Effects of the fungicide pyrimethanil on biofilm and organic matter processing in outdoor lentic mesocosms.

The effect of the fungicide pyrimethanil (0.7 mg L(−1)) on biofilm development and alder leaf litter decomposition in aquatic ecosystems was assessed in outdoor lentic mesocosms immediately and 274 days after pyrimethanil application. Pyrimethanil decreased ergosterol concentrations (an indicator of fungal biomass) and the abundance and richness of the macroinvertebrate community associated with decomposing leaves. However, because neither fungi nor macroinvertebrates were main factors contributing to decomposition in this particular system, organic matter processing rates were not affected. After 274 days, pyrimethanil concentration in the water column was ≤0.004 mg L(−1) but richness, biomass and composition of the invertebrate community associated with decomposing leaf-litter still showed the effect. The comparison of ergosterol (a molecule existing on both algae and fungal cell membranes), with chlorophyll (an indicator of algal biomass) associated with biofilm suggests that pyrimethanil may decrease fungal biomass and alter the relative abundance of algae and fungi on biofilm developing in control- and treated-mesocosms.

Unraveling the interactive effects of climate change and oil contamination on laboratory-simulated estuarine benthic communities.

There is growing concern that modifications to the global environment such as ocean acidification and increased ultraviolet radiation may interact with anthropogenic pollutants to adversely affect the future marine environment. Despite this, little is known about the nature of the potential risks posed by such interactions. Here, we performed a multifactorial microcosm experiment to assess the impact of ocean acidification, ultraviolet B (UV-B) radiation and oil hydrocarbon contamination on sediment chemistry, the microbial community (composition and function) and biochemical marker response of selected indicator species. We found that increased ocean acidification and oil contamination in the absence of UV-B will significantly alter bacterial composition by, among other things, greatly reducing the relative abundance of Desulfobacterales, known to be important oil hydrocarbon degraders. Along with changes in bacterial composition, we identified concomitant shifts in the composition of oil hydrocarbons in the sediment and an increase in oxidative stress effects on our indicator species. Interestingly, our study identifies UV-B as a critical component in the interaction between these factors, as its presence alleviates harmful effects caused by the combination of reduced pH and oil pollution. The model system used here shows that the interactive effect of reduced pH and oil contamination can adversely affect the structure and functioning of sediment benthic communities, with the potential to exacerbate the toxicity of oil hydrocarbons in marine ecosystems.

A short-term laboratory and in situ sediment assay based on the postexposure feeding of the estuarine isopod Cyathura carinata.

In situ assays based on feeding depression have been proposed as sublethal assays able to assess immediate contaminant effects on key ecosystem functions, long before effects on life-history traits can be detected. The in situ peculiarities provide more realistic exposure scenarios than laboratory-controlled conditions, which is particularly relevant for estuarine ecosystems where environmental conditions are highly variable. In this context, we developed a short-term cost-effective in situ assay based on the postexposure feeding of the estuarine species, Cyathura carinata, through the following steps: i) develop a methodology to quantify feeding under laboratory conditions; ii) quantify the sensitivity of the postexposure feeding response under laboratory conditions; iii) deploy the in situ assay at various field sites covering different environmental variables in a reference estuary and degrees of contamination in a contaminated estuary to, respectively, derive a model to predict postexposure feeding rates across sites varying in environmental variables and evaluate the potential of this in situ tool to assess sediment contamination in estuaries. A quantity of 100 defrosted nauplii of Artemia franciscana per isopod given during a short period of 30 min in darkness was adequate to allow estimating precise postexposure feeding rates. Laboratory toxicity tests showed the high sensitivity of postexposure feeding to Cd as a sublethal endpoint for C. carinata (7-fold lower than the corresponding 48-h LC50). However, this species appears to be able to face high concentrations of non-essential metals as Cd, with 96-h LC50 values (37 mg Cd/L) higher than values reported for other isopods, and among the highest concentrations reported for saltwater invertebrates. Results of in situ deployments at reference sites indicated that sediment with size fraction between 63-125 µm (very fine sand) influences feeding rates. The in situ assay across contaminated sites was able to discriminate between unpolluted and polluted estuaries, and also to detect degrees of toxicity among sampling sites within an estuary, after statistically adjusting feeding rates on the basis of the proportion of the sediment very fine sand influencing baseline feeding rates. The in situ postexposure feeding assay with C. carinata was found to be, therefore, a potential useful cost-effective tool for estuarine sediment toxicity assessments.

Contaminants as habitat disturbers: PAH-driven drift by Andean paramo stream insects.

Contaminants can behave as toxicants, when toxic effects are observed in organisms, as well as habitat disturbers and fragmentors, by triggering avoidance responses and generating less- or uninhabited zones. Drift by stream insects has long been considered a mechanism to avoid contamination by moving to most favorable habitats. Given that exploration and transportation of crude oil represent a threat for surrounding ecosystems, the key goal of the present study was to assess the ability of autochthonous groups of aquatic insects from the Ecuadorian paramo streams to avoid by drift different concentrations of polycyclic aromatic hydrocarbons (PAH) contained in the soluble fraction of locally transported crude oil. In the laboratory, different groups of insects were exposed to PAH for 12h. Three different assays, which varied in taxa and origin of the organisms, concentrations of PAH (0.6-38.8µgL(-1)), and environment settings (different levels of refuge and flow) were performed. For Anomalocosmoecus palugillensis (Limnephilidae), drift was a major cause of population decline in low concentration treatments but at higher concentrations mortality dominated. PAH was highly lethal, even at lower concentrations, for Chironomidae, Grypopterygidae (Claudioperla sp.) and Hydrobiosidae (Atopsyche sp.), and, therefore, no conclusion about drift can be drawn for these insects. Contamination by PAH showed to be a threat for benthic aquatic insects from Ecuadorian paramo streams as it can cause a population decline due to avoidance by drift and mortality.

Ethoprophos fate on soil-water interface and effects on non-target terrestrial and aquatic biota under Mediterranean crop-based scenarios.

The present study aimed to assess the environmental fate of the insecticide and nematicide ethoprophos in the soil-water interface following the pesticide application in simulated maize and potato crops under Mediterranean agricultural conditions, particularly of irrigation. Focus was given to the soil-water transfer pathways (leaching and runoff), to the pesticide transport in soil between pesticide application (crop row) and non-application areas (between crop rows), as well as to toxic effects of the various matrices on terrestrial and aquatic biota. A semi-field methodology mimicking a "worst-case" ethoprophos application (twice the recommended dosage for maize and potato crops: 100% concentration v/v) in agricultural field situations was used, in order to mimic a possible misuse by the farmer under realistic conditions. A rainfall was simulated under a slope of 20° for both crop-based scenarios. Soil and water samples were collected for the analysis of pesticide residues. Ecotoxicity of soil and aquatic samples was assessed by performing lethal and sublethal bioassays with organisms from different trophic levels: the collembolan Folsomia candida, the earthworm Eisenia andrei and the cladoceran Daphnia magna. Although the majority of ethoprophos sorbed to the soil application area, pesticide concentrations were detected in all water matrices illustrating pesticide transfer pathways of water contamination between environmental compartments. Leaching to groundwater proved to be an important transfer pathway of ethoprophos under both crop-based scenarios, as it resulted in high pesticide concentration in leachates from Maize (130µgL(-1)) and Potato (630µgL(-1)) crop scenarios, respectively. Ethoprophos application at the Potato crop scenario caused more toxic effects on terrestrial and aquatic biota than at the Maize scenario at the recommended dosage and lower concentrations. In both crop-based scenarios, ethoprophos moved with the irrigation water flow to the soil between the crop rows where no pesticide was applied, causing toxic effects on terrestrial organisms. The two simulated agricultural crop-based scenarios had the merit to illustrate the importance of transfer pathways of pesticides from soil to groundwater through leaching and from crop rows to the surrounding soil areas in a soil-water interface environment, which is representative for irrigated agricultural crops under Mediterranean conditions.

Active avoidance from a crude oil soluble fraction by an Andean paramo copepod.

Several oil spills due to ruptures in the pipeline oil systems have occurred at the Andean paramo. A sample of this crude oil was mixed with water from a nearby Andean lagoon and the toxicity of the soluble fraction was assessed through lethal and avoidance assays with a locally occurring copepod (Boeckella occidentalis intermedia). The integration of mortality and avoidance aimed at predicting the immediate decline of copepod populations facing an oil leakage. The 24-h median lethal PAH concentration was 42.7 (26.4-91.6) µg L(-1). In the 12-h avoidance assay, 30% avoidance was recorded at the highest PAH concentration (19.4 µg L(-1)). The mortality at this PAH concentration would be of 25% and, thus, the population immediate decline would be of 55%. The inclusion of non-forced exposure testing with the quantification of the avoidance response in environmental risk assessments is, therefore, supported due to underestimation of the lethal assays.

Ecological risk assessment of the Paraíba River estuary (Brazil): Plan for a tier 1 screening phase for the ecotoxicological line of evidence.

The knowledge produced in temperate regions is still used today to evaluate environmental problems in the tropics, without taking into consideration environmental differences, such as local conditions, the sensitivity and ecology of species, and exposure routes of contaminants, essential for understanding and determining the fate and toxicity of chemicals. Considering that Environmental Risk Assessment (ERA) studies for tropical systems are scarce and need to be adjusted, the present study aims at contributing to the awareness and development of tropical ecotoxicology. The estuary of the Paraíba River in Northeast Brazil was selected as a model study-case, as it is a large estuary, subjected to a heavy human pressure due to the numerous social, economic and industrial activities. The present study describes the framework for the problem formulation phase of the ERA process; it first includes a detailed integration of the scientific information available on the study area, to then derive the conceptual model, and finally present the analysis plan for the tier 1 screening phase. The latter is designed to be fundamentally supported by the ecotoxicological line of evidence, to ascertain, without delay, where and why there are environmental problems (i.e., adverse biological effects); ecotoxicological tools developed in temperate climates will be optimized to assess water quality in a tropical system. In addition to its intrinsic value for protecting the study area, the results of the present study are expected to provide an important baseline for conducting ERA in similar tropical aquatic systems across the globe.

Appraisal on the role of passive sampling for more integrative frameworks on the environmental risk assessment of contaminants.

Over time multiple lines of research have been integrated as important components of evidence for assessing the ecological quality status of water bodies within the framework of Environmental Risk Assessment (ERA) approaches. One of the most used integrative approaches is the triad which combines, based on the weight-of-evidence, three lines of research, the chemical (to identify what is causing the effect), the ecological (to identify the effects at the ecosystem level) and the ecotoxicological (to ascertain the causes of ecological damage), with the agreement between the different lines of risk evidence increasing the confidence in the management decisions. Although the triad approach has proven greatly strategic in ERA processes, new assessment (and monitoring) integrative and effective tools are most welcome. In this regard, the present study is an appraisal on the boost that passive sampling, by allowing to increase information reliability, can give within each of the triad lines of evidence, for more integrative ERA frameworks. In parallel to this appraisal, examples of works that used passive samplers within the triad are presented providing support for the use of these devices in a complementary form to generate holistic information for ERA and ease the process of decision-making.

Amendment of soils with metal-rich sludge: Potential water toxicity due to metal release via over-time slaking.

Industrial metal-rich sludge can improve soil properties, but it is potentially toxic to soils and adjacent aquatic systems. The soil-sludge-water interactions influence metals bioavailability over time, a phenomenon mostly regulated by the still debatable "sludge physical protection" or "sludge delayed release" hypotheses. The present study aimed to investigate: (1) whether sludge increases soil aggregate stability against slaking, (2) which hypothesis mostly regulates metal release from soils to water and (3) the ecotoxicity of the metals released during soil slaking for aquatic organisms. Under a realistic field scale, soils amended with an industrial sludge or spiked with equivalent metal solutions (of Cr, Cu, Ni, Zn) were collected over three months to test soil aggregate stability, the ecotoxicity of the slaking water and metal contents in soil and water. The "sludge physical protection" was verified for all metals, though for Cu the "sludge delayed release" hypothesis appears plausible after three months. Soil amendment with sludge did not lead to effects on the growth of the microalga Raphidocelis subcapitata, contrarily to the observed for the metal-spiked soil. Criteria regulating soils sludge-amendment management should thus include doses not hazardous to biota, and not only metal threshold levels.

Ecotoxicological characterization of a tropical soil after diazinon spraying.

The impact of diazinon spraying in an agricultural tropical soil through the evaluation of both the habitat and retention functions of the soil system was never reported. To fill this gap, five times the recommended dose of a commercial diazinon formulation was sprayed in an agricultural area of Costa Rica, and dilution gradients of the sprayed soil were prepared in the laboratory. Avoidance and reproduction tests with soil organisms (Eisenia andrei, Enchytraeus crypticus and Folsomia candida) to evaluate losses in terrestrial habitat function, and growth and reproduction tests with aquatic organisms (Chlorella vulgaris and Daphnia magna, respectively) to evaluate the retention function of soil were performed. Results demonstrated that regarding habitat function, F. candida reproduction was the most sensitive endpoint (EC(50) = 0.288 mg a.i./kg), followed by avoidance behaviour of E. andrei (EC(20) = 1.75 mg a.i./kg). F. candida avoidance and the reproduction of E. andrei and E. crypticus were not affected by diazinon. The toxicity tests with aquatic organisms showed that the soil retention function was insufficient to prevent effects of diazinon either on microalgae growth (EC(50) ≤ 0.742 mg/L and EC(20) ≤ 0.223 mg/L) and on the reproduction of the cladoceran (EC(50) ≤ 0.00771 mg/L and EC(20) ≤ 0.00646 mg/L). Results suggested that diazinon exerted toxic effects even at the dilution corresponding to the recommended dose, fact which makes its misuse an issue of environmental concern. Furthermore, the present study highlighted the importance and complementary nature of the assessment of both habitat and retention functions to an ecological risk assessment in tropical systems.

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.

Suitability of five cladoceran species from Mexico for in situ experimentation.

Although they are highly suited for site-specific risk assessment, little attention has been given to in situ ecotoxicological experimentation in tropical regions. This study intended to assess the suitability of five freshwater cladocerans from Mexico for in situ experiments. Juveniles of Ceriodaphnia cornuta, Macrothrix triserialis group, Simocephalus vetulus, Diaphanosoma birgei, and Pseudosida cf. ramosa, from two populations, were used to perform seven day laboratory and in situ experiments. Pseudosida cf. ramosa was very sensitive to handling and was discarded from further evaluations. Ceriodaphnia cornuta reproduced significantly more neonates, with smaller coefficients of variation, than M. triserialis and S. vetulus. Also due to its worldwide distribution in the tropics, C. cornuta is suggested to be highly suitable for in situ experiments.

Evaluating formulation and storage of Arthrobacter aurescens strain TC1 as a bioremediation tool for terbuthylazine contaminated soils: Efficacy on abatement of aquatic ecotoxicity.

Recently, a bioremediation tool consisting of freshly grown cells of the Arthrobacter aurescens strain TC1 proved successful in the cleanup of terbuthylazine (TBA) contaminated soil, with potential to prevent TBA dispersal via the soil-water pathway. The present work aimed at examining the feasibility of preparing and formulating bioaugmentation bacterial cells in advance which retain adequate viability and herbicide-degrading activity during storage and transport to contaminated sites. Three different types of formulation of A. aurescens TC1 cells were performed, namely cell paste, lyophilized, and adsorbed onto the mineral carrier vermiculite. Vermiculite-based cell material and cell paste offered survival rates significantly higher (≥78% of cells recovered viable, when compared to the initial numbers) than lyophilized cells (≥65%) after storage at 4 °C for up to a maximum period of 30 days. Inocula prepared from the three types of formulated and stored cells supported adequate levels of herbicide-biodegradation activity, ultimately allowing an almost complete removal of TBA from either liquid buffer or soil. In soil microcosms, the efficacy of bioremediation of TBA-contaminated soil in terms of decontamination (%) based on the ecotoxicity assessment of soil eluates towards a freshwater microalga was found to differ within formulations in the order: lyophilized cells (~100% efficacy at the end of one or two weeks of bioremediation treatment with an initial inoculum of ~2.5 × 107 viable cells/g of soil; similar to fresh cells) > vermiculite-adsorbed cells (81% with ~2.4 × 108 cells/g) > cell paste (68% with ~2.4 × 108 cells/g). The limitations and advantages of each type of bacterial formulation, together with the prediction of their applicability in the field are discussed.

Long-term effects of the fungicide pyrimethanil on aquatic primary producers in macrophyte-dominated outdoor mesocosms in two European ecoregions.

Even though empirical data supporting sound ecoregion-specific ecotoxicological evaluations are still scarce, the differences of environmental (including climatic) conditions in specific ecoregions are already currently being regulated for environmental risk assessment of pesticides in Europe. To shed new light on the ecotoxicological effects of pesticides on aquatic communities across ecoregions, the model pollutant pyrimethanil (fungicide) was tested in an outdoor mesocosm study with macrophyte-dominated communities in the European "South" (Portugal) and "Centre" (Germany) regulatory zones. Phytoplankton indicators monitored over 12 months indicated a low risk of the fungicide (0.73 or 0.77 mg pyrimethanil L-1, single application) to phytoplankton functioning; as expected since exposure simulated worst-case scenarios. However, the growth of key structural macroalgae and macrophytes was affected by the fungicide and negative effects occurred, especially in the Central zone experiment. Such effects were not detected earlier than approximately nine months post single pyrimethanil application. The presence or absence of such extremely long-lasting/delayed pyrimethanil effects depended on species, competitive situation, and ecoregion-specific physico-chemical environment. The present findings suggest that a better understanding of both direct and indirect effects of fungicide pollution on aquatic flora in two European ecoregions helps to consolidate the environmental risk assessment of pesticides in specific regulatory zones.

Avoidance tests with small fish: determination of the median avoidance concentration and of the lowest-observed-effect gradient.

The purpose of the present study was to develop sensitive, rapid, and easily quantified avoidance tests for small fish (Danio rerio) in order to provide important ecological information during toxicity assessments. Fish were exposed in three replicate linear flow-through chambers consisting of five compartments. The test system was found to provide a linear contamination gradient, with mean dilutions in each compartment of 90, 70, 50, 30, and 10%. Also, in the absence of a toxic gradient, the fish were uniformly distributed along the five-compartment chambers. Then the apparatus was evaluated by exposing fish to a concentration gradient of copper and a dilution gradient of a field sample contaminated with acid mine drainage (AMD). Avoidance was monitored at 24-h intervals up to 96 h of exposure. The avoidance of copper and AMD by D. rerio was confirmed. The apparatus enabled quantification of median avoidance effect concentrations or dilutions (EC50 or EDil50) and also lowest-observed-effect gradients, which express the minimum toxicant gradient eliciting avoidance, a parameter increasing the ecological relevance of the laboratory avoidance responses. For quantifying avoidance, a 24-h exposure was sufficient, as the 24- to 96-h EC50 and EDil50 values were similar. The avoidance response was easy and rapid to quantify, leading this test to routine use in environmental risk assessment.

Stressor-driven emigration and recolonisation patterns in disturbed habitats.

Although essential to conservation, little is known about how stress intensity can provoke emigration from disturbed habitats and allow recolonisation of those same environments. To demonstrate the applicability of laboratory experiments, we tested two hypotheses empirically using zebrafish response to artificially polluted environments that exhibited a linear gradient of stressor (acid mine drainage) levels. We hypothesized that emigration is distance-independent but time-correlated (spacelessness hypothesis). Additionally, we hypothesized that stressor-driven emigration could predict the extent of population growth in recovering habitats (avoidance-recolonisation hypothesis). For example, if half the organisms emigrate at a given stressor level, then the remainder should be able to recolonise a habitat experiencing the same stressor intensity. Comparisons of the small-scale experiment with a larger-scale simulation suggested that controlled laboratory results can be extrapolated to field populations (although time to perceive the contamination gradient may pose differential individual effects) because AC50 (median avoidance concentration) values of an acid mine drainage sample (AMD) were not statistically different when fish were exposed to the same gradient in 3-m long [0.50% (0.43-0.57)] or 30-m long [0.73% (0.30-2.2)] systems. Regarding the avoidance-recolonisation hypothesis, the number of recolonisers was inversely proportional to the number of avoiders (ACx = RC100-x). In particular, the similar distribution of fish along the 0-3% AMD gradient in both avoidance and recolonisation experiments resulted in identical AC50 and RC50 values: 0.55% (0.34-0.87) and 0.55% (0.45-0.67) AMD, respectively. The inclusion of avoidance and recolonisation responses in the environmental risk assessments provides a novel perspective of risk based on the emigration of organisms and contributes to the understanding and prediction of biological invasions and ecosystem recovery after restoration.

Stress-driven emigration in complex field scenarios of habitat disturbance: The heterogeneous multi-habitat assay system (HeMHAS).

Lines of evidence used in ecological risk assessment (ERA) are essentially three: chemistry, biology and ecotoxicology. Until now, the fundamental assumption made when measuring ecological risks is that organisms are forcedly exposed to stressors. However, when organisms can avoid disturbed habitats by escaping to less stressful areas, the assumption that exposure is mandatory to pose risk may not match field disturbance scenarios. A non-forced exposure approach using a linear free-choice multi-compartmented system has been proposed previously as a complementary tool to assess the effects on organisms' spatial avoidance/preference responses. Yet, the linearity of the latter system limits avoidance measurements to one spatial dimension. A novel, heterogeneous multi-habitat assay system (HeMHAS) consisting of 18 connected circular compartments (3 compartments on a vertical axis in each one of 6 zones on a longitudinal axis; a 2D system) is put forward here to be used in heterogeneous-habitat selection studies, as it makes it possible to assess the ability of organisms to detect contamination and other stressors and select more favorable habitats. In the present study, the avoidance to copper by zebrafish (Danio rerio) was tested after exposing organisms to a copper gradient in the HeMHAS and compared with that in the linear system. Avoidance occurred for all copper concentrations: 43% in the lowest (21 µg·L-1) to 72% in the highest (221 µg·L-1). Results obtained within the HeMHAS (AC50: 60 µg·L-1) were statistically (p = 0.72) similar to avoidance of copper by D. rerio in the linear non-forced system (AC50: 89 µg·L-1). In summary, the simulation of a copper gradient in the HeMHAS (2D system) allowed to assess the potential repellency of copper to zebrafish and to corroborate the ability of organisms to detect and avoid potentially toxic concentrations.

Habitat fragmentation caused by contaminants: Atrazine as a chemical barrier isolating fish populations.

Information on how atrazine can affect the spatial distribution of organisms is non-existent. As this effect has been observed for some other contaminants, we hypothesized that atrazine-containing leachates/discharges could trigger spatial avoidance by the fish Poecilia reticulata and form a chemical barrier isolating upstream and downstream populations. Firstly, guppies were exposed to an atrazine gradient in a non-forced exposure system, in which organisms moved freely among the concentrations, to assess their ability to avoid atrazine. Secondly, a chemical barrier formed by atrazine, separating two clean habitats (extremities of the non-forced system), was simulated to assess whether the presence of the contaminant could prevent guppies from migrating to the other side of the system. Fish were able to avoid atrazine contamination at environmentally relevant concentrations (0.02 µg L-1), below those described to cause sub-lethal effects. The AC50 (atrazine concentration causing avoidance to 50% of the population) was 0.065 µg L-1. The chemical barrier formed by atrazine at 150 µg L-1 (concentration that should produce an avoidance around 82%) caused a reduction in the migratory potential of the fish by 47%; while the chemical barrier at 1058 µg L-1 (concentration that produces torpidity) caused a reduction in the migratory potential of the fish by 91%. Contamination by atrazine, besides driving the spatial distribution of fish populations, has potential to act as a chemical barrier by isolating fish populations. This study includes a novel approach to be integrated in environmental risk assessment schemes to assess high-tier contamination effects such as habitat fragmentation and population displacement and isolation.

A novel approach to assessing environmental disturbance based on habitat selection by zebra fish as a model organism.

Aquatic ecotoxicity assays used to assess ecological risk assume that organisms living in a contaminated habitat are forcedly exposed to the contamination. This assumption neglects the ability of organisms to detect and avoid contamination by moving towards less disturbed habitats, as long as connectivity exists. In fluvial systems, many environmental parameters vary spatially and thus condition organisms' habitat selection. We assessed the preference of zebra fish (Danio rerio) when exposed to water samples from two western Ecuadorian rivers with apparently distinct disturbance levels: Pescadillo River (highly disturbed) and Oro River (moderately disturbed). Using a non-forced exposure system in which water samples from each river were arranged according to their spatial sequence in the field and connected to allow individuals to move freely among samples, we assayed habitat selection by D. rerio to assess environmental disturbance in the two rivers. Fish exposed to Pescadillo River samples preferred downstream samples near the confluence zone with the Oro River. Fish exposed to Oro River samples preferred upstream waters. When exposed to samples from both rivers simultaneously, fish exhibited the same pattern of habitat selection by preferring the Oro River samples. Given that the rivers are connected, preference for the Oro River enabled us to predict a depression in fish populations in the Pescadillo River. Although these findings indicate higher disturbance levels in the Pescadillo River, none of the physical-chemical variables measured was significantly correlated with the preference pattern towards the Oro River. Non-linear spatial patterns of habitat preference suggest that other environmental parameters like urban or agricultural contaminants play an important role in the model organism's habitat selection in these rivers. The non-forced exposure system represents a habitat selection-based approach that can serve as a valuable tool to unravel the factors that dictate organisms' spatial distribution in connected ecosystems.

In situ assays with tropical cladocerans to evaluate edge-of-field pesticide runoff toxicity.

Tropical regions' economy is usually based on agriculture, which involves an intensive use, and even frequent overuse, of pesticides. Nevertheless, not much research has been done on the impact of pesticides on tropical aquatic ecosystems, which are often contaminated by runoff-related pesticide inputs due to unpredictable and torrential rainfalls. This study aimed to: (i) adapt and evaluate a short-term sublethal in situ assay using post-exposure feeding as an endpoint, to the tropical cladoceran species Diaphanosoma brachyurum (collected at the Pedra do Cavalo dam in the Paraguaçu River basin, Bahia, Brazil), and, (ii) assess the role of the standard species Daphnia magna as an adequate laboratory surrogate. Lethal and sublethal (post-exposure feeding) responses were assessed for the two species. To evaluate these responses under environmentally realistic exposure conditions, a runoff event was simulated in an agricultural area previously contaminated with different deltamethrin concentrations. The resultant runoff water was used to set up microcosms with different dilutions, simulating the entrance of runoff water in an adjacent lentic system. An in situ assay with D. brachyurum was performed inside the microcosms, allowing to discriminate the effects due to deltamethrin from those due to other potential stressors associated with the experimental design (e.g. organism handling, load of suspended particles, microcosm design). Water samples were collected from microcosms to conduct a laboratory assay with D. magna. The in situ methodologies revealed to be suitable to conduct assays with D. brachyurum under tropical conditions, since all exposed organisms were successfully retrieved from the chambers. Furthermore, none of the potential stressors associated with the experimental design influenced the daphnids' performance. The tropical cladoceran species, exposed under more realistic conditions, revealed to be more sensitive than the laboratory standard species: lethal effects were only observed for D. brachyurum and sublethal effects were noticed at a lower deltamethrin concentration for this species than for D. magna.