Environmental Hypercapnia Research Articles

Welfare effects of environmental hypercapnia quantified by indicators based on morphology and allostatic load in Atlantic salmon (Salmo salar)

Water supply is a limited resource in most salmon hatcheries, which is compensated by reduced water flow and oxygenation. However, reduced water exchange can lead to accumulation of CO2, resulting in environmental hypercapnia, which may have negative impacts on fish welfare. Thus, environmental hypercapnia can be a common welfare problem for salmon in hatcheries, and particularly in recirculating systems (RAS).In this experiment, Atlantic salmon were exposed to chronic environmental hypercapnia during the last 68 days of the freshwater phase, whereupon effects on physiological stress coping mechanisms and morphological welfare indicators were investigated. Effects on stress coping mechanisms were quantified by measuring changes in brain serotonergic chemistry and plasma cortisol at basal levels and in response to a standardized acute stress test. The results show that exposure to elevated CO2 saturation in the water compromised stress responsiveness of brainstem serotonergic activity, altered osmotic homeostasis, and suppressed growth indicating that fish experience allostatic overload. However, no effects on morphological welfare indicators were observed. This accentuates the need for physiological measures, including physiological responses to controlled challenges to activate the stress axis, when investigating the welfare status of fish reared in systems with potential high CO2.

Specialized adaptations allow vent-endemic crabs (Xenograpsus testudinatus) to thrive under extreme environmental hypercapnia

Shallow hydrothermal vent environments are typically very warm and acidic due to the mixing of ambient seawater with volcanic gasses (> 92% CO2) released through the seafloor making them potential ‘natural laboratories’ to study long-term adaptations to extreme hypercapnic conditions. Xenograpsus testudinatus, the shallow hydrothermal vent crab, is the sole metazoan inhabitant endemic to vents surrounding Kueishantao Island, Taiwan, where it inhabits waters that are generally pH 6.50 with maximum acidities reported as pH 5.50. This study assessed the acid–base regulatory capacity and the compensatory response of X. testudinatus to investigate its remarkable physiological adaptations. Hemolymph parameters (pH, [HCO3−], {text{P}}_{{{text{CO}}_{2} }}, [NH4+], and major ion compositions) and the whole animal’s rates of oxygen consumption and ammonia excretion were measured throughout a 14-day acclimation to pH 6.5 and 5.5. Data revealed that vent crabs are exceptionally strong acid–base regulators capable of maintaining homeostatic pH against extreme hypercapnia (pH 5.50, 24.6 kPa {text{P}}_{{{text{CO}}_{2} }}) via HCO3−/Cl− exchange, retention and utilization of extracellular ammonia. Intact crabs as well as their isolated perfused gills maintained {text{P}}_{{{text{CO}}_{2} }}tensions below environmental levels suggesting the gills can excrete CO2 against a hemolymph-directed {text{P}}_{{{text{CO}}_{2} }} gradient. These specialized physiological mechanisms may be amongst the adaptations required by vent-endemic animals surviving in extreme conditions.

Fossorial giant Zambian mole-rats have blunted ventilatory responses to environmental hypoxia and hypercapnia

Fossorial giant Zambian mole-rats are believed to live in a hypoxic and hypercapnic subterranean environment but their physiological responses to these challenges are entirely unknown. To investigate this, we exposed awake and freely-behaving animals to i) 6 h of normoxia, ii) acute graded normocapnic hypoxia (21, 18, 15, 12, 8, and 5% O2, 0% CO2, balance N2; 1 h each), or iii) acute graded normoxic hypercapnia (0, 2, 5, 7, 9, and 10% CO2, 21% O2, balance N2; 1 h each), followed by a 1 h normoxic normocapnic recovery period, while non-invasively measuring ventilation, metabolic rate, and body temperature (Tb). We found that these mole-rats had a blunted hypoxic ventilatory response that manifested at 12% inhaled O2, a robust hypoxic metabolic response (up to a 68% decrease, starting at 15% O2), and decreased Tb (at or below 8% O2). Upon reoxygenation, metabolic rate increased 52% above normoxic levels, suggesting the paying off of an O2 debt. Ventilation was less sensitive to environmental hypercapnia than to environmental hypoxia and animals also exhibited a blunted hypercapnic ventilatory response that did not manifest below 9% inhaled CO2. Conversely, metabolism and Tb were not affected by hypercapnia. Taken together, these results indicate that, like most other fossorial rodents, giant Zambian mole-rats have blunted hypoxic and hypercapnic ventilatory responses and employ metabolic suppression to tolerate acute hypoxia. Blunted physiological responses to hypoxia and hypercapnia likely reflect the subterranean lifestyle of this mammal, wherein intermittent but severe hypoxia and/or hypercapnia may be common challenges.

Neurokinin-1 receptor activation is sufficient to restore the hypercapnic ventilatory response in the Substance P-deficient naked mole-rat.

Naked mole-rats (NMRs) live in large colonies within densely populated underground burrows. Their collective respiration generates significant metabolic carbon dioxide (CO2) that diffuses slowly out of the burrow network, creating a hypercapnic environment. Currently, the physiological mechanisms that underlie the ability of NMRs to tolerate environmental hypercapnia are largely unknown. To address this, we used whole-body plethysmography and respirometry to elucidate the hypercapnic ventilatory and metabolic responses of awake, freely behaving NMRs to 0%-10% CO2. We found that NMRs have a blunted hypercapnic ventilatory response (HCVR): ventilation increased only in 10% CO2. Conversely, metabolism was unaffected by hypercapnia. NMRs are insensitive to cutaneous acid-based pain caused by modified substance P (SP)-mediated peripheral neurotransmission, and SP is also an important neuromodulator of ventilation. Therefore, we re-evaluated physiological responses to hypercapnia in NMRs after an intraperitoneal injection of exogenous substance P (2 mg/kg) or a long-lived isoform of substance P {[pGlu5-MePhe8-MeGly9]SP(5-11), DiMe-C7; 40-400 μg/kg}. We found that both drugs restored hypercapnia sensitivity and unmasked an HCVR in animals breathing 2%-10% CO2. Taken together, our findings indicate that NMRs are remarkably tolerant of hypercapnic environments and have a blunted HCVR; however, the signaling network architecture required for a "normal" HCVR is retained but endogenously inactive. This muting of chemosensitivity likely suits the ecophysiology of this species, which presumably experiences hypercapnia regularly in their underground niche.

Increased intestinal carbonate precipitate abundance in the sea bream (Sparus aurata L.) in response to ocean acidification.

Marine fish contribute to the carbon cycle by producing mineralized intestinal precipitates generated as by-products of their osmoregulation. Here we aimed at characterizing the control of epithelial bicarbonate secretion and intestinal precipitate presence in the gilthead sea bream in response to predicted near future increases of environmental CO2. Our results demonstrate that hypercapnia (950 and 1800 μatm CO2) elicits higher intestine epithelial HCO3- secretion ex vivo and a subsequent parallel increase of intestinal precipitate presence in vivo when compared to present values (440 μatm CO2). Intestinal gene expression analysis in response to environmental hypercapnia revealed the up-regulation of transporters involved in the intestinal bicarbonate secretion cascade such as the basolateral sodium bicarbonate co-transporter slc4a4, and the apical anion transporters slc26a3 and slc26a6 of sea bream. In addition, other genes involved in intestinal ion uptake linked to water absorption such as the apical nkcc2 and aquaporin 1b expression, indicating that hypercapnia influences different levels of intestinal physiology. Taken together the current results are consistent with an intestinal physiological response leading to higher bicarbonate secretion in the intestine of the sea bream paralleled by increased luminal carbonate precipitate abundance and the main related transporters in response to ocean acidification.

Fossorial Damaraland mole rats do not exhibit a blunted hypercapnic ventilatory response.

Damaraland mole rats (DMRs, Fukomys damarensis) are a eusocial fossorial species that spend the majority of their life in densely populated underground burrows, in which they likely experience intermittent periods of elevated CO2 (i.e. hypercapnia). The primary physiological response to hypercapnia in most mammals is to increase depth and rate of breathing (i.e. hyperpnoea), but this response is often blunted in species that inhabit hypercapnic environments. In their natural habitat, DMRs putatively experience a gaseous environment ranging from normocapnic (0.1% CO2) to hypercapnic (6.0% CO2) conditions (Roper et al. 2001 J. Zool. 254, 101-107). As such, we hypothesized that DMRs would exhibit blunted hypercapnic ventilatory and metabolic responses, relative to those of non-fossorial rodent species. To test this hypothesis, we exposed awake, freely behaving DMRs to normoxic normocapnia (21% O2, 0% CO2, balance N2) or graded normoxic hypercapnia (21% O2, 0, 2, 5, 7 and 10% CO2, balance N2), and measured ventilation and metabolism using whole-body plethysmography and indirect calorimetry, respectively. We found that ventilation and metabolism were unchanged during prolonged normocapnia, whereas during graded hypercapnia, ventilation was elevated at 2% CO2 and above. As a result, O2 extraction efficiency at the lungs decreased with increasing hyperpnoea. Conversely, metabolic rate did not increase until 10% CO2, presumably due to the metabolic cost of hyperpnoea. Taken together, our results suggest that despite their fossorial lifestyle, DMRs do not exhibit adaptations in their ventilatory or metabolic responses to environmental hypercapnia.

Divergent gene expression in the gills of juvenile turbot (Psetta maxima) exposed to chronic severe hypercapnia indicates dose-dependent increase in intracellular oxidative stress and hypoxia

Elevated concentrations of carbon dioxide are a common stressor for fish and other aquatic animals. In particular, intensive aquaculture can impose prolonged periods of severe environmental hypercapnia, manifold exceeding CO2 concentrations of natural habitats. In order to cope with this stressor, gills are essential and constitute the primary organ in the acclimatization process. Yet, despite a general understanding of changes in ion regulation, not much is known with regard to other cellular mechanisms. In this study, we apply RT-qPCR to investigate changes in the expression of several genes associated with metabolism, stress and immunity within gills of juvenile turbot (Psetta maxima) after an eight-week exposure to different concentrations of CO2 (low = ∼3000 μatm, medium = ∼15,000 μatm and high = ∼25,000 μatm CO2). Histological examination of the gill tissue only found a significant increase of hypertrophied secondary lamella in the highest tested treatment level. gene expression results, on the other hand, implied both, mutual and dose-dependent transcriptional adjustments. Comparable up-regulation of IL-1ß, LMP7 and Grim19 at medium and high hypercapnia indicated an increase of reactive oxygen species (ROS) within gill cells. Simultaneous increase in Akirin and PRDX transcripts at medium CO2 indicated enhanced anti-oxidant activity and regulation of transcription, while reduced mRNA concentrations of COX, EF1α and STAT2 at high CO2 denoted suppressed protein synthesis and reduced metabolic capacity. In addition to upregulated DFAD and ApoE expression, implying compensating repair measures, gills exposed to the highest tested treatment level seemed to operate close to or even beyond their maximum capacity. Thus, fitting the model of capacity limitation, our results provide evidence for accretive intracellular hypoxia and oxidative stress in the gills of turbot, dependent on the level of environmental hypercapnia. Further, genes, such as COX, may be valuable biomarkers when attempting to discriminate between a successful and an overpowered stress response.

Impact of environmental hypercapnia on fertilization success rate and the early embryonic development of the clam Limecola balthica (Bivalvia, Tellinidae) from the southern Baltic Sea – A potential CO2 leakage case study

Carbon capture and storage technology was developed as a tool to mitigate the increased emissions of carbon dioxide by capture, transportation, injection and storage of CO2 into subterranean reservoirs. There is, however, a risk of future CO2 leakage from sub-seabed storage sites to the sea-floor sediments and overlying water, causing a pH decrease. The aim of this study was to assess effects of CO2-induced seawater acidification on fertilization success and early embryonic development of the sediment-burrowing bivalve Limecola balthica L. from the Baltic Sea. Laboratory experiments using a CO2 enrichment system involved three different pH variants (pH 7.7 as control, pH 7.0 and pH 6.3, both representing environmental hypercapnia). The results showed significant fertilization success reduction under pH 7.0 and 6.3 and development delays at 4 and 9 h post gamete encounter. Several morphological aberrations (cell breakage, cytoplasm leakages, blastomere deformations) in the early embryos at different cleavage stages were observed.

Behavioural responses to environmental hypercapnia in two eusocial species of African mole rats.

Damaraland and naked mole rat are eusocial mammals that live in crowded burrows in which CO2 is elevated. These species are thought to be highly tolerant of CO2 but their behavioural responses to hypercapnia are poorly understood. We hypothesized that Damaraland and naked mole rats would exhibit blunted behavioural responses to hypercapnia and predicted that their activity levels would be unaffected at low to moderate (2-5%) CO2 but increased at > 7% CO2. To test this, we exposed Damaraland and naked mole rats to stepwise increases in environmental CO2 (0-10%) and measured activity, exploratory behaviour, and body temperature. Surprisingly, we found that both species exhibited no differences in movement velocity, distance travelled, zone transitions (exploration), or body temperature at any level of environmental hypercapnia. Conversely, when carbonic anhydrase was inhibited with acetazolamide (50mg kg-1 intraperitonially, to increase whole-animal acidosis), exploration was significantly elevated relative to hypercapnic controls in both species at all levels of inhaled CO2, and naked mole rat body temperature decreased in > 7% CO2. We conclude that both species are largely non-responsive to environmental CO2, and that this tolerance may be dependent on bicarbonate buffering at the level of the kidney or within the blood.

Ventilatory responses of the clown knifefish, Chitala ornata, to hypercarbia and hypercapnia.

The aim of the present study was to determine the roles of externally versus internally oriented CO2/H+-sensitive chemoreceptors in promoting cardiorespiratory responses to environmental hypercarbia in the facultative air-breathing fish, Chitala ornata (the clown knifefish). Fish were exposed to environmental acidosis (pH ~ 6.0) or hypercarbia (≈ 30torr PCO2) that produced changes in water pH equal to the pH levels of the acidotic water to distinguish the relative roles of CO2 versus H+. We also injected acetazolamide to elevate arterial levels of PCO2 and [H+] in fish in normocarbic water to distinguish between internal and external stimuli. We measured changes in gill ventilation frequency, air breathing frequency, heart rate and arterial blood pressure in response to each treatment as well as the changes produced in arterial PCO2 and pH. Exposure to normocarbic water of pH 6.0 for 1h did not produce significant changes in any measured variable. Exposure to hypercarbic water dramatically increased air breathing frequency, but had no effect on gill ventilation. Hypercarbia also produced a modest bradycardia and fall in arterial blood pressure. Injection of acetazolamide produced similar effects. Both hypercarbia and acetazolamide led to increases in arterial PCO2 and falls in arterial pH although the changes in arterial PCO2/pH were more modest following acetazolamide injection as were the increases in air breathing frequency. The acetazolamide results suggest that the stimulation of air breathing was due, at least in part, to stimulation of internally oriented CO2/H+ chemoreceptors monitoring blood gas changes.

Native and exotic oysters in Brazil: Comparative tolerance to hypercapnia

Environmental hypercapnia in shallow coastal marine ecosystems can be exacerbated by increasing levels of atmospheric CO2. In these ecosystems organisms are expected to become increasingly subjected to pCO2 levels several times higher than those inhabiting ocean waters (e.g.: 10,000µatm), but still our current understanding on different species capacity to respond to such levels of hypercapnia is limited. Oysters are among the most important foundation species inhabiting these coastal ecosystems, although natural oyster banks are increasingly threatened worldwide. In the present study we studied the effects of hypercapnia on two important oyster species, the pacific oyster C. gigas and the mangrove oyster C. brasiliana, to bring new insights on different species response mechanisms towards three hypercapnic levels (ca. 1,000; 4,000; 10,000 µatm), by study of a set of biomarkers related to metabolic potential (electron transport system - ETS), antioxidant capacity (SOD, CAT, GSH), cellular damage (LPO) and energetic fitness (GLY), in two life stages (juvenile and adult) after 28 days of exposure.Results showed marked differences between each species tolerance capacity to hypercapnia, with contrasting metabolic readjustment strategies (ETS), different antioxidant response capacities (SOD, CAT, GSH), which generally allowed to prevent increased cellular damage (LPO) and energetic impairment (GLY) in both species. Juveniles were more responsive to hypercapnia stress in both congeners, and are likely to be most sensitive to extreme hypercapnia in the environment. Juvenile C. gigas presented more pronounced biochemical alterations at intermediate hypercapnia (4,000µatm) than C. brasiliana. Adult C. gigas showed biochemical alterations mostly in response to high hypercapnia (10,000µatm), while adult C. brasiliana were less responsive to this environmental stressor, despite presenting decreased metabolic potential.Our data bring new insights on the biochemical performance of two important oyster species, and suggest that the duration of extreme hypercapnia events in the ecosystem may pose increased challenges for these organisms as their tolerance capacity may be time limited.

The effect of environmental hypercapnia and size on nitrite toxicity in the striped catfish (Pangasianodon hypophthalmus)

Striped catfish (Pangasianodon hypophthalmus) are farmed intensively at high stocking densities in Vietnam where they are likely to encounter environmental hypercapnia as well as occasional high levels of aquatic nitrite. Nitrite competes with Cl− for uptake at the branchial HCO3−/Cl− exchanger, causing a drastic reduction in the blood oxygen carrying capacity through the formation of methaemoglobin and nitrosylhaemoglobin. Environmental hypercapnia induces a respiratory acidosis where the branchial HCO3−/Cl− exchange activity is reduced in order to retain HCO3− for pH recovery, which should lead to a reduced nitrite uptake. To assess the effect of hypercapnia on nitrite uptake, fish were cannulated in the dorsal aorta, allowing repeated blood sampling for measurements of haemoglobin derivatives, plasma ions and acid-base status during exposure to 0.9mM nitrite alone and in combination with acute and 48h acclimated hypercapnia over a period of 72h. Nitrite uptake was initially reduced during the hypercapnia-induced acidosis, but after pH recovery the situation was reversed, resulting in higher plasma nitrite concentrations and lower functional haemoglobin levels that eventually caused mortality. This suggests that branchial HCO3−/Cl− exchange activity is reduced only during the initial acid-base compensation, but subsequently increases with the greater availability of internal HCO3− counter-ions as pH is compensated. The data further suggest that branchial Na+/H+ exchange plays a significant role in the initial phase of acid-base compensation. Overall, longer term environmental hypercapnia does not protect against nitrite uptake in P. hypophthalmus, but instead enhances it. In addition, we observed a significant size effect in nitrite accumulation, where large fish attained plasma [nitrite] above the ambient concentration, while small fish did not. Small P. hypophthalmus instead had significantly higher plasma [nitrate], and haemoglobin concentrations, revealing greater capacity for detoxifying nitrite by oxidising it to nitrate.

Deficit in digestive capabilities of bamboo shark early stages under climate change

Little empirical information is currently available on the potential effects of acidification and/or warming in sharks, but none exist about digestive capabilities under such future conditions. Here, we investigated the impact of both acidification (ΔpH = 0.5) and warming (+4; 30 °C) on the digestive enzyme levels of recently hatched tropical bamboo shark (Chiloscyllium punctatum). Thirty days post-hatching, juvenile sharks revealed a significant increase in pancreatic trypsin levels under warming, but also a significant decrease under acidification, namely a 42 % drop under present-day temperature and 44 % drop under the warming condition. A similar trend was recorded for the alkaline phosphatase activity in shark’s intestine, i.e. the impact of environmental hypercapnia was also quite notorious—a 50 % drop under present-day temperature and 49 % drop under the warming condition. Thus, our present findings suggest that acidification and warming acted antagonistically, leading to similar enzyme activities under present and future conditions. Future directions on this topic of research should point towards the study of other types of sharks, namely pelagic ones with quite higher energetic demands.

Interactive effects of copper exposure and environmental hypercapnia on immune functions of marine bivalves Crassostrea virginica and Mercenaria mercenaria.

Estuarine organisms such as bivalves are commonly exposed to trace metals such as copper (Cu) and hypercapnia (elevated CO2 levels) in their habitats, which may affect their physiology and immune function. This study investigated the combined effects of elevated CO2 levels (∼800-2000μatm PCO2, such as predicted by the near-future scenarios of global climate change) and Cu (50μgl(-1)) on immune functions of the sediment dwelling hard clams Mercenaria mercenaria and an epifaunal bivalve, the eastern oyster Crassostrea virginica. Clams and oysters were exposed for 4 weeks to different CO2 and Cu levels, and tissue Cu burdens and immune parameters were assessed to test the hypothesis that hypercapnia will enhance Cu uptake due to the higher bioavailability of free Cu(2+) and increase the immunomodulatory effects of Cu. Exposure to Cu stimulated key immune parameters of clams and oysters leading to increased number of circulating hemocytes, higher phagocytosis and adhesion ability of hemocytes, as well as enhanced antiparasitic and antibacterial properties of the hemolymph reflected in higher activities of lysozyme and inhibitors of cysteine proteases. Lysozyme activation by Cu exposure was most prominent in normocapnia (∼400μatm PCO2) and an increase in the levels of the protease inhibitors was strongest in hypercapnia (∼800-2000μatm PCO2), but other immunostimulatory effects of Cu were evident in all PCO2 exposures. Metabolic activity of hemocytes of clams and oysters (measured as routine and mitochondrial oxygen consumption rates) was suppressed by Cu exposure likely reflecting lower rates of ATP synthesis and/or turnover. However, this metabolic suppression had no negative effects of the studied immune functions of hemocytes such as phagocytosis or adhesion capacity. Hypercapnia (∼800-2000μatm PCO2) slightly but significantly enhanced accumulation of Cu in hemocytes, consistent with higher Cu(2+) bioavailability in CO2-acidified water, but had little effect on cellular and humoral immune traits of clams and oysters. These findings indicate that low levels of Cu contamination may enhance immunity of estuarine bivalves while moderate hypercapnia (such as predicted by the near future scenarios of the global climate change) does not strongly affect their immune parameters.

Ocean acidification does not alter grazing in the calanoid copepods Calanus finmarchicus and Calanus glacialis

AbstractIt is currently under debate whether organisms that regulate their acid–base status under environmental hypercapnia demand additional energy. This could impair animal fitness, but might be compensated for via increased ingestion rates when food is available. No data are yet available for dominant Calanus spp. from boreal and Arctic waters. To fill this gap, we incubated Calanus glacialis at 390, 1120, and 3000 µatm for 16 d with Thalassiosira weissflogii (diatom) as food source on-board RV Polarstern in Fram Strait in 2012. Every 4 d copepods were subsampled from all CO2 treatments and clearance and ingestion rates were determined. During the SOPRAN mesocosm experiment in Bergen, Norway, 2011, we weekly collected Calanus finmarchicus from mesocosms initially adjusted to 390 and 3000 µatm CO2 and measured grazing at low and high pCO2. In addition, copepods were deep frozen for body mass analyses. Elevated pCO2 did not directly affect grazing activities and body mass, suggesting that the copepods did not have additional energy demands for coping with acidification, neither during long-term exposure nor after immediate changes in pCO2. Shifts in seawater pH thus do not seem to challenge these copepod species.

Acclimation to hypercarbia protects cardiac contractility and alters tissue carbohydrate metabolism in the Amazonian armored catfish Pterygoplichthys pardalis

The armored catfish Pterygoplichthys pardalis tolerates environmental hypercarbia, high partial pressures of CO2 (\(P_{{{\text{CO}}_{ 2} }}\)), by preferentially protecting intracellular pH (pHi) in the face of extracellular acidosis. This response is associated with ionic changes which may disrupt contractility in cardiac muscle, and it is not known whether acclimation to hypercarbia provides protection against these changes. We studied the influence of different \(P_{{{\text{CO}}_{ 2} }}\) acclimation histories on cardiac muscle function using isometrically contracting ventricular strip preparations. Fish were held for >4 months at 21 mmHg \(P_{{{\text{CO}}_{ 2} }}\) and then exposed to normocarbia (6 mmHg \(P_{{{\text{CO}}_{ 2} }}\)) for either 15 h or 5–6 days. Acclimation to chronic hypercarbia eliminated the negative inotropic effects of in vitro hypercarbia, decreased extracellular Ca2+ sensitivity, and reduced maximum pacing frequency in ventricular strip preparations. Fish acclimated to chronic hypercarbia also exhibited hepatic glycogen and plasma glucose accumulation, and lower plasma lactate levels compared to fish acclimated to normocarbia for 5–6 days. We suggest chronic hypercarbia may induce cardiac remodeling to protect contractility and reduce the energetic demands of pHi regulation. The activation of HCO3− synthesis pathways may decrease glucose utilization and enhance carbohydrate stores, potentially providing protection against hypoxia, a stressor frequently encountered in conjunction with hypercarbia in the Amazon.

Elevated pCO2 drives lower growth and yet increased calcification in the early life history of the cuttlefish Sepia officinalis (Mollusca: Cephalopoda)

AbstractOcean acidification is an escalating environmental issue and associated changes in the ocean carbonate system have implications for many calcifying organisms. The present study followed the growth of Sepia officinalis from early-stage embryos, through hatching, to 7-week-old juveniles. Responses of cuttlefish to elevated pCO2 (hypercapnia) were investigated to test the impacts of near-future and extreme ocean acidification conditions on growth, developmental time, oxygen consumption, and yolk utilization as proxies for individual fitness. We further examined gross morphological characteristics of the internal calcareous cuttlebone to determine whether embryonically secreted shell lamellae are impacted by environmental hypercapnia. Embryonic growth was reduced and hatching delayed under elevated pCO2, both at environmentally relevant levels (0.14 kPa pCO2 similar to predicted ocean conditions in 2100) and extreme conditions (0.40 kPa pCO2). Comparing various metrics from control and intermediate treatments generally showed no significant difference in experimental measurements. Yet, results from the high pCO2 treatment showed significant changes compared with controls and revealed a consistent general trend across the three treatment levels. The proportion of animal mass contributed by the cuttlebone increased in both elevated pCO2 treatments. Gross cuttlebone morphology was affected under such conditions and cuttlebones of hypercapnic individuals were proportionally shorter. Embryonic shell morphology was maintained consistently in all treatments, despite compounding hypercapnia in the perivitelline fluid; however, post-hatching, hypercapnic animals developed denser cuttlebone laminae in shorter cuttlebones. Juvenile cuttlefish in acidified environments thus experience lower growth and yet increased calcification of their internal shell. The results of this study support recent findings that early cuttlefish life stages are more vulnerable towards hypercapnia than juveniles and adults, which may have negative repercussions on the biological fitness of cuttlefish hatchlings in future oceans.

Interactive effects of metal pollution and ocean acidification on physiology of marine organisms

Abstract Changes in the global environment such as ocean acidification (OA) may interact with anthropogenic pollutants including trace metals threatening the integrity of marine ecosystems. We analyze recent studies on the interactive effects of OA and trace metals on marine organisms with a focus on the physiological basis of these interactions. Our analysis shows that the responses to elevated CO2 and metals are strongly dependent on the species, developmental stage, metal biochemistry and the degree of environmental hypercapnia, and cannot be directly predicted from the CO2-induced changes in metal solubility and speciation. The key physiological functions affected by both the OA and trace metal exposures involve acid-base regulation, protein turnover and mitochondrial bioenergetics, reflecting the sensitivity of the underlying molecular and cellular pathways to CO2and metals. Physiological interactions between elevated CO2 and metals may impact the organisms’ capacity to maintain acid-base homeostasis and reduce the amount of energy available for fitness-related functions such as growth, development and reproduction thereby affecting survival and performance of estuarine populations. Environmental hypercapnia may also affect the marine food webs by altering predator-prey interactions and the trophic transfer of metals in the food chain. However, our understanding of the degree to which these effects can impact the function and integrity of marine ecosystems is limited due the scarcity of the published research and its bias towards certain taxonomic groups. Future research priorities should include studies of metal x PCO2 interactions focusing on critical physiological functions (including acid-base, protein and energy homeostasis) in a greater range of ecologically and economically important marine species, as well as including the field populations naturally exposed (and potentially adapted) to different levels of metals and CO2 in their environments.

Effects of environmental hypercapnia and metal (Cd and Cu) exposure on acid-base and metal homeostasis of marine bivalves

Elevated CO2 levels reduce seawater pH and may affect bioavailability of trace metals in estuaries. We studied the interactive effects of common metal pollutants (50μgl−1 Cd or Cu) and PCO2 (~395, 800 and 2000μatm) on metal levels, intracellular pH, expression of metal binding proteins and stress biomarkers in estuarine bivalves Crassostrea virginica (oysters) and Mercenaria mercenaria (hard clams). Cd (but not Cu or hypercapnia) exposure affected the acid-base balance of hemocytes resulting in elevated intracellular pH. Cd and Cu exposure led to the increase in the tissue metal burdens, and metal accumulation was reduced by elevated PCO2 in the mantle but not hemocytes. No change was found in the intracellular free Cd2+, Cu2+ or Fe2+ during Cu or Cd exposure indicating that these metals are bound to intracellular ligands. Free Zn2+ content in oyster hemocytes was suppressed by Cd and Cu exposure and below the detection limits in clam hemocytes, which went hand-in-hand with the elevated mRNA expression of metallothioneins and ferritin in Cd- and Cu-exposed bivalves, enhanced by hypercapnia. The metal-binding and antioxidant mechanisms of oysters and clams were sufficient to effectively maintain intracellular redox status, even though metal exposure combined with moderate hypercapnia (~800μatm PCO2) led to the elevated production of reactive oxygen species in hemocytes. Overall, while hypercapnia modulates metal accumulation, binding capacity and oxidative stress in estuarine bivalves, the physiological effects of elevated CO2 are mild compared to the effects of other common stressors.

Effects of elevated CO2 levels on eggs and larvae of a North Pacific flatfish

AbstractThe Bering Sea and Gulf of Alaska support a number of commercially important flatfish fisheries. These high latitude ecosystems are predicted to be most immediately impacted by ongoing ocean acidification, but the range of responses by commercial fishery species has yet to be fully explored. In this study, we examined the growth responses of northern rock sole (Lepidopsetta polyxystra) eggs and larvae across a range of CO2 levels (ambient to 1500 µatm) to evaluate the potential sensitivity to ocean acidification. Laboratory-spawned eggs and larvae were reared at 8°C in a flow-through culture system in which CO2 levels were maintained via computer-controlled injection of CO2 into a seawater conditioning tank. Overall, we observed only minor effects of elevated CO2 level on sizes of northern rock sole larvae. Size at hatch differed among offspring from four different females, but there was no significant effect of CO2 level on egg survival or size at hatch. In three separate larval growth trials, there was little effect of CO2 level on growth rates through the first 28 d post-hatch (DPH). However, in the one trial extended to 60 DPH, fish reared at the highest CO2 level had lower condition factors after 28 DPH, suggesting that larvae undergoing metamorphosis may be more sensitive to environmental hypercapnia than earlier pre-flexion stages. These results suggest that while early life stages of northern rock sole are less sensitive to ocean acidification than previously examined flatfish, they may be more sensitive to elevated CO2 levels than a previously studied gadid with a similar geographic range.