Table_1_Copper mediates life history responses of Daphnia pulex to predation threat.DOCX

A key challenge for ecological and ecotoxicological risk assessment is to predict the risk of organisms when exposed simultaneously to multiple stressors in sub-lethal concentrations. Here, we assessed whether sub-lethal concentrations of an anthropogenic stressors, the heavy metal copper (Cu), mediates the impacts of a natural ecological threat to species, predation risk, among six distinct Daphnia pulex clones. We investigated the interaction between the two stressors on morphological defenses and on several life-history traits including maturation time, size at maturity, somatic growth rate and survival rates. Combining a life table experiment on a response surface design, we found no evidence that the heavy metal copper mediates the effects of predator cue induced morphological responses in the tested D. pulex clones. However, our data indicate that copper can mediate several key life-history responses to predation risk. For age at maturity, we found also clear evidence that the observed interaction between predation risk and copper varied by whether clones were strong or weak morphological responders. Specific exploration of the relationship between morphological responses and life history traits under predation risk and copper suggest a strong hypothesis for multiple strategies to deal with multiple stressors. While interactions between different stressors make it harder to predict their outcomes, and ultimately assess water quality regulations about the effects of such stressors, our study provides evidence that life history theory can aid in understanding and predicting their impacts.</p

Table_2_Copper mediates life history responses of Daphnia pulex to predation threat.DOCX

A key challenge for ecological and ecotoxicological risk assessment is to predict the risk of organisms when exposed simultaneously to multiple stressors in sub-lethal concentrations. Here, we assessed whether sub-lethal concentrations of an anthropogenic stressors, the heavy metal copper (Cu), mediates the impacts of a natural ecological threat to species, predation risk, among six distinct Daphnia pulex clones. We investigated the interaction between the two stressors on morphological defenses and on several life-history traits including maturation time, size at maturity, somatic growth rate and survival rates. Combining a life table experiment on a response surface design, we found no evidence that the heavy metal copper mediates the effects of predator cue induced morphological responses in the tested D. pulex clones. However, our data indicate that copper can mediate several key life-history responses to predation risk. For age at maturity, we found also clear evidence that the observed interaction between predation risk and copper varied by whether clones were strong or weak morphological responders. Specific exploration of the relationship between morphological responses and life history traits under predation risk and copper suggest a strong hypothesis for multiple strategies to deal with multiple stressors. While interactions between different stressors make it harder to predict their outcomes, and ultimately assess water quality regulations about the effects of such stressors, our study provides evidence that life history theory can aid in understanding and predicting their impacts.</p

Image_3_Copper mediates life history responses of Daphnia pulex to predation threat.jpg

A key challenge for ecological and ecotoxicological risk assessment is to predict the risk of organisms when exposed simultaneously to multiple stressors in sub-lethal concentrations. Here, we assessed whether sub-lethal concentrations of an anthropogenic stressors, the heavy metal copper (Cu), mediates the impacts of a natural ecological threat to species, predation risk, among six distinct Daphnia pulex clones. We investigated the interaction between the two stressors on morphological defenses and on several life-history traits including maturation time, size at maturity, somatic growth rate and survival rates. Combining a life table experiment on a response surface design, we found no evidence that the heavy metal copper mediates the effects of predator cue induced morphological responses in the tested D. pulex clones. However, our data indicate that copper can mediate several key life-history responses to predation risk. For age at maturity, we found also clear evidence that the observed interaction between predation risk and copper varied by whether clones were strong or weak morphological responders. Specific exploration of the relationship between morphological responses and life history traits under predation risk and copper suggest a strong hypothesis for multiple strategies to deal with multiple stressors. While interactions between different stressors make it harder to predict their outcomes, and ultimately assess water quality regulations about the effects of such stressors, our study provides evidence that life history theory can aid in understanding and predicting their impacts.</p

Image_4_Copper mediates life history responses of Daphnia pulex to predation threat.jpg

A key challenge for ecological and ecotoxicological risk assessment is to predict the risk of organisms when exposed simultaneously to multiple stressors in sub-lethal concentrations. Here, we assessed whether sub-lethal concentrations of an anthropogenic stressors, the heavy metal copper (Cu), mediates the impacts of a natural ecological threat to species, predation risk, among six distinct Daphnia pulex clones. We investigated the interaction between the two stressors on morphological defenses and on several life-history traits including maturation time, size at maturity, somatic growth rate and survival rates. Combining a life table experiment on a response surface design, we found no evidence that the heavy metal copper mediates the effects of predator cue induced morphological responses in the tested D. pulex clones. However, our data indicate that copper can mediate several key life-history responses to predation risk. For age at maturity, we found also clear evidence that the observed interaction between predation risk and copper varied by whether clones were strong or weak morphological responders. Specific exploration of the relationship between morphological responses and life history traits under predation risk and copper suggest a strong hypothesis for multiple strategies to deal with multiple stressors. While interactions between different stressors make it harder to predict their outcomes, and ultimately assess water quality regulations about the effects of such stressors, our study provides evidence that life history theory can aid in understanding and predicting their impacts.</p

Image_2_Copper mediates life history responses of Daphnia pulex to predation threat.jpg

A key challenge for ecological and ecotoxicological risk assessment is to predict the risk of organisms when exposed simultaneously to multiple stressors in sub-lethal concentrations. Here, we assessed whether sub-lethal concentrations of an anthropogenic stressors, the heavy metal copper (Cu), mediates the impacts of a natural ecological threat to species, predation risk, among six distinct Daphnia pulex clones. We investigated the interaction between the two stressors on morphological defenses and on several life-history traits including maturation time, size at maturity, somatic growth rate and survival rates. Combining a life table experiment on a response surface design, we found no evidence that the heavy metal copper mediates the effects of predator cue induced morphological responses in the tested D. pulex clones. However, our data indicate that copper can mediate several key life-history responses to predation risk. For age at maturity, we found also clear evidence that the observed interaction between predation risk and copper varied by whether clones were strong or weak morphological responders. Specific exploration of the relationship between morphological responses and life history traits under predation risk and copper suggest a strong hypothesis for multiple strategies to deal with multiple stressors. While interactions between different stressors make it harder to predict their outcomes, and ultimately assess water quality regulations about the effects of such stressors, our study provides evidence that life history theory can aid in understanding and predicting their impacts.</p

Image_6_Copper mediates life history responses of Daphnia pulex to predation threat.jpg

A key challenge for ecological and ecotoxicological risk assessment is to predict the risk of organisms when exposed simultaneously to multiple stressors in sub-lethal concentrations. Here, we assessed whether sub-lethal concentrations of an anthropogenic stressors, the heavy metal copper (Cu), mediates the impacts of a natural ecological threat to species, predation risk, among six distinct Daphnia pulex clones. We investigated the interaction between the two stressors on morphological defenses and on several life-history traits including maturation time, size at maturity, somatic growth rate and survival rates. Combining a life table experiment on a response surface design, we found no evidence that the heavy metal copper mediates the effects of predator cue induced morphological responses in the tested D. pulex clones. However, our data indicate that copper can mediate several key life-history responses to predation risk. For age at maturity, we found also clear evidence that the observed interaction between predation risk and copper varied by whether clones were strong or weak morphological responders. Specific exploration of the relationship between morphological responses and life history traits under predation risk and copper suggest a strong hypothesis for multiple strategies to deal with multiple stressors. While interactions between different stressors make it harder to predict their outcomes, and ultimately assess water quality regulations about the effects of such stressors, our study provides evidence that life history theory can aid in understanding and predicting their impacts.</p

Image_5_Copper mediates life history responses of Daphnia pulex to predation threat.jpg

A key challenge for ecological and ecotoxicological risk assessment is to predict the risk of organisms when exposed simultaneously to multiple stressors in sub-lethal concentrations. Here, we assessed whether sub-lethal concentrations of an anthropogenic stressors, the heavy metal copper (Cu), mediates the impacts of a natural ecological threat to species, predation risk, among six distinct Daphnia pulex clones. We investigated the interaction between the two stressors on morphological defenses and on several life-history traits including maturation time, size at maturity, somatic growth rate and survival rates. Combining a life table experiment on a response surface design, we found no evidence that the heavy metal copper mediates the effects of predator cue induced morphological responses in the tested D. pulex clones. However, our data indicate that copper can mediate several key life-history responses to predation risk. For age at maturity, we found also clear evidence that the observed interaction between predation risk and copper varied by whether clones were strong or weak morphological responders. Specific exploration of the relationship between morphological responses and life history traits under predation risk and copper suggest a strong hypothesis for multiple strategies to deal with multiple stressors. While interactions between different stressors make it harder to predict their outcomes, and ultimately assess water quality regulations about the effects of such stressors, our study provides evidence that life history theory can aid in understanding and predicting their impacts.</p

Image_1_Copper mediates life history responses of Daphnia pulex to predation threat.jpg

A key challenge for ecological and ecotoxicological risk assessment is to predict the risk of organisms when exposed simultaneously to multiple stressors in sub-lethal concentrations. Here, we assessed whether sub-lethal concentrations of an anthropogenic stressors, the heavy metal copper (Cu), mediates the impacts of a natural ecological threat to species, predation risk, among six distinct Daphnia pulex clones. We investigated the interaction between the two stressors on morphological defenses and on several life-history traits including maturation time, size at maturity, somatic growth rate and survival rates. Combining a life table experiment on a response surface design, we found no evidence that the heavy metal copper mediates the effects of predator cue induced morphological responses in the tested D. pulex clones. However, our data indicate that copper can mediate several key life-history responses to predation risk. For age at maturity, we found also clear evidence that the observed interaction between predation risk and copper varied by whether clones were strong or weak morphological responders. Specific exploration of the relationship between morphological responses and life history traits under predation risk and copper suggest a strong hypothesis for multiple strategies to deal with multiple stressors. While interactions between different stressors make it harder to predict their outcomes, and ultimately assess water quality regulations about the effects of such stressors, our study provides evidence that life history theory can aid in understanding and predicting their impacts.</p

Additional file 3: of The transcriptomic and proteomic responses of Daphnia pulex to changes in temperature and food supply comprise environment-specific and clone-specific elements

Figure S2. Two-dimensional protein gels from the D. pulex clone G under starvation stress. The 2D gels, which are fusion (averaged) images from a varying number (n) of gels (biological replicates, 25-30 animals each), show changes in protein expression in the D. pulex clone G after the acute exposure of control animals (20 °C, ad libitum feeding) (blue spots; n = 5) to (a) 24 h (orange spots; n = 4) or (c) 48 h (orange spots; n = 4) of starvation (T = 20 °C). Red or green spot IDs mark significantly up- or downregulated proteins (t-tests, P < 0.05; see Table 4). The scatter plots show changes in expression level (Vrelative, relative spot volume) between control and starving animals (b, 24 h; d, 48 h) of significantly (large circles and letters) or non-significantly (small circles) up- or down-regulated proteins (data from a or c). Proteins, which were upregulated under starvation stress, are found above the diagonal line. (PDF 126 kb

Additional file 2: of The transcriptomic and proteomic responses of Daphnia pulex to changes in temperature and food supply comprise environment-specific and clone-specific elements

Figure S1. Two-dimensional protein gels from the 20 °C-acclimated D. pulex clones G and M. The RuBPs-stained 2D gels from control animals (20 °C, ad libitum feeding) of (a) the D. pulex clone G [fusion image from n = 5 gels (biological replicates)] and (b) the D. pulex clone M [fusion image from n = 4 gels (biological replicates, 25-30 animals each)] served as reference for the excision of protein spots for mass spectrometry (pI: 4.5-6, molecular mass: 20-150 or 20-100 kDa). Red characters are spot identifiers (IDs). (PDF 118 kb