Aquatic community structure in Mediterranean edge-of-field waterbodies as explained by environmental factors and the presence of pesticide mixtures.

Studies addressing the predicted effects of pesticides in combination with abiotic and biotic factors on aquatic biota in ditches associated with typical Mediterranean agroecosystems are scarce. The current study aimed to evaluate the predicted effects of pesticides along with environmental factors and biota interactions on macroinvertebrate, zooplankton and phytoplankton community compositions in ditches adjacent to Portuguese maize and tomato crop areas. Data was analysed with the variance partitioning procedure based on redundancy analysis (RDA). The total variance in biological community composition was divided into the variance explained by the multi-substance potentially affected fraction [(msPAF) arthropods and primary producers], environmental factors (water chemistry parameters), biotic interactions, shared variance, and unexplained variance. The total explained variance reached 39.4% and the largest proportion of this explained variance was attributed to msPAF (23.7%). When each group (phytoplankton, zooplankton and macroinvertebrates) was analysed separately, biota interactions and environmental factors explained the largest proportion of variance. Results of this study indicate that besides the presence of pesticide mixtures, environmental factors and biotic interactions also considerably influence field freshwater communities. Subsequently, to increase our understanding of the risk of pesticide mixtures on ecosystem communities in edge-of-field water bodies, variations in environmental and biological factors should also be considered.

Evaluation of FOCUS surface water pesticide concentration predictions and risk assessment of field-measured pesticide mixtures-a crop-based approach under Mediterranean conditions.

FOCUS models are used in the European regulatory risk assessment (RA) to predict individual pesticide concentrations in edge-of-field surface waters. The scenarios used in higher tier FOCUS simulations were mainly based on Central/North European, and work is needed to underpin the validity of simulated exposure profiles for Mediterranean agroecosystems. In addition, the RA of chemicals are traditionally evaluated on the basis of single substances although freshwater life is generally exposed to a multitude of pesticides. In the present study, we monitored 19 pesticides in surface waters of five locations in the Portuguese 'Lezíria do Tejo' agricultural area. FOCUS step 3 simulations were performed for the South European scenarios to estimate predicted environmental concentrations (PECs). We verified that 44% of the PECs underestimated the measured environmental concentrations (MEC) of the pesticides, showing a non-compliance with the field data. Risk was assessed by comparing the environmental quality standards (EQS) and regulatory acceptable concentrations with their respective MECs. Risk of mixtures was demonstrated in 100% of the samples with insecticides accounting for 60% of the total risk identified. The overall link between the RA and the actual situation in the field must be considerably strengthened, and field studies on pesticide exposure and effects should be carried out to assist the improvement of predictive approaches used for regulatory purposes.

Toxicity of environmentally realistic concentrations of chlorpyrifos and terbuthylazine in indoor microcosms.

Few studies have been conducted into the evaluation of environmentally realistic pesticide mixtures using model ecosystems. In the present study, the effects of single and combined environmentally realistic concentrations of the herbicide terbuthylazine and the insecticide chlorpyrifos were evaluated using laboratory microcosms. Direct toxic effects of chlorpyrifos were noted on copepod nauplii and cladocerans and the recovery of the latter was likely related with the decrease observed in rotifer abundances. Terbuthylazine potentiated the effect of chlorpyrifos on feeding rates of Daphnia magna, presumably by triggering the transformation of chlorpyrifos to more toxic oxon-analogs. Possible food-web interactions resulting from multiple chemical (and other) stressors likely to be present in edge-of-field water bodies need to be further evaluated.

Predicting the aquatic risk of realistic pesticide mixtures to species assemblages in Portuguese river basins.

Although pesticide regulatory tools are mainly based on individual substances, aquatic ecosystems are usually exposed to multiple pesticides from their use on the variety of crops within the catchment of a river. This study estimated the impact of measured pesticide mixtures in surface waters from 2002 and 2008 within three important Portuguese river basins ('Mondego', 'Sado' and 'Tejo') on primary producers, arthropods and fish by toxic pressure calculation. Species sensitivity distributions (SSDs), in combination with mixture toxicity models, were applied. Considering the differences in the responses of the taxonomic groups as well as in the pesticide exposures that these organisms experience, variable acute multi-substance potentially affected fractions (msPAFs) were obtained. The median msPAF for primary producers and arthropods in surface waters of all river basins exceeded 5%, the cut-off value used in the prospective SSD approach for deriving individual environmental quality standards. A ranking procedure identified various photosystem II inhibiting herbicides, with oxadiazon having the relatively largest toxic effects on primary producers, while the organophosphorus insecticides, chlorfenvinphos and chlorpyrifos, and the organochloride endosulfan had the largest effects on arthropods and fish, respectively. These results ensure compliance with European legislation with regard to ecological risk assessment and management of pesticides in surface waters.

Aquatic risk assessment of priority and other river basin specific pesticides in surface waters of Mediterranean river basins.

To meet good chemical and ecological status, Member States are required to monitor priority substances and chemicals identified as substances of concern at European Union and local/river-basin/national level, respectively, in surface water bodies, and to report exceedances of the environmental quality standards (EQSs). Therefore, standards have to be set at national level for river basin specific pollutants. Pesticides used in dominant crops of several agricultural areas within the catchment of Mediterranean river basins ('Mondego', 'Sado' and 'Tejo', Portugal) were selected for monitoring, in addition to the pesticides included in priority lists defined in Europe. From the 29 pesticides and metabolites selected for the study, 20 were detected in surface waters of the river basins, seven of which were priority substances: alachlor, atrazine, chlorfenvinphos, chlorpyrifos, endosulfan, simazine and terbutryn, all of which exceeded their respective EQS values. QSs for other specific pollutants were calculated using different extrapolation techniques (i.e. deterministic or probabilistic) largely based on the method described in view of the Water Framework Directive. Non-acceptable aquatic risks were revealed for molinate, oxadiazon, pendimethalin, propanil, terbuthylazine, and the metabolite desethylatrazine. Implications of these findings for the classification of the ecological status of surface water bodies in Portugal and at the European level are discussed.

Concentration addition-based approach for aquatic risk assessment of realistic pesticide mixtures in Portuguese river basins.

A two-tiered outline for the predictive environmental risk assessment of chemical mixtures with effect assessments based on concentration addition (CA) approaches as first tier and consideration of independent action (IA) as the second tier was applied based on realistic pesticide mixtures measured in surface waters from 2002 to 2008 within three important Portuguese river basins ('Mondego', 'Sado' and 'Tejo'). The CA-based risk quotients, based on acute data and an assessment factor of 100, exceeded 1 in more than 39 % of the 281 samples, indicating a potential risk for the aquatic environment, namely to algae. Seven herbicide compounds and three insecticides were the most toxic compounds in the pesticide mixtures and provided at least 50 % of the mixture's toxicity in almost 100 % of the samples with risk quotients based on the sum of toxic units (RQSTU) above 1. In eight samples, the maximum cumulative ratio (MCR) and the Junghan's ratio values indicated that a chemical-by-chemical approach underestimated the toxicity of the pesticide mixtures, and CA predicted higher mixture toxicity than that of IA. From a risk management perspective, the results pointed out that, by deriving appropriate programmes of measures to a limited number of pesticides with the highest contribution to the total mixture toxicity, relevant benefits also on mixture impact could be produced.

Comparing ecotoxicological standards of plant protection products potentially toxic to groundwater life with their measured and modelled concentrations.

Trigger values (TVs) for groundwater ecosystems in the European Union (EU), as elsewhere, are not based on toxicity data for the biota of that ecosystem. At present, very few toxicity tests have been conducted with groundwater organisms so the true sensitivity of groundwater ecosystems is largely unknown. In a previous published study, we set groundwater TVs for all plant protection products (PPPs) allowed for use at the time of the study based on toxicity data for surface water organisms as surrogates for groundwater organisms and calculated TVs lower than the current EU standard of 0.1 µg/L for 16 PPPs. This thus reveals that the effect assessment of these PPPs may not be fully adequate, but would still only indicate risk if the (expected) concentrations of these PPPs are greater than their calculated TVs. The present study was therefore initiated to evaluate whether predicted and measured concentrations of these PPPs are higher than the previously calculated TVs lower than 0.1 µg/L. To this end, predicted environmental concentrations (PECs) were calculated using the PELMO and SCI-GROW models that are currently used for this purpose in the EU and USA, respectively, and measured concentrations (MECs) were obtained from the open literature. In addition, the empirical PERPEST model was used to assess the severity and probability of effects that may be expected at these concentrations on taxonomic groups known to be well represented in groundwater ecosystems. In addition, only for dimethoate a PEC greater than 0.1 µg/L was calculated. However, when considering concentrations actually measured in the field, 99.7% showed risk quotients (RQ, as MEC/TV) values higher than 1 and 36.7% even higher than 100. Future field monitoring studies are needed to validate and eventually calibrate the way PEC values are currently calculated with the different models and scenarios currently in use. Such studies would also aid in the question to what extent the high MEC values may be attributed to diffuse or point-source pollution.

Preliminary aquatic risk assessment of imidacloprid after application in an experimental rice plot.

The potential aquatic risk of application of the neonicotinoid insecticide imidacloprid for aphid control in rice was assessed. To this end, imidacloprid was applied as Confidor(®) 200 SC at the recommended field dose of 100g a.i./ha to a Portuguese rice plot. Subsequently, fate of the test compound in water and potential effects of water samples on a battery of test species were determined. As compared to the first-tier predicted environmental concentrations (PECs) calculated using MED-Rice (around 30µg/L depending on the scenario used) and US-EPA (78µg/L) simulations, the actual peak concentration measured in the paddy water (52µg/L) was higher and lower, respectively. As was anticipated based on 50% effect concentrations (EC50 values) for Daphnia magna published in the open literature and that calculated in the present study (48h-EC50 immobility=84mg/L), no effects were observed of field water samples on daphnids. The sediment-dwelling ostracod Heterocypris incongruens, however, appeared relatively sensitive towards imidacloprid (6d-EC50 growth inhibition=0.01-0.015mg/L) and a slight effect was indeed noted in field samples taken the first week after application. Species sensitivity distributions based on published EC50 and NOEC values also revealed that other species are likely to be affected at the peak and time-weighted average imidacloprid concentrations, respectively. By applying the relative tolerance approach (i.e. by dividing the EC50 value of a certain species with that of Daphnia magna), ostracods appear to contain the most sensitive taxa to imidacloprid, followed by EPT (Ephemeroptera, Plecoptera and Trichoptera) taxa. Future field studies into (higher-tier) fate modelling of pesticides in rice paddies and effect assessment on field communities are required to ensure protection of aquatic life and wildlife (e.g. birds) from pesticide stress.

Evaluation of chemical parameters and ecotoxicity of a soil developed on gossan following application of polyacrylates and growth of Spergularia purpurea.

The aim of this study was to evaluate the chemical characteristics and ecotoxicity of a mine soil developed on gossan materials and amended with hydrophilic polyacrylate polymers after a growth cycle of Spergularia purpurea. Different acute bioassays (Daphnia magna immobilization; microalgae growth inhibition; germination and growth of lettuce and oat) were carried out with simulated leachates, pore water and soil samples. The germination and growth of native shrubs (Cistus ladanifer and Lavandula sampaioana) were also evaluated in the lysimeters where S. purpurea had grown. The soil had high total concentrations (g/kg) of Al (3.50-8.60), As (2.55-2.73), Cu (0.13-0.91) and Pb (4.48-6.16). However, the percentages of elements in aqueous extracts (simulating leachates, pore water, and the conditions of the rhizosphere soil) were small when compared to their total soil concentrations (less than 9% except for Na in leachates). Growth of S. purpurea and other natural colonization of plant species (Poaceae, Fabaceae and Asteraceae families) improved chemical characteristics but the application of the polyacrylate polymers contributed to a further improvement of soil quality. However, this was not sufficient to ensure the growth of a large number of shrubs despite a great germination rate. Among the several species used on the ecotoxicological assessment, the D. magna test was the only bioassay that showed a clear toxicity of soil leachates, suggesting the importance of using several ecotoxicological tests to assess the environmental risk of soil contamination and its rehabilitation. Although the studied soil can be considered contaminated taking into account the total soil concentrations of Al, As, Cu and Pb, the low concentrations of the same chemical elements in extractable solutions, that simulated the fractions really available for organisms, did not demonstrate a substantial toxic effects in the organisms and, consequently, negative impact on the environment.

Assessing the quality of freshwaters in a protected area within the Tagus River basin district (central Portugal).

Water-sediment quality was assessed in an agricultural zone of a protected area within the Tagus River basin district (central Portugal) combining chemical analysis to 12 pesticide compounds and whole toxicity testing using the bacterium , the algae , the crustacean , and the midge . The herbicides alachlor, atrazine ethofumesate, metolachlor, terbuthylazine, the insecticides chlorfenvinphos and chlorpyrifos, and the metabolite 3,4-dichloroaniline were detected in surface water samples at four sites and in groundwater samples from six wells, during four sampling occasions. Measured concentrations were compared with parametric values for human consumption, groundwater quality standards, and environmental quality standards applicable to surface water established in European Union legislation. Most severe adverse effects were noted on the growth of and lethality of in nondiluted water samples. Taking into account the values calculated by the method of toxic unit summation for pesticide mixtures, it was not possible to link the pesticides found to the toxicity detected in the water samples. Conducting this study with chemical analyses and biotests provided a more comprehensive quality assessment and realistic picture of the environmental samples analyzed, although additional studies are needed to evaluate the performance of mixture models for predicting mixture toxicity. This study underlines the importance of chemical analysis and whole toxicity testing as tools for assessing the impact of human activity on the status of water, mainly in protected zones.

Comparing the sensitivity of soil invertebrates to pesticides with that of Eisenia fetida.

The sole routine testing of the standard earthworm Eisenia fetida for the terrestrial risk assessment of pesticides has been under much debate since other soil invertebrates may be more sensitive than this standard test species. However, the very low availability of laboratory toxicity data for taxa other than E. fetida has greatly hampered sensitivity comparisons. In the present study, the relative tolerance (T(rel)) approach was used to enable comparing toxicity thresholds obtained from the US-EPA ECOTOX database, for main terrestrial taxonomic groups and pesticidal types of action (insecticides, fungicides, herbicides, and other) separately. Analyses confirmed previously reported lower and higher sensitivity of collembolans to fungicides and insecticides, respectively. However, various other discrepancies in susceptibility relative to E. fetida were encountered as indicated by species sensitivity distributions and/or calculated 95% confidence intervals of T(rel) values. Arachnids and isopods were found to be more sensitive to insecticides, and nematodes to fungicides, as compared to E. fetida. Implications of study findings for the terrestrial risk assessment of pesticides are discussed.

Exposure assessment of pesticides in a shallow groundwater of the Tagus vulnerable zone (Portugal): a multivariate statistical approach (JCA).

PURPOSE: To assess groundwater exposure to pesticides, in agricultural areas of 'Ribatejo' region (Portugal), and the influence of some key factors in that exposure, field, laboratory and modelling studies were carried out. METHODS: The study was performed in maize, potato, sugar beet, tomato and vegetables agricultural areas, located in a shallow aquifer, with pesticides use and, in most cases, with irrigation practices. Pesticides used in the studied agricultural areas and having leaching potential were selected, being considered also other pesticides included in priority lists, defined in Europe. Evaluation of groundwater exposure to pesticides was carried out by successively: (1) groundwater sampling in seven campaigns over the period 2004-2006; (2) pesticide analysis [including isolation and concentration from the groundwater samples and further determination by gas chromatography-mass spectrometry (GC-MS) of 14 herbicides, four insecticides and two metabolites]; and (3) analysis and discussion of the results by applying joint correspondence analysis (JCA). RESULTS: From the 20 pesticides and metabolites selected for the study, 11 were found in groundwater. Pesticides and metabolites most frequently detected were atrazine, alachlor, metolachlor, desethylatrazine, ethofumesate, α-endosulfan, metribuzine, lindane and ß-endosulfan. The results showed that groundwater exposure to pesticides is influenced by local factors-either environmental or agricultural, as precipitation, soil, geology, crops and irrigation practices. Spring and autumn were more associated with the detection of pesticides being more likely to observe mixtures of these compounds in a groundwater sample in these transition seasons. CONCLUSIONS: This work evidences the importance of models, which evaluate pesticides environmental behaviour, namely their water contamination potential (as Mackay multicompartimental fugacity model) and, specially, groundwater contamination potential (as GUS and Bacci and Gaggi leaching indices), in pesticide selection. Moreover, it reveals the importance to adapt proper statistical methods according to level of left-censored data. Using JCA was still possible to establish relations between pesticides and their temporal trend in a case study where there were more than 80% of data censored. This study will contribute to the Tagus river basin management plan with information on the patterns of pesticide occurrence in the alluvial aquifer system.

Integrated approach for the quality assessment of freshwater resources in a vineyard area (South Portugal).

An integrated chemical and biological approach for the quality assessment of freshwater resources in a vineyard area of the 'Alentejo' region (South Portugal) is presented. This includes analysis to 11 pesticide compounds and whole toxicity testing on algae and crustaceans. Simazine, terbuthylazine, terbutryn, desethylatrazine and chlorpyrifos were the most frequently detected pesticides in water collected from wells and drainage channels. Mixtures of up to three compounds in different qualitative combinations were also found. The quality standards for individual pesticides (0.1 µg L(-1)) and pesticides-total (0.5 µg L(-1)) were exceeded in some samples. However, their maximum concentrations were lower than the WHO guidelines, the USEPA health advisory values and the environmental quality standards for priority substances applicable to surface water. In five samples, the herbicides terbuthylazine and terbutryn and the insecticide chlorpyrifos did not pass the toxicity exposure ratio (TER) trigger values specified for aquatic organisms (algae, Daphnia and fish). Maximum toxic effects on Daphnia magna (100%) and Pseudokirchneriella subcapitata (82.56%) were determined in groundwater samples, while in surface water, no toxicity was observed. Concerning effects on Heterocypris incongruens in sediment samples collected at the drainage channels, mortality and growth inhibition values were below 38%. Pro-active management of the use of pesticides is recommended for implementing at the farm and catchment level to reduce inputs into ground- and surface water.

Does the actual standard of 0.1 microg/L overestimate or underestimate the risk of plant protection products to groundwater ecosystems?

The present groundwater standard of 0.1 microg/L for plant protection products (PPPs) has been under much debate because an ecotoxicological base is missing. In the present study, groundwater threshold values were calculated for all PPPs currently included in Annex I to Directive 91/414/EEC using three different approaches: (1) first-tier (Daphnia magna and Vibrio fisheri); (2) species sensitivity distributions, constructed for surrogate freshwater organisms for the truncated groundwater biodiversity; (3) the case-based model PERPEST. For the majority of the PPPs, the trigger value of 0.1 microg/L appears to be sufficiently protective. However, it may not fully protect groundwater life from several insecticides. Implications for the environmental risk assessment of groundwater and recommendations for future research are discussed.

Calibration and validation of a dynamic water model in agricultural scenarios.

A dynamic aquatic model (DynA model) was previously developed to predict the fate of a chemical in aquatic scenarios characterized by daily or periodic changes in several input parameters. DynA model is here calibrated with data obtained from the literature in specific unsteady state scenarios, such as those of rice fields. The results obtained for two herbicides (cinosulfuron and pretilachlor) in rice paddy scenarios revealed the capability of the model to accurately predict water and sediment concentrations, as shown by some statistical indicators. Modelling efficiency (EF) values of 0.86-0.99 for the water compartment and of 0.77-0.84 for sediment show the good agreement between predicted and measured concentrations. An "external validation" was performed using measured data for a different herbicide (molinate) applied in a Portuguese paddy rice scenario. A sensitivity analysis for this volatile chemical revealed the influence of some climatic parameters (e.g. temperature) to the model outcomes, such as water and sediment concentrations. This confirmed the capability of DynA model as an efficient tool for the pesticide risk assessment in dynamic scenarios.