Description: 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.

Description: The in vitro oxygen-binding characteristics of haemocyanin were investigated in whole blood of two species of pelagic squid, Illex illecebrosus and Loligo pealei. pH-independent Haldane coefficients (ΔHCO3−/ΔHcyO2) (where HcyO2 is haemocyanin-bound oxygen) slightly smaller than —1 were found in both species. Oxygen-linked CO2 binding was not present. Buffer values ranged between 5 and 5.8 m mol l−1pH unit−1. For further analyses a pH/saturation diagram was selected to show the effect of pH on oxygen binding at constant POO2 in a continuous plot. The slopes of the resulting oxygen isobars (ΔHcyO2/ΔpH or ΔS/ΔpH) (where S is oxygen saturation) depend on pH. The diagram allows evaluation of both the Bohr coefficients (ΔlogP50/ΔpH) and the Hill coefficients (n50) at specific pH values. It provides an integrated illustration of the importance of the Bohr effect and cooperativity for oxygen binding. In accordance with Wyman's linkage equation, Bohr and Haldane coefficients are found to be identical. Both are pH-independent between pH7 and 8. The changing slopes of the oxygen isobars are likely to reflect changes in cooperativity with pH. Maximum values of n50 coincide with maximum steepness of the oxygen isobars in the physiological range of pH and POO2. Assuming that the haemocyanin acts as a buffer for venous POO2, this maximum in pH sensitivity and its decrease in the higher and lower pH ranges are discussed in the light of the maintenance of pigment function in vivo.

Description: The health of the ocean, central to human well-being, has now reached a critical point. Most fish stocks are overexploited, climate change and increased dissolved carbon dioxide are changing ocean chemistry and disrupting species throughout food webs, and the fundamental capacity of the ocean to regulate the climate has been altered. However, key technical, organizational, and conceptual scientific barriers have prevented the identification of policy levers for sustainability and transformative action. Here, we recommend key strategies to address these challenges, including (1) stronger integration of sciences and (2) ocean-observing systems, (3) improved science-policy interfaces, (4) new partnerships supported by (5) a new ocean-climate finance system, and (6) improved ocean literacy and education to modify social norms and behaviors. Adopting these strategies could help establish ocean science as a key foundation of broader sustainability transformations.

Description: 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 FST = 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 FST = 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 FST = 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.

Description: In the eurythermal cuttlefish Sepia officinalis, performance greatly 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 supporting thermal acclimation that could be critical for the cuttlefish's competitive 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 activity. 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 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, cold adapted English Channel cuttlefish had larger hearts with lower mitochondrial capacities than warm adapted Adriatic cuttlefish. The changes observed for substrate oxidation, mitochondrial complexes, 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.

Description: 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 O2 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 O2 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 CO2. 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.

Description: In this study, we describe a de novo sequencing and assembly of the spleen transcriptome of Lepidonotothen nudifrons, a notothenioid fish widely distributed around the Antarctic Peninsula and the Scotia Arc. Sequences were generated on an Illumina MiSeq system and assembled to a total of 112,477 transcripts. Putative functional annotation was possible for more than 34% of the transcripts. This data will be relevant for future studies targeting the erythrocyte turnover, oxygen transport mechanism and immune system, which are key functional traits to investigate cold adaptation and thermal sensitivity of Antarctic notothenioids.

Description: Abstract 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.

Description: The interaction between ocean warming, hypoxia and hypercapnia, suggested by climate projections, may push an organism earlier to the limits of its thermal tolerance window. In a previous study on juveniles of green abalone (Haliotis fulgens), combined exposure to hypoxia and hypercapnia during heat stress induced a lowered critical thermal maximum (CTmax), indicated by constrained oxygen consumption, muscular spams and loss of attachment. Thus, the present study investigated the cell physiology in foot muscle of H. fulgens juveniles exposed to acute warming (18 °C to 32 °C at +3 °C day−1) under hypoxia (50% air saturation) and hypercapnia (~1000 μatm PCO2), alone and in combination, to decipher the mechanisms leading to functional loss in this tissue. Under exposure to either hypoxia or hypercapnia, citrate synthase (CS) activity decreased with initial warming, in line with thermal compensation, but returned to control levels at 32 °C. The anaerobic enzymes lactate and tauropine dehydrogenase increased only under hypoxia at 32 °C. Under the combined treatment, CS overcame thermal compensation and remained stable overall, indicating active mitochondrial regulation under these conditions. Limited accumulation of anaerobic metabolites indicates unchanged mode of energy production. In all treatments, upregulation of Hsp70 mRNA was observed already at 30 °C. However, lack of evidence for Hsp70 protein accumulation provides only limited support to thermal denaturation of proteins. We conclude that under combined hypoxia and hypercapnia, metabolic depression allowed the H. fulgens musculature to retain an aerobic mode of metabolism in response to warming but may have contributed to functional loss. Keywords: 1H NMR spectroscopy; Citrate synthase; CTmax; Hsp70; Lactate dehydrogenase; Tauropine; Tauropine dehydrogenase