Nathan A. Miller

Impact of ocean acidification on metabolism and energetics during early life stages of the intertidal porcelain crab Petrolisthes cinctipes

SUMMARY Absorption of elevated atmospheric CO2 is causing surface ocean pH to decline, a process known as ocean acidification (OA). To date, few studies have assessed the physiological impacts of OA on early life-history stages of intertidal organisms, which transition from habitats with fluctuating pH (intertidal zone) to relatively stable (pelagic zone) pH environments. We used the intertidal crab Petrolisthes cinctipes to determine whether metabolic responses to year 2300 predictions for OA vary among early developmental stages and to examine whether the effects were more pronounced in larval stages developing in the open ocean. Oxygen consumption rate, total protein, dry mass, total lipids and C/N were determined in late-stage embryos, zoea I larvae and newly settled juveniles reared in ambient pH (7.93±0.06) or low pH (7.58±0.06). After short-term exposure to low pH, embryos displayed 11% and 6% lower metabolism and dry mass, respectively, which may have an associated bioenergetic cost of delayed development to hatching. However, metabolic responses appeared to vary among broods, suggesting significant parental effects among the offspring of six females, possibly a consequence of maternal state during egg deposition and genetic differences among broods. Larval and juvenile metabolism were not affected by acute exposure to elevated CO2. Larvae contained 7% less nitrogen and C/N was 6% higher in individuals reared at pH 7.58 for 6 days, representing a possible switch from lipid to protein metabolism under low pH; the metabolic switch appears to fully cover the energetic cost of responding to elevated CO2. Juvenile dry mass was unaffected after 33 days exposure to low pH seawater. Increased tolerance to low pH in zoea I larvae and juvenile stages may be a consequence of enhanced acid–base regulatory mechanisms, allowing greater compensation of extracellular pH changes and thus preventing decreases in metabolism after exposure to elevated PCO2. The observed variation in responses of P. cinctipes to decreased pH in the present study suggests the potential for this species to adapt to future declines in near-shore pH.

Effects of ocean acidification on early life-history stages of the intertidal porcelain crab Petrolisthes cinctipes.

SUMMARY Intertidal zone organisms naturally experience daily fluctuations in pH, presently reaching values beyond what is predicted for open ocean surface waters from ocean acidification (OA) by the year 2100, and thus present an opportunity to study the pH sensitivity of organisms that are presumably adapted to an acidified environment. The intertidal zone porcelain crab, Petrolisthes cinctipes, was used to study physiological responses to low pH in embryonic, larval and newly recruited juvenile life-history stages. In these crabs, embryonic development occurs in the pH-variable intertidal zone (pH 6.9–9.5), larvae mature in the more stable pelagic environment (pH 7.9–8.2), and juvenile crabs settle back into the pH-variable intertidal zone. We examined survival, cardiac performance, energetics and morphology in embryonic, larval and juvenile crabs exposed to two pH conditions (pH 7.9 and 7.6). Embryos and larvae were split by brood between the pH treatments for 9 days to examine brood-specific responses to low pH. Hatching success did not differ between pH conditions, but ranged from 30% to 95% among broods. Larval survival was not affected by acidification, but juvenile survival was reduced by ~30% after longer (40 days) exposure to low pH. Embryonic and larval heart rates were 37% and 20% lower at low pH, and there was a brood-specific response in embryos. Embryos did not increase in volume under acidified conditions, compared with a 15% increase in ambient conditions. We conclude that sustained exposure to low pH could be detrimental to P. cinctipes embryos and larvae despite the fact that embryos are regularly exposed to naturally fluctuating hypercapnic water in the intertidal zone. Importantly, our results indicate that early life-history stage responses to OA may be brood specific through as yet undetermined mechanisms.

Temperature and acidification variability reduce physiological performance in the intertidal zone porcelain crab Petrolisthes cinctipes

We show here that increased variability of temperature and pH synergistically negatively affects the energetics of intertidal zone crabs. Under future climate scenarios, coastal ecosystems are projected to have increased extremes of low tide-associated thermal stress and ocean acidification-associated low pH, the individual or interactive effects of which have yet to be determined. To characterize energetic consequences of exposure to increased variability of pH and temperature, we exposed porcelain crabs, Petrolisthes cinctipes, to conditions that simulated current and future intertidal zone thermal and pH environments. During the daily low tide, specimens were exposed to no, moderate or extreme heating, and during the daily high tide experienced no, moderate or extreme acidification. Respiration rate and cardiac thermal limits were assessed following 2.5 weeks of acclimation. Thermal variation had a larger overall effect than pH variation, though there was an interactive effect between the two environmental drivers. Under the most extreme temperature and pH combination, respiration rate decreased while heat tolerance increased, indicating a smaller overall aerobic energy budget (i.e. a reduced O2 consumption rate) of which a larger portion is devoted to basal maintenance (i.e. greater thermal tolerance indicating induction of the cellular stress response). These results suggest the potential for negative long-term ecological consequences for intertidal ectotherms exposed to increased extremes in pH and temperature due to reduced energy for behavior and reproduction.

Ocean acidification exerts negative effects during warming conditions in a developing Antarctic fish

Polar areas are experiencing some of the most rapid impacts of climate change, yet we have a limited understanding of biological vulnerability to multiple stressors. Increased temperature broadly affected early embryo physiology in the naked dragonfish, while ocean acidification interacted synergistically with temperature to decrease survival and alter developmental rate.

The role of oxygen in determining upper thermal limits in lottia digitalis under air exposure and submersion

Oxygen limitation of aerobic metabolism is hypothesized to drive organismal thermal tolerance limits. Differences in oxygen availability in air and water may underlie observed differences in upper thermal tolerance of intertidal limpets if oxygen is limiting in submerged environments. We explored how cardiac performance (heart rate, breakpoint temperature [BPT], flat-line temperature [FLT], and temperature sensitivity) was affected by hyperoxia and hypoxia in the finger limpet, Lottia digitalis, under air exposure and submersion. Upper thermal tolerance limits were unchanged by increasing availability of oxygen, although air-exposed limpets were able to maintain cardiac function to higher temperatures than submerged limpets. Maximum heart rate did not increase with greater partial pressure of oxygen (Po2), suggesting that tissue Po2 levels are likely maximized during normoxia. Hypoxia reduced breakpoint BPTs and FLTs in air-exposed and submerged limpets and accentuated the difference in BPTs between the two groups through greater reductions in BPT in submerged limpets. Differences in respiratory structures and the degree to which thermal limits are already maximized may play significant roles in determining how oxygen availability influences upper temperature tolerance.

Neural Thermal Performance in Porcelain Crabs, Genus Petrolisthes

Neurons are highly temperature sensitive. Temperature-induced nerve failure may play an important role in determining organismal thermal tolerance limits and distribution patterns. To expand our understanding of the role of neuronal thermal performance in setting thermal limits, we compared the thermal performance of neurons from five porcelain crab (genus Petrolisthes) congeners that differ in thermal habitat. In experiment 1, neuronal performance of sensory neurons was determined by extracellular recording of spontaneous action potentials during thermal ramps. Arrhenius break temperatures of action potential generation were used to calculate maximum critical temperature (CTmax) and minimum critical temperature (CTmin) for neuronal performance. CTmax and CTmin were related to habitat temperature across the five species and were found to respond to acclimation temperature. In experiment 2, we assessed the performance of neurons from Petrolisthes cinctipes acclimated at 8°, 18°, and 25°C when placed at 30°C (near the whole-organism CTmax of this species) and demonstrated that neural performance near whole-organism CTmax increases with increasing acclimation temperature. In experiment 3, we compared the thermal limits of sensory afferents and pacemaker efferents and found that they were correlated, although pacemaker efferents tended to have a higher CTmax and reduced plasticity. Our final analysis, which was of transcriptomic data in cardiac tissue, leads us to hypothesize that nerve membrane K+ conductance may underlie variation in nerve thermal tolerance.

Juvenile Antarctic rockcod, Trematomus bernacchii, are physiologically robust to CO2–acidified seawater

ABSTRACT To date, numerous studies have shown negative impacts of CO2-acidified seawater (i.e. ocean acidification, OA) on marine organisms, including calcifying invertebrates and fishes; however, limited research has been conducted on the physiological effects of OA on polar fishes and even less on the impact of OA on early developmental stages of polar fishes. We evaluated aspects of aerobic metabolism and cardiorespiratory physiology of juvenile emerald rockcod, Trematomus bernacchii, an abundant fish in the Ross Sea, Antarctica, to elevated partial pressure of carbon dioxide (PCO2) [420 (ambient), 650 (moderate) and 1050 (high) μatm PCO2] over a 1 month period. We examined cardiorespiratory physiology, including heart rate, stroke volume, cardiac output and ventilation rate, whole organism metabolism via oxygen consumption rate and sub-organismal aerobic capacity by citrate synthase enzyme activity. Juvenile fish showed an increase in ventilation rate under high PCO2 compared with ambient PCO2, whereas cardiac performance, oxygen consumption and citrate synthase activity were not significantly affected by elevated PCO2. Acclimation time had a significant effect on ventilation rate, stroke volume, cardiac output and citrate synthase activity, such that all metrics increased over the 4 week exposure period. These results suggest that juvenile emerald rockcod are robust to near-future increases in OA and may have the capacity to adjust for future increases in PCO2 by increasing acid-base compensation through increased ventilation. Summary: Juvenile Antarctic emerald rockcod are robust to projected ocean acidification scenarios and may have the capacity to adjust for future increases in PCO2 by increasing acid-base compensation through increased ventilation.

Metabolic physiology of the invasive clam, Potamocorbula amurensis: the interactive role of temperature, salinity, and food availability.

In biological systems energy serves as the ultimate commodity, often determining species distributions, abundances, and interactions including the potential impact of invasive species on native communities. The Asian clam Potamocorbula amurensis invaded the San Francisco Estuary (SFE) in 1986 and is implicated in the decline of native fish species through resource competition. Using a combined laboratory/field study we examined how energy expenditure in this clam is influenced by salinity, temperature and food availability. Measures of metabolism were made at whole organism (metabolic rate) and biochemical (pyruvate kinase (PK) and citrate synthase (CS) enzyme activities) levels. We found in the field, over the course of a year, the ratio of PK to CS was typically 1.0 suggesting that aerobic and fermentative metabolism were roughly equivalent, except for particular periods characterized by low salinity, higher temperatures, and intermediate food availabilities. In a 30-day laboratory acclimation experiment, however, neither metabolic rate nor PK:CS ratio was consistently influenced by the same variables, though the potential for fermentative pathways did predominate. We conclude that in field collected animals, the addition of biochemical measures of energetic state provide little additional information to the previously measured whole organism metabolic rate. In addition, much of the variation in the laboratory remained unexplained and additional variables, including reproductive stage or body condition may influence laboratory-based results. Further study of adult clams must consider the role of organismal condition, especially reproductive state, in comparisons of laboratory experiments and field observations.

Juvenile rockfish show resilience to CO2-acidification and hypoxia across multiple biological scales

The present study showed short-term elevations in cellular metabolism, alterations in behavior and susceptibility to predation in juvenile rockfish after acute acclimation to CO2-acidification and hypoxic conditions. Physiological and behavioral alterations were restored after 3 weeks suggesting that rockfish possess mechanisms to defend rapid changes in PCO2 and oxygen conditions.