Mitochondrial metabolism and respiration adjustments following temperature acclimation in Daphnia magna.

Seasonal temperature changes and local variations in the water column challenge lentic zooplankton in their habitat. At the cellular level, exposure to varying temperatures affects the mitochondrial functional properties of poikilothermic organisms. Metabolic enzymes that supply reduced substrates to the electron transport chain and elements of the oxidative phosphorylation system must therefore adjust their activity and flux rates to the altered temperature conditions. In the present study, Daphnia magna respiration was analyzed in response to acute and chronic changes in ambient temperature. Oxygen consumption as well as substrate and electron flux rates were measured at the animals' acclimation temperature and at two additional acute temperatures. High activity of citrate synthase (CS) in cold-acclimated animals (10 °C) may have resulted from mitochondrial quantitative adjustments. However, thermal sensitivity of the functional properties of mitochondrial enzymes was greater in warm-acclimated animals (30 °C). In whole animals, temperature-induced changes were partly compensated by acclimation, but these changes were promoted by acclimation in the case of the mitochondrial electron transport chain. Thus, respiration realised in whole animals was limited by the provision of reduced substrates in the tricarboxylic acid cycle rather than by restrictions of the respiratory chain complexes. This may minimize production of reactive oxygen species and resulting damage and reduce waste of substrates from the animals' energy reserves. Still, the integrated biomarker response indicated increased defense against oxidative stress at elevated temperatures.

A common temperature dependence of nutritional demands in ectotherms.

In light of ongoing climate change, it is increasingly important to know how nutritional requirements of ectotherms are affected by changing temperatures. Here, we analyse the wide thermal response of phosphorus (P) requirements via elemental gross growth efficiencies of Carbon (C) and P, and the Threshold Elemental Ratios in different aquatic invertebrate ectotherms: the freshwater model species Daphnia magna, the marine copepod Acartia tonsa, the marine heterotrophic dinoflagellate Oxyrrhis marina, and larvae of two populations of the marine crab Carcinus maenas. We show that they all share a non-linear cubic thermal response of nutrient requirements. Phosphorus requirements decrease from low to intermediate temperatures, increase at higher temperatures and decrease again when temperature is excessive. This common thermal response of nutrient requirements is of great importance if we aim to understand or even predict how ectotherm communities will react to global warming and nutrient-driven eutrophication.

Short term fluctuating temperature alleviates Daphnia stoichiometric constraints.

In this study, we analysed how short term temperature fluctuation interacts with nutrient limitation in the vertical migrating Daphnia commutata. We hypothesize that short term (daily) temperature fluctuation will alleviate nutrient limitation. We carried out experiments analysing growth rates, phosphorus and RNA content of D. commutate grown under four different temperature regimes and two P-limited conditions. Our experiments showed that individuals grown under fluctuating temperature grew more than at the mean temperature. We estimated the expected sizes for the 15 °C treatment based on the Q10 and for the fluctuating temperature treatment. These expected sizes for both treatments resulted well below the observed ones. The P and RNA content of individuals grown at 10 °C were significantly higher than those at 20 °C, and when individuals grown at 10 °C were translocated to 20 °C they exerted an increased growth rate. Our results suggest that, under a regime of diel vertical migration, the temperature alternation would allow migrating organisms to alleviate the effect of severe nutrient limitation maintaining population growth. Under a scenario of global warming, where epilimnetic temperatures will increase, lake temperature will interact with nutrient limitation for consumers, but, organisms may be able to face these changes if they can still regularly move from a cold hypolimnion to a warmer epilimnion.

Ex vivo and in vivo effects of arsenite on GST and ABCC2 activity and expression in the middle intestine of the rainbow trout Oncorhynchus mykiss.

In fish of freshwaters environments, the accumulation and toxic effects of arsenite (AsIII) can be attenuated by detoxification proteins such as GST and ABCC transporters. We studied the effects of AsIII on the middle intestine of O. mykiss in ex-vivo and in vivo/ex vivo assays. For the ex vivo assays, we measured the transport rate of the ABCC substrate DNP-SG and GST activity in intestinal strips and everted sacs. AsIII inhibited DNP-SG transport in a concentration-dependent manner, specifically when we applied it on the basolateral side. GST activity increased when we applied a maximum concentration of AsIII. For the in vivo/ex vivo assays, we kept fish in water with or without 7.7 µmol L-1 of AsIII for 48 h. Then, we measured DNP-SG transport rate, GST activity, and PP1 activity in intestine strips during one hour. For PP1 activity, we incubated the strips with or without microcystin-LR (MCLR), a toxin excreted through ABCC2 proteins. We also analyzed Abcc2 and Gst-π mRNA expression in intestine and liver tissue. In the group exposed in vivo to AsIII, DNP-SG transport rate and GST activity were higher and the effect of MCLR over PP1 activity was attenuated. AsIII significantly induced only Abcc2 mRNA expression in both middle intestine and liver. Our results suggest that, in the middle intestine of O. mykiss, AsIII is absorbed mainly at the basolateral side of the enterocytes, excreted to the lumen by ABCC2 transporters, and is capable of modulating Abcc2 mRNA expression by a transcriptional mechanism.

Does the stoichiometric carbon:phosphorus knife edge apply for predaceous copepods?

Recent work has indicated that stoichiometric food quality in terms of the carbon:phosphorus (C:P) ratio affects consumers whether the imbalance involves a deficit or an excess of nutrients; hence, organisms exist on a "stoichiometric knife edge". While previous studies have focused primarily on autotroph-herbivore trophic transfer, nutritional imbalances may also affect the interactions between species at higher trophic levels. Since the foods of carnivores are normally stoichiometrically similar to the body compositions of those carnivores, they may be more severely affected than herbivores if imbalances become pronounced. We analysed the response of the predatory copepod Parabroteas sarsi to monospecific diet treatments consisting of high and low C:P prey items. These dietary treatments strongly affected the predator's elemental composition and growth, although prey selection, excretion, egestion, and respiration rates were not affected. We suggest that, due to their low threshold elemental ratio and a narrow C:P stoichiometric knife edge, these predators are highly vulnerable to stoichiometric imbalances, whether an excess or a deficit of nutrients is involved. Our results demonstrating this high sensitivity to prey C:P ratio show that the stoichiometric knife edge may apply to not only herbivores but also higher trophic levels. Thus, predators such as P. sarsi, with a much narrower range of food quality, may also be strongly affected by fluctuations in the quality of their prey, with negative consequences for their secondary production.

Glacier melting and stoichiometric implications for lake community structure: zooplankton species distributions across a natural light gradient.

Glaciers around the globe are melting rapidly, threatening the receiving environments of the world's fresh water reservoirs with significant changes. The meltwater, carried by rivers, contains large amounts of suspended sediment particles, producing longitudinal gradients in the receiving lakes. These gradients may result in changes in the light : nutrient ratio that affect grazer performance by altering elemental food quality. Thus, glacial melting may induce a shift in the phytoplankton carbon : nutrient ratio and hence influence the dominance of herbivorous zooplankton through stoichiometric mechanisms. To test this hypothesis, we combined field and experimental data, taking advantage of a natural light intensity gradient caused by glacial clay input in a deep oligotrophic Patagonian lake. Across this gradient, we evaluated the abundances of two consumer taxa with different phosphorus requirements, the copepod Boeckella gracilipes and the cladoceran Daphnia commutata, using a six-station transect along the lake. We found significant differences in light : nutrient ratio and stoichiometric food quality of the seston, together with a switch from dominance of P-rich Daphnia in low carbon : nutrient stations to dominance of low-P copepods in high carbon : nutrient stations. The laboratory experiments confirmed that the difference in the carbon : nutrient ratio across the gradient is sufficient to impair Daphnia growth. The overall patterns are consistent with our prediction that shifts in the environmental light : nutrient ratio as a result of glacial melting would contribute to shifts in the dominance of stoichiometrically contrasting taxa in consumer guilds.

Effect of ultraviolet radiation on acetylcholinesterase activity in freshwater copepods.

We analyzed the effects of UV radiation (UVR) effects on acetylcholinesterase (AChE) activity in two calanoid copepods, Boeckella gibbosa and Parabroteas sarsi that inhabit Patagonian shallow lakes. We studied the effect of experimental UVR (UV-B and UV-A) exposure on AChE activity in relation to basal antioxidant capacities of both copepods. Our experiments showed that UVR can effectively depress AChE activity, although with differences between species. In both copepods AChE was affected by UV-B, whereas UV-A only affected AChE in B. gibbosa. Both copepods also differed in body elemental composition (C:N:P), photoprotecting compound content (carotenoids and mycosporine-like amino acids) and enzymatic antioxidant capacity (glutathione S-transferase [GST]). Our results suggest that when exposed to UVR, AChE activity would depend more on the antioxidant capacity (GST) and P availability for enzyme synthesis than on the photoprotective compounds.