Acclimatory gene expression of primed clams enhances robustness to elevated pCO2.

Sublethal exposure to environmental challenges may enhance ability to cope with chronic or repeated change, a process known as priming. In a previous study, pre-exposure to seawater enriched with pCO2 improved growth and reduced antioxidant capacity of juvenile Pacific geoduck Panopea generosa clams, suggesting that transcriptional shifts may drive phenotypic modifications post-priming. To this end, juvenile clams were sampled and TagSeq gene expression data were analysed after (i) a 110-day acclimation under ambient (921 µatm, naïve) and moderately elevated pCO2 (2870 µatm, pre-exposed); then following (ii) a second 7-day exposure to three pCO2 treatments (ambient: 754 µatm; moderately elevated: 2750 µatm; severely elevated: 4940 µatm), a 7-day return to ambient pCO2 and a third 7-day exposure to two pCO2 treatments (ambient: 967 µatm; moderately elevated: 3030 µatm). Pre-exposed geoducks frontloaded genes for stress and apoptosis/innate immune response, homeostatic processes, protein degradation and transcriptional modifiers. Pre-exposed geoducks were also responsive to subsequent encounters, with gene sets enriched for mitochondrial recycling and immune defence under elevated pCO2 and energy metabolism and biosynthesis under ambient recovery. In contrast, gene sets with higher expression in naïve clams were enriched for fatty-acid degradation and glutathione components, suggesting naïve clams could be depleting endogenous fuels, with unsustainable energetic requirements if changes in carbonate chemistry persist. Collectively, our transcriptomic data indicate that pCO2 priming during post-larval periods could, via gene expression regulation, enhance robustness in bivalves to environmental change. Such priming approaches may be beneficial for aquaculture, as seafood demand intensifies concurrent with increasing climate change in marine systems.

Repeat exposure to hypercapnic seawater modifies growth and oxidative status in a tolerant burrowing clam.

Although low levels of thermal stress, irradiance and dietary restriction can have beneficial effects for many taxa, stress acclimation remains little studied in marine invertebrates, even though they are threatened by climate change stressors such as ocean acidification. To test the role of life-stage and stress-intensity dependence in eliciting enhanced tolerance under subsequent stress encounters, we initially conditioned pediveliger Pacific geoduck (Panopea generosa) larvae to ambient and moderately elevated PCO2 (920 µatm and 2800 µatm, respectively) for 110 days. Then, clams were exposed to ambient, moderate or severely elevated PCO2 (750, 2800 or 4900 µatm, respectively) for 7 days and, following 7 days in ambient conditions, a 7-day third exposure to ambient (970 µatm) or moderate PCO2 (3000 µatm). Initial conditioning to moderate PCO2 stress followed by second and third exposure to severe and moderate PCO2 stress increased respiration rate, organic biomass and shell size, suggesting a stress-intensity-dependent effect on energetics. Additionally, stress-acclimated clams had lower antioxidant capacity compared with clams under ambient conditions, supporting the hypothesis that stress over postlarval-to-juvenile development affects oxidative status later in life. Time series and stress intensity-specific approaches can reveal life-stages and magnitudes of exposure, respectively, that may elicit beneficial phenotypic variation.

Metabolic recovery and compensatory shell growth of juvenile Pacific geoduck Panopea generosa following short-term exposure to acidified seawater.

While acute stressors can be detrimental, environmental stress conditioning can improve performance. To test the hypothesis that physiological status is altered by stress conditioning, we subjected juvenile Pacific geoduck, Panopea generosa, to repeated exposures of elevated pCO2 in a commercial hatchery setting followed by a period in ambient common garden. Respiration rate and shell length were measured for juvenile geoduck periodically throughout short-term repeated reciprocal exposure periods in ambient (~550 µatm) or elevated (~2400 µatm) pCO2 treatments and in common, ambient conditions, 5 months after exposure. Short-term exposure periods comprised an initial 10-day exposure followed by 14 days in ambient before a secondary 6-day reciprocal exposure. The initial exposure to elevated pCO2 significantly reduced respiration rate by 25% relative to ambient conditions, but no effect on shell growth was detected. Following 14 days in common garden, ambient conditions, reciprocal exposure to elevated or ambient pCO2 did not alter juvenile respiration rates, indicating ability for metabolic recovery under subsequent conditions. Shell growth was negatively affected during the reciprocal treatment in both exposure histories; however, clams exposed to the initial elevated pCO2 showed compensatory growth with 5.8% greater shell length (on average between the two secondary exposures) after 5 months in ambient conditions. Additionally, clams exposed to the secondary elevated pCO2 showed 52.4% increase in respiration rate after 5 months in ambient conditions. Early exposure to low pH appears to trigger carryover effects suggesting bioenergetic re-allocation facilitates growth compensation. Life stage-specific exposures to stress can determine when it may be especially detrimental, or advantageous, to apply stress conditioning for commercial production of this long-lived burrowing clam.

Temperature-related heart rate in water and air and a comparison to other temperature-related measures of performance in the fiddler crab Leptuca pugilator (Bosc 1802).

Performance in poikilotherms is known to be sensitive to temperature, often with a low-sloping increase with temperature to a peak, and a steep decline with increasing temperature past the peak. We complemented past measures of performance by measuring heartbeat rates of the fiddler crab Leptuca pugilator in water and in air as a function of a range of temperatures previously shown to affect other measures of performance. In water over a range of 20-50 °C, heartbeat increased steadily to a peak at 40 °C and then steeply declined to near zero at 50 °C. In air, heartbeat also increased, but to a peak at 35 °C and then with a gentler decline than was found in water. Part of this different response may be due to evaporative water loss, which reduced body temperature in air, and therefore thermal stress, relative to body temperature when crabs were immersed in water. Increased availability of oxygen from air, according to the oxygen and capacity-limited thermal tolerance hypothesis, likely increased aerobic scope past the thermal peak, relative to within water, where oxygen delivery at higher temperatures may have been curtailed. We compared the heart rate performance relations to two previous measures of performance - endurance on a treadmill and sprint speed, both done in air. The peak performance temperature increased in the order: treadmill endurance time, sprint speed, heart rate in air, and heart rate in water, which demonstrates that different performance measures give different perspectives on the relation of thermal tolerance and fitness to temperature. Endurance may therefore be the limiting upper thermal stress factor in male fiddler crabs, when on hot sand flats. Temperature preference, found to be for temperatures <30 °C in air, could be a bet-hedging evolutionary strategy to avoid aerobic scope affecting endurance.