C (winter) or to 10 °C (spring/summer) at either ambient (0.04 kPa, normocapnia) or predicted future PCO2 levels (0.11 kPa, hypercapnia). Cold-exposed (4 °C) groups revealed thermal stress exacerbated by PCO2 indicated by a high mortality overall and its increase from 55 % under normocapnia to 90 % under hypercapnia. We therefore excluded the 4 °C groups from further experimentation. Scallops at 10 °C showed impaired clapping performance following hypercapnic exposure. Force production was significantly reduced although the number of claps was unchanged between normocapnia- and hypercapnia-exposed scallops. The difference between maximal and resting metabolic rate (aerobic scope) of the hypercapnic scallops was significantly reduced compared with normocapnic animals, indicating a reduction in net aerobic scope. Our data confirm that ocean acidification narrows the thermal tolerance range of scallops resulting in elevated vulnerability to temperature extremes and impairs the animal’s performance capacity with potentially detrimental consequences for its fitness and survival in the ocean of tomorrow.

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.

Marine organisms have to cope with increasing CO2 partial pressures and decreasing pH in the oceans. We elucidated the impacts of an 8-week acclimation period to four seawater pCO2 treatments (39, 113, 243 and 405 Pa/385, 1,120, 2,400 and 4,000 µatm) on mantle gene expression patterns in the blue mussel Mytilus edulis from the Baltic Sea. Based on the M. edulis mantle tissue transcriptome, the expression of several genes involved in metabolism, calcification and stress responses was assessed in the outer (marginal and pallial zone) and the inner mantle tissues (central zone) using quantitative real-time PCR. The expression of genes involved in energy and protein metabolism (F-ATPase, hexokinase and elongation factor alpha) was strongly affected by acclimation to moderately elevated CO2 partial pressures. Expression of a chitinase, potentially important for the calcification process, was strongly depressed (maximum ninefold), correlating with a linear decrease in shell growth observed in the experimental animals. Interestingly, shell matrix protein candidate genes were less affected by CO2 in both tissues. A compensatory process toward enhanced shell protection is indicated by a massive increase in the expression of tyrosinase, a gene involved in periostracum formation (maximum 220-fold). Using correlation matrices and a force-directed layout network graph, we were able to uncover possible underlying regulatory networks and the connections between different pathways, thereby providing a molecular basis of observed changes in animal physiology in response to ocean acidification.

The efficiency of aerial respiration and the overall dependence on water availability during embryonic development have never been tested in semi-terrestrial or terrestrial crabs. In the present experimental study, we show that respiration of the embryos of a semi-terrestrial intertidal crab, Armases miersii from Jamaica, is bimodal. They are able to extract oxygen from both air and water, although a lower air respiration with approximately half O 2 uptake than in water from the stage III onward was observed. Comparing the embryos of ovigerous females that were impeded for 18 h per day to submerge their egg clutches in seawater (n = 8 treatments), with those having unlimited access to water (n = 7 controls), we observed no differences in embryonic respiration or development time, but the lack of water caused complete brood-loss in 5 females and massive developmental problems in the remaining 3 broods, with an average hatching rate of only 28% and deformations in most newly hatched larvae. Although the higher embryonic O 2 uptake in water suggests a reduced embryonic ability to extract oxygen from air, we propose as an alternative hypothesis an adaptive down-regulation of the metabolic rate, which is not linked to oxygen extraction but to the water dependence of vital metabolic pathways, such as the excretion of ammonia and CO 2 . Since bimodal embryonic respiration has only been known from vertebrates, this is the first study demonstrating aerial respiration in brachyuran embryos. However, developmental deficiencies in water-limited egg clutches suggest that the embryos A. miersii still depend on water to avoid desiccation stress and, probably, an impediment of excretory processes.

As CO2 from fossil fuel emissions enters the ocean, it is becoming acidified. Global surface pH has already decreased by more than 0.1 units since preindustrial times and might decrease by 0.10 – 0.35 additional units by 2100. The level of supersaturation of calcium carbonate has decreased and the partial pressure of carbon dioxide PCO2 has increased over much of the upper ocean. We present predicted changes in ocean surface pH from the Canadian Earth System Model (CanESM-1). Results presented at a 2010 workshop in Sendai, Japan, of experiments on marine animals under elevated CO2 conditions indicate that few general conclusions across species can as yet be made. Recommendations include protocols that mimic natural diel and seasonal fluctuations in PCO2 and that extend over multiple generations. Projections from CanESM-1 suggest that other changes associated with climate change have little influence on acidification relative to the addition of CO2 and that freshwater dilution at high northern latitudes exacerbates acidification and results in undersaturation with respect to carbonate minerals at higher pH than in undiluted seawater. Finally, we recommend a framework that considers the effects of multiple stressors associated with a changing climate on the “thermal window” of activity of marine animals.

The combined effects of ocean warming and acidification were compared in larvae from two populations of the cold-eurythermal spider crab Hyas araneus, from one of its southernmost populations (around Helgoland, southern North Sea, 54A degrees N, habitat temperature 3-18A degrees C; collection: January 2008, hatch: January-February 2008) and from one of its northernmost populations (Svalbard, North Atlantic, 79A degrees N, habitat temperature 0-6A degrees C; collection: July 2008, hatch: February-April 2009). Larvae were exposed to temperatures of 3, 9 and 15A degrees C combined with present-day normocapnic (380 ppm CO(2)) and projected future CO(2) concentrations (710 and 3,000 ppm CO(2)). Calcium content of whole larvae was measured in freshly hatched Zoea I and after 3, 7 and 14 days during the Megalopa stage. Significant differences between Helgoland and Svalbard Megalopae were observed at all investigated temperatures and CO(2) conditions. Under 380 ppm CO(2), the calcium content increased with rising temperature and age of the larvae. At 3 and 9A degrees C, Helgoland Megalopae accumulated more calcium than Svalbard Megalopae. Elevated CO(2) levels, especially 3,000 ppm, caused a reduction in larval calcium contents at 3 and 9A degrees C in both populations. This effect set in early, at 710 ppm CO(2) only in Svalbard Megalopae at 9A degrees C. Furthermore, at 3 and 9A degrees C Megalopae from Helgoland replenished their calcium content to normocapnic levels and more rapidly than Svalbard Megalopae. However, Svalbard Megalopae displayed higher calcium contents under 3,000 ppm CO(2) at 15A degrees C. The findings of a lower capacity for calcium incorporation in crab larvae living at the cold end of their distribution range suggests that they might be more sensitive to ocean acidification than those in temperate regions.

Marine brachyuran and anomuran crustaceans are completely absent from the extremely cold (− 1.8 °C) Antarctic continental shelf, but caridean shrimps are abundant. This has at least partly been attributed to low capacities for magnesium excretion in brachyuran and anomuran lithodid crabs ([Mg2+]HL = 20–50 mmol L− 1) compared to caridean shrimp species ([Mg2+]HL = 5–12 mmol L− 1). Magnesium has an anaesthetizing effect and reduces cold tolerance and activity of adult brachyuran crabs. We investigated whether the capacity for magnesium regulation is a factor that influences temperature-dependent activity of early ontogenetic stages of the Sub-Antarctic lithodid crab Paralomis granulosa. Ion composition (Na+, Mg2+, Ca2+, Cl−, SO42−) was measured in haemolymph withdrawn from larval stages, the first and second juvenile instars (crabs I and II) and adult males and females. Magnesium excretion improved during ontogeny, but haemolymph sulphate concentration was lowest in the zoeal stages. Neither haemolymph magnesium concentrations nor Ca2+:Mg2+ ratios paralleled activity levels of the life stages. Long-term (3 week) cold exposure of crab I to 1 °C caused a significant rise of haemolymph sulphate concentration and a decrease in magnesium and calcium concentrations compared to control temperature (9 °C). Spontaneous swimming activity of the zoeal stages was determined at 1, 4 and 9 °C in natural sea water (NSW, [Mg2+] = 51 mmol L− 1) and in sea water enriched with magnesium (NSW + Mg2+, [Mg2+] = 97 mmol L− 1). It declined significantly with temperature but only insignificantly with increased magnesium concentration. Spontaneous velocities were low, reflecting the demersal life style of the zoeae. Heart rate, scaphognathite beat rate and forced swimming activity (maxilliped beat rate, zoea I) or antennule beat rate (crab I) were investigated in response to acute temperature change (9, 6, 3, 1, − 1 °C) in NSW or NSW + Mg2+. High magnesium concentration reduced heart rates in both stages. The temperature–frequency curve of the maxilliped beat (maximum: 9.6 beats s− 1 at 6.6 °C in NSW) of zoea I was depressed and shifted towards warmer temperatures by 2 °C in NSW + Mg2+, but antennule beat rate of crab I was not affected. Magnesium may therefore influence cold tolerance of highly active larvae, but it remains questionable whether the slow-moving lithodid crabs with demersal larvae would benefit from an enhanced magnesium excretion in nature.

Dissolved oxygen (DO) concentration in the water column is an environmental parameter that is crucial for the successful development of many pelagic organisms. Hypoxia tolerance and threshold values are species- and stage-specific and can vary enormously. While some fish species may suffer from oxygen values of less than 3 mL O2 L−1 through impacted growth, development and behaviour, other organisms such as euphausiids may survive DO levels as low as 0.1 mL O2 L−1. A change in the average or the range of DO may have significant impacts on the survival of certain species and hence on the species composition in the ecosystem with consequent changes in trophic pathways and productivity. Evidence for the deleterious effects of oxygen depletion on pelagic species is scarce, particularly in terms of the effect of low oxygen on development, recruitment and patterns of migration and distribution. While planktonic organisms have to cope with variable DOs and exploit adaptive mechanisms, nektonic species may avoid areas of unfavourable DO and develop adapted migration strategies. Planktonic organisms may only be able to escape vertically, above or beneath the Oxygen Minimum Zone (OMZ). In shallow areas only the surface layer can serve as a refuge, but in deep waters many organisms have developed vertical migration strategies to use, pass through and cope with the OMZ. This paper elucidates the role of DO for different taxa in the pelagic realm and the consequences of low oxygen for foodweb structure and system productivity. We describe processes in two contrasting systems, the semi-enclosed Baltic Sea and the coastal upwelling system of the Benguela Current to demonstrate the consequences of increasing hypoxia on ecosystem functioning and services.

While documentation of climate effects on marine ecosystems has a long history, the underlying processes have often been elusive. In this paper we review some of the ecosystem responses to climate variability and discuss the possible mechanisms through which climate acts. Effects of climatological and oceanographic variables, such as temperature, sea ice, turbulence, and advection, on marine organisms are discussed in terms of their influence on growth, distribution, reproduction, activity rates, recruitment and mortality. Organisms tend to be limited to specific thermal ranges with experimental findings showing that sufficient oxygen supply by ventilation and circulation only occurs within these ranges. Indirect effects of climate forcing through effects on the food web are also discussed. Research and data needs required to improve our knowledge of the processes linking climate to ecosystem changes are presented along with our assessment of our ability to predict ecosystem responses to future climate change scenarios.

SUMMARY The concept of oxygen- and capacity-dependent thermal tolerance in aquatic ectotherms has successfully explained climate-induced effects of rising temperatures on species abundance in the field. Oxygen supply to tissues and the resulting aerobic performance characters thus form a primary link between organismal fitness and its role and functioning at the ecosystem level. The thermal window of performance in water breathers matches their window of aerobic scope. Loss of performance reflects the earliest level of thermal stress, caused by hypoxaemia and the progressive mismatch of oxygen supply and demand at the borders of the thermal envelope. Oxygen deficiency elicits the transition to passive tolerance and associated systemic and cellular stress signals like hormonal responses or oxidative stress as well as the use of protection mechanisms like heat shock proteins at thermal extremes. Thermal acclimatization between seasons or adaptation to a climate regime involves shifting thermal windows and adjusting window widths. The need to specialize on a limited temperature range results from temperature-dependent trade-offs at several hierarchical levels, from molecular structure to whole-organism functioning, and may also support maximized energy efficiency. Various environmental factors like CO2 (ocean acidification) and hypoxia interact with these principal relationships. Existing knowledge suggests that these factors elicit metabolic depression supporting passive tolerance to thermal extremes. However, they also exacerbate hypoxaemia, causing a narrowing of thermal performance windows and prematurely leading the organism to the limits of its thermal acclimation capacity. The conceptual analysis suggests that the relationships between energy turnover, the capacities of activity and other functions and the width of thermal windows may lead to an integrative understanding of specialization on climate and, as a thermal matrix, of sensitivity to climate change and the factors involved. Such functional relationships might also relate to climate-induced changes in species interactions and, thus, community responses at the ecosystem level.

Ongoing climate change is predicted to affect individual organisms during all life stages, thereby affecting populations of a species, communities and the functioning of ecosystems. These effects of climate change can be direct, through changing water temperatures and associated phenologies, the lengths and frequency of hypoxia events, through ongoing ocean acidification trends or through shifts in hydrodynamics and in sea level. In some cases, climate interactions with a species will also, or mostly, be indirect and mediated through direct effects on key prey species which change the composition and dynamic coupling of food webs. Thus, the implications of climate change for marine fish populations can be seen to result from phenomena at four interlinked levels of biological organization: (1) organismal-level physiological changes will occur in response to changing environmental variables such as temperature, dissolved oxygen and ocean carbon dioxide levels. An integrated view of relevant effects, adaptation processes and tolerance limits is provided by the concept of oxygen and capacity-limited thermal tolerance (OCLT). (2) Individual-level behavioural changes may occur such as the avoidance of unfavourable conditions and, if possible, movement into suitable areas. (3) Population-level changes may be observed via changes in the balance between rates of mortality, growth and reproduction. This includes changes in the retention or dispersion of early life stages by ocean currents, which lead to the establishment of new populations in new areas or abandonment of traditional habitats. (4) Ecosystem-level changes in productivity and food web interactions will result from differing physiological responses by organisms at different levels of the food web. The shifts in biogeography and warming-induced biodiversity will affect species productivity and may, thus, explain changes in fisheries economies. This paper tries to establish links between various levels of biological organization by means of addressing the effective physiological principles at the cellular, tissue and whole organism levels.

Future ocean acidification has the potential to adversely affect many marine organisms. A growing body of evidence suggests that many species could suffer from reduced fertilization success, decreases in larval- and adult growth rates, reduced calcification rates, and even mortality when being exposed to near-future levels (year 2100 scenarios) of ocean acidification. Little research focus is currently placed on those organisms/taxa that might be less vulnerable to the anticipated changes in ocean chemistry; this is unfortunate, as the comparison of more vulnerable to more tolerant physiotypes could provide us with those physiological traits that are crucial for ecological success in a future ocean. Here, we attempt to summarize some ontogenetic and lifestyle traits that lead to an increased tolerance towards high environmental pCO2. In general, marine ectothermic metazoans with an extensive extracellular fluid volume may be less vulnerable to future acidification as their cells are already exposed to much higher pCO2 values (0.1 to 0.4 kPa, ca. 1000 to 3900 μatm) than those of unicellular organisms and gametes, for which the ocean (0.04 kPa, ca. 400 μatm) is the extracellular space. A doubling in environmental pCO2 therefore only represents a 10% change in extracellular pCO2 in some marine teleosts. High extracellular pCO2 values are to some degree related to high metabolic rates, as diffusion gradients need to be high in order to excrete an amount of CO2 that is directly proportional to the amount of O2 consumed. In active metazoans, such as teleost fish, cephalopods and many brachyuran crustaceans, exercise induced increases in metabolic rate require an efficient ion-regulatory machinery for CO2 excretion and acid-base regulation, especially when anaerobic metabolism is involved and metabolic protons leak into the extracellular space. These ion-transport systems, which are located in highly developed gill epithelia, form the basis for efficient compensation of pH disturbances during exposure to elevated environmental pCO2. Compensation of extracellular acid-base status in turn may be important in avoiding metabolic depression. So far, maintained "performance" at higher seawater pCO2 (>0.3 to 0.6 kPa) has only been observed in adults/juveniles of active, high metabolic species with a powerful ion regulatory apparatus. However, while some of these taxa are adapted to cope with elevated pCO2 during their regular embryonic development, gametes, zygotes and early embryonic stages, which lack specialized ion-regulatory epithelia, may be the true bottleneck for ecological success – even of the more tolerant taxa. Our current understanding of which marine animal taxa will be affected adversely in their physiological and ecological fitness by projected scenarios of anthropogenic ocean acidification is quite incomplete. While a growing amount of empirical evidence from CO2 perturbation experiments suggests that several taxa might react quite sensitively to ocean acidification, others seem to be surprisingly tolerant. However, there is little mechanistic understanding on what physiological traits are responsible for the observed differential sensitivities (see reviews of Seibel and Walsh, 2003; Pörtner et al., 2004; Fabry et al., 2008; Pörtner, 2008). This leads us to the first very basic question of how to define general CO2 tolerance on the species level.

Future scenarios for the oceans project combined developments of CO2 accumulation and global warming and their impact on marine ecosystems. The synergistic impact of both factors was addressed by studying the effect of elevated CO2 concentrations on thermal tolerance of the cold-eurythermal spider crab Hyas araneus from the population around Helgoland. Here ambient temperatures characterize the southernmost distribution limit of this species. Animals were exposed to present day normocapnia (380 ppm CO2), CO2 levels expected towards 2100 (710 ppm) and beyond (3000 ppm). Heart rate and haemolymph PO2 (PeO2) were measured during progressive short term cooling from 10 to 0°C and during warming from 10 to 25°C. An increase of PeO2 occurred during cooling, the highest values being reached at 0°C under all three CO2 levels. Heart rate increased during warming until a critical temperature (Tc) was reached. The putative Tc under normocapnia was presumably >25°C, from where it fell to 23.5°C under 710 ppm and then 21.1°C under 3000 ppm. At the same time, thermal sensitivity, as seen in the Q10 values of heart rate, rose with increasing CO2 concentration in the warmth. Our results suggest a narrowing of the thermal window of Hyas araneus under moderate increases in CO2 levels by exacerbation of the heat or cold induced oxygen and capacity limitation of thermal tolerance.

Tolerance to a changing climate regime and persistence in the natural environment depends on the limited capacity to acclimate to changing temperatures. The present study aimed to identify and characterize thermal limits of the Mediterranean fish Sparus aurata as well as the processes providing heat protection during exposure to high temperatures. Processes studied included heat shock protein expression, protein kinase activity and metabolic adjustments. Molecular responses were addressed through the expression of Hsp70 and Hsp90 and the phosphorylation of stress-activated protein kinases, p38 mitogen-activated protein kinase (p38 MAPK) and cJun-N-terminal kinases (JNKs). Thermal impacts on metabolic capacities were assessed by studying the maximum activities of citrate synthase (CS), malate dehydrogenase (MDH) and 3-hydroxyacyl CoA dehydrogenase (HOAD) as well as pyruvate kinase (PK) and lactate dehydrogenase (L-LDH). The expression of Hsp70 and hsp90 was activated when the fish were exposed to temperatures beyond 20°C. Increased phosphorylation of p38 MAPK and JNKs indicated the parallel activation of MAPK signaling cascades and the potential involvement of MAPKs in the induction of Hsp genes. Exposure to extreme temperatures beyond 24°C caused an increase in the enzymatic activity of PK and LDH indicating an enhanced glycolytic potential.

Acidification of ocean surface waters by anthropogenic carbon dioxide (CO(2)) emissions is a currently developing scenario that warrants a broadening of research foci in the study of acid-base physiology. Recent studies working with environmentally relevant CO(2) levels, indicate that some echinoderms and molluscs reduce metabolic rates, soft tissue growth and calcification during hypercapnic exposure. In contrast to all prior invertebrate species studied so far, growth trials with the cuttlefish Sepia officinalis found no indication of reduced growth or calcification performance during long-term exposure to 0.6 kPa CO(2). It is hypothesized that the differing sensitivities to elevated seawater pCO(2) could be explained by taxa specific differences in acid-base regulatory capacity. In this study, we examined the acid-base regulatory ability of S. officinalis in vivo, using a specially modified cannulation technique as well as (31)P NMR spectroscopy. During acute exposure to 0.6 kPa CO(2), S. officinalis rapidly increased its blood [HCO(3)(-)] to 10.4 mM through active ion-transport processes, and partially compensated the hypercapnia induced respiratory acidosis. A minor decrease in intracellular pH (pH(i)) and stable intracellular phosphagen levels indicated efficient pH(i) regulation. We conclude that S. officinalis is not only an efficient acid-base regulator, but is also able to do so without disturbing metabolic equilibria in characteristic tissues or compromising aerobic capacities. The cuttlefish did not exhibit acute intolerance to hypercapnia that has been hypothesized for more active cephalopod species (squid). Even though blood pH (pHe) remained 0.18 pH units below control values, arterial O(2) saturation was not compromised in S. officinalis because of the comparatively lower pH sensitivity of oxygen binding to its blood pigment. This raises questions concerning the potentially broad range of sensitivity to changes in acid-base status amongst invertebrates, as well as to the underlying mechanistic origins. Further studies are needed to better characterize the connection between acid-base status and animal fitness in various marine species.

SUMMARYExposure of ectothermic organisms to variations in temperatures causes a transient mismatch between energy supply and demand, which needs to be compensated for during acclimation. Adenosine accumulation from ATP breakdown indicates such an imbalance and its reversal reflects a restoration of energy status. We monitored adenosine levels in blood serum and liver of common eelpout (Zoarces viviparus) during cold exposure in vivo. Furthermore, we tested its effect on the pattern of thermal acclimation in hepatocytes isolated from cold- (4°C) versus warm- (11°C) exposed fish. Adenosine levels increased during cold exposure in vivo and reached a transient maximum after 24 h in serum, but remained permanently elevated in liver. Whole animal cold acclimation induced a rise of liver citrate synthase activity by 44±15%, but left cytochrome c oxidase activity (COX) and RNA expression of the respective genes unchanged. Cold incubation of hepatocytes from warm-acclimated fish failed to cause an increase of mitochondrial enzyme activities despite increased COX4 mRNA levels. Conversely, warm acclimation of hepatocytes from cold-acclimated fish reduced both enzyme activities and COX2 and COX4 mRNA levels by 26–37%. Adenosine treatment of both warm- and cold-acclimated hepatocytes suppressed COX activities but activated COX mRNA expression. These effects were not receptor mediated. The present findings indicate that adenosine has the potential to regulate mitochondrial functioning in vivo, albeit the pathways resulting in the contrasting effects on expression and activity need to be identified.

Ocean warming and acidification occur at global scales and, in the case of temperature, have already caused shifts in marine ecosystem composition and function. In the case of CO2-induced ocean hypercapnia and acidification, however, effects may still be so small that evidence for changes in the field is largely lacking. Future scenarios indicate that marine life forms are threatened by the specific or synergistic effects of factors involved in these processes. The present paper builds on the view that development of a cause and effect understanding is required beyond empirical observations, for a more accurate projection of ecosystem effects and for quantitative scenarios. Identification of the mechanisms through which temperatureand CO2-related ocean physicochemistry affect organism fitness, survival and success, is crucial with this research strategy. I suggest operation of unifying physiological principles, not only of temperature but also CO2 effects, across animal groups and phyla. Thermal windows of optimized performance emerge as a basic character defining species fitness and survival, including their capacity to interact with other species. Through effects on performance at the level of reproduction, behaviour and growth, ocean acidification acts especially on lower marine invertebrates, which are characterized by a low capacity to compensate for disturbances in extracellular ion and acid–base status and sensitivity of metabolism to such disturbances. Available data suggest that one key consequence of these features is a narrowing of thermal tolerance windows, as well as a reduced scope for performance at ecosystem level. These changes in bioenvelopes may have major implications for the ranges of geographical distribution of these organisms and in species interactions.

Low temperature limits the rate of biochemical reactions and aerobic scopes of cold water ectotherms. To compensate for this limiting effect, animals living in cold environments often possess physiological or morphological adaptations to maintain vital functions. Cross-latitudinal comparison of aerobic capacities is one method to test which factors constrain activity in thermally distinct environments particularly when congeneric studies are carried out on related species with conservative ecology and habitat. Burrowing is a major aerobic activity of bivalve molluscs that is described here for the first time for the tropical mangrove species Laternula truncata and Laternula boschasina and then compared with their Antarctic congener Laternula elliptica. About 80% of L. truncata (16.3–46.1 mm shell length) and 63% of L. boschasina (11.3–27.7 mm shell length) buried within 24 h at 28°C. The burrowing rate index (BRI = [3√wet weight/time to bury]×104) ranged between 1.1 and 20.2 for L. boschasina and 1.1–32.9 for L. truncata. These values are 2–3 orders of magnitude less than other tropical bivalve molluscs and are amongst the lowest recorded for any bivalve. Comparisons with the Antarctic L. elliptica showed little or no differences in BRI (Q10 of 1.0–1.2 for specimens of the same size). This is contrary to the general pattern over a wide range of bivalves, where BRI increases with a Q10 of between 2.9 and 6.4 between high latitudes and the equator. L. elliptica has 25–30% longer relative foot length than tropical congeners of the same size, which could be a morphological adaptation compensating for reduced burrowing speeds in a colder environment. Burrowing rates within the genus Laternula could, however, also be maintained by differing habitat, ecological and physiological constraints on burrowing capability.

We compared physiological ageing parameters in 2 scallops, the temperate Aequipecten opercularis and the Antarctic Adamussium colbecki. These 2 species are phylogeneti- cally closely related and display a similar lifestyle but have distinctly different maximum lifespans (MLSP). A. opercularis does not live longer than 8 to 10 yr, whereas A. colbecki lives over 18 yr. The development of several physiological ageing parameters over time, chosen according to the 'free rad- ical theory of ageing', was compared in the 2 species to identify differences in the ageing process. In the shorter-lived A. opercularis, activities of the mitochondrial enzymes citrate synthase and cytochrome c oxidase and of the antioxidant enzyme catalase showed a more pronounced decrease with increasing age than in the longer-lived A. colbecki. In line with this finding, lipofuscin accumu- lation increased more distinctly in A. opercularis than in A. colbecki, while tissue protein content decreased in A. opercularis but increased in A. colbecki. Its better preservation of mitochondrial and antioxidant enzyme activities and the avoidance of waste accumulation may enable A. colbecki to live longer than A. opercularis. Mitochondrial function investigated in A. opercularis showed only minor changes with age, and mitochondrial H2O2 generation rates were low at all ages. We relate our findings to the 'free radical-rate of living' theory, to the 'uncoupling to survive' hypothesis, and to the particular lifestyle of these scallops.

The levels of heat-shock proteins of the 70 kDa family (Hsp70s) were measured in different soft tissues of Atlantic cod Gadus morhua from different locations and after exposure to various thermal conditions: acute temperature increments (1° C day -1 ), mid-term (73 days at 4-15° C) and long-term thermal acclimation (278 days at 8-15° C), and seasonal and latitudinal temperature variations (field samples). Tissue specific distribution patterns of Hsp70s were observed: liver > gills > red blood cells > brain > white muscle. Thus, different tissues may have required different levels of protection by Hsp70s, and possibly this was related to the rate of protein synthesis. There were no differences in tissue Hsp70s between Arctic cod populations (Arctic, i.e. Barents and White Seas, Norwegian coast, and North or Baltic Seas). No changes in Hsp70s levels were observed in response to temperature variation of any intensity (acute fluctuation or seasonal and latitudinal) within the range of physiological temperatures (4-15° C) in wild and laboratory Atlantic cod. This confirms previous observations that changes in Hsp70 caused by such temperature variation are often small in fishes. Probably, the constitutive level of Hsp70s in Atlantic cod was high enough to overcome potentially harmful effects of temperature variations within the physiological range. A suppressing effect of high temperature (15° C) has already been observed at a systematic level (as reduced rate of somatic growth), whereas it is not reflected in modified Hsp70s. Therefore, Hsp70s apparently played a secondary role in defining thermal tolerance limits in Atlantic cod. These conclusions are in line with a recent concept of thermal tolerance which indicated that the first line of thermal limitation in the cold and warm is a loss in aerobic scope.

Recent records of lithodid crabs in deeper waters off the Antarctic continental slope raised the question of the return of crabs to Antarctic waters, following their extinction in the lower Miocene ;15 million years ago. Antarctic cooling may be responsible for the impoverishment of the marine high Antarctic decapod fauna, presently comprising only five benthic shrimp species. Effects of polar conditions on marine life, including lowered metabolic rates and short seasonal food availability, are discussed as main evolutionary driving forces shaping Antarctic diversity. In particular, planktotrophic larval stages should be vulnerable to the mismatch of prolonged development and short periods of food availability, selecting against complex life cycles. We hypothesize that larval lecithotrophy and cold tolerance, as recently observed in Subantarctic lithodids, represent, together with other adaptations in the adults, key features among the life-history adaptations of lithodids, potentially enabling them to conquer polar ecosystems. The return of benthic top predators to high Antarctic waters under conditions of climate change would considerably alter the benthic communities.

The effect of acute increase in temperature on oxygen partial pressure (Po 2 ) was measured in the gill arches of Atlantic cod Gadus morhua between 10 and 19° C by use of oxygen microoptodes. Oxygen saturation of the gill blood under control conditions varied between 90 and 15% reflecting a variable percentage of arterial or venous blood in accordance with the position of each optode in the gill arch. The data obtained suggested that arterial Po 2 remained more or less constant and arterial oxygen uptake did not become limiting during warming. A progressive drop in venous Po 2 . however, was observed at >10° C indicating that excessive oxygen uptake from the blood is not fully compensated for by circulatory performance, until finally, Po 2 levels fully collapse. In a second set of experiments energy and acid-base status of white muscle of Atlantic cod in vivo was measured by magnetic resonance ( 31 P-NMR) spectroscopy in unanaesthetized and unimmobilized fish in the temperature range between 13 and 21° C. A decrease in white muscle intracellular pH (pH i ) with temperature occurred between 10 and 16° C (ApH per ° C = -0.025 per C). In white muscle temperature changes had no influence on high-energy phosphates such as phosphocreatine (PCr) or ATP except during exposure to high critical temperatures (>16° C), indicating that white muscle energy status appears to be relatively insensitive to thermal stress if compared to the thermal sensitivity of the whole animal. The data were consistent with the hypothesis of an oxygen limitation of thermal tolerance in animals, which is set by limited capacity of oxygen supply mechanisms. In the case of Atlantic cod circulatory rather than ventilatory performance may be the first process to cause oxygen deficiency during heat stress.

Daily rhythms in activity have been reported for marine and terrestrial organisms, including brooding behaviors which supply oxygen to the embryos of marine invertebrates. Considering that oxygen is a limiting factor in embryo masses of Brachyuran crabs and that female crabs play a critical role in supplying oxygen to the embryos, we studied and compared daily patterns of (1) brooding and nonbrooding behaviors in brooding females, (2) behaviors in nonbrooding females, and (3) oxygen provision to the embryos in a Xanthid crab (Homalaspis plana). We also experimentally evaluated whether daily patterns of nonbrooding behaviors were related to food intake. Behaviors were identified using video recordings and correlated with oxygen analyses in the center of the egg masses by use of microoptodes. According to PO2 recordings abdominal flapping was identified as the single, most important behavior supplying oxygen to the center of the egg mass, maintained at a constant rate during both day and night. Furthermore, the lack of a daily pattern in oxygen availability in the center of the embryo mass is consistent with the lack of daily pattern in abdominal flapping. In contrast, locomotor activity (walking) and maxilliped movements remained unrelated to PO2 oscillations and showed a pronounced diurnal pattern, increasing during the night. This strong diurnal rhythmicity in walking behavior decreased as embryos developed. The frequency of locomotor activity and egg ventilation increased as embryo development progressed. The increase in egg ventilation throughout embryo development may be a response to the increase in embryonic oxygen demand during development. The change in locomotor activity was unrelated to feeding activity, but may serve to make the ventilation process more efficient. These findings emphasize that oxygen is a crucial factor during early development, affecting the normal rate of development of embryos. In consequence, females permanently provide oxygen to the brood despite daily cycles in other behaviors.

SUMMARY Mitochondrial respiration, energetic coupling to phosphorylation and the production of reactive oxygen species (ROS) were studied in mitochondria isolated from the eurythermal bivalve Mya arenaria (Myoidea) from a low-shore intertidal population of the German Wadden Sea. Measurements were conducted both within the range of the habitat temperatures (5-15 °C) and when subjected to heat exposure at 20 °C and 25 °C. Experimental warming resulted in an increase in the rate of state 3 and state 4 respiration in isolated mitochondria. The highest respiratory coupling ratios (RCR) were found at 15 °C; at higher temperatures mitochondrial coupling decreased,and release of ROS doubled between 15 and 25 °C. ROS production was 2-3%of total oxygen consumption in state 3 (0.3-0.5 nmol ROS mg-1protein min-1) at the habitat temperature, reaching a maximum of 4.3 % of state 3 respiration and 7 % of oligomycin-induced state 4+respiration under heat stress. Thus, state 4 respiration, previously interpreted exclusively as a measure of proton leakage, included a significant contribution from ROS formation in this animal, especially under conditions of heat stress. Oxygen radical formation was directly dependent on temperature-controlled respiration rates in states 3 and 4 and inversely related to mitochondrial coupling (RCR+) in state 4. Mitochondrial ROS formation is therefore involved in cellular heat stress in this eurythermal marine ectotherm.

SUMMARYIncreased CO2 partial pressures (hypercapnia) as well as hypoxia are natural features of marine environments like the intertidal zone. Nevertheless little is known about the specific effects of CO2 on metabolism, except for the well-described effects on acid—base variables and regulation. Accordingly, the sediment-dwelling worm Sipunculus nudus was used as an experimental model to investigate the correlation of acid—base-induced metabolic depression and protein/amino acid catabolism, by determining the rates of oxygen consumption, ammonia excretion and O/N ratios in non-perfused preparations of body wall musculature at various levels of extra- and intracellular pH, PCO2 and [HCO3-]. A decrease in extracellular pH from control level (7.9) to 6.7 caused a reduction in aerobic metabolic rate of both normocapnic and hypercapnic tissues by 40-45 %. O/N ratios of 4.0-4.5 under control conditions indicate that amino acid catabolism meets the largest fraction of aerobic energy demand. A significant 10-15 % drop in ammonia excretion, a simultaneous reduction of O/N ratios and a transient accumulation of intracellular bicarbonate during transition to extreme acidosis suggest a reduction in net amino acid catabolism and a shift in the selection of amino acids used,favouring monoamino dicarboxylic acids and their amines (asparagine,glutamine, aspartic and glutamic acids). A drop in intracellular pH was identified as mediating this effect. In conclusion, the present data provide evidence for a regulatory role of intracellular pH in the selection of amino acids used by catabolism.

Mitochondrial mechanisms, which may define and adjust the thermal tolerance window to the environmental temperature regime, were studied in two intertidal populations of the polychaete worm Arenicola marina from the North Sea (boreal) and the White Sea (subpolar).Adaptation to lower mean annual temperatures in White Sea animals (4 versus 10 °C) is reflected by a 2.4 times higher mitochondrial volume density in the muscle tissue of subpolar animals. In White Sea worms acclimated to 6 °C a 10 times higher cytochrome c-oxidase (CYTOX) activity is seen and the value of activation energy (Ea) for the oxidation of cytochrome c is reduced compared with boreal specimens acclimated to 11 °C. Moreover, mitochondria from White Sea lugworms are characterised by a 2.7 times higher succinate oxidation rate in state 3 respiration, a reduced Ea-value for mitochondrial state 3 respiration at low temperatures, as well as a higher activity of NADP dependent isocitrate dehydrogenase (IDH) compared to North Sea animals, even when acclimated to the same temperature of 11 °C. All of these patterns reflect an overall rise in the capacity of aerobic energy production with cold adaptation. This explains the downward shift of the low critical temperature, beyond which anaerobic metabolism sets in. However, the higher mitochondrial density is likely to induce the rise in standard metabolic rate seen in White Sea lugworms, thereby causing a concomitant shift of the high Tc to a lower value. An increase in the Ea value for the decarboxylation of isocitrate in White Sea specimens may help to minimize the increment in standard metabolic rate induced by the higher mitochondrial density and capacity, at the expense of a higher thermal sensitivity of metabolism in the warm.Key Words: Cold adaptation, Mitochondria, Aerobic capacity, Critical temperature, Arenicola marina

The study by Grans et al. ([Grans et al., 2014][1]) investigated growth performance and oxygen demand at rest and during recovery from fatiguing exercise in Atlantic halibut ( Hippoglossus hippoglossus ) under simulated scenarios of ocean warming and acidification. The authors claim that their

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.