Katja Anttila

Responses to temperature and hypoxia as interacting stressors in fish: implications for adaptation to environmental change.

Anthropogenic environmental change is exposing animals to changes in a complex array of interacting stressors and is already having important effects on the distribution and abundance of species. However, despite extensive examination of the effects of stressors in isolation, knowledge of the effects of stressors in combination is limited. This lack of information makes predicting the responses of organisms to anthropogenic environmental change challenging. Here, we focus on the effects of temperature and hypoxia as interacting stressors in fishes. A review of the available evidence suggests that temperature and hypoxia act synergistically such that small shifts in one stressor could result in large effects on organismal performance when a fish is exposed to the 2 stressors in combination. Although these stressors pose substantial challenges for fish, there also is substantial intraspecific variation in tolerance to these stressors that could act as the raw material for the evolution of improved tolerance. However, the potential for adaptive change is, in part, dependent on the nature of the correlations among traits associated with tolerance. For example, negative genetic correlations (or trade-offs) between tolerances to temperature and hypoxia could limit the potential for adaptation to the combined stressors, while positive genetic correlations might be of benefit. The limited data currently available suggest that tolerances to hypoxia and to high-temperature may be positively correlated in some species of fish, suggesting the possibility for adaptive evolution in these traits in response to anthropogenic environmental change.

Variation in temperature tolerance among families of Atlantic salmon (Salmo salar) is associated with hypoxia tolerance, ventricle size and myoglobin level

SUMMARY In fishes, performance failure at high temperature is thought to be due to a limitation on oxygen delivery (the theory of oxygen and capacity limited thermal tolerance, OCLTT), which suggests that thermal tolerance and hypoxia tolerance might be functionally associated. Here we examined variation in temperature and hypoxia tolerance among 41 families of Atlantic salmon (Salmo salar), which allowed us to evaluate the association between these two traits. Both temperature and hypoxia tolerance varied significantly among families and there was a significant positive correlation between critical maximum temperature (CTmax) and hypoxia tolerance, supporting the OCLTT concept. At the organ and cellular levels, we also discovered support for the OCLTT concept as relative ventricle mass (RVM) and cardiac myoglobin (Mb) levels both correlated positively with CTmax (R2=0.21, P<0.001 and R2=0.17, P=0.003, respectively). A large RVM has previously been shown to be associated with high cardiac output, which might facilitate tissue oxygen supply during elevated oxygen demand at high temperatures, while Mb facilitates the oxygen transfer from the blood to tissues, especially during hypoxia. The data presented here demonstrate for the first time that RVM and Mb are correlated with increased upper temperature tolerance in fish. High phenotypic variation between families and greater similarity among full- and half-siblings suggests that there is substantial standing genetic variation in thermal and hypoxia tolerance, which could respond to selection either in aquaculture or in response to anthropogenic stressors such as global climate change.

Using maximum heart rate as a rapid screening tool to determine optimum temperature for aerobic scope in Pacific salmon Oncorhynchus spp.

The mean ±s.e. optimum temperature (T(opt)) for aerobic scope in juvenile coho salmon Oncorhynchus kisutch was determined to be 17·0 ± 0·7° C. The repeated measures protocol took 3 weeks to complete the T(opt) determination using 12 fish tested at five temperatures separated by 2° C increments. This experiment also demonstrated that the T(opt) was associated with maximum heart rate (f(H)) failing to maintain a Q(10) -related increase with temperature. When maximum f(H) was produced in anaesthetized fish with pharmacological stimulation and f(H) measured from electrocardiogram recordings during acute warming, the Arrhenius break temperature (ABT) for Q(10) discontinuities in maximum f(H) (mean ±s.e. = 17·1 ± 0·5° C for 15 ppm clove oil and 16·5 ± 0·2° C for 50 ppm MS-222) was statistically indistinguishable from the T(opt) measured using aerobic scope. Such a determination took only 3 days rather than 3 weeks. Therefore, it is proposed that determining ABT for discontinuities in maximum f(H) in anaesthetized fish presents itself as a valuable, high-throughput screening tool to assess T(opt) in fishes, a metric that has become recognized as being extremely valuable in fish biology and fisheries management.

Atlantic salmon show capability for cardiac acclimation to warm temperatures

Increases in environmental temperature predicted to result from global warming have direct effects on performance of ectotherms. Moreover, cardiac function has been observed to limit the tolerance to high temperatures. Here we show that two wild populations of Atlantic salmon originating from northern and southern extremes of its European distribution have strikingly similar cardiac responses to acute warming when acclimated to common temperatures, despite different local environments. Although cardiac collapse starts at 21-23 °C with a maximum heart rate of ~150 beats per min (bpm) for 12 °C-acclimated fish, acclimation to 20 °C considerably raises this temperature (27.5 °C) and maximum heart rate (~200 bpm). Only minor population differences exist and these are consistent with the warmer habitat of the southern population. We demonstrate that the considerable cardiac plasticity discovered for Atlantic salmon is largely independent of natural habitat, and we propose that observed cardiac plasticity may aid salmon to cope with global warming.

Optimum Temperature in Juvenile Salmonids: Connecting Subcellular Indicators to Tissue Function and Whole-Organism Thermal Optimum

Temperature affects processes at all levels of biological organization, but it is unclear whether processes at different levels have similar thermal optima (Topt). Here, we compare the Topt for aerobic scope, a whole-organism measure of performance, with both the Arrhenius breakpoint temperature for maximum heart rate (HR-ABT), a measure of tissue level performance, and the temperature at which AMP-activated protein kinase (AMPK) is phosphorylated in the heart, an indicator of an increase in dependence on anaerobic energy metabolism at the cellular level in juvenile rainbow trout Oncorhynchus mykiss. The Topt for aerobic scope was 19°C, with aerobic scope being maintained at ≥90% of maximum (termed a “Topt window”) from 16.5° to 20.5°C. HR-ABT occurred at , while the profile of AMPK phosphorylation started to change from baseline at 19°C, suggesting that these processes have similar thermal sensitivities as a fish is warmed to Topt. The effects of temperature on AMPK phosphorylation were also measured in coho salmon Oncorhynchus kisutch hearts and compared with previously published values for HR-ABT and aerobic scope Topt. AMPK phosphorylation in coho hearts began to change at temperatures above 17°C, which again is comparable with the published Topt for aerobic scope (17°C) and HR-ABT () in these individuals. Thus, the thermal sensitivity of these subcellular, tissue, and whole-organism functions are highly correlated in both rainbow trout and coho salmon and may depend on each other.

A heart to heart on temperature: impaired temperature tolerance of triploid rainbow trout (Oncorhynchus mykiss) due to early onset of cardiac arrhythmia.

Triploid (3N) salmonids are of interest to aquaculture and sport fishing industries, however it has been shown that 3N fish have impaired tolerance to high temperatures. To test the hypothesis that poor high temperature tolerance in 3N salmonids is related to impaired O2 delivery to the body, maximum heart rate (fH) was measured in 2N (diploid) and 3N rainbow trout (Oncorhynchus mykiss) during an incremental temperature challenge. fH of both ploidies was similar at 10°C. However, a significant effect of ploidy on the response of fH to temperature from 10 to 22°C was reflected in a lower Q10 for 3N individuals. Additionally, all 3N trout developed a cardiac arrhythmia by 22°C, where 30% of 2N trout continued to maintain a rhythmic heartbeat. These findings suggest that reduced 3N high temperature tolerance could be due to early collapse of the cardiovascular systems ability to deliver O2 to the body during warming.

Thermal optima and tolerance in the eurythermic goldfish (Carassius auratus): relationships between whole-animal aerobic capacity and maximum heart rate.

The wide thermal tolerance range of a eurythermic fish (goldfish, Carassius auratus) was used to evaluate how temperature performance curves derived from maximum heart rate (fH) related to those for aerobic scope. For acclimation temperatures of 12°, 20°, and 28°C, optimum temperatures derived from aerobic scope curves (Topt) were 19.9° ± 0.4°, 19.3° ± 0.8°, and 28.7° ± 0.8°C, respectively. The Arrhenius breakpoint temperatures (TAB) for maximum fH were 21.5° ± 0.6°, 23.8° ± 0.9°, and 24.6° ± 0.5°C, respectively. The TQB (temperature where the incremental Q10 of maximum fH decreased abruptly below 1.9) was 24.0° ± 0.7° and 29.8° ± 0.6°C for the 12° and 28°C acclimation temperatures, respectively, and was within the Topt window (11.5°–30.3° and 26.9°–30.5°C, respectively), but TQB for the 20°C acclimation temperature (27.3° ± 0.6°C) was higher than the Topt window (15.4°–23.2°C). Warm acclimation increased the upper critical temperature (Tcrit; from 37.2° ± 0.7° to 44.7° ± 11.8°C) as well as the temperature that triggered a cardiac arrhythmia (Tarr; from 31.1° ± 0.7° to 39.3° ± 0.4°C). In conclusion, we propose that maximum fH and its associated rate transition temperatures (TAB, TQB, and Tarr) can be used to estimate the upper thermal tolerance of eurythermic as well as stenothermic fish independent of acclimation temperature. All the same, great care is needed with such evaluations. For the goldfish, while TAB and TQB were always within the Topt window for 90% of maximum aerobic scope and Topt was closely associated with TAB for 12°C-acclimated fish, TQB had the closest association after 28°C acclimation, and both TAB and TQB were above the Topt window after 20°C acclimation.

Warm acclimation and oxygen depletion induce species-specific responses in salmonids

ABSTRACT Anthropogenic activities are greatly altering the habitats of animals, whereby fish are already encountering several stressors simultaneously. The purpose of the current study was to investigate the capacity of fish to respond to two different environmental stressors (high temperature and overnight hypoxia) separately and together. We found that acclimation to increased temperature (from 7.7±0.02°C to 14.9±0.05°C) and overnight hypoxia (daily changes from normoxia to 63–67% oxygen saturation), simulating climate change and eutrophication, had both antagonistic and synergistic effects on the capacity of fish to tolerate these stressors. The thermal tolerance of Arctic char (Salvelinus alpinus) and landlocked salmon (Salmo salar m. sebago) increased with warm acclimation by 1.3 and 2.2°C, respectively, but decreased when warm temperature was combined with overnight hypoxia (by 0.2 and 0.4°C, respectively). In contrast, the combination of the stressors more than doubled hypoxia tolerance in salmon and also increased hypoxia tolerance in char by 22%. Salmon had 1.2°C higher thermal tolerance than char, but char tolerated much lower oxygen levels than salmon at a given temperature. The changes in hypoxia tolerance were connected to the responses of the oxygen supply and delivery system. The relative ventricle mass was higher in cold- than in warm-acclimated salmon but the thickness of the compact layer of the ventricle increased with the combination of warm and hypoxia acclimation in both species. Char had also significantly larger hearts and thicker compact layers than salmon. The results illustrate that while fish can have protective responses when encountering a single environmental stressor, the combination of stressors can have unexpected species-specific effects that will influence their survival capacity. Summary: Overnight hypoxia combined with warm acclimation increases the hypoxia tolerance of salmonids while upper critical temperature is reduced in a species-specific manner.

Indirect genetic effects underlie oxygen-limited thermal tolerance within a coastal population of chinook salmon

With global temperatures projected to surpass the limits of thermal tolerance for many species, evaluating the heritable variation underlying thermal tolerance is critical for understanding the potential for adaptation to climate change. We examined the evolutionary potential of thermal tolerance within a population of chinook salmon (Oncorhynchus tshawytscha) by conducting a full-factorial breeding design and measuring the thermal performance of cardiac function and the critical thermal maximum (CTmax) of offspring from each family. Additive genetic variation in offspring phenotype was mostly negligible, although these direct genetic effects explained 53% of the variation in resting heart rate (fH). Conversely, maternal effects had a significant influence on resting fH, scope for fH, cardiac arrhythmia temperature and CTmax. These maternal effects were associated with egg size, as indicated by strong relationships between the mean egg diameter of mothers and offspring thermal tolerance. Because egg size can be highly heritable in chinook salmon, our finding indicates that the maternal effects of egg size constitute an indirect genetic effect contributing to thermal tolerance. Such indirect genetic effects could accelerate evolutionary responses to the selection imposed by rising temperatures and could contribute to the population-specific thermal tolerance that has recently been uncovered among Pacific salmon populations.

Upper thermal tolerance of closely related Danio species.

The main finding of this study was that measuring maximum heart rate during incremental warming was an effective tool to estimate upper thermal limits in three small cyprinid Danio species, which differed significantly. Arrhenius breakpoint temperature for maximum heart rate, purportedly an index of optimum temperature, was 21·2 ± 0·4, 20·1 ± 0·4 and 18·9 ± 0·8° C (mean ± s.e.) for zebrafish Danio rerio, pearl danio Danio albolineatus and glowlight danio Danio choprae, respectively. The temperature where cardiac arrhythmias were first induced during warming (T(arr)) was 36·6 ± 0·7, 36·9 ± 0·8 and 33·2 ± 0·8° C (mean ± s.e.) and critical thermal maximum (T(Cm)) was 39·9 ± 0·1, 38·9 ± 0·1 and 37·2 ± 0·1° C (mean ± s.e.) for D. rerio, D. albolineatus and D. choprae, respectively. The finding that T(arr) was consistently 3-4° C lower than T(Cm) suggests that collapse of the cardiac life support system may be a critical trigger for upper temperature tolerance. The upper thermal limits established here, which correlate well with a broad natural environmental temperature range for D. rerio and a narrow one for D. choprae, suggest that upper thermal tolerance may be a genetic trait even among closely related species acclimated to common temperatures.