Plastic debris in lakes and reservoirs.

Plastic debris is thought to be widespread in freshwater ecosystems globally1. However, a lack of comprehensive and comparable data makes rigorous assessment of its distribution challenging2,3. Here we present a standardized cross-national survey that assesses the abundance and type of plastic debris (>250 µm) in freshwater ecosystems. We sample surface waters of 38 lakes and reservoirs, distributed across gradients of geographical position and limnological attributes, with the aim to identify factors associated with an increased observation of plastics. We find plastic debris in all studied lakes and reservoirs, suggesting that these ecosystems play a key role in the plastic-pollution cycle. Our results indicate that two types of lakes are particularly vulnerable to plastic contamination: lakes and reservoirs in densely populated and urbanized areas and large lakes and reservoirs with elevated deposition areas, long water-retention times and high levels of anthropogenic influence. Plastic concentrations vary widely among lakes; in the most polluted, concentrations reach or even exceed those reported in the subtropical oceanic gyres, marine areas collecting large amounts of debris4. Our findings highlight the importance of including lakes and reservoirs when addressing plastic pollution, in the context of pollution management and for the continued provision of lake ecosystem services.

Diet and food selection by fish larvae in turbid and clear water shallow temperate lakes.

Fish larvae play an important structuring role for their prey and show ontogenetic shifts in diet. Changes in diet differ between species and habitats and may also be affected by turbidity (eutrophication). We investigated the diet (stomach content) and the food selection (ratio of ingested prey and prey availability) of roach and perch larvae in a clear lake and of roach, perch and pikeperch larvae in a turbid lake multiple times during spring to autumn. The diet of the fish larvae changed with size, and for roach and perch larvae between the lakes. Coexisting species of fish larvae had different diets in the two lakes, pointing to resource partitioning; yet, in the clear lake, medium-sized larvae had a high diet overlap, suggesting a competitive relationship at this developmental stage. In the clear lake, roach larvae showed diel differentiation in diet, while perch demonstrated diet shifts between habitats, which probably aided in reducing competition and also evidenced an effect of light on the larval prey capture and/or predator-fish larvae interactions. In the turbid lake, roach and perch larvae did not reveal differences in diet between habitats or time of the day, owing to homogeneity of food items and poor light conditions. However, the diet of pikeperch larvae differed between day and night following daily variations in the abundance of its preferred prey. The roach larvae were highly selective for Bosmina, Daphnia and benthic cladocerans, perch larvae generally consumed what was available, while pikeperch primarily preyed on cyclopoid copepodites. We conclude that turbidity acted as a cover for fish larvae in the turbid lake. Under eutrophication-induced turbidity scenarios the effects of fish larvae on their prey are stronger (i.e., high selectivity for several resources) than that of larvae in clear waters, creating a negative feedback on the path to restore water clarity.

A European Multi Lake Survey dataset of environmental variables, phytoplankton pigments and cyanotoxins.

Under ongoing climate change and increasing anthropogenic activity, which continuously challenge ecosystem resilience, an in-depth understanding of ecological processes is urgently needed. Lakes, as providers of numerous ecosystem services, face multiple stressors that threaten their functioning. Harmful cyanobacterial blooms are a persistent problem resulting from nutrient pollution and climate-change induced stressors, like poor transparency, increased water temperature and enhanced stratification. Consistency in data collection and analysis methods is necessary to achieve fully comparable datasets and for statistical validity, avoiding issues linked to disparate data sources. The European Multi Lake Survey (EMLS) in summer 2015 was an initiative among scientists from 27 countries to collect and analyse lake physical, chemical and biological variables in a fully standardized manner. This database includes in-situ lake variables along with nutrient, pigment and cyanotoxin data of 369 lakes in Europe, which were centrally analysed in dedicated laboratories. Publishing the EMLS methods and dataset might inspire similar initiatives to study across large geographic areas that will contribute to better understanding lake responses in a changing environment.

Life history response of Daphnia magna to a mixotrophic golden alga, Poterioochromonas sp., at different food levels.

The toxicity of Poterioochromonas to Daphnia magna was investigated at different food (Scenedesmus acutus) levels. Poterioochromonas alone of 0.4-20 mg C L(-1) was not acutely toxic to D. magna, but did not support D. magna growth, either. When fed mixed diets (2 mg C L(-1) in total), D. magna's survival and reproduction were significantly depressed when Poterioochromonas comprised above 50%, likely due to the inhibition of food ingestion. Large juveniles were less sensitive to poor quality food than neonates. Therefore, Poterioochromonas may affect D. magna living to various extents depending on its concentration, age structure of D. magna populations and availability of other food.

Early signs of lethal effects in Daphnia magna (Branchiopoda, Cladocera) exposed to the insecticide cypermethrin and the fungicide azoxystrobin.

This study presents the effects of sublethal concentrations of pesticides cypermethrin and azoxystrobin on the activity of several physiological parameters of egg-carrying Daphnia magna studied using a video-image technique. Single tethered daphnids were continuously recorded for 24 h of pesticide exposure, and the activity of the heart, the filtering limbs, the mandibles, and the focal spine were subsequently analyzed. Acute toxicity tests based on the criteria of immobilization were performed on egg-carrying D. magna, and sublethal concentrations of 0.1, 1.0, and 10 µg/L cypermethrin and 0.5, 1.0, and 2.0 mg/L azoxystrobin were established. At a concentration as low as 0.1 µg/L cypermethrin, the 5% effective concentration after 24 h of exposure (EC5,24h), the activity of the focal spine increased and the filtering limb activity decreased. The activity of the mandibles was reduced by exposure to 1.0 (EC18,24 h) and 10 µg/L (EC41,24 h) cypermethrin, whereas heart activity increased at a concentration of 10 µg/L (EC41,24 h). With regard to azoxystrobin, the activity of all response parameters except the focal spine decreased by exposure to 0.5 mg/L (EC4,24h) azoxystrobin. The focal spine was not affected by azoxystrobin. The results show that physiological mechanisms important for ingestion of food in D. magna may be impaired by low concentrations of commonly used pesticides.

Life history responses of Daphnia magna feeding on toxic Microcystis aeruginosa alone and mixed with a mixotrophic Poterioochromonas species.

The toxic effects of a mixotrophic golden alga (Poterioochromonas sp. strain ZX1) and a cyanobacterium Microcystis aeruginosa grazed by Poterioochromonas to a cladoceran were investigated through life history experiments using Daphnia magna. Poterioochromonas cultured in two ways (fed M. aeruginosa in an inorganic medium or grown in an organic medium) both induced starvation-like effects on D. magna, indicating that Poterioochromonas is neither acutely toxic nor a good food for D. magna. Despite a microcystin-LR content of 10(-8)microgcell(-1) in M. aeruginosa, no toxins were accumulated in Poterioochromonas fed the cyanobacterium. The toxic effect of M. aeruginosa to D. magna was significantly reduced in the presence of Poterioochromonas, which may be performed in two ways: decrease M. aeruginosa cells ingestion of D. magna by grazing on M. aeruginosa; and decrease the toxicity of the medium by degrading the toxins released by M. aeruginosa. This study provides new information on the interactions between a cyanobacterium and its grazer under laboratory conditions and may increase our understanding of the ecological significance of such interactions in the aquatic food webs.

Feeding characteristics of a golden alga (Poterioochromonas sp.) grazing on toxic cyanobacterium Microcystis aeruginosa.

Microcystis aeruginosa has quickly risen in infamy as one of the most universal and toxic bloom-forming cyanobacteria. Here we presented a species of golden alga (Poterioochromonas sp. strain ZX1), which can feed on toxic M. aeruginosa without any adverse effects from the cyanotoxins. Using flow cytometry, the ingestion and maximal digestion rates were estimated to be 0.2 approximately 1.2 and 0.2 M. aeruginosa cells (ZX1 cell)(-1)h(-1), respectively. M. aeruginosa in densities below 10(7)cells mL(-1) could be grazed down by ZX1, but no significant decrease was observed when the initial density was 3.2 x 10(7)cells mL(-1). ZX1 grazing was a little influenced by the light intensity (0.5 approximately 2500l x) and initial pH of the medium (pH=5.0 approximately 9.5). ZX1 could not survive in continuous darkness for longer than 10 days. The pH value was adjusted to 8 by ZX1 while to 10 by M. aeruginosa. This study may shed light on understanding the ecological interactions between M. aeruginosa and mixotrophic Poterioochromonas sp. in aquatic ecosystems.

Clonal variation in physiological responses of Daphnia magna to the strobilurin fungicide azoxystrobin.

Because of its high grazing potential, Daphnia magna is an ecologically important species in aquatic food webs. This is especially true in small, shallow ponds lacking fish, where grazing by D. magna may have a relatively higher impact on water clarity as compared to larger lakes. Thus, a reduction in daphnid abundance may have dramatic ecological consequences for shallow ponds. At the same time, shallow ponds in close proximity to agricultural areas likely experience higher concentrations of pesticides because of runoff, spray drift, and drain flow. In the present study, the acute and chronic physiological effects of the strobilurin fungicide azoxystrobin on three clones of D. magna originating from different Danish lakes were evaluated. Significant clonal variation in the sensitivity of D. magna toward azoxystrobin was demonstrated. One clone had a 48-h median lethal concentration (LC50) of 0.277 mg/L (95% confidence limits [CL], 0.145 and 0.427 mg/L), which is comparable to the value widely used in risk assessments (0.259 mg/L). The two remaining clones were far more sensitive, however, and had LC50s of 0.071 mg/L (95% CL, 0.034 and 0.126 mg/L) and 0.098 mg/L (95% CL, 0.066 and 0.139 mg/L), respectively. Furthermore, through respiration measurements and life-table experiments, sublethal stress was shown to exist at exposure to an ecologically relevant concentration (0.026 microg/L). Based on these results, we may expect changes in daphnid populations at azoxystrobin concentrations much lower than previously thought. Thus, ponds in the agricultural areas may experience changes in food-web structure even at very low concentrations of azoxystrobin.

Inter-annual growth of Arctic charr (Salvelinus alpinus, L.) in relation to climate variation.

BACKGROUND: Major changes in climate have been observed in the Arctic and climate models predict further amplification of the enhanced greenhouse effect at high-latitudes leading to increased warming. We propose that warming in the Arctic may affect the annual growth conditions of the cold adapted Arctic charr and that such effects can already be detected retrospectrally using otolith data. RESULTS: Inter-annual growth of the circumpolar Arctic charr (Salvelinus alpinus, L.) was analysed in relation to climatic changes observed in the Arctic during the last two decades. Arctic charr were sampled from six locations at Qeqertarsuaq in West Greenland, where climate data have been recorded since 1990. Two fish populations met the criteria of homogeny and, consequently, only these were used in further analyses. The results demonstrate a complex coupling between annual growth rates and fluctuations in annual mean temperatures and precipitation. Significant changes in temporal patterns of growth were observed between cohorts of 1990 and 2004. CONCLUSION: Differences in pattern of growth appear to be a consequence of climatic changes over the last two decades and we thereby conclude that climatic affects short term and inter-annual growth as well as influencing long term shifts in age-specific growth patterns in population of Arctic charr.

Frequency of inhibitors of daphnid trypsin in the widely distributed cyanobacterial genus Planktothrix.

Recent findings showed that inhibition of the digestive enzyme trypsin by cyanobacterial metabolites can result in the death of the microcrustacean Daphnia. Compounds that are active against daphnid trypsin can therefore be considered as potentially toxic to Daphnia. Here we reported on the frequency of such compounds in the widely distributed cyanobacterial genus Planktothrix. Of the 89 Planktothrix strains analysed, about 70% produced inhibitors of daphnid trypsin. The strains tested positive represented three common Planktothrix species and were isolated from diverse localities and geographical regions. Our findings suggest therefore that inhibitors of daphnid trypsin are common in Planktothrix and maybe other cyanobacterial genera. These compounds should therefore be considered in future studies on the chemical ecology of cyanobacteria.

Cyanobacterial protease inhibitor microviridin J causes a lethal molting disruption in Daphnia pulicaria.

Laboratory experiments identified microviridin J as the source of a fatal molting disruption in Daphnia species organisms feeding on Microcystis cells. The molting disruption was presumably linked to the inhibitory effect of microviridin J on daphnid proteases, suggesting that hundreds of further cyanobacterial protease inhibitors must be considered potentially toxic to zooplankton.

Isolation, characterization, and quantitative analysis of Microviridin J, a new Microcystis metabolite toxic to Daphnia.

This paper describes the purification and characterization of microviridin J. a newly discovered metabolite of Microcystis that causes a lethal molting disruption in Daphnia spp., upon ingestion of living cyanobacterial cells. Microviridin J consists of an acetylated chain of 13 amino acids arranged in three rings and two side chains. Unlike other known isoforms of microviridin, microviridin J contains arginine that imparts a unique solution conformation characterized by proximal hydrophobic interactions between Arg and other regions of the molecule. This eventually results in the formation and stabilization of an additional ring system. Microviridin J potently inhibits porcine trypsin, bovine chymotrypsin, and daphnid trypsin-like proteases. The activity against trypsin is most likely due to Arg and its distinctive conformational interactions. Overall, the data presented for microviridin J emphasize once again the ability of cyanobacteria to produce numerous and potent environmental toxins.

Laboratory studies of dissolved radiolabelled microcystin-LR in lake water.

The fate of dissolved microcystin-LR was studied in laboratory experiments using surface water taken from a eutrophic lake. Based on initial range finding, a concentration of 50 microg l(-1) dissolved 14C-microcystin-LR was selected for subsequent time-course experiments. The first was performed in May before the cyanobacterial bloom season and low increases in the radioactivity of particulate fractions occurred with an approx. halving of the cyano-toxin during 4 days. The radioactivity of the dissolved fraction remained stable and there was no significant formation of radiolabelled inorganic carbon. A second time-course experiment was performed in September during the cyanobacterial bloom season. At the end of the four-day incubation period, the microcystin-LR concentration had decreased to an undetectable level and 24% of the added radiolabelled substance was found in different particulate fractions. The study demonstrated that biodegradation of dissolved microcystin-LR occurred in water collected at a lake surface with carbon dioxide as a major end-product.

Effects of the pyrethroid insecticide, cypermethrin, on a freshwater community studied under field conditions. I. Direct and indirect effects on abundance measures of organisms at different trophic levels.

The effects of the pyrethroid insecticide cypermethrin on a natural freshwater community were studied in small in situ enclosures over an 11-day period. The experiment was conducted in a eutrophic lake using a regression design that included three untreated controls and a gradient of six unreplicated cypermethrin concentrations, ranging from 0.01 to 6.1 microg/l. This paper is the first in a series of two, and describes the fate of cypermethrin and its effects on the abundance of crustaceans, rotifers, protozoans (cilliates and heterotrophic nanoflagellates (HNF)) and bacteria and the biomass of periphytic and planktonic algae. The concentration of cypermethrin decreased quickly during the experiment, with a half-life of 48 h for the total and 25 h for the dissolved fractions of cypermethrin, respectively. Cypermethrin proved to be acutely toxic to crustaceans in enclosures receiving nominal cypermethrin concentrations of >/=0.13 microg/l. No Effect Concentration (NEC) and median Effect Concentration (EC(50)) for the total crustacean community and cladoceran and copepod subgroups ranged between 0.02-0.07 and 0.04-0.17 microg/l, respectively, with copepods being less sensitive than cladocerans. The abundance of rotifers, protozoans and bacteria and the chlorophyll-a concentration of planktonic and periphytic algae was significantly related to the concentration of cypermethrin. All groups proliferated within 2-7 days after the cypermethrin application in those enclosures where the abundance of crustaceans was seriously affected by cypermethrin (i.e. >/=0.13 microg/l). We hypothesise that the proliferation of rotifers, protozoans, bacteria and algae was due to a reduced grazer control from crustaceans and thereby mediated indirectly by cypermethrin. The results of this experiment provide knowledge on how an entire microplankton community may respond to pyrethroids in nature, and the indirect effects observed on the community clearly demonstrates the necessity of multispecies field experiments in ecotoxicological risk assessment.

Effects of the pyrethroid insecticide cypermethrin on a freshwater community studied under field conditions. II. Direct and indirect effects on the species composition.

The effects of cypermethrin, a commonly used pyrethroid insecticide, were studied in small in situ enclosures situated in an eutrophic lake over an 11-day period. The experimental design used a regression principle that included three untreated controls and a gradient of six unreplicated cypermethrin concentrations, ranging from 0.01 to 6 microg/l. This paper is the second in a series of two and describes the effects on the species composition of the crustacean, rotifer, periphyton and phytoplankton communities. Multivariate ordination technique (redundancy analysis (RDA) combined with Monte Carlo permutation tests) showed that exposure to cypermethrin caused significant changes in the species composition of the communities. Changes in the structure of the communities were observed following exposure to a nominal concentration of 0.13 microg cypermethrin per litre above. The direct acute effect of exposure to cypermethrin was a rapid decrease of many species of crustacean zooplankton. The alterations in crustacean species composition were probably due to variations in susceptibility to the direct toxic effects of cypermethrin. No effects concentration (NEC) for individual zooplankton species were calculated using inverse regression and revealed that copepod nauplii were the most sensitive (NEC=0.01 microg/l) of the crustacean groups examined. The observed alterations of the species composition of the autotrophic communities as well as of the rotifers were most likely caused indirectly by cypermethrin, mediated through the direct negative effects of the insecticide on the crustacean grazers. The results of this experiment provide further knowledge about the direct and indirect effects of pesticide stress on the ecosystem level. They also show that there is a variation in sensitivity between different species of zooplankton under natural conditions and thus exemplify the necessity of multispecies approaches in the risk assessment of pesticides.

Influence of LAS on marine calanoid copepod population dynamics and potential reproduction.

The toxicity of linear alkyl benzene sulfonate (LAS) to marine invertebrates is well documented under laboratory conditions using single-species tests. It is less known how LAS affects natural populations of aquatic organisms. We hypothesised that LAS was more toxic to the calanoid copepod Acartia sp. under natural conditions than Acartia tonsa under cultured conditions in the laboratory. This hypothesis was checked by a direct comparison of LAS toxicity in single-species and model (mesocosm) studies. The acute and sublethal effects of LAS on the survival and egg production of laboratory reared A. tonsa were examined by standard test, i.e. incubation with LAS without food for 24-72 h. The LC(50) and EC(50) values averaged 1.23 and 0.74 mg l(-1) for survival and egg production, respectively. These values are comparable to previous reports. The effects of LAS on a natural copepod community were also investigated under in situ conditions. A series of seven mesocosms (holding approx. 3 m(3) of seawater each) was established with two mesocosms being controls without LAS and five mesocosms with increasing concentrations of LAS ranging from 0.1 to 6.5 mg l(-1) applied as a single dose. The indigenous copepod community, dominated by Acartia sp. and Centropages sp., responded clearly to LAS concentrations above 0.1 mg l(-1). The calculated no effect value was 0.14 mg LAS l(-1) (95% CI=0.08-1.82 mg LAS l(-1)) for the entire copepod community including all development stages after 24 h exposure. The increased sensitivity under in situ conditions was probably promoted by the suboptimal growth conditions, e.g. no saturated food concentration or inadequate nutritive values of the food. The amount of food expressed as chlorophyll concentration was low (around 2 microg chl. a l(-1)) but was not affected by LAS. It appeared that the naupliar stages of Acartia and Centropages were the least affected by LAS and that new cohorts were able to develop 15 days after the dosing with LAS.

Comparing sensitivity of ecotoxicological effect endpoints between laboratory and field.

Extrapolating toxicant effects with a fixed application factor (AF) approach or one of the species sensitivity distribution (SSD) models presumes that toxicant effects on single, individual-level endpoints reflect effects at the ecosystem level. Measured effect concentrations on plankton from multispecies field tests using tributyltin (TBT) and linear alkylbenzene sulfonates (LAS) were compared with published laboratory single-species test results and measured in situ concentrations. Extrapolation methods were evaluated by comparing predicted no-effect concentrations (PNECs), calculated by AF and SSD models with NOECs and E(L)C(50)s obtained from field studies. Overall, structural parameters were more sensitive than functional ones. Measured effect concentrations covered approximately the same range between laboratory and field experiments. Both SSD and AF approaches provide PNECs that appear to be protective for ecosystems. The AF approach is simpler to apply than the SSD models and results in PNECs that are no less conservative. Calculated PNEC values and the lowest field effect concentrations were lower than measured environmental concentrations for both substances, indicating that they may pose a risk to marine ecosystems.