Sensitivity ranking for freshwater invertebrates towards hydrocarbon contaminants.

Hydrocarbons have an utmost economical importance but may also cause substantial ecological impacts due to accidents or inadequate transportation and use. Currently, freshwater biomonitoring methods lack an indicator that can unequivocally reflect the impacts caused by hydrocarbons while being independent from effects of other stressors. The aim of the present study was to develop a sensitivity ranking for freshwater invertebrates towards hydrocarbon contaminants, which can be used in hydrocarbon-specific bioindicators. We employed the Relative Sensitivity method and developed the sensitivity ranking S hydrocarbons based on literature ecotoxicological data supplemented with rapid and mesocosm test results. A first validation of the sensitivity ranking based on an earlier field study has been conducted and revealed the S hydrocarbons ranking to be promising for application in sensitivity based indicators. Thus, the first results indicate that the ranking can serve as the core component of future hydrocarbon-specific and sensitivity trait based bioindicators.

Pesticide impact on aquatic invertebrates identified with Chemcatcher® passive samplers and the SPEAR(pesticides) index.

Pesticides negatively affect biodiversity and ecosystem function in aquatic environments. In the present study, we investigated the effects of pesticides on stream macroinvertebrates at 19 sites in a rural area dominated by forest cover and arable land in Central Germany. Pesticide exposure was quantified with Chemcatcher® passive samplers equipped with a diffusion-limiting membrane. Ecological effects on macroinvertebrate communities and on the ecosystem function detritus breakdown were identified using the indicator system SPEARpesticides and the leaf litter degradation rates, respectively. A decrease in the abundance of pesticide-vulnerable taxa and a reduction in leaf litter decomposition rates were observed at sites contaminated with the banned insecticide Carbofuran (Toxic Units≥-2.8), confirming the effect thresholds from previous studies. The results show that Chemcatcher® passive samplers with a diffusion-limiting membrane reliably detect ecologically relevant pesticide pollution, and we suggest Chemcatcher® passive samplers and SPEARpesticides as a promising combination to assess pesticide exposure and effects in rivers and streams.

Environmental stressors can enhance the development of community tolerance to a toxicant.

Ecosystems are subject to a combination of recurring anthropogenic and natural disturbances, such as climate change and pesticide exposure. Biological communities are known to develop tolerance to recurring disturbances due to successive changes at both the community and organismal levels. However, information on how additional stressors may affect the development of such community tolerance is scarce to date. We studied the influence of hydrological disturbance on the reaction of zooplankton communities to repeated insecticide pulses in outdoor microcosms. The communities were exposed to three successive pulses of the insecticide esfenvalerate (0.03, 0.3, and 3 µg/L) and to the gradual removal of water and its subsequent replacement over three cycles or to a constant water level. Except at the highest esfenvalerate concentration, the communities developed tolerance to the toxicant, as indicated by their decreasing reaction to subsequent insecticide applications, and this development was enhanced by hydrological disturbance. The pronounced decline of the key taxa Daphnia spp. through the combined action of the two stressors was identified as the main mechanism responsible for the increase in community tolerance under a fluctuating water level. Under a constant water level, the abundance of Daphnia spp. did not decrease significantly without the insecticide treatment, indicating that other mechanisms were responsible for the observed community tolerance. The present study shows that additional stressors can facilitate the development of community tolerance and that such facilitation is propagated through community-level mechanisms.

Statistics matter: data aggregation improves identification of community-level effects compared to a commonly used multivariate method.

The identification of the effects of toxicants on biological communities is hampered by the complexity and variability of communities. To overcome these challenges, the trait-based SPEAR approach has been developed. This approach is based on (i) identifying the vulnerable taxa using traits and (ii) aggregating these taxa into a group to reduce the between-replicate differences and scattered low-abundance distribution, both of which are typical for biological communities. This approach allows for reduction of the noise and determination of the effects of toxicants at low concentrations in both field and mesocosm studies. However, there is a need to quantitatively investigate its potential for mesocosm data evaluations and application in the ecological risk assessment of toxicants. In the present study, we analysed how the aggregation of the sensitive taxa can facilitate the identification of the effects. We used empirical data from a long-term mesocosm experiment with stream invertebrates and an insecticide as well as a series of simulated datasets characterised by different degrees of data matrix saturation (corresponding to different sampling efforts), numbers of replicates, and between-replicate differences. The analyses of both the empirical and simulated data sets revealed that the taxa aggregation approach allows for the detection of effects at a lower saturation of the data matrices, smaller number of replicates, and higher between-replicate differences when compared to the multivariate statistical method redundancy analysis. These improvements lead to a higher sensitivity of the analysed systems, as long-term effects were detected at lower concentrations (up to 1,000 times). These outcomes suggest that methods based on taxa aggregation have a strong potential for use in mesocosm data evaluations because mesocosm studies are usually poorly replicated, have high between-replicate variability, and cannot be exhaustively sampled due to technical and financial constraints.

How to characterize chemical exposure to predict ecologic effects on aquatic communities?

Reliable characterization of exposure is indispensable for ecological risk assessment of chemicals. To deal with mixtures, several approaches have been developed, but their relevance for predicting ecological effects on communities in the field has not been elucidated. In the present study, we compared nine metrics designed for estimating the total toxicity of mixtures regarding their relationship with an effect metric for stream macroinvertebrates. This was done using monitoring data of biota and organic chemicals, mainly pesticides, from five studies comprising 102 streams in several regions of Europe and South-East Australia. Mixtures of less than 10 pesticides per water sample were most common for concurrent exposure. Exposure metrics based on the 5% fraction of a species sensitivity distribution performed best, closely followed by metrics based on the most sensitive species and Daphnia magna as benchmark. Considering only the compound with the highest toxicity and ignoring mixture toxicity was sufficient to estimate toxicity in predominantly agricultural regions with pesticide exposure. The multisubstance Potentially Affected Fraction (msPAF) that combines concentration and response addition was advantageous in the study where further organic toxicants occurred. We give recommendations on exposure metric selection depending on data availability and the involved compounds.

Pesticides reduce regional biodiversity of stream invertebrates.

The biodiversity crisis is one of the greatest challenges facing humanity, but our understanding of the drivers remains limited. Thus, after decades of studies and regulation efforts, it remains unknown whether to what degree and at what concentrations modern agricultural pesticides cause regional-scale species losses. We analyzed the effects of pesticides on the regional taxa richness of stream invertebrates in Europe (Germany and France) and Australia (southern Victoria). Pesticides caused statistically significant effects on both the species and family richness in both regions, with losses in taxa up to 42% of the recorded taxonomic pools. Furthermore, the effects in Europe were detected at concentrations that current legislation considers environmentally protective. Thus, the current ecological risk assessment of pesticides falls short of protecting biodiversity, and new approaches linking ecology and ecotoxicology are needed.

Elevated temperature prolongs long-term effects of a pesticide on Daphnia spp. due to altered competition in zooplankton communities.

Considerable research efforts have been made to predict the influences of climate change on species composition in biological communities. However, little is known about how changing environmental conditions and anthropogenic pollution can affect aquatic communities in combination. We investigated the influence of short warming periods on the response of a zooplankton community to the insecticide esfenvalerate at a range of environmentally realistic concentrations (0.03, 0.3 and 3 µg L(-1) ) in 55 outdoor pond microcosms. Warming periods increased the cumulative water temperature, but did not exceed the maximum temperature measured under ambient conditions. Under warming conditions alone the abundance of some zooplankton taxa increased selectively compared to ambient conditions. This resulted in a shift in the community composition that had not recovered by the end of the experiment, 8 weeks after the last warming period. Regarding the pesticide exposure, short-term effects of esfenvalerate on the community structure and the sensitive taxa Daphnia spp. did not differ between the two temperature regimes. In contrast, long-term effects of esfenvalerate on Daphnia spp., a taxon that did not benefit from elevated temperatures, were observed twice as long under warming than under ambient conditions. This resulted in long-term effects on Daphnia spp. until 4 months after contamination at 3 µg L(-1) esfenvalerate. Under both temperature regimes, we identified strength of interspecific competition as the mechanism determining the time until recovery. However, enhanced interspecific competition under warming conditions was prolonged and explained the delayed recovery of Daphnia spp. from esfenvalerate. These results show that, for realistic prediction of the combined effects of changing environmental factors and toxicants on sensitive taxa, the impacts of stressors on the biotic interactions within the community need to be considered.

Two stressors and a community: effects of hydrological disturbance and a toxicant on freshwater zooplankton.

Climate change models predict an increase in the frequency and intensity of extreme fluctuations in water level in aquatic habitats. Therefore, it is necessary to understand the combined effects of hydrological fluctuations and toxicants on aquatic biological communities. We investigated the individual and combined effects of the insecticide esfenvalerate and recurring fluctuations in water level on zooplankton communities in a system of 55 outdoor pond microcosms. The communities were exposed to esfenvalerate contamination as a single pulse (at 0.03, 0.3, or 3µg/L) and gradual removal of water and its subsequent replacement over three cycles and monitored until 84 days after contamination. The results showed that the sensitivities of the community and its constituent populations to the toxicant were increased by the hydrological stress. Specifically, for both the community structure and abundance of Daphnia spp. the lowest-observed-effect concentrations (LOEC) were 0.03 and 0.3µg/L for the series with fluctuating and constant water levels, respectively. Despite these differences in sensitivity, the interactive effects of the two stressors were found to be additive for both the community structure and the abundance of the most affected species. Presumably, it was not possible to detect synergism due to the strong individual effects of the water level fluctuations. Recovery times in the series exposed to the highest pesticide concentration were 64 and 55 days under fluctuating and constant water level regimes, respectively. Competition and water quality are suggested to be the major factors that underlie the observed effects of fluctuations in the water level. For the ecological risk assessment of toxicants, the present results suggest that (i) community sensitivity may vary substantially, depending on the environmental context, and (ii) this variability can be assessed experimentally to derive safety factors (coefficients used to avoid unexpected effects and define safe concentrations of toxicants) based on empirical findings. This contrasts with the current approach where such factors are usually defined arbitrarily.

Intraspecific competition increases toxicant effects in outdoor pond microcosms.

Competition is a ubiquitous factor in natural populations and has been reported to alter the ecological impact of xenobiotics. We investigated conditions that mirror the natural variation of environmental factors. For this, different treatments were applied to 96 outdoor pond microcosms by shading the ponds and harvesting the communities. Then, the effect of esfenvalerate (0.03, 0.3, and 3 µg/L) on populations of Daphnia spp. was investigated. The pesticide effect and the sensitivity of Daphnia spp. in the context of a zooplankton community was increased by intraspecific competition 11 days after contamination. This relationship was most pronounced at 0.03 and 0.3 µg/L esfenvalerate, which were the concentrations that led to partial mortality. In contrast, interspecific interaction did not significantly alter the effect of the toxicant on Daphnia spp. Modelled concentration-response curves showed that the negative effects of the pesticide differed by a factor of up to 100 depending on the strength of intraspecific competition. In addition, a wider range of concentrations led to negative effects at high levels of intraspecific competition than at low levels. We argue that increased intraspecific competition reduces the availability of resources at the individual level and thereby increases the effect of contaminants. This knowledge about the interaction between competition and the response to toxicants is important in assessing the effects of these factors under field conditions.

Thresholds for the effects of pesticides on invertebrate communities and leaf breakdown in stream ecosystems.

We compiled data from eight field studies conducted between 1998 and 2010 in Europe, Siberia, and Australia to derive thresholds for the effects of pesticides on macroinvertebrate communities and the ecosystem function leaf breakdown. Dose-response models for the relationship of pesticide toxicity with the abundance of sensitive macroinvertebrate taxa showed significant differences to reference sites at 1/1000 to 1/10,000 of the median acute effect concentration (EC50) for Daphnia magna, depending on the model specification and whether forested upstream sections were present. Hence, the analysis revealed effects well below the threshold of 1/100 of the EC50 for D. magna incorporated in the European Union Uniform Principles (UP) for registration of single pesticides. Moreover, the abundances of sensitive macroinvertebrates in the communities were reduced by 27% to 61% at concentrations related to 1/100 of the EC50 for D. magna. The invertebrate leaf breakdown rate was positively linearly related to the abundance of pesticide-sensitive macroinvertebrate species in the communities, though only for two of the three countries examined. We argue that the low effect thresholds observed were not mainly because of an underestimation of field exposure or confounding factors. From the results gathered we derive that the UP threshold for single pesticides based on D. magna is not protective for field communities subject to multiple stressors, pesticide mixtures, and repeated exposures and that risk mitigation measures, such as forested landscape patches, can alleviate effects of pesticides.

Interspecific competition delays recovery of Daphnia spp. populations from pesticide stress.

Xenobiotics alter the balance of competition between species and induce shifts in community composition. However, little is known about how these alterations affect the recovery of sensitive taxa. We exposed zooplankton communities to esfenvalerate (0.03, 0.3, and 3 µg/L) in outdoor microcosms and investigated the long-term effects on populations of Daphnia spp. To cover a broad and realistic range of environmental conditions, we established 96 microcosms with different treatments of shading and periodic harvesting. Populations of Daphnia spp. decreased in abundance for more than 8 weeks after contamination at 0.3 and 3 µg/L esfenvalerate. The period required for recovery at 0.3 and 3 µg/L was more than eight and three times longer, respectively, than the recovery period that was predicted on the basis of the life cycle of Daphnia spp. without considering the environmental context. We found that the recovery of sensitive Daphnia spp. populations depended on the initial pesticide survival and the related increase of less sensitive, competing taxa. We assert that this increase in the abundance of competing species, as well as sub-lethal effects of esfenvalerate, caused the unexpectedly prolonged effects of esfenvalerate on populations of Daphnia spp. We conclude that assessing biotic interactions is essential to understand and hence predict the effects and recovery from toxicant stress in communities.

Ecotoxicology and macroecology--time for integration.

Despite considerable progress in ecotoxicology, it has become clear that this discipline cannot answer its central questions, such as, "What are the effects of toxicants on biodiversity?" and "How the ecosystem functions and services are affected by the toxicants?". We argue that if such questions are to be answered, a paradigm shift is needed. The current bottom-up approach of ecotoxicology that implies the use of small-scale experiments to predict effects on the entire ecosystems and landscapes should be merged with a top-down macroecological approach that is directly focused on ecological effects at large spatial scales and consider ecological systems as integral entities. Analysis of the existing methods in ecotoxicology, ecology, and environmental chemistry shows that such integration is currently possible. Therefore, we conclude that to tackle the current pressing challenges, ecotoxicology has to progress using both the bottom-up and top-down approaches, similar to digging a tunnel from both ends at once.

Ultraviolet radiation increases sensitivity to pesticides: synergistic effects on population growth rate of Daphnia magna at low concentrations.

In the present study we aimed to investigate whether UV-B radiation can exacerbate effects of pesticides fenoxycarb, pirimicarb, and tebufenpyrad on the survival, reproduction, and population growth rate of the standard test species Daphnia magna. We applied sublethal pesticides' concentrations and UV doses and observed no effects on survival. However, we observed synergistic effects of UV and pesticides on both cumulative reproduction and population growth rate (21 days) for fenoxycarb (100 µg/L) and pirimicarb (10 µg/L), but a less-than-additive effect for tebufenpyrad (5-10 µg/L). In the series exposed to UV and fenoxycarb or pirimicarb, the population growth rate dropped down to 0.1, while in the control series it was around 0.3. The results indicate that concentrations of some toxicants that are nontoxic in standard tests can cause harmful population-level effects when combined with UV.

Environmental context determines community sensitivity of freshwater zooplankton to a pesticide.

The environment is currently changing worldwide, and ecosystems are being exposed to multiple anthropogenic pressures. Understanding and consideration of such environmental conditions is required in ecological risk assessment of toxicants, but it remains basically limited. In the present study, we aimed to determine how and to what extent alterations in the abiotic and biotic environmental conditions can alter the sensitivity of a community to an insecticide, as well as its recovery after contamination. We conducted an outdoor microcosm experiment in which zooplankton communities were exposed to the insecticide esfenvalerate (0.03, 0.3, and 3 µg/L) under different regimes of solar radiation and community density, which represented different levels of food availability and competition. We focused on the sensitivity of the entire community and analysed it using multivariate statistical methods, such as principal response curves and redundancy analysis. The results showed that community sensitivity varied markedly between the treatments. In the experimental series with the lowest availability of food and strongest competition significant effects of the insecticide were found at the concentration of 0.03 µg/L. In contrast, in the series with relatively higher food availability and weak competition such effects were detected at 3 µg/L only. However, we did not find significant differences in the community recovery rates between the experimental treatments. These findings indicate that environmental context is more important for ecotoxicological evaluation than assumed previously.

What environmental factors are important determinants of structure, species richness, and abundance of mosquito assemblages?

Despite numerous ecological studies with mosquitoes, it remains unclear what environmental factors are the most important determinants of structure, species richness, and abundance of mosquito assemblages. In the current study, we investigated relations between these characteristics of mosquito larvae assemblages and environmental factors in a large set of different habitats. Particular objectives were (1) to rank the factors regarding their explanatory power, and (2) to quantify the contribution of major sets of factors such as habitat spatial/hydrological (H), water physico-chemical (W), and aquatic vegetation characteristics (V). Variance partitioning and forward selection based on ordinations and multiple regressions were applied to the data set on 79 water-bodies in southwestern Siberia covering a wide gradient of environmental characteristics and diverse mosquito assemblages. The results showed that richness and abundance inter-correlated poorly (r2 = 0.21), and assemblage structure, richness, and abundance depended on different sets of predictors. Explanatory importance of the three sets of environmental factors differed among the three assemblage variables: H, W, and V had equal importance for assemblage structure, while richness and abundance depended on H and V more than on W. The study showed that contradiction between the aims of conservation (support biodiversity) and mosquito control (reduce mosquito abundance) can be avoided, as relevant environmental factors can be used to define habitats with high richness and low abundance (i.e., high conservation value and low nuisance and disease transmission risk) for conservation activities, and conversely for control measures.

Variability of pesticide exposure in a stream mesocosm system: macrophyte-dominated vs. non-vegetated sections.

For flowing water bodies no information is available about patterns of contaminant distribution in flowing water compared to macrophyte-dominated structures. The aim of the study was to examine temporal dynamic and spatial cross-channel variability of pulse exposure of the insecticide thiacloprid in outdoor stream mesocosms. Two distinct cross-channel sections have been considered: macrophyte-dominated littoral and non-vegetated midstream. Median disappearance time ranged from 17 to 43 h (water phase, midstream). We showed that during the exposure pulse (10h) thiacloprid concentrations in the macrophyte-dominated section were 20-60% lower than those in the non-vegetated section. This suggests that spatial variability in contaminant concentrations, particularly in streams containing macrophytes, should be taken into account to enable a more realistic assessment of (i) exposure and associated effects and (ii) mass transport of pesticides and other chemicals into river systems (e.g. losses with surface runoff).

Long-term stream invertebrate community alterations induced by the insecticide thiacloprid: effect concentrations and recovery dynamics.

In pesticide risk assessment, effect concentrations and dynamics of long-term community-level effects caused by pulse exposures remain to be investigated. This is because long-term experiments are exceptionally rare, and most of the previously investigated communities had low proportions of sensitive long-living species. The aim of the present study was to investigate the effect of a single pulse contamination with the insecticide thiacloprid on invertebrates. We employed mesocosms designed to realistically mimic communities in small streams within the agricultural landscape. Specifically, the objectives were to (i) compare the community Lowest-Observed-Effect Concentration (LOEC) with organism-level median lethal concentrations (LC50), and (ii) to assess recovery dynamics with special focus on short- and long-living taxa. The contamination resulted in long-term alteration of the overall invertebrate community structure (7 months, until the end of the experiment). Long-term community LOEC was 3.2 microg/L (Redundancy Analysis), slightly below the acute LC50s known for sensitive invertebrates relevant to the mesocosm community. However, one species (stonefly Nemoura cinerea) was affected at the lowest tested concentration, 70 times below the lowest known LC50. Concerning time to recovery from the effect, we found that the duration depends on the life-cycle characteristics of species, but not on the toxicant concentration: short-living (mulivoltine) species recovered after 10 weeks following contamination, whereas long-living (uni- and semivoltine) species did not recover until the end of the experiment (7 months). The present example shows that concentrations of pesticides at which majority of the species is affected can be predicted by acute organism-level toxicity tests with sensitive species. However, tests with longer observation periods, as well as consideration of environmental factors and inter-taxon variability in sensitivity are required to predict effects on all species comprising a community. Realistic prediction of community recovery dynamics requires consideration of the species' life-cycle traits.

An indicator for effects of organic toxicants on lotic invertebrate communities: Independence of confounding environmental factors over an extensive river continuum.

Distinguishing between effects of natural and anthropogenic environmental factors on ecosystems is a fundamental problem in environmental science. In river systems the longitudinal gradient of environmental factors is one of the most relevant sources of dissimilarity between communities that could be confounded with anthropogenic disturbances. To test the hypothesis that in macroinvertebrate communities the distribution of species' sensitivity to organic toxicants is independent of natural longitudinal factors, but depends on contamination with organic toxicants, we analysed the relationship between community sensitivity SPEAR(organic) (average community sensitivity to organic toxicants) and natural and anthropogenic environmental factors in a large-scale river system, from alpine streams to a lowland river. The results show that SPEAR(organic) is largely independent of natural longitudinal factors, but strongly dependent on contamination with organic toxicants (petrochemicals and synthetic surfactants). Usage of SPEAR(organic) as a stressor-specific longitude-independent measure will facilitate detection of community disturbance by organic toxicants.

Acute and delayed effects of the neonicotinoid insecticide thiacloprid on seven freshwater arthropods.

Ecotoxicological risk assessment of contaminants often is based on toxicity tests with continuous-exposure profiles. However, input of many contaminants (e.g., insecticides) to surface waters typically occurs in pulses rather than continuously. Neonicotinoids are a new group of insecticides, and little is known about their toxicity to nontarget freshwater organisms and potential effects on freshwater ecosystems. The aim of the present research was to assess effects of short-term (24-h) exposure to the neonicotinoid insecticide thiacloprid, including a postexposure observation period. A comparison of several freshwater insect and crustacean species showed an increase of sensitivity by three orders of magnitude in the following order: Daphnia magna < Asellus aquaticus = Gammarus pulex < Simpetrum striolatum < Culex pipiens = Notidobia ciliaris = Simulium latigonium, with median lethal concentrations (LC50s) of 4,400, 153, 190, 31.2, 6.78, 5.47, and 5.76 mug/L, respectively (postexposure observation 11-30 d). Thiacloprid caused delayed lethal and sublethal effects, which were observed after 4 to 12 d following exposure. Reduction in LC50s found when postexposure observation was extended from 1 d to a longer period (11-30 d) was up to >50-fold. Hence, delayed effects occurring after short-term exposure should be considered in risk assessment. The 5% hazardous concentration (HC5) of thiacloprid obtained in the present study (0.72 microg/L) is more than one order of magnitude below the currently predicted worst-case environmental concentrations in surface water. Concerning the selection of test organisms, we observed that the widely employed test organism D. magna is least sensitive among the arthropods tested and that, for neonicotinoid insecticides, an insect like the mosquito C. pipiens would be more suitable for predicting effects on sensitive species.

Potential of 11 pesticides to initiate downstream drift of stream macroinvertebrates.

Downstream drift of lotic macroinvertebrates induced by toxicants is a well-known ecologically relevant phenomenon. However, little is known about which toxicants can initiate drift, and potential drift-initiating effects of contaminants are not taken into account in ecotoxicological risk assessment. The aim of the present study was to evaluate potential drift-initiating action of 11 pesticides having different target groups and modes of action. Sublethal concentrations of the pesticides were tested in stream microcosms with amphipods (Gammarus pulex), blackfly larvae (Simulium latigonium), and mayfly larvae (Baetis rhodani). The results show that 6 out of 11 pesticides tested can initiate drift of macroinvertebrates at sublethal concentrations 7-22 times lower than acute LC(50)s (thiacloprid, imidacloprid, acetamiprid, iprodione, fenvalerate, and indoxacarb). All the toxicants that exhibited drift-initiating action are neurotoxic insecticides belonging to the groups of pyrethroids and neonicotinoids except the fungicide iprodione. The pesticides that did not initiate drift are fungicides (cyprodinil, prochloraz, and azoxystrobin), a juvenile-hormone mimic (fenoxycarb), and a pyrazole insecticide (tebufenpyrad) affecting cell energy production. Remarkably, for all the drift-initiating toxicants, drift of the tested animals was detected within 2 h after contamination. This shows that macroinvertebrate drift can be induced even by short-term pulse exposures to neurotoxic insecticides, at field-relevant concentrations. The present results imply that the possibility of drift-initiating effects of pesticides should be considered within the risk assessment framework for pesticides, as all neurotoxic substances that were investigated did initiate drift at sublethal concentrations.