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Volatile organic compounds (VOCs) are known to potentially cause a severe change in the respiratory metabolism of freshwater species, however the effect of these contaminants on groundwater-obligate species has not been investigated to date. Tetrachloroethylene (TCE) is a VOC frequently found in the groundwater bodies of industrialized areas, even years after a contamination event because TCE degradation takes several decades to occur. Contamination from TCE is considered persistent and difficult to remediate. Its high density favors a gravity-driven vertical infiltration into groundwater bodies. The TCE threshold value is 1.1 μg/L in groundwater bodies of Italy. TCE concentration in many Italian groundwater bodies is widely over this legal limit. In this study, we investigated the effect of 1.1 μg/L TCE on the survival, oxygen consumption, and locomotory activities of a groundwater-obligate copepod species. The specimens required for the trials were collected in the Antro del Corchia Cave (Tuscany). We measured the individual-based oxygen consumption of this species as a proxy of possible metabolic reactions to long-term (5 days) exposures to TCE at 8.0°C that is the mean annual temperature of groundwater flowing in the cave. To this end, we used a sealed glass microplate equipped with 24-planar oxygen sensor spots with optical isolation glued onto the bottom of 80-μL wells (Loligo Systems, Denmark) integrated with a 24-channel fluorescence-based respirometry system (SDR Sensor Dish Reader, PreSens, Germany). The system allows simultaneous measurement of 20 replicates and 4 controls. Survival and locomotory activity assessments were performed by counting the number of alive individuals and measuring the number of moving animals in 5 mL glass vials each containing 20 individuals. Preliminary results showed a decreasing in oxygen consumption of the organisms exposed to 1.1 μg/L TCE for 5 days at 8°C respect to the control. However, neither survival nor locomotory activities appeared to have been affected by exposure to TCE. See Suppl. material 1.

The organophosphorus herbicide glyphosate (GLY) and the organophosphate insecticide chlorpyriphos (CPF) are key pesticides in modern management cultures worldwide. Sublethal toxicity of the commercial herbicide formulation Roundup® and the insecticide formulation Terfos® were evaluated on Eisenia fetida coelomocytes exposed under in vivo and ex vivo laboratory conditions. Induction of DNA single-strand breaks evaluated by the single cell gel electrophoresis assay and coelomocyte viability as well as alterations in coelomocyte trophic indexes were employed as endpoints for genotoxicity and cytotoxicity, respectively. Specimens were exposed at concentrations corresponding to recommended pesticide field application rate, and endpoints were evaluated after 7 and 14 days of treatment (in vivo exposure). In addition, coelomocytes were exposed to aqueous leachate of pesticide-contaminated soils during 1 h (ex vivo exposure). Earthworms exposed to Roundup® and Terfos® showed an increased frequency of DNA damage. Also, a decrease of coelomocyte viability and decrease of trophic indexes were observed in all treatments. The results demonstrate that either GLY- and CPF-based formulations exerted genotoxic as well as cytotoxic effects in coelomocytes of E. fetida exposed in vivo and ex vivo. Fil: Curieses, Silvana Patricia. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Sáenz, María Elena. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Alberdi, José Luis. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina Fil: Martinez, Ricardo Santiago. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Larramendy, Marcelo Luis. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina Fil: Di Marzio, Walter Dario. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

The alkaline single-cell gel electrophoresis assay (comet assay) was used for the study of the genotoxic effects of insecticide Chlorpyrifos and fungicide Tebuconazole (commercial formulations) on two freshwater green algae species, Pseudokirchneriella subcapitata and Nannocloris oculata, after 24 h of exposure. The percentage of DNA in tail of migrating nucleoids was taken as an endpoint of DNA impairment. Cell viability was measured by fluorometric detection of chlorophyll “a” in vivo and the determination of cell auto-fluorescence. Only the higher concentration of Chlorpyrifos tested resulted to affect significantly the cell viability of P. subcapitata, whereas cells of N. oculata were not affected. Tebuconazole assayed concentrations (3 and 6 mg/l) did not affect cell viability of both species. The results of comet assay on P. subcapitata showed that Chlorpyrifos concentration evaluated (0.8 mg/l) exerted a genotoxic effects; while for the other specie a concentration of 10 mg/l was needed. Tebuconazole was genotoxic at 3 and 6 mg/l for both species. The comet assay evidenced damage at the level of DNA simple strains molecule at pesticide concentrations were cytotoxicity was not evident, demonstrating that algae are models to take into account in ecological risk assessments for aquatic environments. Fil: Martinez, Ricardo Santiago. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Di Marzio, Walter Dario. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina Fil: Sáenz, María Elena. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Luján. Departamento de Ciencias Básicas. Programa de Investigación en Ecotoxicología; Argentina

The management and final disposal of industrial wastes are a matter of considerable human concern. The present study evaluates the cyto/genotoxic effects and changes of the coelomic cell formulas exerted by aqueous leachates and solid waste (SW) of two industrial residues using coelomocytes extruded from Eisenia fetida. The assayed wastes corresponded to industrial foundry and cosmetic activities. After 14 days of exposure, we obtained a group of endpoints that reflect the toxicity/genotoxicity, coelomocyte formula and indexes; and the mortality classical value (LC50-14d). Among the variables measured, total coelomocytes formula (eleocytes + amebocytes + granulocytes) appears as a single and easy parameter to assess the toxicity of eluates at short exposure times. We applied a set of assays using earthworms as test organism that would allow evaluating SW as well as its aqueous leachates. It is easy to run trials combining exposures of 1 h to 14 days, which can be integrated into the implementation of the traditional test for evaluating acute toxicity., Facultad de Ciencias Naturales y Museo

Aquatic environments of the pampasic region of Argentina are severely affected by agricultural contamination due to an increase in a glyphosate tolerant transgenic variety of soybean crops. The present study is aimed to determine the effects of a commonly used Cyfluthrin commercial formulation (CCF) on growth, some physiological and biochemical parameter of four species of green algae. Significant inhibition of algal growth was observed from 0.1 mg Cyf/l. 96 h IC 50 were between 0.92 and 4.85 mg Cyf/l. CCF caused algicidal effects. Photosynthesis was stimulated by 50% in Scenedesmus quadricauda cultures exposed to the lowest concentration (hormesis). Algal photosynthesis inhibition was observed at higher concentrations with IC 50 values between 1.7 and 8.9 mg Cyf/l. Similar toxicity endpoints were found as a consequence of applying the traditional methodology of short-term chronic toxicity test of 96 h of exposition and the methodology developed using the Clark type photosynthetic oxygen evolution method. CAT activity was significantly increased between 23% and 33% considering the four species, at a lower concentration than those affecting algal growth and photosynthesis, indicating a potential biomarker. Taking into account that the extent of the soybean crops in the region is about fourteen million hectares, the improvement and extension of environmental tools for early detection of the action of pesticides on this essential group of organisms are discussed.

Quantitative structure–activity relationships (QSARs) with aromatic hydrocarbons were obtained. Biological response was measured by acute toxicity of several aquatic trophic levels. The chemicals assayed were benzene, toluene, ethylbenzene, o-xylene, m-xylene, p-xylene, isopropylbenzene, n-propylbenzene, and butylbenzene. Acute toxicity tests were carried out with Scenedesmus quadricauda, as representative of primary producers; Daphnia spinulata, a zooplanctonic cladoceran; Hyalella curvispina, a benthic macroinvertebrate; and Bryconamericus iheringii, an omnivorous native fish. The EC50 or LC50 was calculated from analytical determinations of aromatic hydrocarbons. Nonlinear regression analysis between the logarithm of the octanol–water partition coefficient (log Kow) of each compounds and the toxicity end points was performed. QSARs were positively related to increases in log Kow at all trophic levels. Intertaxonomic differences were found in comparisons of algae with animals and of invertebrates with vertebrates. We observed that these differences were not significant with a log Kow higher than 3 for all organisms. Aromatic hydrocarbons with log Kow values of less than 3 showed different toxicity responses, with algae more resistant than fish and invertebrates. We concluded that this was a result of the narcotic mode of action related to liposolubility and the ability of the compound to reach its target site in the cell. The bioconcentration factor (BCF) achieved to start nonpolar narcosis fell almost 1 order of magnitude below the BCF expected from the log Kow. Predicted critical body residues for nonpolar narcosis ranged between 2 and 1 mM. © 2006 Wiley Periodicals, Inc. Environ Toxicol 21: 118–124, 2006.

The earthworm (Eisenia foetida) is an attractive sentinel species for detecting genotoxicity in soil. In this study, an improved single-cell gel electrophoresis (SCGE) assay was developed for detecting DNA damage in the coelomocytes (lymphocytes) of earthworms. Coelomocytes were obtained from the coelomic fluid using a modified extrusion medium that did not include the mucolytic agent guaiacol. The extruded coelomocytes contained at least three types of cells: eleocytes (75% of the total), amoebocytes, and granulocytes. The DNA migration parameters were determined for untreated cells of each type in order that the assay could be performed with minimum inter- and intra-individual variation. In addition, lysis time was reduced to 10 min, and only one neutralization step was used. DNA damage was detected in isolated eleocytes treated with hydrogen peroxide and cadmium, and in eleocytes from earthworms exposed for up to 21 days to soil containing polycyclic aromatic hydrocarbons. The SCGE assay using earthworm eleocytes appears to be a sensitive biomarker for evaluating exposure to genotoxic compounds.