Climate warming restructures an aquatic food web over 28 years.

Climate warming can restructure lake food webs if trophic levels differ in their thermal responses, but evidence for these changes and their underlying mechanisms remain scarce in nature. Here we document how warming lake temperatures by up to 2°C, rather than changes in trophic state or fishing effort, have restructured the pelagic food web of a large European lake (Lake Maggiore, Italy). Our approach exploited abundance and biomass data collected weekly to yearly across five trophic levels from 1981 to 2008. Temperature generally had stronger effects on taxa than changes in fish predation or trophic state mediated through primary productivity. Consequently, we found that, as the lake warmed, the food web shifted in numerical abundance towards predators occupying middle trophic positions. Of these taxa, the spiny water flea (Bythotrephes longimanus) most prospered. Bythotrephes strongly limited abundances of the keystone grazer Daphnia, strengthening top-down structuring of the food web. Warmer temperatures partly restructured the food web by advancing peak Bythotrephes densities by approximately 60 days and extending periods of positive population growth by three times. Nonetheless, our results suggested that advances in the timing and size of peak Bythotrephes densities could not outpace changes in the timing and size of peak densities in their Daphnia prey. Our results provide rare evidence from nature as to how long-term warming can favour higher trophic levels, with the potential to strengthen top-down control of food webs.

Road Salt Impacts Freshwater Zooplankton at Concentrations below Current Water Quality Guidelines.

Widespread use of NaCl for road deicing has caused increased chloride concentrations in lakes near urban centers and areas of high road density. Chloride can be toxic, and water quality guidelines have been created to regulate it and protect aquatic life. However, these guidelines may not adequately protect organisms in low-nutrient, soft water lakes such as those underlain by the Precambrian Shield. We tested this hypothesis by conducting laboratory experiments on six Daphnia species using a soft water culture medium. We also examined temporal changes in cladoceran assemblages in the sediments of two small lakes on the Canadian Shield: one near a highway and the other >3 km from roads where salt is applied in the winter. Our results showed that Daphnia were sensitive to low chloride concentrations with decreased reproduction and increased mortality occurring between 5 and 40 mg Cl-/L. Analysis of cladoceran remains in lake sediments revealed changes in assemblage composition that coincided with the initial application of road salt in this region. In contrast, there were no changes detected in the remote lake. We found that 22.7% of recreational lakes in Ontario have chloride concentrations between 5 and 40 mg/L suggesting that cladoceran zooplankton in these lakes may already be experiencing negative effects of chloride.

Ca2+ levels in Daphnia hemolymph may explain occurrences of daphniid species along recent Ca gradients in Canadian soft-water lakes.

Calcium levels are declining in eastern North American and western European lakes. This widespread issue is affecting the composition of crustacean zooplankton communities, as the presence and abundance of several calcium-rich daphniid species are declining, while two other daphniids, D. catawba and D. ambigua, that apparently tolerate low calcium environments, are prospering. The physiological basis for low calcium tolerance of these daphniids is unknown. In this study the presence of one Ca-rich (D. pulicaria) and one Ca-poor (D. ambigua) daphniid species in Canadian Shield lakes is assessed in relation to lake water Ca levels. The occurrence of D. ambigua was independent of Ca levels in Ontario lakes, whereas D. pulicaria was more likely to occur in lakes with relatively more Ca. In the laboratory, D. ambigua maintained lower levels of hemolymph Ca2+ across a range of low Ca levels (0.7 to 7 mg l-1) compared with D. pulicaria. The hemolymph pH remained steady across this Ca gradient in D. ambigua while it was significantly more acidic in D. pulicaria in the two lowest Ca treatments. While Ca2+ uptake was observed adjacent to the surface of D. ambigua individuals, Ca2+ loss was observed for D. pulicaria assayed under moderately high Ca levels. Based on these observations we propose that D. ambigua is able to survive in low Ca lakes by maintaining low free ionic Ca2+ levels in the hemolymph which minimizes the Ca gradient across the body wall in low Ca water thus limiting overall Ca loss and facilitating Ca2+ uptake.

The jellification of north temperate lakes.

Calcium (Ca) concentrations are decreasing in softwater lakes across eastern North America and western Europe. Using long-term contemporary and palaeo-environmental field data, we show that this is precipitating a dramatic change in Canadian lakes: the replacement of previously dominant pelagic herbivores (Ca-rich Daphnia species) by Holopedium glacialis, a jelly-clad, Ca-poor competitor. In some lakes, this transformation is being facilitated by increases in macro-invertebrate predation, both from native (Chaoborus spp.) and introduced (Bythotrephes longimanus) zooplanktivores, to which Holopedium, with its jelly coat, is relatively invulnerable. Greater representation by Holopedium within cladoceran zooplankton communities will reduce nutrient transfer through food webs, given their lower phosphorus content relative to daphniids, and greater absolute abundances may pose long-term problems to water users. The dominance of jelly-clad zooplankton will likely persist while lakewater Ca levels remain low.

Consequences of calcium decline on the embryogenesis and life history of Daphnia magna.

Ambient calcium is declining in thousands of soft-water lake habitats in temperate regions as a consequence of unsustainable forestry practices, decreased atmospheric calcium deposition and acidic deposition. As their exoskeleton is heavily reinforced with calcium, freshwater crustaceans have a high specific calcium requirement relative to other aquatic organisms. Daphnia, in particular, is an ideal crustacean for investigating the consequences of calcium decline because it is an abundant and important member of freshwater zooplankton communities. Although it has been established that adult and juvenile Daphnia have different tolerances to low ambient calcium as a result of their different life stage-specific calcium requirements, the consequences of declining calcium on embryonic development have never been investigated. Here, we describe the distribution of calcium in embryonic stages of D. magna and introduce a novel and easy to use staging scheme. We tested whether calcium can be traced from mothers to their offspring. Finally, we assessed the fitness consequences of maternal provisioning in limiting calcium environments. We found that while embryos require calcium for their development and moulting, they do not equilibrate with environmental calcium levels. Instead, we were able to trace calcium from mothers to their offspring. Furthermore, our data strongly suggest that females are faced with an allocation trade-off between providing calcium to their offspring and using it for growth and moulting. Together, these data provide novel insights into the consequences of calcium decline for freshwater zooplankton.

Synergistic interactions of biotic and abiotic environmental stressors on gene expression.

Understanding the response of organisms to multiple stressors is critical for predicting if populations can adapt to rapid environmental change. Natural and anthropogenic stressors often interact, complicating general predictions. In this study, we examined the interactive and cumulative effects of two common environmental stressors, lowered calcium concentration, an anthropogenic stressor, and predator presence, a natural stressor, on the water flea Daphnia pulex. We analyzed expression changes of five genes involved in calcium homeostasis - cuticle proteins (Cutie, Icp2), calbindin (Calb), and calcium pump and channel (Serca and Ip3R) - using real-time quantitative PCR (RT-qPCR) in a full factorial experiment. We observed strong synergistic interactions between low calcium concentration and predator presence. While the Ip3R gene was not affected by the stressors, the other four genes were affected in their transcriptional levels by the combination of the stressors. Transcriptional patterns of genes that code for cuticle proteins (Cutie and Icp2) and a sarcoplasmic calcium pump (Serca) only responded to the combination of stressors, changing their relative expression levels in a synergistic response, while a calcium-binding protein (Calb) responded to low calcium stress and the combination of both stressors. The expression pattern of these genes (Cutie, Icp2, and Serca) were nonlinear, yet they were dose dependent across the calcium gradient. Multiple stressors can have complex, often unexpected effects on ecosystems. This study demonstrates that the dominant interaction for the set of tested genes appears to be synergism. We argue that gene expression patterns can be used to understand and predict the type of interaction expected when organisms are exposed simultaneously to natural and anthropogenic stressors.

Food quantity affects the sensitivity of Daphnia to road salt.

Road deicing operations have raised chloride (Cl) levels in many temperate lakes in Europe and North America. These lakes vary widely in trophic status, but to date, no one has quantified the interaction between food quantity and road salt toxicity. We examined the effects of food quantity (particulate algal C concentration (C)) on the chronic toxicity of Cl to Daphnia in soft-water bioassays. There was a strong positive linear relationship (r(2) = 0.92 for NaCl and r(2) = 0.96 for CaCl2) between food quantity and Cl LC50. As food quantity increased from 0.2 to 1.0 mg C/L (levels characteristic of oligotrophic to eutrophic lakes, respectively), the chronic Cl LC50 increased from 55.7 to 284.8 mg Cl/L. Salt type (NaCl or CaCl2) did not affect the Cl LC50, Daphnia life history parameters, or the intrinsic rate of population increase (r). The life history parameter most sensitive to Cl was neonate production. Cl did not inhibit egg production, nor was the maternal lipid investment in eggs changed, but egg viability and the subsequent release of live neonates decreased as Cl levels increased and food decreased. Our results suggest the trophic status of lakes should be considered when assessing ecological threat from Cl.

Changes in water chemistry can disable plankton prey defenses.

The effectiveness of antipredator defenses is greatly influenced by the environment in which an organism lives. In aquatic ecosystems, the chemical composition of the water itself may play an important role in the outcome of predator-prey interactions by altering the ability of prey to detect predators or to implement defensive responses once the predator's presence is perceived. Here, we demonstrate that low calcium concentrations (<1.5 mg/L) that are found in many softwater lakes and ponds disable the ability of the water flea, Daphnia pulex to respond effectively to its predator, larvae of the phantom midge, Chaoborus americanus. This low-calcium environment prevents development of the prey's normal array of induced defenses, which include an increase in body size, formation of neck spines, and strengthening of the carapace. We estimate that this inability to access these otherwise effective defenses results in a 50-186% increase in the vulnerability of the smaller juvenile instars of Daphnia, the stages most susceptible to Chaoborus predation. Such a change likely contributes to the observed lack of success of daphniids in most low-calcium freshwater environments, and will speed the loss of these important zooplankton in lakes where calcium levels are in decline.

Application of Biotic Ligand and Toxic Unit modeling approaches to predict improvements in zooplankton species richness in smelter-damaged lakes near Sudbury, Ontario.

Using a 30-year record of biological and water chemistry data collected from seven lakes near smelters in Sudbury (Ontario, Canada) we examined the link between reductions of Cu, Ni, and Zn concentrations and zooplankton species richness. The toxicity of the metal mixtures was assessed using an additive Toxic Unit (TU) approach. Four TU models were developed based on total metal concentrations (TM-TU); free ion concentrations (FI-TU); acute LC50s calculated from the Biotic Ligand Model (BLM-TU); and chronic LC50s (acute LC50s adjusted by metal-specific acute-to-chronic ratios, cBLM-TU). All models significantly correlated reductions in metal concentrations to increased zooplankton species richness over time (p < 0.01) with a rank based on r(2) values of cBLM-TU > BLM-TU = FI-TU > TM-TU. Lake-wise comparisons within each model showed that the BLM-TU and cBLM-TU models provided the best description of recovery across all seven lakes. These two models were used to calculate thresholds for chemical and biological recovery using data from reference lakes in the same region. A threshold value of TU = 1 derived from the cBLM-TU provided the most accurate description of recovery. Overall, BLM-based TU models that integrate site-specific water chemistry-derived estimates of toxicity offer a useful predictor of biological recovery.

Communities contain closely related species during ecosystem disturbance.

Predicting community and species responses to disturbance is complicated by incomplete knowledge about species traits. A phylogenetic framework should partially solve this problem, as trait similarity is generally correlated with species relatedness, closely related species should have similar sensitivities to disturbance. Disturbance should thus result in community assemblages of closely related species. We tested this hypothesis with 18 disturbed and 16 reference whole-lake, long-term zooplankton data sets. Regardless of disturbance type, communities generally contained more closely related species when disturbed. This effect was independent of species richness, evenness, and abundance. Communities already under stress (i.e., those in acidic lakes) changed most when disturbed. Species sensitivities to specific disturbances were phylogenetically conserved, were independent of body size, and could be predicted by the sensitivities of close relatives within the same community. Phylogenetic relatedness can effectively act as a proxy for missing trait information when predicting community and species responses to disturbance.

Environmental stability and lake zooplankton diversity - contrasting effects of chemical and thermal variability.

Environmental variability in space and time is a primary mechanism allowing species that share resources to coexist. Fluctuating conditions are a double edged sword for diversity, either promoting coexistence through temporal niche partitioning or excluding species by stochastic extinctions. The net effect of environmental variation on diversity is largely unknown. We examined the association between zooplankton species richness in lakes and environmental variability on interannual, seasonal and shorter time scales, as well as long-term average conditions. We analyzed data on physical, chemical and biological limnology in 53 temperate zone lakes in North America and Europe sampled over a combined 1042 years. Large fluctuations in pH, phosphorus and dissolved organic carbon concentration on different time scales were associated with reduced zooplankton species richness. More species were found in lakes that showed greater temperature variation on all time scales. Environmental variability on different time scales showed similar or, in some cases, stronger associations with zooplankton species richness compared with long-term average conditions. Our results suggest that temporal fluctuations in the chemical environment tend to exclude zooplankton species while temperature variability promotes greater richness. The results indicate that anthropogenic increases in temporal variability of future climates may have profound effects on biodiversity.

Accumulated organic debris in catch basins improves the efficacy of S-methoprene against mosquitoes in Toronto, Ontario, Canada.

To control West Nile Virus in the greater Toronto area of Ontario, Canada, S-methoprene (Altosid XRbriquets 2.1% AI) is applied each year to storm water catch basins. Because the efficacy of the XRbriquets to reduce adult mosquito populations had not been evaluated locally and was influenced by organic debris in a pilot study, we compared the efficacy of the briquets in 17 sediment and debris-filled catch basins versus 20 catch basins that were vacuumed free of debris. Emergence rates approached 100% in the 5 untreated control catch basins. Emergence rates were significantly lower, and S-methoprene was detected more often and at higher levels, in debris-filled basins versus cleaned catch basins. Overall, 20% of pupae emerged from clean catch basins versus only 3% from debris-filled ones, the difference between treatments becoming significant after 26 days. S-methoprene and total organic carbon concentrations in the catch basins were positively correlated (P < 0.001). We hypothesize that S-methoprene is binding to the organic fraction in the water and sediment in the debris-filled basins, prolonging S-methoprene doses, which are reflected in lower mosquito emergence rates.

Diversity-stability relationship varies with latitude in zooplankton.

Analyses of temporal patterns of diversity across a wide range of taxa have found that more diverse communities often show smaller compositional changes over time. This generality indicates that high diversity is associated with greater temporal stability in species composition. We examined patterns of diversity and community stability in zooplankton time series data from 36 lakes sampled over a combined 483 years. The species-time relationship was flatter in more species-rich lakes in the temperate zone. However, high-latitude lakes had both low richness and low turnover. These patterns were consistent for turnover both within and among years. Daily, annual and long-term richness were all higher in large lakes while turnover was unaffected by the surface area. Richness on all time scales, as well as turnover within and among years, all declined at high latitude. Species-area relations and latitudinal gradients in richness therefore reflect different temporal components of diversity. Our results suggest that diversity shows strong associations with compositional stability that vary qualitatively across biogeographical provinces. Community stability increases with diversity among lakes in the temperate zone; however, the two are negatively correlated across latitudinal gradients. These patterns indicate that either the direct effects of diversity on stability or their covariance with environmental fluctuations vary with latitude.

Metabolomics confirms that dissolved organic carbon mitigates copper toxicity.

Reductions in atmospheric emissions from the metal smelters in Sudbury, Canada, produced major improvements in acid and metal contamination of local lakes and indirectly increased dissolved organic carbon (DOC) concentrations. Metal toxicity, however, has remained a persistent problem for aquatic biota. Integrating high-throughput, nontargeted mass spectrometry metabolomics with conventional toxicological measures elucidated the mediating effects of dissolved organic matter (DOM) on the toxicity of Cu to Daphnia pulex-pulicaria, a hybrid isolated from these soft water lakes. Two generations of daphniids were exposed to Cu (0-20 µg/L) at increasing levels of natural DOM (0-4 mg DOC/L). Added DOM reduced Cu toxicity monotonically with median lethal concentration values increasing from 2.3 µg/L Cu without DOM to 22.7 µg/L Cu at 4 mg DOC/L. Reproductive output similarly benefited, increasing with DOM, yet falling with increases in Cu. Second generation reproduction was more impaired than the first generation. Dissolved organic matter had a greater influence than Cu on the metabolic status of the daphniids. Putative identification of metabolite peaks indicated that DOM elevation increased the metabolic energy status of the first generation animals, but this benefit was reduced in the second generation, although evidence of increased oxidative stress was detected. These results indicate that Sudbury's terrestrial ecosystems should be managed to increase aquatic DOM supply to enable daphniid colonists to both survive and foster stable populations.

Predicting chronic copper and nickel reproductive toxicity to Daphnia pulex-pulicaria from whole-animal metabolic profiles.

The emergence of omics approaches in environmental research has enhanced our understanding of the mechanisms underlying toxicity; however, extrapolation from molecular effects to whole-organism and population level outcomes remains a considerable challenge. Using environmentally relevant, sublethal, concentrations of two metals (Cu and Ni), both singly and in binary mixtures, we integrated data from traditional chronic, partial life-cycle toxicity testing and metabolomics to generate a statistical model that was predictive of reproductive impairment in a Daphnia pulex-pulicaria hybrid that was isolated from an historically metal-stressed lake. Furthermore, we determined that the metabolic profiles of organisms exposed in a separate acute assay were also predictive of impaired reproduction following metal exposure. Thus we were able to directly associate molecular profiles to a key population response - reproduction, a key step towards improving environmental risk assessment and management.

The Temporal Coherence of Zooplankton Population Abundances in Neighboring North-Temperate Lakes.

We investigated the temporal coherence (i.e., the correlation or synchrony between time series) of annual abundances among populations of freshwater zooplankton in eight lakes in Ontario, Canada, from 1980 to 1992. We estimated temporal coherence using the intraclass correlation coefficient (ri). While values of ri were relatively low among comparisons of all eight lakes, they were statistically significant for three of the seven common cladoceran and copepod taxa (Bosmina longirostris, Leptodiaptomus minutus, and Mesocyclops edax). These significant positive correlations imply that a portion of the interannual variation in abundance was produced by factors operating on a scale larger than the individual lake catchments. Because the eight-lake analysis might obscure strong, but conflicting, patterns among lakes in the region, we identified homogeneous and temporally coherent subsets of lakes for each species using an exploratory stepwise deletion procedure. The resultant homogeneous subsets exhibited much greater temporal coherence, accounting for 47% (Eubosmina) to 84% (Leptodiaptomus) of the interannual variation in abundance. Our results suggest that the factors affecting annual variation in zooplankton abundance must be sought both within lakes and beyond their watersheds.

Regulation of Zooplankton Community Structure of an Acidified Lake by Chaoborus.

To test the hypothesis that the structure of zooplankton communities of acidified, fish-free lakes can be regulated by predation from larval Chaoborus, the rates of production of the principal prey species of Chaoborus in Swan Lake, a small, fish-less, acid lake near Sudbury, Canada, were compared with their estimated rates of consumption by Chaoborus. The production of Bosmina longirostris, the major crustacean zooplankter in the lake, rarely exceeded its apparent rate of loss to Chaoborus. In contrast, the production rate of Keratella taurocephala, the dominant rotifer in Chaoborus diets, virtually always exceeded the rate at which it was consumed. The unusually small contribution of Crustacea to total zooplankton biomass in the lake could be attributed to predation by Chaoborus. While Chaoborus can regulate zooplankton community structure in acidified, fish-free lakes, the frequency of occurrence of such control remains uncertain.

Recovery of crustacean zooplankton communities from acidification in Killarney Park, Ontario, 1971-2000: pH 6 as a recovery goal.

Despite reductions in atmospheric SO4(2-) deposition and resultant decreases in surface water acidity, widespread biological recovery from acidification has not yet been documented. Temporal trends in crustacean zooplankton species richness (number of species) and composition were examined between 1971-2000 in 46 Killarney Park lakes, Ontario, Canada, to assess the degree of biological recovery in lakes with significant water quality improvements, i.e. pH now > 6, compared to 2 other groups: i) lakes which never acidified; and ii) lakes which are still acidified (pH < 6). Time trends in species richness could not be distinguished among the 3 groups of lakes, nor did changes in species richness indicate recovery. In contrast, the zooplankton community composition of lakes in which the pH increased to above 6, as measured by a multivariate index of species abundances, changed from a "damaged" state to one typical of neutral lakes. Some recovery in composition was also documented for the acidic lakes. While still acidic, the pH levels of these lakes have risen. The extent and pace of recovery in Killarney Provincial Park bodes well for the future of other acidified regions in North America and Europe.

UVR sensitivity of Chaoborus larvae.

In small temperate lakes, predation by fish generally regulates the species structure and abundance of larval Chaoborus. Yet, Chaoborus abundance may also vary appreciably among lakes with no fish. Many fishless lakes in Sudbury, Ontario, have transparent waters. This raises the possibility that low abundance of Chaoborus in such lakes may be attributable to UVR-induced mortality. To determine whether UVR affects Chaoborus survival, we performed 6 in situ experiments over 2 to 4 day periods at 4 depths in Ruth-Roy Lake (a clear fishless lake with few Chaoborus). Third and fourth instar C. punctipennis were randomly allocated to 3 treatments: quartz (UVR+ PAR), OP3 acrylite (PAR only) and dark controls. Survival under UVR+PAR was significantly reduced in comparison with the other treatments. Survival under PAR only was high, and did not differ from the dark controls. Time to death increased with incubation depth and larval stage. These results suggest that the small Chaoborus population in Ruth-Roy Lake, and perhaps in other fishless, clear lakes may be attributed to UVR-induced mortality.

Littoral microcrustacean (Cladocera and Copepoda) indicators of acidification in Canadian Shield lakes.

We identify littoral microcrustacean indicators of acidification in 2 surveys of Canadian Shield lakes conducted 10 years apart. We found a total of 90 cladoceran and copepod species with richness increasing severalfold from acidic to nonacidic lakes. The fauna of the nonacidic lakes differed between the surveys. The 1987 survey employed activity traps, and caught more littoral taxa than the more recent, net-haul-based survey. Similar faunas were identified in the acidified lakes in both surveys, and several good indicator species were identified. For example, Acanthocycops vernalis was restricted to lakes with pH < 6. Sinobosmina sp. was very common but only in lakes with pH > 4.8. Tropocyclops extensus, Mesocyclops edax, and Sida crystallina were commonly found but only at pH > 5, and Chydorus faviformis only at pH > 5.9. These indicators showed promise in gauging the early stages of recovery from acidification in 3 lakes that were included in both surveys.