Impact of the antidepressant Bupropion on the Dynamic Energy Budget of Daphnia magna.

Psychiatric drugs are considered among the emerging contaminants of concern in ecological risk assessment, due to their potential to disrupt homeostasis in aquatic organisms. Bupropion is an antidepressant that acts by selective reuptake inhibition of norepinephrine and dopamine. Little is known about this compound's effects on aquatic organisms, despite being detected in significant concentrations in both water and biota close to waste-water treatment plants and densely populated areas. Dynamic Energy Budget (DEB) models are flexible mechanistic tools that can be applied to understand toxic effects and extrapolate individual responses to higher biological levels and under untested environmental conditions. In this work, we used the stdDEB-TKTD (an application of the DEB theory to ecotoxicology) approach to investigate the possible physiological mode of action of Bupropion on the model organism Daphnia magna. Next, Dynamic Energy Budget Individual-Based Models (DEB-IBM) were used to extrapolate the results to the population level and to predict the combined effects of Bupropion exposure and food availability on the daphnids. Our results revealed an increasing negative effect of this antidepressant on the reproduction and survival of the animals with increasing concentration (0.004, 0.058, 0.58 and 58 µM). At the population level, we found that even the lowest used doses of Bupropion could reduce the population density and its reproductive output. The impacts are predicted to be stronger under limited food conditions.

Divergence in rates of phenotypic plasticity among ectotherms.

An individual's fitness cost associated with environmental change likely depends on the rate of adaptive phenotypic plasticity, and yet our understanding of plasticity rates in an ecological and evolutionary context remains limited. We provide the first quantitative synthesis of existing plasticity rate data, focusing on acclimation of temperature tolerance in ectothermic animals, where we demonstrate applicability of a recently proposed analytical approach. The analyses reveal considerable variation in plasticity rates of this trait among species, with half-times (how long it takes for the initial deviation from the acclimated phenotype to be reduced by 50% when individuals are shifted to a new environment) ranging from 3.7 to 770.2 h. Furthermore, rates differ among higher taxa, being higher for amphibians and reptiles than for crustaceans and fishes, and with insects being intermediate. We argue that a more comprehensive understanding of phenotypic plasticity will be attained through increased focus on the rate parameter.

Inferring temperature adaptation from thermal performance curves of somatic growth rate: The importance of growth measurements and mortality.

When comparing somatic growth thermal performance curves (TPCs), higher somatic growth across experimental temperatures is often observed for populations originating from colder environments. Such countergradient variation has been suggested to represent adaptation to seasonality, or shorter favourable seasons in colder climates. Alternatively, populations from cold climates may outgrow those from warmer climates at low temperature, and vice versa at high temperature, representing adaptation to temperature. Using modelling, we show that distinguishing between these two types of adaptation based on TPCs requires knowledge about (i) the relationship between somatic growth rate and population growth rate, which in turn depends on the scale of somatic growth (absolute or proportional), and (ii) the relationship between somatic growth rate and mortality rate in the wild. We illustrate this by quantifying somatic growth rate TPCs for three populations of Daphnia magna where population growth scales linearly with proportional somatic growth. For absolute somatic growth, the northern population outperformed the two more southern populations across temperatures, and more so at higher temperatures, consistent with adaptation to seasonality. In contrast, for the proportional somatic growth TPCs, and hence population growth rate, TPCs tended to converge towards the highest temperatures. Thus, if the northern population pays an ecological mortality cost of rapid growth in the wild, this may create crossing population growth TPCs consistent with adaptation to temperature. Future studies within this field should be more explicit in how they extrapolate from somatic growth in the lab to fitness in the wild.

Insights into the quantification and reporting of model-related uncertainty across different disciplines.

Quantifying uncertainty associated with our models is the only way we can express how much we know about any phenomenon. Incomplete consideration of model-based uncertainties can lead to overstated conclusions with real-world impacts in diverse spheres, including conservation, epidemiology, climate science, and policy. Despite these potentially damaging consequences, we still know little about how different fields quantify and report uncertainty. We introduce the "sources of uncertainty" framework, using it to conduct a systematic audit of model-related uncertainty quantification from seven scientific fields, spanning the biological, physical, and political sciences. Our interdisciplinary audit shows no field fully considers all possible sources of uncertainty, but each has its own best practices alongside shared outstanding challenges. We make ten easy-to-implement recommendations to improve the consistency, completeness, and clarity of reporting on model-related uncertainty. These recommendations serve as a guide to best practices across scientific fields and expand our toolbox for high-quality research.

Evolution of population dynamics following invasion by a non-native predator.

Invasive predatory species are frequently observed to cause evolutionary responses in prey phenotypes, which in turn may lead to evolutionary shifts in the population dynamics of prey. Research has provided a link between rates of predation and the evolution of prey population growth in the lab, but studies from natural populations are rare. Here, we tested for evolutionary changes in population dynamics parameters of zooplankton Daphnia pulicaria following invasion by the predator Bythotrephes longimanus into Lake Kegonsa, Wisconsin, US. We used a resurrection ecological approach, whereby clones from pre- and post-invasive periods were hatched from eggs obtained in sediment cores and were used in a 3-month growth experiment. Based on these data, we estimated intrinsic population growth rates (r), the shape of density dependence (θ) and carrying capacities (K) using theta-logistic models. We found that post-invasion Daphnia maintained a higher r and K under these controlled, predation-free laboratory conditions. Evidence for changes in θ was weaker. Whereas previous experimental evolution studies of predator-prey interactions have demonstrated that genotypes that have evolved under predation have inferior competitive ability when the predator is absent, this was not the case for the Daphnia. Given that our study was conducted in a laboratory environment and the possibility for genotype-by-environment interactions, extrapolating these apparent counterintuitive results to the wild should be done with caution. However, barring such complications, we discuss how selection for reduced predator exposure, either temporally or spatially, may have led to the observed changes. This scenario suggests that complexities in ecological interactions represents a challenge when predicting the evolutionary responses of population dynamics to changes in predation pressure in natural systems.

Stochastic variation in gut bacterial community affects reproductive rates in the water flea Daphnia magna.

It is well-documented that perturbation of the gut bacterial community can influence the reproductive rates of the host. Less is known about how natural ecological processes can change the bacterial composition in the gut and how such changes influence the reproductive rate of the host. Here, we provide novel experimental insights into such processes using the clonally reproducing water flea, Daphnia magna. A total of 20 replicate cultures were reared for 5 weeks (Phase 1) to allow for divergence of bacterial communities through stochastic processes (i.e. drift, founder effects, and/or colonization). Duplicate cultures created from each of these were reared for 21 days (Phase 2) while recording reproductive rates. There was a significant repeatability in reproductive rates between these duplicates, suggesting that divergence of the bacterial communities during Phase 1 translated into reproductive rate effects during Phase 2. This was further supported by significant differences in the relative abundance of gut bacteria (investigated by amplicon sequencing of a part of the 16S rRNA gene) between cultures with high and low reproductive rate in Phase 2. These results are consistent with the hypothesis that stochastic processes can cause natural variation in the bacterial composition in the gut, which in turn affect host reproductive rates.

Evolutionary change in metabolic rate of Daphnia pulicaria following invasion by the predator Bythotrephes longimanus.

Metabolic rate is a trait that may evolve in response to the direct and indirect effects of predator-induced mortality. Predators may indirectly alter selection by lowering prey densities and increasing resource availability or by intensifying resource limitation through changes in prey behavior (e.g., use of less productive areas). In the current study, we quantify the evolution of metabolic rate in the zooplankton Daphnia pulicaria following an invasive event by the predator Bythotrephes longimanus in Lake Mendota, Wisconsin, US. This invasion has been shown to dramatically impact D. pulicaria, causing a ~60% decline in their biomass. Using a resurrection ecology approach, we compared the metabolic rate of D. pulicaria clones originating prior to the Bythotrephes invasion with that of clones having evolved in the presence of Bythotrephes. We observed a 7.4% reduction in metabolic rate among post-invasive clones compared to pre-invasive clones and discuss the potential roles of direct and indirect selection in driving this change.

Environmental change and the rate of phenotypic plasticity.

With rapid and less predictable environmental change emerging as the 'new norm', understanding how individuals tolerate environmental stress via plastic, often reversible changes to the phenotype (i.e., reversible phenotypic plasticity, RPP), remains a key issue in ecology. Here, we examine the potential for better understanding how organisms overcome environmental challenges within their own lifetimes by scrutinizing a somewhat overlooked aspect of RPP, namely the rate at which it can occur. Although recent advances in the field provide indication of the aspects of environmental change where RPP rates may be of particular ecological relevance, we observe that current theoretical models do not consider the evolutionary potential of the rate of RPP. Whilst recent theory underscores the importance of environmental predictability in determining the slope of the evolved reaction norm for a given trait (i.e., how much plasticity can occur), a hitherto neglected possibility is that the rate of plasticity might be a more dynamic component of this relationship than previously assumed. If the rate of plasticity itself can evolve, as empirical evidence foreshadows, rates of plasticity may have the potential to alter the level predictability in the environment as perceived by the organism and thus influence the slope of the evolved reaction norm. However, optimality in the rate of phenotypic plasticity, its evolutionary dynamics in different environments and influence of constraints imposed by associated costs remain unexplored and may represent fruitful avenues of exploration in future theoretical and empirical treatments of the topic. We conclude by reviewing published studies of RPP rates, providing suggestions for improving the measurement of RPP rates, both in terms of experimental design and in the statistical quantification of this component of plasticity.

Maternal dopamine exposure provides offspring starvation resistance in Daphnia.

The neurotransmitter dopamine has been shown to play an important role in modulating behavioral, morphological, and life history responses to food abundance. However, costs of expressing high dopamine levels remain poorly studied and are essential for understanding the evolution of the dopamine system. Negative maternal effects on offspring size from enhanced maternal dopamine levels have previously been documented in Daphnia. Here, we tested whether this translates into fitness costs in terms of lower starvation resistance in offspring. We exposed Daphnia magna mothers to aqueous dopamine (2.3 or 0 mg/L for the control) at two food levels (ad libitum vs. 30% ad libitum) and recorded a range of maternal life history traits. The longevity of their offspring was then quantified in the absence of food. In both control and dopamine treatments, mothers that experienced restricted food ration had lower somatic growth rates and higher age at maturation. Maternal food restriction also resulted in production of larger offspring that had a superior starvation resistance compared to ad libitum groups. However, although dopamine exposed mothers produced smaller offspring than controls at restricted food ration, these smaller offspring survived longer under starvation. Hence, maternal dopamine exposure provided an improved offspring starvation resistance. We discuss the relative importance of proximate and ultimate causes for why D. magna may not evolve toward higher endogenous dopamine levels despite the fitness benefits this appears to have.

Quantitative mismatch between empirical temperature-size rule slopes and predictions based on oxygen limitation.

In ectotherms, adult body size commonly declines with increasing environmental temperature, a pattern known as the temperature-size rule. One influential hypothesis explaining this observation is that the challenge of obtaining sufficient oxygen to support metabolism becomes greater with increasing body size, and more so at high temperatures. Yet, previous models based on this hypothesis do not account for phenotypic plasticity in the physiology of organisms that counteracts oxygen limitation at high temperature. Here, we model the predicted strength of the temperature-size response using estimates of how both the oxygen supply and demand is affected by temperature when allowing for phenotypic plasticity in the aquatic ectotherm Daphnia magna. Our predictions remain highly inconsistent with empirical temperature-size responses, with the prior being close to one order of magnitude stronger than the latter. These results fail to provide quantitative support for the hypothesis that oxygen limitation drives temperature-size clines in aquatic ectotherms. Future studies into the role of oxygen limitation should address how the strength of the temperature-size response may be shaped by evolution under fluctuating temperature regimes. Finally, our results caution against applying deterministic models based on the oxygen limitation hypothesis when predicting future changes in ectotherm size distributions under climate change.

Population dynamics and resting egg production in Daphnia: Interactive effects of mercury, population density and temperature.

Toxicity studies on freshwater organisms are commonly conducted by quantifying effects on asexual (clonal) reproductive rates in Daphnia, whereas studies of effects on sexual reproductive rates remain relatively rare. Sexual reproduction in Daphnia and the associated production of resting eggs allows them to survive unfavorable environmental conditions and is thus a crucial component of their long-term fitness. It also maintains genetic diversity within Daphnia populations and hence their potential for adaptation to new environmental conditions. This aspect of their biology may therefore be important to consider in toxicity studies. The aim of this study was to investigate for the first time how mercury (Hg) affects sexual versus asexual reproduction in Daphnia under varying environmental conditions. Specifically, we experimentally tested the interactive effects of Hg and temperature on the population dynamics of Daphnia magna. For this purpose, we exposed D. magna to environmentally relevant concentrations (0 µg/L, 0.5 µg/L and 2 µg/L) of Hg (in the form of mercury (II) chloride) found in stream water and measured biomass growth rate resulting from asexual reproduction, and resting egg production resulting from sexual reproduction. This was done at both 17 °C and 24 °C. Biomass growth rate did not vary across Hg treatments and depended mainly on temperature and population density. Density dependence of biomass growth rate was indeed more pronounced at 24 °C than at 17 °C, as resource limitation from intraspecific competition was further exacerbated by the rise in feeding rates with temperature. Density dependence of resting egg production was unaffected by Hg and temperature, but resting egg production was higher under Hg exposure at low temperature. These findings show that depending on environmental conditions, rates of sexual reproduction in D. magna may respond to metal exposure at lower concentrations than those impacting population growth during the asexual phase.

Dopamine mediates life-history responses to food abundance in Daphnia.

Expression of adaptive reaction norms of life-history traits to spatio-temporal variation in food availability is crucial for individual fitness. Yet little is known about the neural signalling mechanisms underlying these reaction norms. Previous studies suggest a role for the dopamine system in regulating behavioural and morphological responses to food across a wide range of taxa. We tested whether this neural signalling system also regulates life-history reaction norms by exposing the zooplankton Daphnia magna to both dopamine and the dopamine reuptake inhibitor bupropion, an antidepressant that enters aquatic environments via various pathways. We recorded a range of life-history traits across two food levels. Both treatments induced changes to the life-history reaction norm slopes. These were due to the effects of the treatments being more pronounced at restricted food ration, where controls had lower somatic growth rates, higher age and larger size at maturation. This translated into a higher population growth rate (r) of dopamine and bupropion treatments when food was restricted. Our findings show that the dopamine system is an important regulatory mechanism underlying life-history trait responses to food abundance and that bupropion can strongly influence the life history of aquatic species such as D. magna. We discuss why D. magna do not evolve towards higher endogenous dopamine levels despite the apparent fitness benefits.

On the use of the coefficient of variation to quantify and compare trait variation.

Meaningful comparison of variation in quantitative trait requires controlling for both the dimension of the varying entity and the dimension of the factor generating variation. Although the coefficient of variation (CV; standard deviation divided by the mean) is often used to measure and compare variation of quantitative traits, it only accounts for the dimension of the former, and its use for comparing variation may sometimes be inappropriate. Here, we discuss the use of the CV to compare measures of evolvability and phenotypic plasticity, two variational properties of quantitative traits. Using a dimensional analysis, we show that contrary to evolvability, phenotypic plasticity cannot be meaningfully compared across traits and environments by mean-scaling trait variation. We further emphasize the need of remaining cognizant of the dimensions of the traits and the relationship between mean and standard deviation when comparing CVs, even when the scales on which traits are expressed allow meaningful calculation of the CV.

The old and the large may suffer disproportionately during episodes of high temperature: evidence from a keystone zooplankton species.

Widespread declines in the body size of aquatic ectotherms have been attributed to the poorer ability of older, larger individuals to tolerate high temperature. Here, using the thermal death time curve framework, we investigate the relationship between temperature tolerance and size/age by measuring the change in heat tolerance of the keystone zooplankton species Daphnia magna across a range of temperature intensities (and hence exposures of varying duration) among individuals that differed up to 3-fold in size and thus varied in age also. Across the gradient of exposure temperatures, younger, smaller individuals were more tolerant than older, larger individuals. This suggests that the young and the small may be better equipped to withstand temperature challenges that are both intense/brief and more moderate/prolonged. Our study generalizes results obtained from more acute tolerance assays, providing physiological evidence consistent with the observed reductions in ectotherm body size as a response to warming in aquatic systems.

Biofilms grown in aquatic microcosms affect mercury and selenium accumulation in Daphnia.

Experiments examining mercury (Hg) toxicity in Daphnia are usually conducted in highly standardized conditions that prevent the formation of biofilm. Although such standardization has many advantages, extrapolation of results to natural conditions and inference of ecological effects is challenging. This is especially true since biofilms can accumulate metals/metalloids and play a key role in their transfer to higher trophic level organisms. In this study, we experimentally tested the effects of spontaneously appearing biofilm in Daphnia cultures on accumulation of Hg and its natural antagonist selenium (Se) in Daphnia magna. We added Hg (in the form of mercury (II) chloride) at two concentrations (0.2 µg/L and 2 µg/L) to experimental microcosms and measured the uptake of Hg and Se by D. magna in the presence and absence of biofilm. To test for consistent and replicable results, we ran two identical experimental sets one week apart. Biofilm presence significantly reduced the accumulation of Hg, while increasing the tissue Se content in D. magna, and these findings were reproducible across experimental sets. These findings indicate that highly standardized tests may not be adequate to predict the bioaccumulation and potential toxicity of metals/metalloids under natural conditions.

Acclimation capacity and rate change through life in the zooplankton Daphnia.

When a change in the environment occurs, organisms can maintain an optimal phenotypic state via plastic, reversible changes to their phenotypes. These adjustments, when occurring within a generation, are described as the process of acclimation. While acclimation has been studied for more than half a century, global environmental change has stimulated renewed interest in quantifying variation in the rate and capacity with which this process occurs, particularly among ectothermic organisms. Yet, despite the likely ecological importance of acclimation capacity and rate, how these traits change throughout life among members of the same species is largely unstudied. Here we investigate these relationships by measuring the acute heat tolerance of the clonally reproducing zooplankter Daphnia magna of different size/age and acclimation status. The heat tolerance of individuals completely acclimated to relatively warm (28°C) or cool (17°C) temperatures diverged during development, indicating that older, larger individuals had a greater capacity to increase heat tolerance. However, when cool acclimated individuals were briefly exposed to the warm temperature (i.e. were 'heat-hardened'), it was younger, smaller animals with less capacity to acclimate that were able to do so more rapidly because they obtained or came closer to obtaining complete acclimation of heat tolerance. Our results illustrate that within a species, individuals can differ substantially in how rapidly and by how much they can respond to environmental change. We urge greater investigation of the intraspecific relationship between acclimation and development along with further consideration of the factors that might contribute to these enigmatic patterns of phenotypic variation.

Genetic and environmental effects on the scaling of metabolic rate with body size.

Metabolic rate (MR) often scales with body mass (BM) following a power function of the form MR=aBM b , where log(a) is the allometric intercept and b is the allometric exponent (i.e. slope on a log-log scale). The variational properties of b have been debated, but very few studies have tested for genetic variance in b, and none have tested for a genotype-by-environment (G×E) interaction in b Consequently, the short-term evolutionary potentials of both b and its phenotypic plasticity remain unknown. Using 10 clones of a population of Daphnia magna, we estimated the genetic variance in b and assessed whether a G×E interaction affected b We measured MR on juveniles of different sizes reared and measured at three temperatures (17, 22 and 28°C). Overall, b decreased with increasing temperature. We found no evidence of genetic variance in b at any temperature, and thus no G×E interaction in b However, we found a significant G×E interaction in size-specific MR. Using simulations, we show how this G×E interaction can generate genetic variation in the ontogenetic allometric slope of animals experiencing directional changes in temperature during growth. This suggests that b can evolve despite having limited genetic variation at constant temperatures.

How to quantify thermal acclimation capacity?

The capacity of organisms to acclimate will influence their ability to cope with ongoing global changes in thermal regimes. Here we highlight methodological issues associated with recent attempts to quantify variation in acclimation capacity among taxa and environments, and describe how these may introduce bias to conclusions. We then propose a measure of thermal acclimation capacity that more directly quantifies the process of acclimation. Future studies of variation in acclimation capacity should critically evaluate whether their chosen empirical metric accurately reflects the theoretical concept of acclimation.

Can variation in standard metabolic rate explain context-dependent performance of farmed Atlantic salmon offspring?

Escaped farmed Atlantic salmon interbreed with wild Atlantic salmon, leaving offspring that often have lower success in nature than pure wild salmon. On top of this, presence of farmed salmon descendants can impair production of wild-type recruits. We hypothesize that both these effects connect with farmed salmon having acquired higher standard metabolic rates (SMR, the energetic cost of self-maintenance) during domestication. Fitness-related advantages of phenotypic traits associated with both high SMR and farmed salmon (e.g., social dominance) depend on environmental conditions, such as food availability. We hypothesize that farmed offspring have an advantage at high food availability due to, for example, dominance behavior but suffer increased risks of starvation when food is scarce because this behavior is energy-demanding. To test these hypotheses, we first compare embryo SMR of pure farmed, farmed-wild hybrids and pure wild offspring. Next, we test early-life performance (in terms of survival and growth) of hybrids relative to that of their wild half-siblings, as well as their competitive abilities, in semi-natural conditions of high and low food availability. Finally, we test how SMR affects early-life performance at high and low food availability. We find inconclusive support for the hypothesis that domestication has induced increased SMR. Further, wild and hybrid juveniles had similar survival and growth in the semi-natural streams. Yet, the presence of hybrids led to decreased survival of their wild half-siblings. Contrary to our hypothesis about context-dependency, these effects were not modified by food availability. However, wild juveniles with high SMR had decreased survival when food was scarce, but there was no such effect at high food availability. This study provides further proof that farmed salmon introgression may compromise the viability of wild salmon populations. We cannot, however, conclude that this is connected to alterations in the metabolic phenotype of farmed salmon.

Automated measurement of upper thermal limits in small aquatic animals.

We present a method for automating the measurement of upper thermal limits in small aquatic organisms. Upper thermal limits are frequently defined by the cessation of movement at high temperature, with measurement being performed by manual observation. Consequently, estimates of upper thermal limits may be subject to error and bias, both within and among observers. Our method utilises video-based tracking software to record the movement of individuals when exposed to high, lethal temperatures. We develop an algorithm in the R computing language that can objectively identify the loss of locomotory function from tracking data. Using independent experimental data, we validate our approach by demonstrating the expected response in upper thermal limits to acclimation temperature.