D. magna ephippia were obtained from three populations: a pond in Værøy, Norway (67.687°N 12.672°E), a pond in Park Midden-Limburg, Zonhoven, Belgium (50.982°N 5.318°E), and a rice field which is flooded and dries out annually in the Delta del Ebro, Riet Vell, Spain (40.659°N 0.775°E). In the following, these three populations are referred to as the Norway, Belgium and Spain populations, respectively. We used 10 clones (originating from 10 different ephippia) from each population in the experiments, and these were reared at 17°C with a 16L:8D photoperiod for three to four parthenogenetic generations prior to the experiment. During this period, individuals were fed three times a week with Shellfish Diet 1800 (Reed Mariculture Inc, USA) at final concentration of algae 4 × 105 cells/ml, and the ADaM medium was changed once a week. For the experiment, second or later clutch neonates were collected and photographed less than 24 hours after birth. After photographing, neonates were placed individually in 50 ml tubes containing 17°C ADaM medium. Each tube was placed in a Memmert Peltier-cooled incubator IPP 260plus (Memmert, Germany). We used a 16L:8D photoperiod and the temperature in different cabinets was set to 12.0, 15.0, 17.0, 19.0, 22.0, 24.0 and 26.0 °C. Each temperature treatment received eight individuals from each of the 10 clones. Animals were fed every second day with concentrations that had previously been established to represent ad lib rations. Due to logistic constraints, the different temperature treatments were run simultaneously for one population at a time (Norway May-June 2015, Spain December-February 2018, Belgium July-September 2018). All individuals were checked daily for mortality and sexual maturity (presence of eggs in the brood chamber). Tubes were rotated daily within the climate cabinets during the maturity checks to avoid positional effects. Upon maturation individuals were photographed and terminated.

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 favorable 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.