A guideline to frame stressor effects in freshwater ecosystems.

Environmental policies fall short in protecting freshwater ecosystems, which are heavily threatened by human pressures and their associated stressors. One reason is that stressor effects depend on the context in which they occur and it is difficult to extrapolate patterns to predict the effect of stressors without these being contextualized in a general frame. This study aims at improving existing decision-making frameworks such as the DPSIR approach (Driver-Pressure-State-Impact-Response) or ERA (Environmental Risk Assessment) in the context of stressors. Here, we delve into stressor-impact relationships in freshwater ecosystems and develop a guideline which includes key characteristics such as stressor type, stressor duration, location, the natural levels of environmental variables to which each ecosystem is used to, among others. This guideline is intended to be useful in a wide range of ecosystem conditions and stressors. Incorporating these guidelines may favor the comparability of scientific results and may lead to a substantial advancement in the efficacy of diagnosis and predictive approaches of impacts.

Identifying ecological production functions for use in ecosystem services-based environmental risk assessment of chemicals.

There is increasing research interest in the application of the ecosystem services (ES) concept in the environmental risk assessment of chemicals to support formulating and operationalising regulatory environmental protection goals and making environmental risk assessment more policy- and value-relevant. This requires connecting ecosystem structure and processes to ecosystem function and henceforth to provision of ecosystem goods and services and their economic valuation. Ecological production functions (EPFs) may help to quantify these connections in a transparent manner and to predict ES provision based on function-related descriptors for service providing species, communities, ecosystems or habitats. We review scientific literature for EPFs to evaluate availability across provisioning and regulation and maintenance services (CICES v5.1 classification). We found quantitative production functions for nearly all ES, often complemented with economic valuation of physical or monetary flows. We studied the service providing units in these EPFs to evaluate the potential for extrapolation of toxicity data for test species obtained from standardised testing to ES provision. A broad taxonomic representation of service providers was established, but quantitative models directly linking standard test species to ES provision were extremely scarce. A pragmatic way to deal with this data gap would be the use of proxies for related taxa and stepwise functional extrapolation to ES provision and valuation, which we conclude possible for most ES. We suggest that EPFs may be used in defining specific protection goals (SPGs), and illustrate, using pollination as an example, the availability of information for the ecological entity and attribute dimensions of SPGs. Twenty-five pollination EPFs were compiled from the literature for biological entities ranging from 'colony' to 'habitat', with 75% referring to 'functional group'. With about equal representation of the attributes 'function', 'abundance' and 'diversity', SPGs for pollination therefore would seem best substantiated by EPFs at the level of functional group.

Lower tier toxicity risk assessment of agriculture pesticides detected on the Río Madre de Dios watershed, Costa Rica.

Costa Rica is a tropical country with one of the highest biodiversity on Earth. It also has an intensive agriculture, and pesticide runoff from banana and pineapple plantations may cause a high toxicity risk to non-target species in rivers downstream the plantations. We performed a first tier risk assessment of the maximum measured concentrations of 32 pesticides detected over 4 years in the River Madre de Dios (RMD) and its coastal lagoon on the Caribbean coast of Costa Rica. Species sensitivity distributions (SSDs) were plotted in order to derive HC5 values for each pesticide, i.e., hazard concentrations for 5 % of the species, often used as environmental criteria values in other countries. We also carried out toxicity tests for selected pesticides with native Costa Rican species in order to calculate risk coefficients according to national guidelines in Costa Rica. The concentrations of herbicides diuron and ametryn and insecticides carbofuran, diazinon, and ethoprophos exceeded either the HC5 value or the lower limit of its 90 % confidence interval suggesting toxic risks above accepted levels. Risk coefficients of diuron and carbofuran derived using local guidelines indicate toxicity risks as well. The assessed fungicides did not present acute toxic risks according to our analysis. Overall, these results show a possible toxicity of detected pesticides to aquatic organisms and provide a comparison of Costa Rican national guidelines with more refined methods for risk assessment based on SSDs. Further higher tier risk assessments of pesticides in this watershed are also necessary in order to consider pesticide water concentrations over time, toxicity from pesticide mixtures, and eventual effects on ecosystem functions.

Additive effects of the herbicide glyphosate and elevated temperature on the branched coral Acropora formosa in Nha Trang, Vietnam.

The combined effects of the herbicide glyphosate and elevated temperature were studied on the tropical staghorn coral Acropora formosa, in Nha Trang bay, Vietnam. The corals were collected from two different reefs, one close to a polluted fish farm and one in a marine-protected area (MPA). In the laboratory, branches of the corals were exposed to the herbicide glyphosate at ambient (28 °C) and at 3 °C elevated water temperatures (31 °C). Effects of herbicide and elevated temperature were studied on coral bleaching using photography and digital image analysis (new colorimetric method developed here based on grayscale), chlorophyll a analysis, and symbiotic dinoflagellate (Symbiodinium, referred to as zooxanthellae) counts. All corals from the MPA started to bleach in the laboratory before they were exposed to the treatments, indicating that they were very sensitive, as opposed to the corals collected from the more polluted site, which were more tolerant and showed no bleaching response to temperature increase or herbicide alone. However, the combined exposure to the stressors resulted in significant loss of color, proportional to loss in chlorophyll a and zooxanthellae. The difference in sensitivity of the corals collected from the polluted site versus the MPA site could be explained by different symbiont types: the resilient type C3u and the stress-sensitive types C21 and C23, respectively. The additive effect of elevated temperatures and herbicides adds further weight to the notion that the bleaching of coral reefs is accelerated in the presence of multiple stressors. These results suggest that the corals in Nha Trang bay have adapted to the ongoing pollution to become more tolerant to anthropogenic stressors, and that multiple stressors hamper this resilience. The loss of color and decrease of chlorophyll a suggest that bleaching is related to concentration of chloro-pigments. The colorimetric method could be further fine-tuned and used as a precise, non-intrusive tool for monitoring coral bleaching in situ.

The minimum detectable difference (MDD) and the interpretation of treatment-related effects of pesticides in experimental ecosystems.

In the European registration procedure for pesticides, microcosm and mesocosm studies are the highest aquatic experimental tier to assess their environmental effects. Evaluations of microcosm/mesocosm studies rely heavily on no observed effect concentrations (NOECs) calculated for different population-level endpoints. Ideally, a power analysis should be reported for the concentration-response relationships underlying these NOECs, as well as for measurement endpoints for which significant effects cannot be demonstrated. An indication of this statistical power can be provided a posteriori by calculated minimum detectable differences (MDDs). The MDD defines the difference between the means of a treatment and the control that must exist to detect a statistically significant effect. The aim of this paper is to expand on the Aquatic Guidance Document recently published by the European Food Safety Authority (EFSA) and to propose a procedure to report and evaluate NOECs and related MDDs in a harmonised way. In addition, decision schemes are provided on how MDDs can be used to assess the reliability of microcosm/mesocosm studies and for the derivation of effect classes used to derive regulatory acceptable concentrations. Furthermore, examples are presented to show how MDDs can be reduced by optimising experimental design and sampling techniques.

Theoretically exploring direct and indirect chemical effects across ecological and exposure scenarios using mechanistic fate and effects modelling.

Predicting ecosystem response to chemicals is a complex problem in ecotoxicology and a challenge for risk assessors. The variables potentially influencing chemical fate and exposure define the exposure scenario while the variables determining effects at the ecosystem level define the ecological scenario. In absence of any empirical data, the objective of this paper is to present simulations by a fugacity-based fate model and a differential equation-based ecosystem model to theoretically explore how direct and indirect effects on invertebrate shallow pond communities vary with changing ecological and exposure scenarios. These simulations suggest that direct and indirect effects are larger in mesotrophic systems than in oligotrophic systems. In both trophic states, interaction strength (quantified using grazing rates) was suggested a more important driver for the size and recovery from direct and indirect effects than immigration rate. In general, weak interactions led to smaller direct and indirect effects. For chemicals targeting mesozooplankton only, indirect effects were common in (simple) food-chains but rare in (complex) food-webs. For chemicals directly affecting microzooplankton, the dominant zooplankton group in the modelled community, indirect effects occurred both in food-chains and food-webs. We conclude that the choice of the ecological and exposure scenarios in ecotoxicological modelling efforts needs to be justified because of its influence on the prevalence and magnitude of the predicted effects. Overall, more work needs to be done to empirically test the theoretical expectations formulated here.

Using additive modelling to quantify the effect of chemicals on phytoplankton diversity and biomass.

Environmental authorities require the protection of biodiversity and other ecosystem properties such as biomass production. However, the endpoints listed in available ecotoxicological datasets generally do not contain these two ecosystem descriptors. Inferring the effects of chemicals on such descriptors from micro- or mesocosm experiments is often hampered by inherent differences in the initial biodiversity levels between experimental units or by delayed community responses. Here we introduce additive modelling to establish the effects of a chronic application of the herbicide linuron on 10 biodiversity indices and phytoplankton biomass in microcosms. We found that communities with a low (high) initial biodiversity subsequently became more (less) diverse, indicating an equilibrium biodiversity status in the communities considered here. Linuron adversely affected richness and evenness while dominance increased but no biodiversity indices were different from the control treatment at linuron concentrations below 2.4 µg/L. Richness-related indices changed at lower linuron concentrations (effects noticeable from 2.4 µg/L) than other biodiversity indices (effects noticeable from 14.4 µg/L) and, in contrast to the other indices, showed no signs of recovery following chronic exposure. Phytoplankton biomass was unaffected by linuron due to functional redundancy within the phytoplankton community. Comparing thresholds for biodiversity with conventional toxicity test results showed that standard ecological risk assessments also protect biodiversity in the case of linuron.

Functional redundancy and food web functioning in linuron-exposed ecosystems.

An extensive data set describing effects of the herbicide linuron on macrophyte-dominated microcosms was analysed with a food web model to assess effects on ecosystem functioning. We showed that sensitive phytoplankton and periphyton groups in the diets of heterotrophs were gradually replaced by more tolerant phytoplankton species as linuron concentrations increased. This diet shift--showing redundancy among phytoplankton species--allowed heterotrophs to maintain their functions in the contaminated microcosms. On an ecosystem level, total gross primary production was up to hundred times lower in the treated microcosms but the uptake of dissolved organic carbon by bacteria and mixotrophs was less sensitive. Food web efficiency was not consistently lower in the treated microcosms. We conclude that linuron predominantly affected the macrophytes but did not alter the overall functioning of the surrounding planktonic food web. Therefore, a risk assessment that protects macrophyte growth also protects the functioning of macrophyte-dominated microcosms.

Effects of four fungicides on nine non-target submersed macrophytes.

We tested the sensitivity of nine submersed macrophyte species to the fungicides chlorothalonil, pentachlorophenol, fluazinam, and carbendazim. Endpoints determined 3 weeks after the start of the treatment were based on shoot and root growth in water. Carbendazim proved not or only moderately toxic to these macrophytes. Pentachlorophenol and chlorothalonil were more toxic than fluazinam. Taking all endpoints into consideration, toxicity levels differed very substantially. Although root endpoints reflecting root growth were in some cases more sensitive than shoot endpoints, shoot growth endpoints like relative growth turned out to be more reliable than the root growth endpoints. Due to the large differences in the type of mode of action between fungicides, it is very difficult to predict their potential effect in the environment or even to predict whether non-target organisms like macrophytes are likely to be sensitive. Ideally, therefore, the registration of fungicides requires an extensive risk-assessment procedure, which also covers non-target groups like macrophytes.

Effects of chlorpyrifos, carbendazim, and linuron on the ecology of a small indoor aquatic microcosm.

To validate the use of small indoor microcosms for the risk assessment of pesticides, the fate and effects of chlorpyrifos, carbendazim, and linuron were studied in 8.5-liter indoor freshwater microcosms. Functional and structural responses to selected concentrations were evaluated and compared with responses observed in larger-scale model ecosystem studies. Overall, the microcosms adequately displayed the chain of effects resulting from the application, although they did not always predict the exact fate and responses that were observed in larger semifield studies. Because closed systems were used that did not contain sediment and macrophytes, pesticides were relatively persistent in the present study. Consequently, calculated toxicity values were generally more comparable with those reported in studies with long- than with short-term exposure. Carbendazim had a higher overall no-observed-effect concentration (NOEC) compared with experiments performed in larger systems because macroinvertebrate taxa, the most sensitive species group to this fungicide, were not abundant or diverse. Future refinements to the test system could include the addition of a sediment compartment and sensitive macroinvertebrate taxa. However, the simple design offers the potential to perform experiments under more controlled conditions than larger and, consequently, more complex model ecosystems, while maintaining relatively high ecologic realism compared with standard laboratory tests. Further implications for risk-assessment studies are discussed in an ecotoxicologic and methodologic context.

Type of disturbance and ecological history determine structural stability.

This study aims to reveal whether complexity, namely, community and trophic structure, of chronically stressed soil systems is at increased risk or remains stable when confronted with a subsequent disturbance. Therefore, we focused on a grassland with a history of four centuries of patchy contamination. Nematodes were used as model organisms because they are an abundant and trophically diverse group and representative of the soil food web and ecosystem complexity. In a field survey, a relationship between contaminants and community structures was established. Following, two groups of soil mesocosms from the field that differed in contamination level were exposed to different disturbance regimes, namely, to the contaminant zinc and a heat shock. The zinc treatment revealed that community structure is stable, irrespective of soil contamination levels. This implies that centuries of exposure to contamination led to adaptation of the soil nematode community irrespective of the patchy distribution of contaminants. In contrast, the heat shock had adverse effects on species richness in the highly contaminated soils only. The total nematode biomass was lower in the highly contaminated field samples; however, the biomass was not affected by zinc and heat treatments of the mesocosms. This means that density compensation occurred rapidly, i.e., tolerant species quickly replaced sensitive species. Our results support the hypothesis that the history of contamination and the type of disturbance determine the response of communities. Despite that ecosystems may be exposed for centuries to contamination and communities show adaptation, biodiversity in highly contaminated sites is at increased risk when exposed to a different disturbance regime. We discuss how the loss of higher trophic levels from the entire system, such as represented by carnivorous nematodes after the heat shock, accompanied by local biodiversity loss at highly contaminated sites, may result in detrimental effects on ecosystem functions.

Impact of triphenyltin acetate in microcosms simulating floodplain lakes. II. Comparison of species sensitivity distributions between laboratory and semi-field.

The study objectives were to shed light on the types of freshwater organism that are sensitive to triphenyltin acetate (TPT) and to compare the laboratory and microcosm sensitivities of the invertebrate community. The responses of a wide array of freshwater taxa (including invertebrates, phytoplankton and macrophytes) from acute laboratory Single Species Tests (SST) were compared with the concentration-response relationships of aquatic populations in two types of freshwater microcosms. Representatives of several taxonomic groups of invertebrates, and several phytoplankton and vascular plant species proved to be sensitive to TPT, illustrating its diverse modes of toxic action. Statistically calculated ecological risk thresholds (HC5 values) based on 96 h laboratory EC50 values for invertebrates were 1.3 microg/l, while these values on the basis of microcosm-Species Sensitivity Distributions (SSD) for invertebrates in sampling weeks 2-8 after TPT treatment ranged from 0.2 to 0.6 microg/l based on nominal peak concentrations. Responses observed in the microcosms did not differ between system types and sampling dates, indicating that ecological threshold levels are not affected by different community structures including taxa sensitive to TPT. The laboratory-derived invertebrate SSD curve was less sensitive than the curves from the microcosms. Possible explanations for the more sensitive field response are delayed effects and/or additional chronic exposure via the food chain in the microcosms.

Ecological effects of spring and late summer applications of lambda-cyhalothrin on freshwater microcosms.

The aim of the study was to compare the effects of the pyrethroid insecticide lambda-cyhalothrin (treated at 10, 25, 50, 100, 250 ng active ingredient a.i./L) on a drainage ditch ecosystem in spring and late summer. Microcosms (water volume approximately 430 L) were established using enclosures in a 50-cm-deep experimental ditch system containing communities typical of macrophyte-dominated freshwater ecosystems. Effects on macroinvertebrates, zooplankton, phytoplankton, macrophytes, and community metabolism were assessed and evaluated using univariate and multivariate statistical techniques. The macroinvertebrate community responded most clearly to treatment and, as anticipated, insects and crustaceans were among the most sensitive organisms. Statistical analysis showed that the underlying community structure was significantly different between the spring and summer experiments. However, the most sensitive species (Chaoborus obscuripes and Gammarus pulex) were abundant in spring as well as in late summer. In spring and late summer, only slight and transient effects were observed at the community level in the 10-ng/L treatment. Overall, the study did not show substantial differences in the responses of sensitive taxa between spring and late summer treatments, and effects thresholds were similar irrespective of season of treatment.

Confirming the species-sensitivity distribution concept for endosulfan using laboratory, mesocosm, and field data.

In Australia, water-quality trigger values for toxicants are derived using protective concentration values based on species-sensitivity distribution (SSD) curves. SSD curves are generally derived from laboratory data with an emphasis on using local or site-specific data. In this study, Australian and non-Australian laboratory-species based SSD curves were compared and the concept of species protection confirmed by comparison of laboratory-based SSD curves with local mesocosm experiments and field monitoring data. Acute LC50 data for the organochlorine pesticide endosulfan were used for these comparisons; SSD curves were fitted using the Burr type III distribution. SSD curves indicated that the sensitivities of Australian fish and arthropods were not significantly different from those of corresponding non-Australian taxa. Arthropod taxa in the mesocosm were less sensitive than taxa in laboratory tests, which suggests that laboratory-generated single-species data may be used to predict concentrations protective of semifield (mesocosm) systems. SSDs based on laboratory data were also protective of field populations.

Comparison of laboratory single species and field population-level effects of the pyrethroid insecticide lambda-cyhalothrin on freshwater invertebrates.

The toxicity of the pyrethroid insecticide lambda-cyhalothrin to freshwater invertebrates has been investigated using data from short-term laboratory toxicity tests and in situ bioassays and population-level effects in field microcosms. In laboratory tests, patterns of toxicity were consistent with previous data on pyrethroids. The midge Chaoborus obscuripes was most sensitive (48- and 96-h EC50 = 2.8 ng/L). Other insect larvae (Hemiptera, Ephemeroptera) and macrocrustacea (Amphipoda, Isopoda) were also relatively sensitive, with 48- and 96-h EC50 values between 10 and 100 ng/L. Generally, microcrustacea (Cladocera, Copepoda) and larvae of certain insect groups (Odonata and Chironomidae) were less sensitive, with 48-h EC50 values higher than 100 ng/L. Mollusca and Plathelminthes were insensitive and were unaffected at concentrations at and above the water solubility (5 microg/L). Generally, the EC50 values based on initial population responses in field enclosures were similar to values derived from laboratory tests with the same taxa. Also, the corresponding fifth and tenth percentile hazard concentrations (HC5 and HC10) were similar (laboratory HC5 = 2.7 ng/L and field HC5 = 4.1 ng/L; laboratory and field HC10 = 5.1 ng/L), at least when based on the same sensitive taxonomic groups (insects and crustaceans) and when a similar concentration range was taken into account. In the three field enclosure experiments and at a treatment level of 10 ng/L, consistent effects were observed for only one population (Chaoborus obscuripes), with recovery taking place within 3 to 6 weeks. The laboratory HC5 (2.7 ng/L) and HC10 (5.1 ng/L) based on acute EC50 values of all aquatic arthropod taxa were both lower than this 10 ng/L, a concentration that might represent the "regulatory acceptable concentration." The HC5 and HC10 values in this study in The Netherlands (based on static laboratory tests with freshwater arthropods) were very similar to those derived from a previous study in the United Kingdom (1.4 and 3.3 ng/L). This suggests that for pesticides like lambda-cyhalothrin, HC5 values based on static laboratory tests may provide a conservative estimate of the potential for community-level effects under field conditions. While these HC5 values are conservative for initial effects, they do not provide information on recovery potential, which may be important for regulatory decision-making.

The effects of a pesticide mixture on aquatic ecosystems differing in trophic status: responses of the macrophyte Myriophyllum spicatum and the periphytic algal community.

The effects of a pesticide mixture (asulam, fluazinam, lambda-cyhalothrin, and metamitron) on aquatic ecosystems were investigated in 20 outdoor aquatic microcosms. Ten of the microcosms simulated mesotrophic aquatic ecosystems dominated by submerged macrophytes (Elodea). The others simulated eutrophic ecosystems with a high Lemna surface coverage (Lemna). This paper describes the fate of the chemicals as well as their effects on the growth of Myriophyllum spicatum and the periphytic algal community. In the Elodea-dominated microcosms significant increase in the biomass and alterations of species composition of the periphytic algae were observed, but no effect on M. spicatum growth could be recorded in response to the treatment. The opposite was found in the Lemna-dominated microcosms, in which decreased growth of M. spicatum was observed but no alterations could be found in the periphytic community. In the Elodea-dominated microcosms the species composition of the periphytic algae diverged from that of the control following treatment with 0.5% spray drift emission of the label-recommended rate (5% for lambda-cyhalothrin), while reduced growth of M. spicatum in the Lemna-dominated microcosms was recorded at 2% drift (20% for lambda-cyhalothrin). This study shows that the structure of the ecosystem influences the final effect of pesticide exposure.

Effects of zinc contamination on a natural nematode community in outdoor soil mesocosms.

The effect of zinc on the indigenous nematode fauna of a sandy soil was determined in an experimentally contaminated outdoor field plot. The aims of the study were to describe and quantify the changes in density of separate nematode taxa and total nematodes, and the changes in the number of taxa, species diversity, community maturation, and species composition in response to zinc exposure with time, and to compare the observed responses with benchmark concentrations for soil as derived from the species sensitivity distribution (SSD) for zinc toxicity. Speciation of zinc in pore water was considered and CaCl2-exchangeable zinc concentrations were used as a measure of the bioavailable zinc fraction. After contamination, a reduction of total zinc and an increase of labile sorbed zinc over time occurred, concurrent with various changes in soil and biological characteristics. Data analyses on the nematode species revealed different sensitivity levels for several population and community endpoints to zinc exposure. Based on no observed effect concentration (NOEC) values, the most sensitive community-level response was obtained with principal response curve (PRC) analysis, which incorporates all density data in a single analysis. The PRC-based community NOECs were 56, 100, and 100 mg total Zn/kg dry soil after 3, 10, and 22 months of exposure, respectively. Based on 0.01 M CaCl2-exchangeable zinc, the community response appeared to increase, as NOECs were 4.9, 4.4, and 0.67 mg exchangeable Zn/kg dry weight. Total density was least sensitive, followed by diversity of taxa and the Shannon-Wiener index. NOECs for separate species covered a broad range from sensitive to tolerant species. This range of sensitivities was similar to the one found for other species groups tested in the field plot soil. A comparison was made between benchmark concentrations HC5 and HC50 derived from the general SSD of soil organisms for zinc and the nematode response data. These comparisons roughly confirm the predictions of the SSD model, that is, the community NOEC is in agreement with the benchmark that should protect the soil ecosystem's integrity, and large adverse effects were found at the benchmark derived for setting remediation urgency.

Sensitivity of macrophyte-dominated freshwater microcosms to chronic levels of the herbicide linuron. I. Primary producers.

Effects of chronic concentrations of linuron (0, 0.5, 5, 15, 50, and 150 micrograms/L) were studied in indoor, macrophyte dominated, freshwater microcosms. The concentrations were kept at a constant level for 4 weeks. This paper is the first in a series of two and summarizes the course of the linuron concentrations in time and its effects on macrophytes, periphyton, and phytoplankton. These endpoints were studied from 3 weeks before the start of the treatment until 11 weeks after the start. The degradation of linuron in the water was lower at higher treatment levels, probably due to a decrease in pH. Linuron treatment resulted in a decrease in biomass of the macrophyte Elodea nuttallii and a clear decrease in abundance of the algae Cocconeis, Chroomonas, and Phormidium foveolarum. It was found that Cocconeis first decreased in biovolume and after 2 weeks also in abundance. The alga Chlamydomonas increased in abundance at the two highest doses, resulting in higher chlorophyll-a levels. The NOECs of 0.5 micrograms/L for the inhibition of the growth and photosynthesis of Elodea nuttallii, the abundance of Cocconeis and Chroomonas, and the oxygen and pH levels were the lowest recorded in the microcosms. The safety factors adopted by the EU in the Uniform Principles appeared to ensure adequate protection for the ecosystem in the case of chronic exposure to linuron.

Sensitivity of macrophyte-dominated freshwater microcosms to chronic levels of the herbicide linuron. II. Community metabolism and invertebrates.

Effects of a chronic application of the herbicide Afalon (active ingredient linuron) on physicochemical conditions, decomposition of plant litter, and densities of zooplankton and macroinvertebrates were studied in indoor microcosms intended to model drainage ditches. For 28 days, concentrations of 0, 0.5, 5, 15, 50, and 150 micrograms/L linuron were maintained, each in two replicates. The microcosms were dominated by the macrophyte Elodea nuttallii. The functional response of the ecosystem is discussed in relation to shifts in community structure. Treatment effects of linuron on community metabolism, as a direct effect of the inhibition of the photosynthesis of macrophytes and algae, resulted in a decrease in dissolved oxygen and pH, and an increase in alkalinity and conductivity (NOEC 0.5 microgram/L). During the posttreatment period, differences between controls and highest dose fell gradually, but were still significant 7 weeks after the start of linuron application. Decomposition of particulate organic material in litter bags was not affected, despite decreases in DO. The negative effect of linuron on several algae (cryptophytes, diatoms) and the positive effect on the green alga Chlamydomonas resulted in a decrease of several Rotatoria and an increase in Copepoda, and, to a lesser extent, Cladocera. The complete disappearance of the macrophyte E. nuttallii in the 150 micrograms/L microcosms and a 50% reduction of its biomass in the 50 micrograms/L microcosms reduced the numbers of the snail Physella acuta, which normally inhabits macrophytes. Artificial substrates indicated a significant increase in the isopod Asellus aquaticus in the 50 and 150 micrograms/L microcosms during the post-treatment period. This, however, was counteracted by a significant decrease in A. aquaticus at the final harvest. Changes in the ecosystem structure (decline in macrophyte biomass) made the artificial substrates more attractive.

Ordination techniques for analysing response of biological communities to toxic stress in experimental ecosystems.

The ordination techniques principal component analysis (PCA) and redundancy analysis (RDA) are considered to be useful tools for evaluating community responses in experimental ecotoxicology. Concepts and interpretation of these techniques are summarized. Application of PCA and RDA is illustrated in a case study. In this study, the effects of a single application of the insecticide Dursban(®) 4E (a.i. chlorpyrifos) on an aquatic macroinvertebrate community in microcosms were analysed. Four treatment (nominal chlorpyrifos concentration: 35 µg l(-1)) and four control microcosms were used. PCA visualized a change in species composition with time. Immediately after treatment, a major shift in species composition occurred in treated microcosms. RDA demonstrated that this shift was due to the treatment. RDA also showed that non-arthropods were generally insusceptible to chlorpyrifos; most arthropods were affected. Dynamics of separate taxa were visualized, giving indications of possible primary and secondary effects for these taxa. A Monte Carlo permutation test was used to decide whether treatment had a significant effect on the species composition and to investigate the state of recovery in time. In general, the RDA results gave an adequate condensation of detailed information on abundance and effects obtained by more conventional univariate statistical analysis for some individual taxa of the community. In combination with toxicity and ecological data, ordination techniques can provide insight into effects of toxic substances in complex biological communities.