Olanzapine for young PEople with aNorexia nervosa (OPEN): A protocol for an open-label feasibility study.

INTRODUCTION: Antipsychotics are routinely prescribed off-label for anorexia nervosa (AN) despite limited evidence. This article presents a protocol of a study aiming to assess the feasibility of a future definitive trial on olanzapine in young people with AN. METHODS AND ANALYSIS: In an open-label, one-armed feasibility study, 55 patients with AN or atypical AN, aged 12-24, receiving outpatient, inpatient or day-care treatment who are considered for olanzapine treatment will be recruited from NHS sites based in England. Assessments will be conducted at screening, baseline and at 8-, 16 weeks, 6- and 12 months. Primary feasibility parameters will be proportions of patients who agree to take olanzapine and who adhere to treatment and complete study assessments. Qualitative methods will be used to explore acceptability of the intervention and study design. Secondary feasibility parameters will be changes in body mass index, psychopathology, side effects, health-related quality of life, carer burden and proportion of participants who would enrol in a future randomised controlled trial. The study is funded by the National Institute for Health Research via Health Technology Assessment programme. DISCUSSION: Olanzapine for young PEople with aNorexia nervosa will inform a future randomised controlled trial on the efficacy and safety of prescribing olanzapine in young people with AN.

Threshold shifts and developmental temperature impact trade-offs between tolerance and plasticity.

Mounting evidence suggests that ectotherms are already living close to their upper physiological thermal limits. Phenotypic plasticity has been proposed to reduce the impact of climate change in the short-term providing time for adaptation, but the tolerance-plasticity trade-off hypothesis predicts organisms with higher tolerance have lower plasticity. Empirical evidence is mixed, which may be driven by methodological issues such as statistical artefacts, nonlinear reaction norms, threshold shifts or selection. Here, we examine whether threshold shifts (organisms with higher tolerance require stronger treatments to induce maximum plastic responses) influence tolerance-plasticity trade-offs in hardening capacity for desiccation tolerance and critical thermal maximum (CTMAX) across Drosophila species with varying distributions/sensitivity to desiccation/heat stress. We found evidence for threshold shifts in both traits; species with higher heat/desiccation tolerance required longer hardening treatments to induce maximum hardening responses. Species with higher heat tolerance also showed reductions in hardening capacity at higher developmental acclimation temperatures. Trade-off patterns differed depending on the hardening treatment used and the developmental temperature flies were exposed to. Based on these findings, studies that do not consider threshold shifts, or that estimate plasticity under a narrow set of environments, will have a limited ability to assess trade-off patterns and differences in plasticity across species/populations more broadly.

Vulnerability to climate change increases with trophic level in terrestrial organisms.

The resilience of ecosystem function under global climate change is governed by individual species vulnerabilities and the functional groups they contribute to (e.g. decomposition, primary production, pollination, primary, secondary and tertiary consumption). Yet it remains unclear whether species that contribute to different functional groups, which underpin ecosystem function, differ in their vulnerability to climate change. We used existing upper thermal limit data across a range of terrestrial species (N = 1701) to calculate species warming margins (degrees distance between a species upper thermal limit and the maximum environmental temperature they inhabit), as a metric of climate change vulnerability. We examined whether species that comprise different functional groups exhibit differential vulnerability to climate change, and if vulnerability trends change across geographic space while considering evolutionary history. Primary producers had the broadest warming margins across the globe (µ = 18.72 °C) and tertiary consumers had the narrowest warming margins (µ = 9.64 °C), where vulnerability tended to increase with trophic level. Warming margins had a nonlinear relationship (second-degree polynomial) with absolute latitude, where warming margins were narrowest at about 33°, and were broader at lower and higher absolute latitudes. Evolutionary history explained significant variation in species warming margins, as did the methodology used to estimate species upper thermal limits. We investigated if variation in body mass across the trophic levels could explain why higher trophic level organisms had narrower warming margins than lower trophic level organisms, however, we did not find support for this hypothesis. This study provides a critical first step in linking individual species vulnerabilities with whole ecosystem responses to climate change.

Phylogenetic and environmental patterns of sex differentiation in physiological traits across Drosophila species.

Sex-based differences in physiological traits may be influenced by both evolutionary and environmental factors. Here we used male and female flies from >80 Drosophila species reared under common conditions to examine variance in a number of physiological traits including size, starvation, desiccation and thermal tolerance. Sex-based differences for desiccation and starvation resistance were comparable in magnitude to those for size, with females tending to be relatively more resistant than males. In contrast thermal resistance showed low divergence between the sexes. Phylogenetic signal was detected for measures of divergence between the sexes, such that species from the Sophophora clade showed larger differences between the sexes than species from the Drosophila clade. We also found that sex-based differences in desiccation resistance, body size and starvation resistance were weakly associated with climate (annual mean temperature/precipitation seasonality) but the direction and association with environment depended on phylogenetic position. The results suggest that divergence between the sexes can be linked to environmental factors, while an association with phylogeny suggests sex-based differences persist over long evolutionary time-frames.

Climate change and invasive species: a physiological performance comparison of invasive and endemic bees in Fiji.

Anthropogenic climate change and invasive species are two of the greatest threats to biodiversity, affecting the survival, fitness and distribution of many species around the globe. Invasive species are often expected to have broad thermal tolerance, be highly plastic, or have high adaptive potential when faced with novel environments. Tropical island ectotherms are expected to be vulnerable to climate change as they often have narrow thermal tolerance and limited plasticity. In Fiji, only one species of endemic bee, Homalictus fijiensis, is commonly found in the lowland regions, but two invasive bee species, Braunsapis puangensis and Ceratina dentipes, have recently been introduced into Fiji. These introduced species pollinate invasive plants and might compete with H. fijiensis and other native pollinators for resources. To test whether certain performance traits promote invasiveness of some species, and to determine which species are the most vulnerable to climate change, we compared the thermal tolerance, desiccation resistance, metabolic rate and seasonal performance adjustments of endemic and invasive bees in Fiji. The two invasive species tended to be more resistant to thermal and desiccation stress than H. fijiensis, while H. fijiensis had greater capacity to adjust their CTmax with season, and H. fijiensis females tended to have higher metabolic rates than B. puangensis females. These findings provide mixed support for current hypotheses for the functional basis of the success of invasive species; however, we expect the invasive bees in Fiji to be more resilient to climate change because of their increased thermal tolerance and desiccation resistance.

Phenotypic Plasticity for Desiccation Resistance, Climate Change, and Future Species Distributions: Will Plasticity Have Much Impact?

AbstractWhile species distribution models (SDMs) are widely used to predict the vulnerability of species to climate change, they do not explicitly indicate the extent to which plastic responses ameliorate climate change impacts. Here we use data on plastic responses of 32 species of Drosophila to desiccation stress to suggest that basal resistance, rather than adult hardening, is relatively more important in determining species differences in desiccation resistance and sensitivity to climate change. We go on to show, using the semimechanistic SDM CLIMEX, that the inclusion of plasticity has some impact on current species distributions and future vulnerability for widespread species but has little impact on the distribution of arguably more vulnerable tropically restricted species.

Does Plasticity Trade Off With Basal Heat Tolerance?

Studies suggest that many species are already living close to their upper physiological thermal limits. Phenotypic plasticity is thought to be an important mechanism for species to counter rapid environmental change, yet the extent to which plastic responses may buffer projected climate change - and what limits the evolution of plasticity - is still unclear. The tolerance-plasticity trade-off hypothesis predicts that the evolution of plasticity may be constrained by a species' thermal tolerance. Empirical evidence is equivocal, but we argue that inconsistent patterns likely reflect problems in experimental design/analysis, limiting our ability to detect and interpret trade-off patterns. Here, we address why we may, or may not see tolerance-plasticity trade-offs and outline a framework addressing current limitations, focusing on understanding the underlying mechanisms.

Evolution and plasticity of thermal performance: an analysis of variation in thermal tolerance and fitness in 22 Drosophila species.

The thermal biology of ectotherms is often used to infer species' responses to changes in temperature. It is often proposed that temperate species are more cold-tolerant, less heat-tolerant, more plastic, have broader thermal performance curves (TPCs) and lower optimal temperatures when compared to tropical species. However, relatively little empirical work has provided support for this using large interspecific studies. In the present study, we measure thermal tolerance limits and thermal performance in 22 species of Drosophila that developed under common conditions. Specifically, we measure thermal tolerance (CTmin and CTmax) as well as the fitness components viability, developmental speed and fecundity at seven temperatures to construct TPCs for each of these species. For 10 of the species, we also measure thermal tolerance and thermal performance following developmental acclimation to three additional temperatures. Using these data, we test several fundamental hypotheses about the evolution and plasticity of heat and cold resistance and thermal performance. We find that cold tolerance (CTmin) varied between the species according to the environmental temperature in the habitat from which they originated. These data support the idea that the evolution of cold tolerance has allowed species to persist in colder environments. However, contrary to expectation, we find that optimal temperature ( Topt) and the breadth of thermal performance ( Tbreadth) are similar in temperate, widespread and tropical species and we also find that the plasticity of TPCs was constrained. We suggest that the temperature range for optimal thermal performance is either fixed or under selection by the more similar temperatures that prevail during growing seasons. As a consequence, we find that Topt and Tbreadth are of limited value for predicting past, present and future distributions of species. This article is part of the theme issue 'Physiological diversity, biodiversity patterns and global climate change: testing key hypotheses involving temperature and oxygen'.

Comparing thermal performance curves across traits: how consistent are they?

Thermal performance curves (TPCs) are intended to approximate the relationship between temperature and fitness, and are commonly integrated into species distributional models for understanding climate change responses. However, TPCs may vary across traits because selection and environmental sensitivity (plasticity) differ across traits or because the timing and duration of the temperature exposure, here termed time scale, may alter trait variation. Yet, the extent to which TPCs vary temporally and across traits is rarely considered in assessments of climate change responses. Using a common garden approach, we estimated TPCs for standard metabolic rate (SMR), and activity in Drosophila melanogaster at three test temperatures (16, 25 and 30°C), using flies from each of six developmental temperatures (16, 18, 20, 25, 28 and 30°C). We examined the effects of time scale of temperature exposure (minutes/hours versus days/weeks) in altering TPC shape and position, and commonly used descriptors of the TPC: thermal optimum (Topt), thermal limits (Tmin and Tmax) and thermal breadth (Tbr). In addition, we collated previously published estimates of TPCs for fecundity and egg-to-adult viability in D. melanogaster We found that the descriptors of the TPCs varied across traits (egg-to-adult viability, SMR, activity and fecundity), but variation in TPCs within these traits was small across studies when measured at the same time scales. The time scale at which traits were measured contributed to greater variation in TPCs than the observed variance across traits, although the relative importance of time scale differed depending on the trait (activity versus fecundity). Variation in the TPC across traits and time scales suggests that TPCs using single traits may not be an accurate predictor of fitness and thermal adaptation across environments.

Evidence for lower plasticity in CTMAX at warmer developmental temperatures.

Understanding the capacity for different species to reduce their susceptibility to climate change via phenotypic plasticity is essential for accurately predicting species extinction risk. The climatic variability hypothesis suggests that spatial and temporal variation in climatic variables should select for more plastic phenotypes. However, empirical support for this hypothesis is limited. Here, we examine the capacity for ten Drosophila species to increase their critical thermal maxima (CTMAX ) through developmental acclimation and/or adult heat hardening. Using four fluctuating developmental temperature regimes, ranging from 13 to 33 °C, we find that most species can increase their CTMAX via developmental acclimation and adult hardening, but found no relationship between climatic variables and absolute measures of plasticity. However, when plasticity was dissected across developmental temperatures, a positive association between plasticity and one measure of climatic variability (temperature seasonality) was found when development took place between 26 and 28 °C, whereas a negative relationship was found when development took place between 20 and 23 °C. In addition, a decline in CTMAX and egg-to-adult viability, a proxy for fitness, was observed in tropical species at the warmer developmental temperatures (26-28 °C); this suggests that tropical species may be at even greater risk from climate change than currently predicted. The combined effects of developmental acclimation and adult hardening on CTMAX were small, contributing to a <0.60 °C shift in CTMAX . Although small shifts in CTMAX may increase population persistence in the shorter term, the degree to which they can contribute to meaningful responses in the long term is unclear.

Plasticity for desiccation tolerance across Drosophila species is affected by phylogeny and climate in complex ways.

Comparative analyses of ectotherm susceptibility to climate change often focus on thermal extremes, yet responses to aridity may be equally important. Here we focus on plasticity in desiccation resistance, a key trait shaping distributions of Drosophila species and other small ectotherms. We examined the extent to which 32 Drosophila species, varying in their distribution, could increase their desiccation resistance via phenotypic plasticity involving hardening, linking these responses to environment, phylogeny and basal resistance. We found no evidence to support the seasonality hypothesis; species with higher hardening plasticity did not occupy environments with higher and more seasonal precipitation. As basal resistance increased, the capacity of species to respond via phenotypic plasticity decreased, suggesting plastic responses involving hardening may be constrained by basal resistance. Trade-offs between basal desiccation resistance and plasticity were not universal across the phylogeny and tended to occur within specific clades. Phylogeny, environment and trade-offs all helped to explain variation in plasticity for desiccation resistance but in complex ways. These findings suggest some species have the ability to counter dry periods through plastic responses, whereas others do not; and this ability will depend to some extent on a species' placement within a phylogeny, along with its basal level of resistance.

How important is thermal history? Evidence for lasting effects of developmental temperature on upper thermal limits in Drosophila melanogaster.

A common practice in thermal biology is to take individuals directly from the field and estimate a range of thermal traits. These estimates are then used in studies aiming to understand broad scale distributional patterns, understanding and predicting the evolution of phenotypic plasticity, and generating predictions for climate change risk. However, the use of field-caught individuals in such studies ignores the fact that many traits are phenotypically plastic and will be influenced by the thermal history of the focal individuals. The current study aims to determine the extent to which estimates of upper thermal limits (CTmax), a frequently used measure for climate change risk, are sensitive to developmental and adult acclimation temperatures and whether these two forms of plasticity are reversible. Examining a temperate and tropical population of Drosophila melanogaster we show that developmental acclimation has a larger and more lasting effect on CTmax than adult acclimation. We also find evidence for an interaction between developmental and adult acclimation, particularly when flies are acclimated for a longer period, and that these effects can be population specific. These results suggest that thermal history can have lasting effects on estimates of CTmax. In addition, we provide evidence that developmental and/or adult acclimation are unlikely to contribute to substantial shifts in CTmax and that acclimation capacity may be constrained at higher temperatures.

Metabolic cold adaptation contributes little to the interspecific variation in metabolic rates of 65 species of Drosophilidae.

Metabolic cold adaptation (MCA) is a controversial hypothesis suggesting that cold adapted species display an elevated metabolic rate (MR) compared to their warm climate relatives. Here we test for the presence of MCA in 65 species of drosophilid flies reared under common garden conditions. MR was measured at both 10 and 20°C for both sexes and data were analyzed in relation to the natural thermal environment of these species. We found considerable interspecific variation in MR ranging from 1.34 to 8.99µWmg-1 at 10°C. As predicted by Bergmann's rule body mass of fly species correlated negatively with annual mean temperature (AMT), such that larger species were found in colder environments. Because larger flies have a higher total MR we found MR to vary with AMT, however, after inclusion of mass as a co-variate we found no significant effect of AMT. Furthermore, we did not find that thermal sensitivity of MR (Q10) varied with AMT. Based on this broad collection of species we therefore conclude that there is no adaptive pattern of metabolic cold adaptation within drosophilid species ranging from sub-arctic to tropical environments.

Incorporating evolutionary adaptation in species distribution modelling reduces projected vulnerability to climate change.

Based on the sensitivity of species to ongoing climate change, and numerous challenges they face tracking suitable conditions, there is growing interest in species' capacity to adapt to climatic stress. Here, we develop and apply a new generic modelling approach (AdaptR) that incorporates adaptive capacity through physiological limits, phenotypic plasticity, evolutionary adaptation and dispersal into a species distribution modelling framework. Using AdaptR to predict change in the distribution of 17 species of Australian fruit flies (Drosophilidae), we show that accounting for adaptive capacity reduces projected range losses by up to 33% by 2105. We identify where local adaptation is likely to occur and apply sensitivity analyses to identify the critical factors of interest when parameters are uncertain. Our study suggests some species could be less vulnerable than previously thought, and indicates that spatiotemporal adaptive models could help improve management interventions that support increased species' resilience to climate change.

Experimental Evolution under Fluctuating Thermal Conditions Does Not Reproduce Patterns of Adaptive Clinal Differentiation in Drosophila melanogaster.

Experimental evolution can be a useful tool for testing the impact of environmental factors on adaptive changes in populations, and this approach is being increasingly used to understand the potential for evolutionary responses in populations under changing climates. However, selective factors will often be more complex in natural populations than in laboratory environments and produce different patterns of adaptive differentiation. Here we test the ability of laboratory experimental evolution under different temperature cycles to reproduce well-known patterns of clinal variation in Drosophila melanogaster. Six fluctuating thermal regimes mimicking the natural temperature conditions along the east coast of Australia were initiated. Contrary to expectations, on the basis of field patterns there was no evidence for adaptation to thermal regimes as reflected by changes in cold and heat resistance after 1-3 years of laboratory natural selection. While laboratory evolution led to changes in starvation resistance, development time, and body size, patterns were not consistent with those seen in natural populations. These findings highlight the complexity of factors affecting trait evolution in natural populations and indicate that caution is required when inferring likely evolutionary responses from the outcome of experimental evolution studies.

Evolutionary capacity of upper thermal limits: beyond single trait assessments.

Thermal tolerance is an important factor influencing the distribution of ectotherms, but we still have limited understanding of the ability of species to evolve different thermal limits. Recent studies suggest that species may have limited capacity to evolve higher thermal limits in response to slower, more ecologically relevant rates of warming. However, these conclusions are based on univariate estimates of adaptive capacity. To test these findings within an explicitly multivariate context, we used a paternal half-sibling breeding design to estimate the multivariate evolutionary potential for upper thermal limits in Drosophila melanogaster. We assessed heat tolerance using static (basal and hardened) and ramping assays. Additive genetic variances were significantly different from zero only for the static measures of heat tolerance. Our G: matrix analysis revealed that any response to selection for increased heat tolerance will largely be driven by static basal and hardened heat tolerance, with minimal contribution from ramping heat tolerance. These results suggest that the capacity to evolve upper thermal limits in nature may depend on the type of thermal stress experienced.

Contemporary climate change and terrestrial invertebrates: evolutionary versus plastic changes.

To forecast the responses of species to future climate change, an understanding of the ability of species to adapt to long-term shifts in temperature is crucial. We present a review on evolutionary adaptation and phenotypic plasticity of temperature-related traits in terrestrial invertebrates. The evidence for adaptive evolution in melanization is good, but we caution that genetic determination needs to be tested in each individual species, and complex genetic correlations may exist. For phenological traits allochronic data sets provide powerful means to track climate-induced changes; however, rarely are responses deconstructed into evolutionary and plastic responses. Laboratory studies suggest climate change responses in these traits will be driven by both. For stress resistance, the evidence for shifts in traits is poor. Studies leaning heavily on Drosophila have demonstrated potential limits to evolutionary responses in desiccation and heat resistance. Quantifying the capacity for these species to respond plastically and extending this work to other taxa will be an important next step. We also note that, although not strictly speaking a species trait, the response of endosymbionts to heat stress requires further study. Finally, while clearly genetic, and possibly adaptive, the anonymous nature of latitudinal shifts in clines of genetic markers in Drosophila prevents further interpretation.

Does environmental robustness play a role in fluctuating environments?

Fluctuating environments are expected to select for individuals that have highest geometric fitness over the experienced environments. This leads to the prediction that genetically determined environmental robustness in fitness, and average fitness across environments should be positively genetically correlated to fitness in fluctuating environments. Because quantitative genetic experiments resolving these predictions are missing, we used a full-sib, half-sib breeding design to estimate genetic variance for egg-to-adult viability in Drosophila melanogaster exposed to two constant or fluctuating temperatures that were above the species' optimum temperature, during development. Viability in two constant environments (25°C or 30°C) was used to estimate breeding values for environmental robustness of viability (i.e., reaction norm slope) and overall viability (reaction norm elevation). These breeding values were regressed against breeding values of viability at two different fluctuating temperatures (with a mean of 25°C or 30°C). Our results based on genetic correlations show that average egg-to-adult viability across different constant thermal environments, and not the environmental robustness, was the most important factor for explaining the fitness in fluctuating thermal environments. Our results suggest that the role of environmental robustness in adapting to fluctuating environments might be smaller than anticipated.

Trait associations across evolutionary time within a drosophila phylogeny: correlated selection or genetic constraint?

Traits do not evolve independently. To understand how trait changes under selection might constrain adaptive changes, phenotypic and genetic correlations are typically considered within species, but these capture constraints across a few generations rather than evolutionary time. For longer-term constraints, comparisons are needed across species but associations may arise because of correlated selection pressures rather than genetic interactions. Implementing a unique approach, we use known patterns of selection to separate likely trait correlations arising due to correlated selection from those reflecting genetic constraints. We examined the evolution of stress resistance in >90 Drosophila species adapted to a range of environments, while controlling for phylogeny. Initially we examined the role of climate and phylogeny in shaping the evolution of starvation and body size, two traits previously not examined in this context. Following correction for phylogeny only a weak relationship between climate and starvation resistance was detected, while all of the variation in the relationship between body size and climate could be attributed to phylogeny. Species were divided into three environmental groups (hot and dry, hot and wet, cold) with the expectation that, if genetic correlations underpin trait correlations, these would persist irrespective of the environment, whereas selection-driven evolution should produce correlations dependent on the environment. We found positive associations between most traits in hot and dry environments coupled with high trait means. In contrast few trait correlations were observed in hot/wet and cold environments. These results suggest trait associations are primarily driven by correlated selection rather than genetic interactions, highlighting that such interactions are unlikely to limit evolution of stress resistance.

Complexity of the genetic basis of ageing in nature revealed by a clinal study of lifespan and methuselah, a gene for ageing, in Drosophila from eastern Australia.

Clinal studies are a powerful tool for understanding the genetic basis of climatic adaptation. However, while clines in quantitative traits and genetic polymorphisms have been observed within and across continents, few studies have attempted to demonstrate direct links between them. The gene methuselah in Drosophila has been shown to have a major effect on stress response and longevity phenotypes based largely on laboratory studies of induced mutations in the mth gene. Clinal patterns in the most common mth haplotype and for lifespan (both increasing with latitude) have been observed in North American populations of D. melanogaster, implicating climatic selection. While these clinal patterns have led some to suggest that mth influences ageing in natural populations, limited evidence on the association between the two has so far been collected. Here, we describe a significant cline in the mth haplotype in eastern Australian D. melanogaster populations that parallel the cline in North America. We also describe a cline in mth gene expression. These findings further support the idea that mth is itself under selection. In contrast, we show that lifespan has a strong nonlinear clinal pattern, increasing southwards from the tropics, but then decreasing again from mid-latitudes. Furthermore, in association studies, we find no evidence for a direct link between mth haplotype and lifespan. Thus, while our data support a role for mth variation being under natural selection, we found no link to naturally occurring variation in lifespan and ageing in Australian populations of D. melanogaster. Our results indicate that the mth locus likely has genetic background and environment-specific effects.