Metal reference values established in Brazilian legislation for terrestrial disposal of dredged sediments and soil quality were derived for temperate regions. To evaluate the adequacy of such metal reference values to tropical soils, the ecotoxicity of a dredged sediment (from the Guanabara bay, Rio de Janeiro, Brazil) was investigated in two local soils (ferralsol and chernosol) by performing avoidance and reproduction tests using Folsomia candida and Enchytraeus crypticus. Test doses consisted of 0 %, 1.25 %, 2.5 %, 5, 10 %, and 20 %. Total and potentially bioavailable metal concentrations were determined in the test mixtures. Although the chernosol mixtures had the highest total metal concentrations, the influence of the expansive clay minerals (with high ability to adsorb metals) and the high contents of nutrients typical from this type of soils, seem to reduce the ecotoxicity. Collembolan avoidance behavior was the least sensitive endpoint. The lowest sediment doses increased the reproduction of F. candida in ferralsol mixtures. E. crypticus reproduction in the ferralsol mixtures were more pronounced at lower concentrations than in chernosol mixtures. Possibly the low nutrient content of the ferralsols, in connection with the addition of small amounts of sediment, created particular conditions that promoted reproduction of the test species. Data obtained in the ecotoxicological tests may support the establishment of a "safe" ecological dose of dredged sediments to be applied in tropical soils, supporting decision-makers in programs of environmental management.
Terrestrial ecotoxicological tests are powerful tools for assessing the ecological risks that pesticides pose to soil invertebrates, but they are rarely used to evaluate seed dressing pesticides. This study investigated the effects of seed dressing pesticides on survival and reproduction of Folsomia candida (Collembola), using standardized ecotoxicological tests (after ISO guidelines with few adaptations for tropical conditions). Commercial formulations of five seed dressing pesticides were tested individually in Tropical Artificial Soil (TAS): the insecticides imidacloprid, fipronil, thiametoxam, and the fungicides captan and carboxin+thiram. Thiametoxam, captan, and carboxin+thiram were only lethal to F. candida at the highest concentration tested (1000mg of active ingredient kg(-1) of dry soil). Imidacloprid and fipronil were lethal at lower concentrations (100 and 10mg a.i. kg(-1) soil d.w, respectively), however, these concentrations were much higher than those predicted (PEC) for soil. Imidacloprid and fipronil were the most toxic pesticides in both tests, reducing significantly collembolan reproduction (EC20=0.02 and 0.12mga.i.kg(-1) soil d.w, respectively). Further studies under more realistic conditions are needed, since imidacloprid and fipronil reduced collembolan reproduction at concentrations below or close to their respective PECs.
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.
Soil contaminant concentration limits for the protection of terrestrial plants and soil invertebrates are commonly based on thresholds derived using data from laboratory ecotoxicity tests. A comprehensive assessment has been made for the derivation of ecological soil screening levels (Eco-SSL) in the United States; however, these limits are conservative because of their focus on high bioavailability scenarios. Here, we explain and evaluate approaches to soil limit derivation taken by 4 jurisdictions, 2 of which allow for correction of data for factors affecting bioavailability among soils, and between spiked and field-contaminated soils (Registration Evaluation Authorisation and Restriction of Chemicals [REACH] Regulation, European Union [EU], and the National Environment Protection Council [NEPC], Australia). Scientifically advanced features from these methods have been integrated into a newly developed method for deriving soil clean-up values (SCVs) within the context of site-specific baseline ecological risk assessment. Resulting site-specific SCVs that account for bioavailability may permit a greater residual concentration in soil when compared to generic screening limit concentrations (e.g., Eco-SSL), while still affording acceptable protection. Two choices for selecting the level of protection are compared (i.e., allowing higher effect levels per species, or allowing a higher percentile of species that are potentially unprotected). Implementation of this new method is presented for the jurisdiction of the United States, with a focus on metal and metalloid contaminants; however, the new method can be used in any jurisdiction. A case study for molybdate shows the large effect of bioavailability corrections and smaller effects of protection level choices when deriving SCVs.
The replacement of native Portuguese forests by Eucalyptus globulus is often associated with deleterious effects on terrestrial and aquatic communities. Several studies have suggested that such a phenomenon is linked with the leaf essential oils released into the environment during the Eucalyptus leaf degradation process. However, to date, the way these compounds affect leaf degradation in terrestrial systems i.e. by direct toxic effects to soil invertebrates or indirectly by affecting food of soil fauna, is still unknown. In order to explore this question, the effect of essential oils extracted from E. globulus leaves on terrestrial systems was investigated. Fungal growth tests with species known as leaf colonizers (Mucor hiemalis, Alternaria alternata, Penicillium sp., Penicillium glabrum and Fusarium roseum) were performed to evaluate the antifungal effect of essential oils. In addition, a reproduction test with the collembolans Folsomia candida was done using a gradient of eucalyptus essential oils in artificial soil. The influence of essential oils on feeding behaviour of F. candida and the isopods Porcellio dilatatus was also investigated through food avoidance and consumption tests. Eucalyptus essential oils were lethal at concentrations between 2.5-20 µL/mL and inhibited growth of all fungal species between 1.25-5 µL/mL. The collembolan reproduction EC50 value was 35.0 (28.6-41.2) mg/kg and both collembola and isopods preferred leaves without oils. Results suggested that the effect of essential oils in leaf processing is related to direct toxic effects on fungi and soil fauna and to indirect effects on the quality and availability of food to soil invertebrates.
Organisms inhabiting metal-contaminated areas may develop metal tolerance, with either phenotypic adjustments or genetic changes (adaptation) or with both. In the present study, three populations of the terrestrial isopod Porcellionides sexfasciatus, collected at an abandoned mine area, were compared to assess the effects of metal contamination on tolerance to lethal and sublethal levels of copper, through comparison of survival, avoidance, and feeding. The effects of metal contamination on genetic diversity were considered using random amplified polymorphic DNA (RAPD) markers. No evidence of increased metal tolerance of the population inhabiting the contaminated site was found. There was no correlation between metal exposure and within-population genetic variance, but the three populations were clearly separated from each other. In conclusion, the populations of P. sexfasciatus in the mine landscape live rather isolated from each other and show no differential tolerance to Cu or indications of genetic erosion. Their phenotypic plasticity provides a means to survive despite exposure to extremely high metal concentrations in the soil.
Changes in saprophytic organisms gut microbial communities may present a threat to organic matter breakdown which can ultimately lead to soil function impairment. In this study, Amplified Ribosomal DNA Restriction Analysis (ARDRA) was evaluated as a potential simple molecular tool to assess shifts in bacterial community structure in hindgut populations of Porcellio dilatatus exposed to contaminated food. This prospective tool can also be used for a variety of purposes and samples prior to the use of more specific and sophisticated methods.
Continuing our interest in tridentate ligands to develop new prototypes of cobalt-based metallodrugs for combating cancer, modifications in the backbone of HL1, [(2-hydroxybenzyl)(2-(pyridil-2-yl)ethyl]amine) were proposed in order to modulate the redox potential of new Co(III) complexes. Three ligands with electron withdrawing groups were synthesized: HL2: [(2-hydroxy-5-nitrobenzyl)(2-(pyridil-2-yl)ethyl]amine); HL3: [(2-hydroxybenzyl)(2-(pyridil-2-yl)ethyl]imine) and HL4: [(2-hydroxy-5-nitrobenzyl)(2-(pyridil-2-yl)ethyl]imine). They were used to obtain the respective mononuclear complexes 2, 3 and 4, which are discussed compared to the previous reported complex 1 (obtained from HL1). The new complexes were characterized and studied by several techniques including X-ray crystallography, elemental and conductimetric analysis, IR, UV-vis and (1)H NMR spectroscopies, and electrochemistry. The substitutions of the group in the para position of the phenol (HL1 and HL2) and the imine instead of the amine (HL3 and HL4), promote anodic shifts in the complexes reduction potentials. The influence of these substitutions in the biological activities of the Co(III) complexes against the murine melanoma cell line (B16F10) was also evaluated. Little effect was observed on cellular viability decrease for all free ligands, however the coordination to Co(III) enhances their activities in the following range: 1>4?2>3. The data suggest that no straight correlation can be addressed between the reduction potential of the Co(III) center and the cell viability.
This work intends to implement the use of native soil nematode communities in ecotoxicological tests using a model pesticide and two geographically nematode communities (Mediterranean and sub-tropical) in order to obtain new perspectives on the evaluation of the toxic potential of chemical substances. The environmental condition of the nematode communities was described using a trait-based approach (grouping the organisms according to their feeding traits) and a traditional taxonomic method (identification to family level). Effects on total nematode abundance, number of families and abundance of nematode feeding groups as well as potential shifts in both trophic and family structure were assessed. Agricultural soils from Curitiba (Brazil) and Coimbra (Portugal) were sampled and the corresponding nematode communities were extracted. Part of the collected soil was defaunated and spiked with four doses of a carbofuran commercial formulation. Afterwards each of the replicates was inoculated with a nematode suspension containing ?200 or 300 nematodes. After 14 and 28 d of exposure the nematodes were extracted, counted and identified at family level and separately classified according to their feeding traits. The patterns of nematode responses revealed a decrease in the total abundance and a reduction in the number of families. Despite the similar effects observed for both communities, statistically significant toxic effects were only found within the Portuguese community. The total nematode abundance was significantly reduced at the highest carbofuran concentrations and significant shifts in the family structure were detected. However, the trophic structure, i.e., the contribution of each feeding group for the overall community structure, did not significantly change along the contamination gradient. Results showed that using such a trait-based approach may increase the ecological relevance of toxicity data, by establishing communalities in the response to a chemical from two different taxonomic communities, although with potential loss of information on biodiversity of the communities.
Patients with Parkinsons disease can benefit from controlled released levodopa dosage forms since there is a clear clinical advantage in obtaining sustained plasma concentrations. The purpose of this study was to obtain a tablet that prolonged the release of levodopa.
A current challenge in soil ecotoxicology is the use of natural soils as test substrates to increase ecological relevance of data. Despite the existence of six natural reference soils (the Euro-soils), some parallel projects showed that these soils do not accurately represent the diversity of European soils. Particularly, Mediterranean soils are not properly represented. To fill this gap, 12 natural soils from the Mediterranean regions of Alentejo, Portugal; Cataluña, Spain; and Liguria, Italy, were selected and used in reproduction and avoidance tests to evaluate the soil habitat function for earthworms (Eisenia andrei) and enchytraeids (Enchytraeus crypticus). Predictive models on the influence of soil properties on the responses of these organisms were developed using generalized linear models. Results indicate that the selected soils can impact reproduction and avoidance behavior of both Oligochaete species. Reproduction of enchytraeids was affected by different soil properties, but the test validity criteria were fulfilled. The avoidance response of enchytraeids was highly variable, but significant effects of texture and pH were found. Earthworms were more sensitive to soil properties. They did not reproduce successfully in three of the 10 soils, and a positive influence of moisture, fine sand, pH, and organic matter and a negative influence of clay were found. Moreover, they strongly avoided soils with extreme textures. Despite these limitations, most of the selected soils are suitable substrates for ecotoxicological evaluations.
Ecotoxicological laboratory tests (lower-tier tests) are fundamental tools for assessing the toxicity of pesticides to soil organisms. In this study, using these tests under tropical conditions, we quantified the impact of the insecticides imidacloprid, fipronil, and thiametoxam, and the fungicides captan and carboxin+thiram, all of which are used in the chemical treatment of crop seeds, on the survival, reproduction, and behavior of Eisenia andrei (Oligochaeta). With the exception of imidacloprid, none of the pesticides tested caused mortality in E. andrei in artificial soils. The LC(50) of imidacloprid was estimated as 25.53 mg active ingredient kg(-1) of dry soil. Earthworm reproduction rates were reduced by imidacloprid (EC(50)=4.07 mgkg(-1)), fipronil (EC(20)=23.16 mgkg(-1)), carboxin+thiram (EC(50)=56.38 mgkg(-1)), captan (EC(50)=334.84 mgkg(-1)), and thiametoxam (EC(50)=791.99 mgkg(-1)). Avoidance behavior was observed in the presence of imidacloprid (AC(50)=0.11 mgkg(-1)), captan (AC(50)=33.54 mgkg(-1)), carboxin+thiram (AC(50)=60.32 mgkg(-1)), and thiametoxam (AC(50)=>20 mgkg(-1)). Earthworms showed a preference for soils with the insecticide fipronil. Imidacloprid was the most toxic of the substances tested for E. andrei. The avoidance test was the most sensitive test for most pesticides studied, but results varied between pesticides. These results offer new insights on the toxicity of pesticides used to treat seeds in tropical regions. However, they should be complemented with higher-tier tests in order to reduce the uncertainties in risk assessment.
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.
Adaptation to environmental toxicants, such as metals, can affect population genetic diversity, both at neutral and selectable loci. At the transcriptional level, evolution of metal tolerance is possible due to the existence of polymorphisms in the cis-regulatory sequences of stress-responsive genes such as the metallothionein gene (mt). This study investigated the influence of cadmium adaptation on genetic diversity of soil-living Orchesella cincta (Collembola) populations in neutral (microsatellites and AFLP) and in functional (mt promoter) markers. Also, the influence of cis- and trans-acting factors on increased tolerance was addressed. No reduced genetic diversity was observed in two tolerant populations compared to five sensitive populations, either in neutral or in selectable markers. Extensive migration along with a large population size may explain the high genetic diversity measured. The metal-tolerant phenotype seems to be mostly influenced by genetic factors acting in cis on mt gene expression. The results suggest that certain promoter genotypes, which are found mainly or exclusively in Cd-tolerant populations, contribute to higher constitutive mt gene expression in individuals from these populations. However, more studies are needed to clearly unravel the influence of cis/trans-regulatory evolution in tolerant populations.
The present study evaluated the bioremediation efficacy of a cleanup tool for atrazine-contaminated soils (Pseudomonas sp. ADP plus citrate [P. ADP?+?CIT]) at a semifield scale, combining chemical and ecotoxicological information. Three experiments representing worst-case scenarios of atrazine contamination for soil, surface water (due to runoff), and groundwater (due to leaching) were performed in laboratory simulators (100 × 40 × 20 cm). For each experiment, three treatments were set up: bioremediated, nonbioremediated, and a control. In the first, the soil was sprayed with 10 times the recommended dose (RD) for corn of Atrazerba and with P. ADP?+?CIT at day 0 and a similar amount of P. ADP at day 2. The nonbioremediated treatment consisted of soil spraying with 10 times the RD of Atrazerba (day 0). After 7 d of treatment, samples of soil (and eluates), runoff, and leachate were collected for ecotoxicological tests with plants (Avena sativa and Brassica napus) and microalgae (Pseudokirchneriella subcapitata) species. In the nonbioremediated soils, atrazine was very toxic to both plants, with more pronounced effects on plant growth than on seed emergence. The bioremediation tool annulled atrazine toxicity to A. sativa (86 and 100% efficacy, respectively, for seed emergence and plant growth). For B. napus, results point to incomplete bioremediation. For the microalgae, eluate and runoff samples from the nonbioremediated soils were extremely toxic; a slight toxicity was registered for leachates. After only 7 d, the ecotoxicological risk for the aquatic compartments seemed to be diminished with the application of P. ADP?+?CIT. In aqueous samples obtained from the bioremediated soils, the microalgal growth was similar to the control for runoff samples and slightly lower than control (by 11%) for eluates.
Most laboratory tests may underestimate adverse effects in real scenarios of contamination because they imply the forced exposure of organisms to contaminants, thus overlooking the possibility of emigration. Avoidance from contaminants has been observed in several aquatic organisms, and avoidance-based tests have been recommended to be included in risk assessment studies. To reduce uncertainty in the extrapolation of laboratory derived results, the first aim of the present study was to compare both the median avoidance concentration and the lowest-observed-effect gradient (LOEG) values of atrazine for the cladoceran Daphnia magna, between an already developed 1.1-m-long system and a scaled-up system, three times longer. Second, the present study aimed at evaluating the population immediate decline--the proportion of the population that disappears (avoids and, if not, dies)--through the integration of the relationships between lethality and avoidance versus contaminant concentration. Daphnia magna significantly avoided atrazine, during 12-h exposures, with similar results in the original and scaled-up systems. The population immediate decline at the 48-h median lethal concentration would be 94%. Even at a concentration eliciting only 5% mortality, the population immediate decline would be over 50%. Achieving a higher pertinence of avoidance results and a better understanding of the LOEG values and their time dependence, scaling up the system further both spatially and temporally, and modeling explicit spatial dynamics in exposure and organism movement in space and time are needed.
The upper limit concentrations of metals established by international legislations for dredged sediment disposal and soil quality do not take into consideration the properties of tropical soils (generally submitted to more intense weathering processes) on metal availability and ecotoxicity. Aiming to perform an evaluation on the suitability of these threshold values in tropical regions, the ecotoxicity of metal-contaminated dredged sediment from the Guanabara Bay (Rio de Janeiro, Brazil) was investigated. Acute and avoidance tests with Eisenia andrei were performed with mixtures of dredged sediment with a ferralsol (0.00, 6.66, 13.12, 19.98, and 33.30 %) and a chernosol (0.00, 6.58, 13.16, 19.74, and 32.90 %). Mercury, lead, nickel, chromium, copper, and zinc concentrations were measured in test mixtures and in tissues of surviving earthworms from the acute tests. While ferralsol test mixtures provoked significant earthworm avoidance response at concentrations ?13.31 %, the chernosol mixtures showed significant avoidance behavior only at the 19.74 % concentration. The acute tests showed higher toxicity in ferralsol mixtures (LC50?=?9.9 %) compared to chernosol mixtures (LC50?=?16.5 %), and biomass increased at the lowest sediment doses in treatments of both test soils. Most probably, the expansive clay minerals present in chernosol contributed to reduce metal availability in chernosol mixtures, and consequently, the ecotoxicity of these treatments. The bioconcentration factors (BCF) for zinc and copper were lower with increasing concentrations of the dredged sediment, indicating the existence of internal regulating processes. Although the BCF for mercury also decreased with the increasing test concentrations, the known no biological function of this metal in the earthworms metabolism lead to suppose that Hg measured was not present in bioaccumulable forms. BCFs estimated for the other metals were generally higher in the highest dredged sediment doses.
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