Effects of multiple but low pesticide loads on aquatic fungal communities colonizing leaf litter.

In the first tier risk assessment (RA) of pesticides, risk for aquatic communities is estimated by using results from standard laboratory tests with algae, daphnids and fish for single pesticides such as herbicides, fungicides, and insecticides. However, fungi as key organisms for nutrient cycling in ecosystems as well as multiple pesticide applications are not considered in the RA. In this study, the effects of multiple low pesticide pulses using regulatory acceptable concentrations (RACs) on the dynamics of non-target aquatic fungi were investigated in a study using pond mesocosm. For that, fungi colonizing black alder (Alnus glutinosa) leaves were exposed to multiple, low pulses of 11 different pesticides over a period of 60days using a real farmer's pesticide application protocol for apple cropping. Four pond mesocosms served as treatments and 4 as controls. The composition of fungal communities colonizing the litter material was analyzed using a molecular fingerprinting approach based on the terminal Restriction Fragment Length Polymorphism (t-RFLP) of the fungal Internal Transcribed Spacer (ITS) region of the ribonucleic acid (RNA) gene(s). Our data indicated a clear fluctuation of fungal communities based on the degree of leaf litter degradation. However significant effects of the applied spraying sequence were not observed. Consequently also degradation rates of the litter material were not affected by the treatments. Our results indicate that the nutrient rich environment of the leaf litter material gave fungal communities the possibility to express genes that induce tolerance against the applied pesticides. Thus our data may not be transferred to other fresh water habitats with lower nutrient availability.

Structural changes in a macrozoobenthos assemblage after imidacloprid pulses in aquatic field-based microcosms.

A field-based microcosm experiment was performed to investigate the effects of repeated pulses of the neonicotinoid insecticide imidacloprid on a lentic benthos assemblage. This specific microcosm method was chosen because it allows for both testing of a wide range of organisms under natural conditions and as well as gaining insight into intraspecific and interspecific interactions. The macrozoobenthos that colonised the microcosms was exposed to three pulses each 1 week apart at nominal concentrations ranging from 0.6 to 40 µg/L. Imidacloprid underwent fast aqueous photolysis due to optimal sunlight conditions during the test phase (half-life = 28 ± 8 h [monitored for 21 days]). Nonetheless, decreased abundance and emergence of Ephemeroptera and decreased survival of chironomid species of the subfamilies Tanypodinae and Orthocladiinae were observed at time-weighted average concentrations of 2.3 µg/L. In contrast, the gastropod Radix sp. became dominant at high imidacloprid concentrations, probably due to decreased competition for food with sensitive species. The results of this study show that repeated short-term contamination of imidacloprid at low concentration levels may affect aquatic ecosystems even under optimal conditions for photodegradation. The microcosm approach, with its simple and field-relevant design, proved to be a useful tool for assessing the effects of imidacloprid contamination.

Herbicide effects of metazachlor on duckweed (Lemna minor and Spirodela polyrhiza) in test systems with different trophic status and complexity.

Growth of common duckweed Lemna minor under optimal standard test conditions was compared to growth of L. minor exposed to nutrient-poor water in both a modified standardised test and in oligo- to mesotrophic indoor pond mesocosms in order to test the impact of trophic conditions and test system complexity on the effect of the herbicide metazachlor (2-chloro-N-(pyrazol-1-ylmethyl)acet-2',6'-xylidide) on aquatic macrophytes. In the mesocosms L. minor was replaced by greater duckweed Spirodela polyrhiza after 3 weeks due to high mortality even in the controls. The pond systems contained other macrophytes and biota as well as sediment and were thus more complex than standard test systems. For L. minor front area, the ErC(50) (50% effective concentration related to growth rate) was 2.8 microg L(-1) metazachlor in the standardised and 4.7 microg L(-1) in the modified laboratory test after 7 days (4.9 microg L(-1) and 52.9 microg L(-1) metazachlor when using front number). In the oligo- to mesotrophic pond mesocosms, similar sensitivities to metazachlor (ErC(50) 4.5-6.4 microg L(-1)) were noted for S. polyrhiza after 21 and 28 days of exposure. In comparison with dicotyledonous macrophytes, duckweed species are more sensitive for irreversible enzyme inhibitors of growth such as metazachlor independent of trophic status and complexity of the test system.

Endpoint 'floating leaves' of potamogeton natans: a new method to evaluate the development of macrophytes in pond mesocosms.

GOAL, SCOPE AND BACKGROUND: One of the advantages of long-term mesocosm experiments as compared to short-term standard toxicity tests in the laboratory is the potential for detecting secondary effects due to the interaction of species and recovery with biomass of macrophytes being an important endpoint. However, generating biomass data by harvesting is often laborious, time-consuming, costly and restricted to the end of the experiment. Moreover, valuable information may get lost, in particular in single application studies, since maximal primary effects and secondary effects or recovery occur per se at different times. Potamogeton natans was used as an example in order to test whether number and area of floating leaves can be reliably measured and be used as intermediate and final end-points in mesocosm effect studies. METHODS: Digital photos, which were taken of the water surface in the course of an indoor pond mesocosm study on herbicide effects, were subjected to image analysis. The results were compared to wet weight and ash-free dry weight of Potamogeton at the end of the herbicide study. RESULTS AND DISCUSSION: Both number and area of floating leaves indicated the same herbicide effects as wet weight and ash-free dry weight of Potamogeton. Error introduced by the different work steps is small and can be further minimised by a number of method improvements. RECOMMENDATIONS AND PERSPECTIVES: In indoor mesocosm studies, errors due to the perspective adjustment may be circumvented by taking the photos perpendicular to the water surface. Correction for lens aberration, identical light conditions and the use of fluorescence images are considered promising. Field applications are proposed.

Enhanced experimental flexibility and control in ecotoxicological mesocosm experiments--a new outdoor and indoor pond and stream system.

SCOPE: The German Federal Environmental Agency has put into operation a new modular mesocosm system consisting of eight outdoor and eight indoor ponds and streams in order to investigate fate and effects of chemicals and municipal wastewater in aquatic ecosystems. General design and special characteristics are given to demonstrate the wide range of possibilities for experimental research. GENERAL DESIGN: Each of the 16 streams with riffle and pool sections can be varied in length up to 106 m. The streams can be run as circular or flow-through systems at a flow velocity of 0.02 to 0.6 m/s. Physico-chemical standard parameters are measured on-line. The 16 ponds, which can be connected to the stream systems, are equipped with drainage and pore water-sampling devices for simulating processes in the littoral zone including influent and effluent ground water flow. PERSPECTIVES: Since the highly flexible and controllable construction also allows treating a wide range of hydrological and ecological experiments external institutions are invited to submit proposals.

Artificial light and nocturnal activity in gammarids.

Artificial light is gaining attention as a potential stressor to aquatic ecosystems. Artificial lights located near streams increase light levels experienced by stream invertebrates and we hypothesized light would depress night drift rates. We also hypothesized that the effect of light on drift rates would decrease over time as the invertebrates acclimated to the new light level over the course of one month's exposure. These hypotheses were tested by placing Gammarus spp. in eight, 75 m × 1 m artificial flumes. One flume was exposed to strong (416 lx) artificial light at night. This strong light created a gradient between 4.19 and 0.04 lx over the neighboring six artificial flumes, while a control flume was completely covered with black plastic at night. Night-time light measurements taken in the Berlin area confirm that half the flumes were at light levels experienced by urban aquatic invertebrates. Surprisingly, no light treatment affected gammarid drift rates. In contrast, physical activity measurements of in situ individually caged G. roeseli showed they increased short-term activity levels in nights of complete darkness and decreased activity levels in brightly lit flumes. Both nocturnal and diurnal drift increased, and day drift rates were unexpectadly higher than nocturnal drift.

Macroinvertebrate community response to repeated short-term pulses of the insecticide imidacloprid.

Small streams in agricultural landscape can experience short and repeated pulses of fluctuating pesticide concentrations. A single pesticide pulse may not have adverse effects on macrozoobenthos species but repeated pulses may have, especially if the organisms have not yet fully recovered when the second pesticide pulse occurs. Against this background, a comprehensive indoor stream mesocosm study was carried out in order to evaluate the cumulative effects of repeated insecticide pulses on a macrozoobenthos community. Weekly 12h pulses of 12 µg/L of the insecticide imidacloprid were set 3 times in 4 stream mesocosms in 2 series, one in spring and one in summer. Another 4 mesocosms served as controls. Prior to each pulse series, the mesocosms were stocked with macroinvertebrates from an uncontaminated reference stream using straw bags as attraction devices. The straw bag method proved suitable for establishing a functional macroinvertebrate community in the stream mesocosms. The caddisfly species Neureclipsis sp. reacted immediately and most sensitively after a single imidacloprid pulse whilst insect larvae such as ephemerids and dipteran larvae were negatively affected only after repeated imidacloprid pulses. Effects on insect larvae were more pronounced in the summer series most likely due to increased temperature. Abundance was a less sensitive endpoint than sublethal endpoints such as emergence. The results of the study underline that pulse effects are driven by a number of variables like pulse height, pulse duration, number of pulses, time in between pulses and by the species and live stage specific ability of temperature dependent detoxification which all should be taken into account in the risk assessment of pesticides.

Effects of repeated insecticide pulses on macroinvertebrate drift in indoor stream mesocosms.

Pesticide contaminations via run-off or spray drift have been reported to result in the mass drift of macroinvertebrates as well as causing structural and functional changes of the corresponding stream sections. However, pesticide pulses in the field are associated with sudden increases in flow velocity, water turbidity, and changes in water temperature, which can also induce drift. Only through replicated community testing under highly controlled conditions can these effects be disentangled. In a stream mesocosm study, 12-h pulses of 12 µg/L imidacloprid were set three times at weekly intervals and are considered a "pulse series". Two pulse series of this neonicotinoid insecticide were run in both spring and summer with 4 treatment and 4 control stream mesocosms used in each pulse series. Prior to the start of the mesocosm experiment, both pulse concentration and duration had been screened for drift responses in larval Baetidae, Chironomidae and adult Gammarus roeseli in laboratory experiments. In the subsequent mesocosm study, each pulse caused a pronounced increase in the drift of insect larvae and gammarids. The drift response was taxon-specific, which was related to preferred habitat and exposure to other stressors like current velocity, in addition to imidacloprid sensitivity. Activity measurements employing a Multispecies Freshwater Biomonitor(®) revealed that in Baetis sp. the diurnal activity pattern became more pronounced even 12h after the pulse though with slightly decreased mean physical activity. Adult G. roeseli showed a drastic pulse by pulse decrease in physical activity which after the 3rd pulse lasted longer than 24h. In conclusion, drift is a sensitive, ecologically relevant endpoint and should be regarded when a specific risk assessment for lotic surface waters is done, e.g. in the context of a spatially explicit risk assessment.

Toxic and accumulative potential of the antifouling biocide and TBT successor irgarol on freshwater macrophytes: a pond mesocosm study.

After the ban of tributyltin (TBT) for vessels not longer than 25 m in 1986, Irgarol has become a commonly used antifouling biocide. Irgarol is highly toxic to autotrophic organisms and has the potential to accumulate in organic material. In the literature, environmental concentrations of Irgarol up to 2.4 microg L(-1) were reported forfreshwater. Within a comprehensive freshwater mesocosm study, experiments were conducted to gain more information on the effects of Irgarol on macrophytes. Six indoor pond mesocosms were contaminated once with concentrations between 0.04 and 5 microgl(-1) Irgarol and monitored for 150 days; two mesocosms served as controls. The mesocosm study revealed that all macrophytes were directly affected by this single application. Myriophyllum verticillatum was the most sensitive macrophyte with an EC50 (Day 150) of 0.21 microg L(-1) Irgarol. The duckweed Spirodela polyrhiza was the least sensitive species tested in the mesocosms and number of fronds even increased with increasing Irgarol concentrations. Time-weighted average calculations yielded high BCF values of up to 10,580 L kg(-1) dry weight for M. verticillatum indicating a high potential for accumulation. The results give cause for concern that natural macrophyte communities are impaired at actual environmental concentrations.