Proteomics reveals toxin tolerance and polysaccharide accumulation in Chlorococcum humicola under high CO2 concentration.

Algae have great application prospects in excess sludge reclamation and recovery of high-value biomass. Chlorococcum humicola was cultivated in this research, using sludge extract (mixed with SE medium) with additions of 10%, 20%, and 30% CO2 (v/v). Results showed that under 20% CO2, the dry weight and polysaccharide yield reached 1.389 ± 0.070 g/L and 313.49 ± 10.77 mg/L, respectively. 10% and 20% CO2 promoted the production of cellular antioxidant molecules to resist the toxic stress and the toxicity of 20% CO2 group decreased from 62.16 ± 3.11% to 33.02 ± 3.76%. 10% and 20% CO2 accelerated the electron transfer, enhanced carbon assimilation, and promoted the photosynthetic efficiency, while 30% CO2 led to photosystem damage and disorder of antioxidant system. Proteomic analysis showed that 20% CO2 mainly affected energy metabolism and the oxidative stress level on the early stage (10 d), while affected photosynthesis and organic substance metabolism on the stable stage (30 d). The up-regulation of PSII photosynthetic protein subunit 8 (PsbA, PsbO), A0A383W1S5 and A0A383VRI4 promoted the efficiency of PSII and chlorophyll synthesis, and the up-regulation of A0A383WH74 and A0A2Z4THB7 led to the accumulation of polysaccharides. The up-regulation of A0A383VDH1, A0A383VX37 and A0A383VA86 promoted respiration. Collectively, this work discloses the regulatory mechanism of high-concentration CO2 on Chlorococcum humicola to overcome toxicity and accumulate polysaccharides.

Supercritical CO2 fluid extract from Stellariae Radix ameliorates 2,4-dinitrochlorobenzene-induced atopic dermatitis by inhibit M1 macrophages polarization via AMPK activation.

Yin chai hu (Radix Stellariae) is a root medicine that is frequently used in Chinese traditional medicine to treat fever and malnutrition. In modern medicine, it has been discovered to have anti-inflammatory, anti-allergic, and anticancer properties. In a previous study, we were able to extract lipids from Stellariae Radix using supercritical CO2 extraction (SRE), and these sterol lipids accounted for up to 88.29% of the extract. However, the impact of SRE on the development of atopic dermatitis (AD) has not yet been investigated. This study investigates the inhibitory effects of SRE on AD development using a 2,4-dinitrochlorobenzene (DNCB)-induced AD mouse model. Treatment with SRE significantly reduced the dermatitis score and histopathological changes compared with the DNCB group. The study found that treatment with SRE resulted in a decrease of pro-inflammatory cytokines TNF-α, CXC-10, IL-12, and IL-1ß in skin lesions. Additionally, immunohistochemical analysis revealed that SRE effectively suppressed M1 macrophage infiltration into the AD lesion. Furthermore, the anti-inflammatory effect of SRE was evaluated in LPS + INF-γ induced bone marrow-derived macrophages (BMDMs) M1 polarization, SRE inhibited the production of TNF-α, CXC-10, IL-12, and IL-1ß and decreased the expression of NLRP3. Additionally, SRE was found to increase p-AMPKT172, but had no effect on total AMPK expression, after administration of the AMPK inhibitor Compound C, the inhibitory effect of SRE on M1 macrophages was partially reversed. The results indicate that SRE has an inhibitory effect on AD, making it a potential therapeutic agent for this atopic disorder.

Response of CRH system in brain and gill of marine medaka to seawater acidification.

Corticotropin-releasing hormone (CRH) is mainly secreted by the hypothalamus to regulate stress when environmental factors change. Gills contact with water directly and may also secrete CRH to maintain local homeostasis. Ocean acidification changes water chemical parameters and is becoming an important environmental stressor for marine fish. The response of brain and gill CRH systems to ocean acidification remains unclear. In this study, marine medaka were exposed to CO2-acidified seawater (440 ppm, 1000 ppm, and 1800 ppm CO2) for 2 h, 4 h, 24 h, and 7 d, respectively. At 2 h and 4 h, the expression of crh mRNA in gills increased with increasing CO2 concentration. Crh protein is expressed mainly in the lamellae cells. crhbp and crhr1 expression also increased significantly. However, at 2 h and 4 h, acidification caused little changes in these genes and Crh protein expression in the brain. At 7 d, Crh-positive cells were detected in the hypothalamus; moreover, Crh protein expression in the whole brain increased. It is suggested that CRH autocrine secretion in gills is responsible for local acid-base regulation rather than systemic mobilization after short-term acidification stress, which may help the rapid regulation of body damage caused by environmental stress.

Acid times in physiology: A systematic review of the effects of ocean acidification on calcifying invertebrates.

The reduction in seawater pH from rising levels of carbon dioxide (CO2) in the oceans has been recognized as an important force shaping the future of marine ecosystems. Therefore, numerous studies have reported the effects of ocean acidification (OA) in different compartments of important animal groups, based on field and/or laboratory observations. Calcifying invertebrates have received considerable attention in recent years. In the present systematic review, we have summarized the physiological responses to OA in coral, echinoderm, mollusk, and crustacean species exposed to predicted ocean acidification conditions in the near future. The Scopus, Web of Science, and PubMed databases were used for the literature search, and 75 articles were obtained based on the inclusion criteria. Six main physiological responses have been reported after exposure to low pH. Growth (21.6%), metabolism (20.8%), and acid-base balance (17.6%) were the most frequent among the phyla, while calcification and growth were the physiological responses most affected by OA (>40%). Studies show that the reduction of pH in the aquatic environment, in general, supports the maintenance of metabolic parameters in invertebrates, with redistribution of energy to biological functions, generating limitations to calcification, which can have severe consequences for the health and survival of these organisms. It should be noted that the OA results are variable, with inter and/or intraspecific differences. In summary, this systematic review offers important scientific evidence for establishing paradigms in the physiology of climate change in addition to gathering valuable information on the subject and future research perspectives.

Possible causes of narcosis-like symptoms in freedivers.

During deep-sea freediving, many freedivers describe symptoms fairly similar to what has been related to inert gas narcosis in scuba divers. This manuscript aims to present the potential mechanisms underlying these symptoms. First, known mechanisms of narcosis are summarized while scuba diving. Then, potential underlying mechanisms involving the toxicity of gases (nitrogen, carbon dioxide and oxygen) are presented in freedivers. As the symptoms are felt during ascent, nitrogen is likely not the only gas involved. Since freedivers are frequently exposed to hypercapnic hypoxia toward the end of the dive, it is proposed that carbon dioxide and oxygen gases both play a major role. Finally, a new "hemodynamic hypothesis" based on the diving reflex is proposed in freedivers. The underlying mechanisms are undoubtedly multifactorial and therefore require further research and a new descriptive name. We propose a new term for these types of symptoms: freediving transient cognitive impairment.

Rapid blood acid-base regulation by European sea bass (Dicentrarchus labrax) in response to sudden exposure to high environmental CO2.

Fish in coastal ecosystems can be exposed to acute variations in CO2 of between 0.2 and 1 kPa CO2 (2000-10,000 µatm). Coping with this environmental challenge will depend on the ability to rapidly compensate for the internal acid-base disturbance caused by sudden exposure to high environmental CO2 (blood and tissue acidosis); however, studies about the speed of acid-base regulatory responses in marine fish are scarce. We observed that upon sudden exposure to ∼1 kPa CO2, European sea bass (Dicentrarchus labrax) completely regulate erythrocyte intracellular pH within ∼40 min, thus restoring haemoglobin-O2 affinity to pre-exposure levels. Moreover, blood pH returned to normal levels within ∼2 h, which is one of the fastest acid-base recoveries documented in any fish. This was achieved via a large upregulation of net acid excretion and accumulation of HCO3- in blood, which increased from ∼4 to ∼22 mmol l-1. While the abundance and intracellular localisation of gill Na+/K+-ATPase (NKA) and Na+/H+ exchanger 3 (NHE3) remained unchanged, the apical surface area of acid-excreting gill ionocytes doubled. This constitutes a novel mechanism for rapidly increasing acid excretion during sudden blood acidosis. Rapid acid-base regulation was completely prevented when the same high CO2 exposure occurred in seawater with experimentally reduced HCO3- and pH, probably because reduced environmental pH inhibited gill H+ excretion via NHE3. The rapid and robust acid-base regulatory responses identified will enable European sea bass to maintain physiological performance during large and sudden CO2 fluctuations that naturally occur in coastal environments.

Different ecological histories of sea urchins acclimated to reduced pH influence offspring response to multiple stressors.

End-of-the-century predictions on carbon dioxide (CO2) driven ocean acidification and the continuous leakage of pesticides from inland to coastal areas are of concern for potential negative effects on marine species' early life stages which are the most vulnerable to environmental changes. Variations in seawater chemistry related to human activities may interfere with the normal development from embryo to juvenile/adult stage. However, transgenerational studies suggest that the parental generation can influence the offspring phenotype, and thus their performances, based on the environment experienced. Here we compared the transgenerational responses to a multiple stressor scenario in sea urchins (Paracentrotus lividus) that experienced different environments since their settlement: i.e., animals from a highly variable environment, such as the Venice lagoon, versus animals from a coastal area with prevailing oligotrophic conditions in the Northern Adriatic Sea. After long-term maintenance (2 and 6 months) of adult sea urchins at natural and -0.4 units reduced pH, the F1 generations were obtained. Embryos were reared under four experimental conditions: natural and -0.4 pH both in the absence and in the presence of an emerging contaminants' mixture (glyphosate and aminomethylphosphonic acid at environmentally relevant concentrations, 100 µg/L). A significant detrimental effect of both the parental and the filial pH was highlighted, affecting embryo development and growth. Nonetheless, sea urchins from both sites were able to cope with ocean acidification. The 6-months F1 response was better than that of the 2-months F1. Conversely, the F1 response of the sea urchins maintained at natural conditions did not change sensibly after more prolonged parental exposure. An additive but mild negative effect of the mixture was observed, mostly in lagoon offspring. Results suggest that long-term exposure to reduced pH leads to transgenerational acclimation but does not affect susceptibility to the tested pollutants.

Impaired cardiorespiratory responses to hypercapnia in neonatal mice lacking PAC1 but not VPAC2 receptors.

The evidence is mounting for a role for abnormal signaling of the stress peptide pituitary adenylate cyclase activating polypeptide (PACAP) and its canonical receptor PAC1 in the pathogenesis of sudden infant death syndrome. In this study, we investigated whether the PACAP receptors PAC1 or VPAC2 are involved in the neonatal cardiorespiratory response to hypercapnic stress. We used head-out plethysmography and surface ECG electrodes to assess cardiorespiratory responses to an 8% hypercapnic challenge in unanesthetized and spontaneously breathing 4-day-old PAC1 or VPAC2 knockout (KO) and wild-type mouse pups. We demonstrate that compared with WTs, breathing frequency (RR) and minute ventilation ([Formula: see text]) in PAC1 KO pups were significantly blunted in response to hypercapnia. Although heart rate was unaltered in PAC1 KO pups during hypercapnia, heart rate recovery posthypercapnia was impaired. In contrast, cardiorespiratory impairments in VPAC2 KO pups were limited to only an overall higher tidal volume (VT), independent of treatment. These findings suggest that PACAP signaling through the PAC1 receptor plays a more important role than signaling through the VPAC2 receptor in neonatal respiratory responses to hypercapnia. Thus deficits in PACAP signaling primarily via PAC1 may contribute to the inability of infants to mount an appropriate protective response to homeostatic stressors in childhood disorders such as SIDS.

Coexposure to Inhaled Aldehydes or Carbon Dioxide Enhances the Carcinogenic Properties of the Tobacco-Specific Nitrosamine 4-Methylnitrosamino-1-(3-pyridyl)-1-butanone in the A/J Mouse Lung.

Tobacco smoke is a complex mixture of chemicals, many of which are toxic and carcinogenic. Hazard assessments of tobacco smoke exposure have predominantly focused on either single chemical exposures or the more complex mixtures of tobacco smoke or its fractions. There are fewer studies exploring interactions between specific tobacco smoke chemicals. Aldehydes such as formaldehyde and acetaldehyde were hypothesized to enhance the carcinogenic properties of the human carcinogen, 4-methylnitrosamino-1-(3-pyridyl)-1-butanone (NNK) through a variety of mechanisms. This hypothesis was tested in the established NNK-induced A/J mouse lung tumor model. A/J mice were exposed to NNK (intraperitoneal injection, 0, 2.5, or 7.5 µmol in saline) in the presence or absence of acetaldehyde (0 or 360 ppmv) or formaldehyde (0 or 17 ppmv) for 3 h in a nose-only inhalation chamber, and lung tumors were counted 16 weeks later. Neither aldehyde by itself induced lung tumors. However, mice receiving both NNK and acetaldehyde or formaldehyde had more adenomas with dysplasia or progression than those receiving only NNK, suggesting that aldehydes may increase the severity of NNK-induced lung adenomas. The aldehyde coexposure did not affect the levels of NNK-derived DNA adduct levels. Similar studies tested the ability of a 3 h nose-only carbon dioxide (0, 5, 10, or 15%) coexposure to influence lung adenoma formation by NNK. While carbon dioxide alone was not carcinogenic, it significantly increased the number of NNK-derived lung adenomas without affecting NNK-derived DNA damage. These studies indicate that the chemicals in tobacco smoke work together to form a potent lung carcinogenic mixture.

Brainstem serotonergic, catecholaminergic, and inflammatory adaptations during chronic hypercapnia in goats.

Despite the prevalence of CO2 retention in human disease, little is known about the adaptive neurobiological effects of chronic hypercapnia. We have recently shown 30-d exposure to increased inspired CO2 (InCO2) leads to a steady-state ventilation that exceeds the level predicted by the sustained acidosis and the acute CO2/H+ chemoreflex, suggesting plasticity within respiratory control centers. Based on data showing brainstem changes in aminergic and inflammatory signaling during carotid body denervation-induced hypercapnia, we hypothesized chronic hypercapnia per se will lead to similar changes. We found that: 1) increased InCO2 increased IL-1ß in the medullary raphe (MR), ventral respiratory column, and cuneate nucleus after 24 h, but not after 30 d of hypercapnia; 2) the number of serotonergic and total neurons were reduced within the MR and ventrolateral medulla following 30 d of increased InCO2; 3) markers of tryptophan metabolism were altered following 24 h, but not 30 d of InCO2; and 4) there were few changes in brainstem amine levels following 24 h or 30 d of increased InCO2. We conclude that these changes may contribute to initiating or maintaining respiratory neuroplasticity during chronic hypercapnia but alone do not account for ventilatory acclimatization to chronic increased InCO2.-Burgraff, N. J., Neumueller, S. E., Buchholz, K. J., LeClaire, J., Hodges, M. R., Pan, L., Forster, H. V. Brainstem serotonergic, catecholaminergic, and inflammatory adaptations during chronic hypercapnia in goats.

Global economic trade-offs between wild nature and tropical agriculture.

Global demands for agricultural and forestry products provide economic incentives for deforestation across the tropics. Much of this deforestation occurs with a lack of information on the spatial distribution of benefits and costs of deforestation. To inform global sustainable land-use policies, we combine geographic information systems (GIS) with a meta-analysis of ecosystem services (ES) studies to perform a spatially explicit analysis of the trade-offs between agricultural benefits, carbon emissions, and losses of multiple ecosystem services because of tropical deforestation from 2000 to 2012. Even though the value of ecosystem services presents large inherent uncertainties, we find a pattern supporting the argument that the externalities of destroying tropical forests are greater than the current direct economic benefits derived from agriculture in all cases bar one: when yield and rent potentials of high-value crops could be realized in the future. Our analysis identifies the Atlantic Forest, areas around the Gulf of Guinea, and Thailand as areas where agricultural conversion appears economically efficient, indicating a major impediment to the long-term financial sustainability of Reducing Emissions from Deforestation and forest Degradation (REDD+) schemes in those countries. By contrast, Latin America, insular Southeast Asia, and Madagascar present areas with low agricultural rents (ARs) and high values in carbon stocks and ES, suggesting that they are economically viable conservation targets. Our study helps identify optimal areas for conservation and agriculture together with their associated uncertainties, which could enhance the efficiency and sustainability of pantropical land-use policies and help direct future research efforts.

CO2 anesthesia on Drosophila survival in aging research.

Carbon dioxide (CO2 ) exposure is a common method of anesthesia in studies of Drosophila melanogaster. A number of negative side effects of CO2 anesthesia have been reported. It is not clear whether the length of CO2 anesthesia time affects Drosophila survival in aging research. Here, we examined the potential effect of the CO2 anesthesia time length of 10-150 min. We found that long CO2 exposure could lead to Drosophila death, more significant in males. The longer the anesthesia time is, the longer it takes for flies to wake up. Long-time CO2 anesthesia can reduce the lifespan. Our stress tests showed that long-time CO2 anesthesia can increase the average survival time in both males and females under starvation conditions, but can only increase female lifespan under H2 O2 oxidative stress. Long-time CO2 anesthesia also significantly affects physiological traits, with spontaneous activity increased in females but decreased in males, and reduced female fecundity. Our study suggests that limiting the CO2 anesthesia time and giving enough recovery time before performing physiological tests are important in Drosophila aging research.

Wound healing in an elasmobranch fish is not impaired by high-CO2 exposure.

The purpose of this study was to test the effects of high CO2 exposure on wound healing rates in an elasmobranch fish (Urobatis jamaicensis). Small dermal injuries (8 mm biopsy) closed by 22 days post wounding with a decrease in haematocrit. High CO2 exposure (ΔpH = 1.4) did not influence healing rate or haematocrit. Combined, these data provide evidence that minimally invasive scientific procedures have short-term impacts on elasmobranch fishes even during exposure to a chronic stressor. Therefore, wound healing rates may not be strongly impacted by ocean acidification (ΔpH = 0.4).

Inhaled H2 or CO2 Do Not Augment the Neuroprotective Effect of Therapeutic Hypothermia in a Severe Neonatal Hypoxic-Ischemic Encephalopathy Piglet Model.

Hypoxic-ischemic encephalopathy (HIE) is still a major cause of neonatal death and disability as therapeutic hypothermia (TH) alone cannot afford sufficient neuroprotection. The present study investigated whether ventilation with molecular hydrogen (2.1% H2) or graded restoration of normocapnia with CO2 for 4 h after asphyxia would augment the neuroprotective effect of TH in a subacute (48 h) HIE piglet model. Piglets were randomized to untreated naïve, control-normothermia, asphyxia-normothermia (20-min 4%O2-20%CO2 ventilation; Tcore = 38.5 °C), asphyxia-hypothermia (A-HT, Tcore = 33.5 °C, 2-36 h post-asphyxia), A-HT + H2, or A-HT + CO2 treatment groups. Asphyxia elicited severe hypoxia (pO2 = 19 ± 5 mmHg) and mixed acidosis (pH = 6.79 ± 0.10). HIE development was confirmed by altered cerebral electrical activity and neuropathology. TH was significantly neuroprotective in the caudate nucleus but demonstrated virtually no such effect in the hippocampus. The mRNA levels of apoptosis-inducing factor and caspase-3 showed a ~10-fold increase in the A-HT group compared to naïve animals in the hippocampus but not in the caudate nucleus coinciding with the region-specific neuroprotective effect of TH. H2 or CO2 did not augment TH-induced neuroprotection in any brain areas; rather, CO2 even abolished the neuroprotective effect of TH in the caudate nucleus. In conclusion, the present findings do not support the use of these medical gases to supplement TH in HIE management.

Effects of Decabromodiphenyl Ether and Elevated Carbon Dioxide on Rice (Oryza sativa L.).

We assessed the effects of carbon dioxide (CO2) and decabromodiphenyl ether (BDE-209, 0, 3 and 30 mg/kg) on rice (Oryza sativa L. cv. Wuyunjing) in field free-air CO2 enrichment system. Rice at elevated (580 ppm) CO2 had increased net photosynthetic rate, intercellular CO2 concentration, shoot biomass, yield and phosphorus content in grains. However, there were no significant changes in such parameters observed on rice at elevated CO2 combined with BDE-209 (3 and 30 mg/kg). Elevated CO2 alone had no significant effects on sugar or starch content in rice grains, whereas its combination with BDE-209 (3 mg/kg) significantly decreased grain sugar and starch content. In conclusion, rice reared in soil polluted by BDE-209 under elevated CO2 modulates the effects in grain feature.

Recoverable impacts of ocean acidification on the tubeworm, Hydroides elegans: implication for biofouling in future coastal oceans.

Ocean uptake of anthropogenic CO2 causes ocean acidification (OA), which not only decreases the calcification rate, but also impairs the formation of calcareous shells or tubes in marine invertebrates such as the dominant biofouling tubeworm species, Hydroides elegans. This study examined the ability of tubeworms to resume normal tube calcification when returned to ambient pH 8.1 from a projected near-future OA level of pH 7.8. Tubeworms produced structurally impaired and mechanically weaker calcareous tubes at pH 7.8 compared to at pH 8.1, but were able to recover when the pH was restored to ambient levels. This suggests that tubeworms can physiologically recover from the impacts of OA on tube calcification, composition, density, hardness and stiffness when returned to optimal conditions. These results help understanding of the progression of biofouling communities dominated by tubeworms in future oceans with low pH induced by OA.

Plant responses to increasing CO2 reduce estimates of climate impacts on drought severity.

Rising atmospheric CO2 will make Earth warmer, and many studies have inferred that this warming will cause droughts to become more widespread and severe. However, rising atmospheric CO2 also modifies stomatal conductance and plant water use, processes that are often are overlooked in impact analysis. We find that plant physiological responses to CO2 reduce predictions of future drought stress, and that this reduction is captured by using plant-centric rather than atmosphere-centric metrics from Earth system models (ESMs). The atmosphere-centric Palmer Drought Severity Index predicts future increases in drought stress for more than 70% of global land area. This area drops to 37% with the use of precipitation minus evapotranspiration (P-E), a measure that represents the water flux available to downstream ecosystems and humans. The two metrics yield consistent estimates of increasing stress in regions where precipitation decreases are more robust (southern North America, northeastern South America, and southern Europe). The metrics produce diverging estimates elsewhere, with P-E predicting decreasing stress across temperate Asia and central Africa. The differing sensitivity of drought metrics to radiative and physiological aspects of increasing CO2 partly explains the divergent estimates of future drought reported in recent studies. Further, use of ESM output in offline models may double-count plant feedbacks on relative humidity and other surface variables, leading to overestimates of future stress. The use of drought metrics that account for the response of plant transpiration to changing CO2, including direct use of P-E and soil moisture from ESMs, is needed to reduce uncertainties in future assessment.

Long term effects of chronic prenatal exposure to hypercarbia on organ growth and cardiovascular responses to adrenaline and hypoxia in common snapping turtles.

Reptilian embryos often face challenging environmental gas compositions during incubation, which may inflict long-lasting effects in the individuals' physiological responses. These conditions can have a lasting effect on the animal into juvenile life as chronic prenatal exposure to hypercarbia results in enlarged hatchling organ size, higher growth rate and resting metabolic rate, although relatively smaller increment in metabolic scope during digestion. Therefore, we wanted to verify whether prenatal hypercarbia exposure would cause persistent effects on morphology and physiological responses in C. serpentina. We measured organ masses and cardiovascular parameters in five years old turtles incubated either under 3.5% hypercarbia (H3.5) or normoxia (N21). We expected that: i) organ masses of H3.5 would be bigger than N21; ii) acute exposure to hypoxia should decrease blood flows in H3.5, since metabolic scope is presumably reduced in this group. As hypoxia exposure elicits catecholamine release, we also tested cardiovascular responses to adrenaline injection. Lungs and stomach exhibited higher growth rates in H3.5. Divergent cardiovascular responses between groups to adrenaline injection were observed for heart rate, pulmonary blood flow, pulmonary mean arterial pressure, blood shunt, systemic stroke volume, and stomach perfusion. Hypoxia caused decreased systemic blood flow and cardiac output, systemic and total stroke volume, and systemic vascular conductance in H3.5. These variables were unaffected in N21, but pulmonary flow and stroke volume, and stomach blood perfusion were reduced. These data support the hypothesis that exposure to hypercarbia during embryonic development has long term effects on organ morphology and cardiovascular responses of C. serpentina.

Hot and bothered: effects of elevated Pco2 and temperature on juvenile freshwater mussels.

Multiple environmental stressors may interact in complex ways to exceed or diminish the impacts of individual stressors. In the present study, the interactive effects of two ecologically relevant stressors [increased temperature and partial pressure of carbon dioxide (Pco2)] were assessed for freshwater mussels, a group of organisms that are among the most sensitive and rapidly declining worldwide. The individual and combined effects of elevated temperature (22°C-34°C) and Pco2 (~230, 58,000 µatm) on juvenile Lampsilis siliquoidea were quantified over a 5- or 14-day period, during which physiological and whole animal responses were measured. Exposure to elevated temperature induced a series of physiological responses, including an increase in oxygen consumption rates following 5 days of exposure at 31°C and an increase in carbonic anhydrase ( ca) and heat shock protein 70 mRNA levels following 14 days of exposure at 28°C and 34°C, respectively. Treatment with elevated Pco2 activated acid-base regulatory responses including increases in CA and Na+-K+-ATPase activity and a novel mechanism for acid-base regulation during Pco2 exposure in freshwater mussels was proposed. Thermal and CO2 stressors also interacted such that responses to the thermal stressor were diminished in mussels exposed to elevated Pco2, resulting in the greatest level of mortality. Additionally, larger mussels were more likely to survive treatment with elevated Pco2 and/or temperature. Together, exposure to elevated Pco2 may compromise the ability of juvenile freshwater mussels to respond to additional stressors, such as increased temperatures, highlighting the importance of considering not only the individual but also the interactive effects of multiple environmental stressors.

Antarctic emerald rockcod have the capacity to compensate for warming when uncoupled from CO2 -acidification.

Increases in atmospheric CO2 levels and associated ocean changes are expected to have dramatic impacts on marine ecosystems. Although the Southern Ocean is experiencing some of the fastest rates of change, few studies have explored how Antarctic fishes may be affected by co-occurring ocean changes, and even fewer have examined early life stages. To date, no studies have characterized potential trade-offs in physiology and behavior in response to projected multiple climate change stressors (ocean acidification and warming) on Antarctic fishes. We exposed juvenile emerald rockcod Trematomus bernacchii to three PCO2 treatments (~450, ~850, and ~1,200 µatm PCO2 ) at two temperatures (-1 or 2°C). After 2, 7, 14, and 28 days, metrics of physiological performance including cardiorespiratory function (heart rate [fH ] and ventilation rate [fV ]), metabolic rate (M˙O2), and cellular enzyme activity were measured. Behavioral responses, including scototaxis, activity, exploration, and escape response were assessed after 7 and 14 days. Elevated PCO2 independently had little impact on either physiology or behavior in juvenile rockcod, whereas warming resulted in significant changes across acclimation time. After 14 days, fH , fV and M˙O2 significantly increased with warming, but not with elevated PCO2 . Increased physiological costs were accompanied by behavioral alterations including increased dark zone preference up to 14%, reduced activity by 12%, as well as reduced escape time suggesting potential trade-offs in energetics. After 28 days, juvenile rockcod demonstrated a degree of temperature compensation as fV , M˙O2, and cellular metabolism significantly decreased following the peak at 14 days; however, temperature compensation was only evident in the absence of elevated PCO2 . Sustained increases in fV and M˙O2 after 28 days exposure to elevated PCO2 indicate additive (fV ) and synergistic (M˙O2) interactions occurred in combination with warming. Stressor-induced energetic trade-offs in physiology and behavior may be an important mechanism leading to vulnerability of Antarctic fishes to future ocean change.