Pesticide Pulse Research Articles

Haematological and biochemical toxicity in freshwater fish Clarias gariepinus and Oreochromis niloticus following pulse exposure to atrazine, mancozeb, chlorpyrifos, lambda-cyhalothrin, and their combination

Transient exposures to high or low concentrations of a single or mixture of pesticides are common in aquatic organisms. Routine toxicity tests disregard transient exposures and the influence of time when examining the toxicity of contaminants. This study investigated the haematological and biochemical responses of juvenile C. gariepinus and O. niloticus to pesticide pulse exposure using three exposure patterns. The patterns include 4-hour pulse exposure to a high pesticide concentration, then 28 days of depuration, continuous exposure to a low pesticide concentration for 28 days, and 4-hour pulse exposure to a high concentration followed by continuous exposure to a low pesticide concentration for 28 days. On days 1, 14, and 28, fish samples were collected for haematological and biochemical analysis. Results showed that red blood cell count, packed cell volume, haemoglobin, platelet count, total protein, and sodium ion decreased, while white blood cell count, total cholesterol, bilirubin, urea, and potassium ion increased in both fish species after pulse, continuous and pulse & continuous exposure to the pesticides (p < 0.05). However, pulse exposure to the pesticides did not significantly affect alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase activity, and creatinine levels. The changes in these biomarkers indicate that 4-hour pulse exposure to high concentration was as hazardous as 24-hour continuous exposure to low pesticide concentration (p > 0.05). The toxic effects of pulse exposure were largely reversible by day 14. Using C. gariepinus and O. niloticus, this study shows that brief exposure to high pesticide pesticides was as hazardous as continuous pesticide exposure.

Responses of signal crayfish Pacifastacus leniusculus to single short-term pulse exposure of pesticides at environmentally relevant concentrations

Although pesticides are often discharged into surface waters in pulses as opposed to a sustained release, the effect of episodic pollution events on freshwater crayfish is largely unknown. We monitored change in heart rate and distance moved to assess the response of signal crayfish Pacifastacus leniusculus to short-term exposure to environmentally relevant concentrations of metazachlor (MTZ), terbuthylazine (TER), and thiacloprid (TCL). Crayfish exposed to 20 µg/L of MTZ exhibited a significant increase in mean heart rate and distance moved. Increased heart rate was detected at 118 ± 74 s post-exposure to MTZ. There were no significant differences in mean heart rate and distance moved in crayfish exposed to 6 µg/L of TCL and 4 µg/L of TER. A significant correlation between heart rate and distance moved was found in all exposed groups. These results suggest that pulse exposure to MTZ impact crayfish physiology and behavior during short-term period. With pulse exposure to TCL and TER, crayfish not exhibiting a locomotor response may continue to be exposed to lower, but potentially harmful, levels of pollutants. Evidence of the impacts of pesticide pulse at environmentally relevant concentrations on crayfish is scarce. Further study is required to determine the ecological effects of such events on freshwater crayfish.

Warming and imidacloprid pulses determine macroinvertebrate community dynamics in experimental streams.

Sustainable management of freshwater and pesticide use is essential for mitigating the impacts of intensive agriculture in the context of a changing climate. To better understand how climate change will affect the vulnerability of freshwater ecosystems to chemical pollutants, more empirical evidence is needed on the combined effects of climatic and chemical stressors in environmentally realistic conditions. Our experiment provides the first empirical evaluation of stream macroinvertebrate community dynamics in response to one of the world's most widely used insecticides, imidacloprid, and increased water temperature. In a 7-week streamside experiment using 128flow-through circular mesocosms, we investigated the effects of pulsed imidacloprid exposure (four environmentally relevant levels between 0 and 4.6µg/L) and raised water temperature (ambient, 3°C above) on invertebrate communities representative of fast- and slow-flowing microhabitats. Invertebrate drift and insect emergence were monitored during three pesticide pulses (10days apart), and benthic invertebrate communities were sampled after 24days of heating and pesticide manipulations. All three manipulated factors strongly affected drift community composition. The first imidacloprid pulse and increased temperature had a greater impact on communities in fast-flowing mesocosms, which contained more pollution-sensitive EPT taxa (mayflies, stoneflies and caddisflies). Heating and imidacloprid caused increased emigration by drift, weak reductions in emergence, and negatively affected the benthic community. The combined effect of stressor manipulations and a 10-day natural heatwave drastically reduced relative abundances of EPT and insects overall and caused a shift to oligochaete-, crustacean- and gastropod-dominated communities. Contrary to our hypothesis, the very high yet realistic water temperatures reached in our experiment meant the negative effects of imidacloprid were clearest at ambient temperatures and fast flow. These findings demonstrate the potential combined impacts of imidacloprid contamination and heatwaves on freshwater invertebrate communities under future climate scenarios and highlight the need for more countries to take regulatory action to control neonicotinoid use.

Daily temperature variation lowers the lethal and sublethal impact of a pesticide pulse due to a higher degradation rate

Daily temperature variation (DTV) is an important warming-related stressor that may magnify pesticide toxicity. Yet, it is unknown whether the pesticide impact under DTV is partly ameliorated by a faster pesticide degradation caused by cyclically higher temperatures under DTV. As synergisms may be more likely under energy-limiting conditions, the impact of the pesticide chlorpyrifos was tested under DTV on the mosquito Culex pipiens in the absence and presence of interspecific competition with the water flea Daphnia magna. Chlorpyrifos exposure at a constant temperature without interspecific competition caused considerable mortality, decreased development time, and increased pupal mass of C. pipiens. Competition with D. magna had negative sublethal effects, but it did not affect the toxicity of chlorpyrifos. In contrast, the presence of C. pipiens decreased the impact of chlorpyrifos on D. magna probably due to corporal absorption of chlorpyrifos by C. pipiens. A key finding was that chlorpyrifos no longer caused lethal effects on C. pipiens under DTV, despite DTV on its own being mildly lethal. Additionally, chlorpyrifos exposure under DTV decreased development time less and had no effect anymore on pupal mass compared to chlorpyrifos exposure at a constant temperature. Similarly, the negative chlorpyrifos impact on adult survival of D. magna was less under DTV than at the constant temperature. This could be explained by a faster chlorpyrifos degradation under DTV. This antagonism between pesticide exposure and DTV is likely widespread because organisms experience DTV, many pesticides are applied in pulses, and pesticide degradation is faster at higher temperatures.

The Exposure Order Strongly Modifies How a Heat Spike Increases Pesticide Toxicity.

The exposure order may strongly affect the impact of stressors, yet is largely ignored for the frequently occurring combinations of toxicants with natural stressors. We tested how exposure order shaped the interactive effects of serial exposure to the pesticide chlorpyrifos and to a heat spike in the larvae of the mosquito Culex pipiens. Notably, the chlorpyrifos-induced mortality was much more magnified by the heat spike and a synergism was already detected at the low concentration when exposure to chlorpyrifos followed the heat spike. This suggests that the preceding heat spike weakened the larvae as reflected in their lower net energy budget, moreover the chlorpyrifos-induced inhibition of its target enzyme (acetylcholinesterase) was only magnified by the heat spike when it was the first stressor. Also the chlorpyrifos-induced reduction in heat tolerance was stronger when the pesticide pulse followed the heat spike, and was buffered by the heat spike when this was the second stressor. Our results provide the first evidence that the exposure order can strongly change the magnifying effect of an important climate change factor on the toxicity of a pesticide. This highlights the importance of exposure order in ecological risk assessment of toxicants under realistic combinations with natural stressors.

Competition magnifies the impact of a pesticide in a warming world by reducing heat tolerance and increasing autotomy.

There is increasing concern that standard laboratory toxicity tests may be misleading when assessing the impact of toxicants, because they lack ecological realism. Both warming and biotic interactions have been identified to magnify the effects of toxicants. Moreover, while biotic interactions may change the impact of toxicants, toxicants may also change the impact of biotic interactions. However, studies looking at the impact of biotic interactions on the toxicity of pesticides and vice versa under warming are very scarce. Therefore, we tested how warming (+4°C), intraspecific competition (density treatment) and exposure to the pesticide chlorpyrifos, both in isolation and in combination, affected mortality, cannibalism, growth and heat tolerance of low- and high-latitude populations of the damselfly Ischnura elegans. Moreover, we addressed whether toxicant exposure, potentially in interaction with competition and warming, increased the frequency of autotomy, a widespread antipredator mechanism. Competition increased the toxicity of chlorpyrifos and made it become lethal. Cannibalism was not affected by chlorpyrifos but increased at high density and under warming. Chlorpyrifos reduced heat tolerance but only when competition was high. This is the first demonstration that a biotic interaction can be a major determinant of 'toxicant-induced climate change sensitivity'. Competition enhanced the impact of chlorpyrifos under warming for high-latitude larvae, leading to an increase in autotomy which reduces fitness in the long term. This points to a novel pathway how transient pesticide pulses may cause delayed effects on populations in a warming world. Our results highlight that the interplay between biotic interactions and toxicants have a strong relevance for ecological risk assessment in a warming polluted world.

Sequential exposure to low levels of pesticides and temperature stress increase toxicological sensitivity of crustaceans

Frequent pesticide-related impacts on ecosystems at concentrations considered environmentally safe indicate that the current risk assessment framework for registration of pesticides is not protective enough. Causes may include difficulties in assessing the effects of sequential pesticide pulses and their interaction with environmental stressors. By contrast to such realistic scenarios, risk assessment for registration of pesticides is typically based on tests of a single exposure period under benign laboratory conditions. Here, we investigated the toxicological sensitivity of Gammarus pulex, an ecologically relevant crustacean, from uncontaminated control streams and pesticide-contaminated agricultural streams by exposing them to pesticide contamination in the laboratory. Individuals from contaminated streams were 2.7-fold more sensitive to pesticide exposure than individuals from the reference streams. We revealed that this increase in sensitivity was the result of a synergistic interaction of sequential pesticide exposure and temperature stress. Such multiple stressor scenarios are typical for agricultural streams. We conclude that the interactive effects of sequential toxicant exposure and additional environmental stressors need to be considered in a realistic risk assessment framework.

Negative effects of pesticides under global warming can be counteracted by a higher degradation rate and thermal adaptation

Summary An alarming finding for biodiversity is that global warming and pesticides often interact synergistically. Yet, this synergism may not capture the full picture because two counteracting processes may reduce the higher impact of pesticides under warming: higher pesticide degradation in the environment and thermal adaptation of populations. We tested for the effects of warming and multiple pulses of the insecticide chlorpyrifos on life history and fitness‐related physiological traits in the damselfly Ischnura elegans. To assess whether thermal adaptation is able to mitigate the impact of the pesticide under future warming, we exposed replicated populations from a colder, high latitude and from a warmer, low latitude in a common garden rearing experiment at 20 and 24 °C (the respective mean summer water temperatures at both latitudes). At the higher temperature pesticide degradation was higher, leading to less accumulation after multiple pulses compared to the lower temperature. Accordingly, the pesticide caused less mortality and less oxidative damage at the higher temperature. This contradicts the general belief (based on studies that kept pesticide concentrations constant) of a higher impact of pesticides under warming. Furthermore, the reduction of the impact of the pesticide at the higher temperature was more pronounced in the warm‐adapted low‐latitude populations, indicating a counteracting role of thermal adaptation. Policy implications. Our findings provide proof‐of‐principle for two key insights that will allow improving ecological risk assessment of pesticides under global warming: (i) higher pesticide degradation in the environment under warming may temper the impact of multiple pesticide pulses; (ii) gradual thermal evolution may further reduce the impact of pesticides at high latitudes under global warming. This knowledge can be very important for policy makers to arrive at a more realistic forecasting of the impacts of chemical pollution and climate change interactions on organisms.

Integrating both interaction pathways between warming and pesticide exposure on upper thermal tolerance in high- and low-latitude populations of an aquatic insect

Global warming and chemical pollution are key anthropogenic stressors with the potential to interact. While warming can change the impact of pollutants and pollutants can change the sensitivity to warming, both interaction pathways have never been integrated in a single experiment. Therefore, we tested the effects of warming and multiple pesticide pulses (allowing accumulation) of chlorpyrifos on upper thermal tolerance (CTmax) and associated physiological traits related to aerobic/anaerobic energy production in the damselfly Ischnura elegans. To also assess the role of latitude-specific thermal adaptation in shaping the impact of warming and pesticide exposure on thermal tolerance, we exposed larvae from replicated high- and low-latitude populations to the pesticide in a common garden rearing experiment at 20 and 24 °C, the mean summer water temperatures at high and low latitudes. As expected, exposure to chlorpyrifos resulted in a lower CTmax. Yet, this pesticide effect on CTmax was lower at 24 °C compared to 20 °C because of a lower accumulation of chlorpyrifos in the medium at 24 °C. The effects on CTmax could partly be explained by reduction of the aerobic scope. Given that these effects did not differ between latitudes, gradual thermal evolution is not expected to counteract the negative effect of the pesticide on thermal tolerance. By for the first time integrating both interaction pathways we were not only able to provide support for both of them, but more importantly demonstrate that they can directly affect each other. Indeed, the warming-induced reduction in pesticide impact generated a lower pesticide-induced climate change sensitivity (in terms of decreased upper thermal tolerance). Our results indicate that, assuming no increase in pesticide input, global warming might reduce the negative effect of multiple pulse exposures to pesticides on sensitivity to elevated temperatures.

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.

Competition impedes the recovery of Daphnia magna from repeated insecticide pulses

The effects of multiple insecticide pulses on non-target organisms have rarely been investigated in combination with relevant biotic interactions, such as competition. In this study, we examined the effects of two repeated pulses of the insecticide pirimicarb (3, 10, 24μg/L) on populations of Daphnia magna with or without competition. To investigate the influence of competition, half of the test systems were supplemented with the pirimicarb-insensitive species Culex pipiens. The pesticide pulses were followed by a recovery period of 28 days, which corresponded to approximately three generation times for D. magna. The one-species setup with the Daphnia populations and the two-species setup with both the Daphnia and Culex populations had a precontamination period of 30 days so that intra- and interspecific competitions were present prior to the insecticide pulse. Short-term effects on the survival of the Daphnia population were observed in both setups immediately after each insecticide pulse at the highest concentration level. In the one-species setup, the short-term effects on population survival were increased by intraspecific competition. However, the Daphnia populations in the one-species setup recovered and reached the control level within approximately two weeks after each insecticide pulse. In contrast, in the two-species setup at the highest concentration, we observed culmination of insecticide effects: the Daphnia populations did not recover and their abundance was below the control level until the end of the observation time. Their recovery was impeded by the presence of the competing species C. pipiens for at least four weeks. At low concentrations, no culmination of effects was observed. We conclude that repeated toxicant pulses on populations that are challenged with interspecific competition may result in a multigenerational culmination of toxicant effects.

Pairing behavior and reproduction in Hyalella azteca as sensitive endpoints for detecting long-term consequences of pesticide pulses

The aim of the present study was to examine acute and delayed effects of pulse exposure of the pyrethroid pesticide, permethrin, on precopulatory pairs of Hyalella azteca. Pairs of H. azteca were exposed to a single 1h pulse of different nominal concentrations of permethrin: 0, 0.3, 0.9 or 2.7μg/L. During exposure, pairing behavior was observed, and during a 56 day post-exposure period the treatments were monitored for pairing behavior, survival and reproductive output. All permethrin-exposed pairs separated within minutes during exposure and shortly thereafter became immobile; however they regained mobility after transfer to clean water. The time to re-form pairs was significantly longer in all tested concentrations compared to the control, although all surviving pairs re-formed within the 56 day test period. Nevertheless not all pairs exposed to 0.9 and 2.7μg/L reproduced. Furthermore the numbers of juveniles produced by pairs exposed to 0.9 and 2.7μg/L, but not 0.3μg/L, were lower throughout the entire post-exposure period compared to the control groups, and the total numbers of juveniles produced during 56 days were significantly lower in organisms exposed to 0.9 and 2.7μg/L, but not 0.3μg/L, compared to the control groups. The long-term effects of short-term exposure on reproductive behavior of pairs could potentially have consequences for the population dynamics of H. azteca. However, since individual-level responses can both overestimate and underestimate effects at the population level, appropriate population models are needed to reduce the uncertainty in extrapolating between these levels of biological organization.

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 12μg/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.

How does a pesticide pulse increase vulnerability to predation? Combined effects on behavioral antipredator traits and escape swimming

An increasing number of studies have documented that sublethal pesticide exposure can change predator–prey interactions. Most of these studies have focused on effects of long-term pesticide exposure on only one type of antipredator traits and have not directly linked changes in these traits to mortality by predation. To get a better mechanistic understanding of how short-term pesticide pulses make prey organisms more vulnerable to predation, we studied effects of 24h exposure to a sublethal concentration of the insecticide endosulfan and the herbicide Roundup on the major antipredator traits and the resulting mortality by predation in larvae of the damselfly Enallagma cyathigerum. A pulse of both pesticides affected antipredator traits involved in avoiding detection by predators as well as traits involved in escape after detection. After a pesticide pulse, larvae increased activity levels and even further increased the number of walks when predation risk was present. Further, an endosulfan pulse tended to reduce escape swimming speed. In contrast, previous exposure to Roundup caused the larvae to swim faster, yet less often when attacked. Importantly, although both studied pesticides induced maladaptive changes in overall activity, only for endosulfan this resulted in an increased mortality by predation. Our study highlights that considering changed predator–prey interactions may improve ecological risk evaluations of short pesticide pulses, yet also underscores the need (1) to consider effects on all important antipredator traits of the prey as trait compensation may occur and (2) to effectively score the outcome of predator–prey interactions in staged encounters.

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 12μg/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.

Competition increases toxicant sensitivity and delays the recovery of two interacting populations

We investigated how persistent competitive pressure alters toxicant sensitivity and recovery from a pesticide pulse at community level. Interacting populations of Daphnia ( Daphnia magna) and Culex larvae ( Culex pipiens molestus) were pulse-exposed (48 h) to the pyrethroid fenvalerate. The abundance and biomass of the populations were monitored by non-invasive image analysis. Shortly after exposure, Daphnia showed a concentration–response relationship with the toxicant with an LC 50 of 0.9 μg/L. Culex larvae were slightly less sensitive with an LC 50 of 1.7 μg/L. For both species, toxicant sensitivity increased with the population biomass of the respective species before exposure, which is explained by intraspecific competition. Several weeks after exposure to the highest treatment concentration of 1 μg/L, the slight differences in sensitivity between the two species were amplified to contrasting long-term effects due to interspecific competition: high interspecific competition impaired the recovery of Daphnia. Subsequently, Culex larvae profited from the slow recovery of Daphnia and showed an increased success of emergence. We conclude that, in natural systems where competition is present, such competitive processes might prolong the recovery of the community structure. Hence, natural communities might be disturbed for a longer period by toxic exposure than predicted from single-species tests alone.

Habitat isolation shapes the recovery of aquatic insect communities from a pesticide pulse

Summary1. The role of spatial processes in ecotoxicology is largely ignored. Yet, together with species‐specific sensitivities to the pollutant, spatial processes may determine community recovery and the resulting community composition after a pollution event.2. We investigated the isolation‐specific potential for internal recovery by reproduction and external recovery by dispersal after a pesticide pulse in experimental aquatic insect communities along an isolation gradient.3. External recovery was important in the univoltine species and in a multivoltine species that went locally extinct but only when a source population was nearby. Internal recovery occurred in all multivoltine species and was unexpectedly stronger in more isolated communities, probably because of release from a dispersal‐limited key predator.4. As a result, community recovery and resulting changes in composition strongly depended on isolation and species differences in the potential for external recovery through dispersal and internal recovery through reproduction.5. Synthesis and applications. Our results indicate that while the immediate impact of a toxicant on natural communities is shaped by species sensitivities, their recovery is primarily dependent on the degree of isolation. Risk assessment and the protection of communities under toxicant threat may improve greatly from considering the spatial context: isolated communities and communities with poor dispersers should receive extra protection to safeguard their member species. Ideally, land use planning should strive to remediate isolation of natural communities under threat in agricultural landscapes. Where this is not possible, spatial regulation of pesticide application may considerably improve protection of extant diversity.

Behavioural changes in three species of freshwater macroinvertebrates exposed to the pyrethroid lambda-cyhalothrin: Laboratory and stream microcosm studies

Pesticides are transported from crop fields to adjacent streams via surface run-off, drains, groundwater, wind drift and atmospheric deposition and give rise to transient pulse contamination. Although the concentrations observed, typically <10 μg L −1, cannot be expected to be acutely lethal, effects in streams at the population and ecosystem level have been reported. One of the most conspicuous phenomena associated with these transient pesticide pulses is drift, where large numbers of freshwater invertebrates are carried along by the current and disappear from the contaminated stretch of the stream. The aim of the present study was to evaluate the feasibility of linking laboratory studies of the sublethal effects of pulse exposure to the pyrethroid lambda-cyhalothrin on the locomotory behaviour of stream invertebrates with effects on drift behaviour under more environmentally realistic conditions in stream microcosms. In the laboratory as well as in the microcosms, the order of sensitivities of the three species tested was (with Leuctra nigra being the most sensitive): L. nigra > Gammarus pulex > Heptagenia sulphurea. The LOECs determined for L. nigra (1 ng L −1), G. pulex (10 ng L −1) and H. sulphurea (100 ng L −1) are all within expected environmental concentrations. For the species of invertebrates investigated, it was possible to extrapolate directly from pyrethroid-induced behavioural changes observed in the laboratory to drift under more realistic conditions in stream microcosms. Consequently, the fast and cost-effective video tracking methodology may be applied for screening for potential effects of a wider range of pesticides and other stressors on the locomotory behaviour of freshwater invertebrates. The results indicate that such behavioural changes may be predictive of effects at the ecosystem level.

Modeling responses of Daphnia magna to pesticide pulse exposure under varying food conditions: intrinsic versus apparent sensitivity

Recent studies showed that limiting food conditions resulted in either increased or decreased sensitivity of Daphnia magna to toxicants. It remained unclear whether these contrasting food-dependent alterations in toxicity resulted from differences in intrinsic sensitivity of the daphnids or from changes in toxicokinetics and resource allocation. It is hypothesized here that, if food level only affects accumulation kinetics and resource allocation, then the intrinsic sensitivity to this toxicant should be the same for all food regimes. This hypothesis was investigated using the DEBtox model, which is based on the theory of Dynamic Energy Budgets. We examined results of two recently conducted life-cycle studies on the combined effects of food level and a pulsed exposure to the pyrethroid insecticide fenvalerate (FV) on D. magna. The model described the effects of the time-varying exposure well, and indicated that when the animals did not die from exposure to FV, full reversibility of toxic effects was possible, allowing a complete recovery. Results revealed furthermore that the data from both studies could be described by the same NECs for survival and assimilation, killing rate and tolerance concentration (132 (49.2-228) x 10(-6) microg/L, 0 (0-1.18 x 10(-5)) microg/L, 74.4 (55.6-96.4) L (microg d)(-1) and 5.39 (2.72-18.5) x 10(-3) microg/L, respectively). It is therefore concluded that food-dependent FV toxicity can be explained by altered toxicokinetics and resource allocation, but not by changes in the intrinsic sensitivity of the daphnids. This study implies that the effect of pesticide application in the field depends on the trophic state of the receiving water body, but also that full recovery of survivors is possible after FV application.

Metabolic effects of dinoseb, diazinon and esfenvalerate in eyed eggs and alevins of Chinook salmon ( Oncorhynchus tshawytscha) determined by 1H NMR metabolomics

Pesticide pulses in the Sacramento River, California, originate from storm-water discharges and non-point source aquatic pollution that can last from a few days to weeks. The Sacramento River and its tributaries have historically supported the majority of California's Chinook salmon ( Oncorhynchus tshawytscha) spawning grounds. Three pesticides currently used in the Sacramento Valley – dinoseb, diazinon, and esfenvalerate – were chosen to model the exposure of salmon embryos to storm-water discharges. Static-renewal (96 h) exposures to eyed eggs and alevins resulted in both toxicity and significant changes in metabolism assessed in whole-embryo extracts by 1H nuclear magnetic resonance (NMR) spectroscopy based metabolomics and HPLC with UV detection (HPLC-UV). The 96-h LC 50 values of eyed eggs and alevins exposed to dinoseb were 335 and 70.6 ppb, respectively, and the corresponding values for diazinon were 545 and 29.5 ppm for eyed eggs and alevins, respectively. The 96-h LC 50 of eyed eggs exposed to esfenvalerate could not be determined due to lack of mortality at the highest exposure concentration, but in alevins was 16.7 ppb. All esfenvalerate exposed alevins developed some degree of lordosis or myoskeletal abnormality and did not respond to stimulus or exhibit normal swimming behavior. ATP concentrations measured by HPLC-UV decreased significantly in eyed eggs due to 250 ppb dinoseb and 10 and 100 ppb esfenvalerate ( p < 0.05). Phosphocreatine, as measured by HPLC-UV, decreased significantly in eyed eggs due to 250 ppb dinoseb, 10 and 100 ppb esfenvalerate, and 100 ppm diazinon ( p < 0.05). Principal components analyses of 1H NMR metabolite fingerprints of eyed egg and alevin extracts revealed both dose-dependent and mechanism of action-specific metabolic effects induced by the pesticides. Furthermore, NMR based metabolomics proved to be more sensitive than HPLC-UV in identifying significant changes in sublethal metabolism of pesticide exposed alevins. In conclusion, we have demonstrated several benefits of a metabolomics approach for chemical risk assessment, when used in conjunction with a fish embryo assay, and have identified significant metabolic perturbations to the early life stages of Chinook salmon by currently used pesticides.