Interindividual behavioural variation in response to elevated CO2 predicts mRNA transcript abundance of genes related to acid-base regulation in medaka (Oryzias latipes).

Rising carbon dioxide (CO2) in aquatic ecosystems due to climate change is a challenge for aquatic ectotherms. We examined whether interindividual variation in behavioural responses to CO2 could predict how a teleost fish would respond to elevated CO2 for multiple phenotypic and molecular traits. To this end, we first quantified behavioural responses of individuals exposed to acute elevated CO2, and used these to assign individuals as either high or low responders relative to the population mean. Subsequently, we exposed both high and low responders to elevated CO2 for 6 weeks and quantified the effect on body condition, behaviour, and mRNA transcript responses of gill and liver genes associated with relevant physiological processes. Generally, we found few relationships between the phenotypic groups and body condition and behaviour following the CO2 exposure period; however, stark differences between the phenotypic groups with respect to gene transcripts from each tissue related to various processes were found, mostly independent of CO2 exposure. The most pronounced changes were in the gill transcripts related to acid-base regulation, suggesting that the observed behavioural variation used to assign fish to phenotypic groups may have an underlying molecular origin. Should the link between behaviour and gene transcripts be shown to have a fitness advantage and be maintained across generations, interindividual variation in behavioural responses to acute CO2 exposure may be a viable and non-invasive tool to predict future population responses to elevated aquatic CO2.

Understanding olfactory and behavioural responses to dietary cues in age-1 lake sturgeon Acipenser fulvescens.

Detection of environmental cues is essential for all vertebrates and is typically established by the olfactory epithelium and olfactory sensory neurons (OSNs). In fishes, microvillous and ciliated OSNs are the principal types, typically detecting amino acids and bile salts, respectively. Activation of OSN receptors by specific ligands initiate downstream signal processing often leading to behavioural responses. In this study we used electrophysiological and behavioural techniques to evaluate olfactory detection and behaviour in juvenile lake sturgeon Acipenser fulvescens in response to hatchery- and natural dietary cues. We hypothesized that electro-olfactogram (EOG) and behavioural responses would be dependent on diet type. We predicted that inhibition of the phospholipase C/inositol 1,4,5-triphosphate (PLC/IP3) secondary transduction pathway would reduce EOG responses to dietary cues and, inhibition of the adenylyl cyclase/adenosine 3,5-cyclic monophosphate (cAMP) pathway, would have no effect. Furthermore, we predicted a strong EOG response would be manifested in a change in behaviour. We observed that both the PLC/IP3 and cAMP pathways were significantly involved in the detection of dietary cues. However, EOG responses did not manifest to behavioural responses, although the foraging activity to the hatchery cue was significantly greater compared to the control. Our results support the notion that lake sturgeon raised in a hatchery and fed a commercial pelleted diet may become accustomed to it prior to release into the wild. Further, this study suggests that, in conservation aquaculture settings, lake sturgeon should be exposed to natural dietary cues prior to release as one strategy to promote food recognition.

Are the effects of catch-and-release angling evident in changes to mRNA abundances related to metabolism, acid-base regulation and stress in lake trout (Salvelinus namaycush) gills?

Catch-and-release (C&R) angling is a conservation-oriented practice intended to reduce the impact recreational angling has on fish populations. Even though most recreationally angled fish are released, little is known about how C&R angling impacts fish at the cellular or tissue level. As the first to explore the impacts of C&R angling on mRNA abundances, our study aimed to identify how the stress of angling influenced metabolism, acid-base regulation and cellular stress in the gills of lake trout (Salvelinus namaycush). Because gills are responsible for metabolic gas exchange, are crucial sites of acid-base homeostasis and respond to stressors quickly, we hypothesized that the relative mRNA abundance of genes related to these three physiological processes would be altered after angling. We took gill samples of live lake trout at 0, 2 or 48 h after fish were angled by rod and reel, and then used quantitative PCR (qPCR) to measure the relative abundance of nine candidate mRNA transcripts. Heat shock protein 70 (hsp70) mRNA levels significantly increased over 5-fold 2 h after angling, indicating a potential activation of a cytoprotective response. However, contrary to our hypothesis, we observed no change in the relative mRNA abundance of genes related to metabolism or acid-base regulation in response to C&R angling within a 48-h period. As C&R angling can negatively impact fish populations, further use of transcript-level studies will allow us to understand the impact C&R has on specific tissues and improve our knowledge of how C&R influences overall fish health.

Elevated CO2 levels did not induce species- or tissue-specific damage in young-of-year salmonids.

OBJECTIVE: The broad objective of our study was to use a histological approach to examine possible tissue-level effects of exposure to elevated CO2 in freshwater young-of-year salmonids. METHODS: To study these effects, young-of-year Arctic Char Salvelinus alpinus, Rainbow Trout Oncorhynchus mykiss, and Brook Trout S. fontinalis were exposed to either control levels of CO2 (1400 µatm) or elevated levels of CO2 (5236 µatm) for 15 days. Fish were then sampled for gill, liver, and heart tissues and histologically analyzed. RESULT: A species effect was observed for the length of secondary lamellae, as Arctic Char had significantly shorter secondary lamellae than the other species. No notable changes within the gills and livers of Arctic Char, Brook Trout, or Rainbow Trout exposed to elevated CO2 were observed. CONCLUSION: Generally, our results indicated that elevated CO2 levels over 15 days do not induce catastrophic tissue damage and it is unlikely that fish health would be seriously impacted. Ongoing research dedicated to examining how elevated CO2 long-term may affect internal tissues of fish will allow for a more comprehensive understanding of how fish may fair with ongoing climate change and in aquaculture facilities.

A 30-year dataset of CO2 in flowing freshwaters in the United States.

Increasing atmospheric carbon dioxide (CO2) concentrations have been linked to effects in a wide range of ecosystems and organisms, with negative effects of elevated CO2 documented for marine organisms. Less is known about the dynamics of CO2 in freshwaters, but the potential exists for freshwater organisms to be challenged by elevated CO2. In flowing freshwaters CO2 exhibits more variability than in lakes or the ocean, yet spatiotemporally extensive direct measures of CO2 in freshwater are rare. However, CO2 can be estimated from pH, temperature, and alkalinity-commonly collected water quality metrics. We used data from the National Water Quality Monitoring Council along with the program PHREEQC to estimate CO2 in flowing freshwaters across 35,000 sites spanning the lower 48 US states from 1990 through 2020. Site data for water chemistry measurements were spatially joined with the National Hydrology Dataset. Our resulting dataset, CDFLOW, presents an opportunity for researchers to add CO2 to their datasets for further investigation.

Resilience of larval wood frogs (Rana sylvatica) to hydrocarbons and other compounds released from naturally weathered diluted bitumen in a boreal lake.

The risks to aquatic wildlife from spills of diluted bitumen (dilbit) into inland waters are poorly understood. In this paper, we describe the response of larval wood frogs (Rana sylvatica) to hydrocarbons and other compounds released from experimental spills of dilbit in a temperate boreal lake. To simulate a wide range of environmentally relevant oil spill scenarios, different volumes of Cold Lake Winter Blend dilbit (0, 1.5, 2.9, 5.5, 18, 42, 82, and 180 L) were added to 10 m diameter in-lake limnocorrals. Larvae (n = 360) were reared (from Gosner Stage (GS) 25 to ∼42) in land-based aquatic microcosms, where they were first exposed to clean water during a 2-week baseline phase, and then (at GS ∼30), to contaminated water withdrawn from the limnocorrals for 3 weeks. We observed no statistically significant trends in survival, growth, or development of larvae as a consequence of exposure to the chemical compounds released from naturally weathered dilbit. Likewise, neither cytochrome P450 1A biomarkers nor levels of thyroid hormones in wood frogs near metamorphic climax were significantly related to volume of the oil spills. However, there was a modest statistically significant decrease in larval activity (up to 8.7% relative to the control), but no change in other behavioral metrics (i.e., sociality or space use). Our work adds to the limited body of literature on the effects of unconventional oils on aquatic wildlife and helps to inform risk assessments regarding pipeline projects.

Fathead Minnows Exposed to Organic Compounds from Oil Sands Tailings as Embryos Have Reduced Survival, Impaired Development, and Altered Behaviors That Persist into Larval Stages.

Our study evaluated whether exposure to naphthenic acid fraction compounds (NAFCs) extracted from oil sands process-affected waters (OSPW) has adverse effects on fish embryos that persist into later life. We exposed fathead minnow (Pimephales promelas) embryos to concentrations of NAFCs found in OSPW (2.5-54 mg/L) for 7 days (1 day postfertilization to hatch), then raised surviving larvae in outdoor mesocosms of uncontaminated lake water for 1 month. Embryos exposed to NAFCs were more likely to exhibit malformations (by up to 8-fold) and had slower heart rates (by up to 24%) compared to controls. Fish raised in uncontaminated lake water following exposure to NAFCs as embryos, were 2.5-fold less likely to survive during the larval stage than control fish. These fish also showed up to a 45% decrease in swim activity and a 36% increase in swim burst events during behavioral tests relative to controls. We conclude that exposure to NAFCs during the embryonic stage can have lasting effects on fish survival, physiology, and behavior that persist at least through the larval stage. These findings of delayed mortalities and persistent sublethal effects of embryonic NAFC exposure are relevant to informing the development of regulations on treated OSPW releases from mining operations. Environ Toxicol Chem 2022;41:1319-1332. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Molecular and physiological responses predict acclimation limits in juvenile brook trout (Salvelinus fontinalis).

Understanding the resilience of ectotherms to high temperatures is essential because of the influence of climate change on aquatic ecosystems. The ability of species to acclimate to high temperatures may determine whether populations can persist in their native ranges. We examined physiological and molecular responses of juvenile brook trout (Salvelinus fontinalis) to six acclimation temperatures (5, 10, 15, 20, 23 and 25°C) that span the thermal distribution of the species to predict acclimation limits. Brook trout exhibited an upregulation of stress-related mRNA transcripts (heat shock protein 90-beta, heat shock cognate 71 kDa protein, glutathione peroxidase 1) and downregulation of transcription factors and osmoregulation-related transcripts (nuclear protein 1, Na+/K+/2Cl- co-transporter-1-a) at temperatures ≥20°C. We then examined the effects of acclimation temperature on metabolic rate (MR) and physiological parameters in fish exposed to an acute exhaustive exercise and air exposure stress. Fish acclimated to temperatures ≥20°C exhibited elevated plasma cortisol and glucose, and muscle lactate after exposure to the acute stress. Fish exhibited longer MR recovery times at 15 and 20°C compared with the 5 and 10°C groups; however, cortisol levels remained elevated at temperatures ≥20°C after 24 h. Oxygen consumption in fish acclimated to 23°C recovered quickest after exposure to acute stress. Standard MR was highest and factorial aerobic scope was lowest for fish held at temperatures ≥20°C. Our findings demonstrate how molecular and physiological responses predict acclimation limits in a freshwater fish as the brook trout in the present study had a limited ability to acclimate to temperatures beyond 20°C.

Sub-lethal temperature thresholds indicate acclimation and physiological limits in brook trout Salvelinus fontinalis.

The upper thermal tolerance of brook trout Salvelinus fontinalis was estimated using critical thermal maxima (CTmax ) experiments on fish acclimated to temperatures that span the species' thermal range (5-25°C). The CTmax increased with acclimation temperature but plateaued in fish acclimated to 20, 23 and 25°C. Plasma lactate was highest, and the hepato-somatic index (IH ) was lowest at 23 and 25°C, which suggests additional metabolic costs at those acclimation temperatures. The results suggest that there is a sub-lethal threshold between 20 and 23°C, beyond which the fish experience reduced physiological performance.

Best practices for non-lethal blood sampling of fish via the caudal vasculature.

Blood sampling through the caudal vasculature is a widely used technique in fish biology for investigating organismal health and physiology. In live fishes, it can provide a quick, easy and relatively non-invasive method for obtaining a blood sample (cf. cannulation and cardiac puncture). Here, a general set of recommendations are provided for optimizing the blood sampling protocol that reflects best practices in animal welfare and sample integrity. This includes selecting appropriate use of anaesthetics for blood sampling as well as restraint techniques for situations where sedation is not used. In addition, ideal sampling environments where the fish can freely ventilate and strategies for minimizing handling time are discussed. This study summarizes the techniques used for extracting blood from the caudal vasculature in live fishes, highlighting the phlebotomy itself, the timing of sampling events and acceptable blood sample volumes. This study further discuss considerations for selecting appropriate physiological metrics when sampling in the caudal region and the potential benefits that this technique provides with respect to long-term biological assessments. Although general guidelines for blood sampling are provided here, it should be recognized that contextual considerations (e.g., taxonomic diversity, legal matters, environmental constraints) may influence the approach to blood sampling. Overall, it can be concluded that when done properly, blood sampling live fishes through the caudal vasculature is quick, efficient and minimally invasive, thus promoting conditions where live release of focal animals is possible.

Repeatability of burst swimming performance in medaka (Oryzias latipes).

Burst swimming performance (Uburst) is a putative indicator of "success" in wild fish. In this study, thirty-five lab-reared medaka (Oryzias latipes) were tested for Uburst using a French press exercise system. Fish were tested once a week for four consecutive weeks and repeatability was estimated in several ways to allow comparisons between studies. Following the initial swimming tests, 50% of fish were either thermally stressed, or not, for 180 s prior to testing Uburst once a week for four consecutive weeks. Burst swimming performance was found to be 24.0 ± 6.7 (s.d.) cm s-1 and repeatability prior to the thermal stress experiment was estimated to be 0.28 (intraclass correlation coefficient) with an upper and lower limit of 0.48 and 0.12, respectively. The measured Uburst and repeatability estimate in the thermal stressor experiment did not significantly differ from the first four trials. Swimming velocities observed match what is known about medaka swimming capabilities and, interestingly, are similar to maximum current velocities observed in their native habitat. Furthermore, our repeatability estimates confirm that burst swimming performance in medaka is a repeatable trait and validate the apparatus and swimming test used.

Swimming performance of a freshwater fish during exposure to high carbon dioxide.

Deterring the spread of invasive fishes is a challenge for managers, and bigheaded carp (including bighead and silver carp, Hypophthalmichthys spp.) are invasive fish that have spread throughout large portions of the Mississippi River basin and threaten to invade the Great Lakes' ecosystem. Studies have shown that elevated levels of carbon dioxide gas (CO2) have the ability to act as a nonphysical fish barrier, but little work has been done on the efficacy of CO2 to deter fish movement in flowing water. An annular swim flume was used to measure Uburst and sprint duration of the model species largemouth bass (Micropterus salmoides) across a range of pCO2 levels (< 400 µatm [ambient]; 10,000 µatm; 50,000 µatm; and 100,000 µatm). This species was tested as a proxy because of the likelihood of a similar CO2 response being produced, as well as constraints in obtaining and housing appropriately sized Asian carp. A significant decrease in Uburst swimming occurred when exposed to 100,000 µatm. No effects on sprint duration were detected. In both swimming tests, 15% of fish lost equilibrium when exposed to 50,000 µatm pCO2, while 50% of fish lost equilibrium when exposed to 100,000 µatm. Together, results define target levels for managers to impede the spread of largemouth bass and potentially other invasive freshwater fishes, helping guide policy to conserve aquatic ecosystems.

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.

Chronic exposure of a freshwater mussel to elevated pCO2 : Effects on the control of biomineralization and ion-regulatory responses.

Freshwater mussels may be exposed to elevations in mean partial pressure of carbon dioxide (pCO2 ) caused by both natural and anthropogenic factors. The goal of the present study was to assess the effects of a 28-d elevation in pCO2 at 15 000 and 50 000 µatm on processes associated with biomineralization, ion regulation, and cellular stress in adult Lampsilis siliquoidea (Barnes, 1823). In addition, the capacity for mussels to compensate for acid-base disturbances experienced after exposure to elevated pCO2 was assessed over a 14-d recovery period. Overall, exposure to 50 000 µatm pCO2 had more pronounced physiological consequences compared with 15 000 µatm pCO2 . Over the first 7 d of exposure to 50 000 µatm pCO2 , the mRNA abundance of chitin synthase (cs), calmodulin (cam), and calmodulin-like protein (calp) were significantly affected, suggesting that shell formation and integrity may be altered during pCO2 exposure. After the removal of the pCO2 treatment, mussels may compensate for the acid-base and ion disturbances experienced during pCO2 exposure, and transcript levels of some regulators of biomineralization (carbonic anhydrase [ca], cs, cam, calp) as well as ion regulation (na+ -k+ -ATPase [nka]) were modulated. Effects of elevated pCO2 on heat shock protein 70 (hsp70) were limited in the present study. Overall, adult L. siliquoidea appeared to regulate factors associated with the control of biomineralization and ion regulation during and/or after the removal of pCO2 exposure. Environ Toxicol Chem 2018;37:538-550. © 2017 SETAC.

Tolerance to Hypercarbia Is Repeatable and Related to a Component of the Metabolic Phenotype in a Freshwater Fish.

Freshwater fish may be exposed to high levels of carbon dioxide (CO2) because of several actions, including anesthesia and high levels of aquatic respiration and potentially as the result of using high-CO2 plumes as a barrier to the movements of invasive fishes. Metabolic phenotype can potentially drive how freshwater fish respond to high CO2. We therefore quantified how tolerance (measured using time to equilibrium loss [ELT]) was driven by metabolic phenotype in a cosmopolitan freshwater fish species, Micropterus salmoides. ELT was repeatable, with 60% of the variance across trials attributable to individual differences. For each fish, standard metabolic rate and maximum metabolic rate were measured using respirometers and time to exhaustion after a chase test was recorded. Fish with high anaerobic performance were less tolerant to elevated CO2, potentially as a result of preexisting metabolic acidosis. Standard metabolic rate and aerobic scope did not predict ELT. Our findings define which fish may be more vulnerable to high CO2, a potential mechanism for this tolerance, and show that tolerance to high CO2 may be acted on by natural selection. Should freshwater ecosystems become elevated in CO2, by either natural means or anthropogenic means, it is possible that there is potential for heritable selection of CO2 tolerance, evidenced by the fact that ELT was found to be repeatable.

Valve movement of three species of North American freshwater mussels exposed to elevated carbon dioxide.

Freshwater mussels are at-risk taxa and may be exposed to high levels of carbon dioxide (CO2) because of the potential use of CO2 to control the movement of invasive aquatic fish species. One potential behavioral response to a change in the partial pressure of CO2 (pCO2) may be altered valve movement. In this study, three species of mussels were fitted with modified sensors and exposed to two regimes of pCO2 to define thresholds of impaired valve movement. The first experiment demonstrated that Pyganodon grandis were much more tolerant to rising pCO2 relative to Lampsilis siliquoidea (acute closure at ∼200,000 µatm in comparison to ∼80,000 µatm). The second experiment consisted of monitoring mussels for 6 days and exposing them to elevated pCO2 (∼70,000 µatm) over a 2-day period. During exposure to high pCO2, Lampsilis cardium were open for nearly the entire high pCO2 period. Conversely, P. grandis were closed for most of the period following exposure to high pCO2. For L. siliquoidea, the number of closures decreased nearly 40-fold during high pCO2. The valve movement responses observed suggest species differences, and exposure to elevated pCO2 requires a reactive response.

Chill out: physiological responses to winter ice-angling in two temperate freshwater fishes.

A large body of research has documented the stress response of fish following angling capture. Nearly all of these studies have taken place during the open-water season, with almost no work focused on the effects of capture in the winter via ice angling. We therefore conducted a study to examine physiological disturbance and reflex impairment following capture by ice-angling in two commonly targeted species, bluegill Lepomis macrochirus and yellow perch Perca flavescens. Fish were captured from a lake in eastern Wisconsin (USA) and sampled either immediately or after being held in tanks for 0.5, 2 or 4 h. Sampling involved the assessment of reflex action mortality predictors (RAMP) and a blood biopsy that was used to measure concentrations of plasma cortisol and lactate. The capture-induced increase in plasma cortisol concentration was delayed relative to responses documented in previous experiments conducted in the summer and reached a relative high point at 4 h post-capture. Reflex impairment was highest at the first post-capture time point (0.5 h) and declined with each successive sampling (2 and 4 h) during recovery. Bluegill showed a higher magnitude stress response than yellow perch in terms of plasma cortisol and RAMP scores, but not when comparing plasma lactate. Overall, these data show that ice-angling induces a comparatively mild stress response relative to that found in previous studies of angled fish. While recovery of plasma stress indicators does not occur within 4 h, declining RAMP scores demonstrate that ice-angled bluegill and yellow perch do recover vitality following capture.

Responses to elevated CO2 exposure in a freshwater mussel, Fusconaia flava.

Freshwater mussels are some of the most imperiled species in North America and are particularly susceptible to environmental change. One environmental disturbance that mussels may encounter that remains understudied is an increase in the partial pressure of CO2 (pCO2). The present study quantified the impacts of acute (6 h) and chronic (up to 32 days) exposures to elevated pCO2 on genes associated with shell formation (chitin synthase; cs) and the stress response (heat shock protein 70; hsp70) in Fusconaia flava. Oxygen consumption (MO2) was also assessed over the chronic CO2 exposure period. Although mussels exhibited an increase in cs following an acute exposure to elevated pCO2, long-term exposure resulted in a decrease in cs mRNA abundance, suggesting that mussels may invest less in shell formation during chronic exposure to elevated pCO2. In response to an acute elevation in pCO2, mussels increased hsp70 mRNA abundance in mantle and adductor muscle and a similar increase was observed in the gill and adductor muscle in response to a chronic elevation in pCO2. A chronic elevation in pCO2 also increased mussel MO2. This overall increase in hsp70 mRNA levels and MO2 in F. flava indicates that exposure to elevated pCO2 initiates activation of the general stress response and an increased energy demand. Together, the results of the present study suggest that freshwater mussels respond to elevated pCO2 by increasing processes necessary to 'deal with' the stressor and, over the long-term, may reduce their investment in non-essential processes such as shell growth.

The response of two species of unionid mussels to extended exposure to elevated carbon dioxide.

Changes in environmental conditions can act as stressors, with potential consequences for the health and fitness of organisms. Rising levels of carbon dioxide (CO2) is one potential environmental stressor that is occurring more frequently in the environment and can be a stressor for aquatic organisms. In this study, the physiological responses of two species of unionid mussel, Lampsilis siliquoidea and Amblema plicata, were assessed in response to exposure to two levels of elevated partial pressure of CO2 (pCO2) (~20,000 and ~55,000µatm) over a 28d period, followed by a subsequent 14d recovery period. Observations were consistent with responses associated with respiratory acidosis, as demonstrated by changes in hemolymph HCO3(-), Ca(2+), Cl(-), and Na(2+). Both species exposed to elevated pCO2 had elevated hemolymph HCO3(-) during the pCO2 treatment period compared to control mussels, but recovered once pCO2 was removed. Similarly, both species had elevated hemolymph Na(+) during exposure to elevated pCO2, and this returned to control levels for A. plicata but remained elevated for L. siliquoidea once the pCO2 stimuli was removed. Changes in hemolymph Ca(2+) and Cl(-) in response to elevated pCO2 were also observed, but these changes were species-specific. Additional physiological responses to elevated pCO2 (e.g., changes in hemolymph glucose and Mg(2+)) were consistent with a stress response in both species. This study demonstrates the importance of considering inter-specific differences in the response of organisms to stress, and also that responses to elevated pCO2 may be transient and can recover once the stress is removed.

Freshwater biota and rising pCO2?

Rising atmospheric carbon dioxide (CO2) has caused a suite of environmental issues, however, little is known about how the partial pressure of CO2 (pCO2) in freshwater will be affected by climate change. Freshwater pCO2 varies across systems and is controlled by a diverse array of factors, making it difficult to make predictions about future levels of pCO2. Recent evidence suggests that increasing levels of atmospheric CO2 may directly increase freshwater pCO2 levels in lakes, but rising atmospheric CO2 may also indirectly impact freshwater pCO2 levels in a variety of systems by affecting other contributing factors such as soil respiration, terrestrial productivity and climate regimes. Although future freshwater pCO2 levels remain uncertain, studies have considered the potential impacts of changes to pCO2 levels on freshwater biota. Studies to date have focused on impacts of elevated pCO2 on plankton and macrophytes, and have shown that phytoplankton nutritional quality is reduced, plankton community structure is altered, photosynthesis rates increase and macrophyte distribution shifts with increasing pCO2. However, a number of key knowledge gaps remain and gaining a better understanding of how freshwater pCO2 levels are regulated and how these levels may impact biota, will be important for predicting future responses to climate change.