La suspensió dels drets constitucionals

relationships (Pearson, R2=0.11) between the ChE and the weight or sex of the birds. The weight of the birds decreased on average 1.54 g after the exposure, possibly due to the stress of capture and the lack of adaptation to captivity. The house sparrow showed to be a species sensitive to the presence of OP compounds in the environment. It is possible that the house sparrow and other birds that co-inhabit the agricultural areas where malathion is applied, as well as other OP insecticides, are prone to develop different levels of intoxication and that in some cases their health condition is compromised.

MO042

Implementation of a worst-case landscape scenario for population modelling of a fungicide applied in cereals

M. Wang, WSC Scientific GmbH / Dept Efate Modelling; T. Preuss, Bayer Ag / Environmental Safety; M. Ebeling, Bayer AG Crop Science Division /

Ecotoxicology - Terrestrial Vertebrates Expert Team

In many cases EU member states prefer the use of country specific scenarios for the exposure or risk assessment of pesticides. This is sometimes requested due to specific geographical features or agricultural practice. In the present example, we demonstrate how the specific agricultural situation of the Netherlands was taken into account for the selection of landscape scenarios for use in population modelling. It is first shown how a country specific landscape scenario is developed. Then, the dose response obtained in a rat reproduction study with an azole fungicide is employed in a population-level risk assessment on small herbivorous mammals (Common vole, Microtus arvalis). The margins of safety obtained in that assessment indicate that no adverse population-level effects would be expected under realistic worst-case field conditions.

MO043

Biomonitoring and validation of non-invasive samples for the analysis of metals in freshwater turtles from mining areas

J. Pareja Carrera, IREC-UCLM / IREC-UCLM; A. Rodríguez-Perez, UCLM-IREC; M. Martinez-Haro, IREC-Instituto de Investigación en Recursos Cinegéticos / Department of Life Sciences; R. Mateo, IREC-CSIC- UCLM / Grupo de Toxicología de Fauna Silvestre; M. Ortiz Santaliestra, Institute for Game and Wildlife Research (IREC) UCLM-CSIC-JCCM

The mobilization of metals present in the earth’s crust due to anthropogenic activities, such as mining, is one of the main contamination routes for wildlife. With the objective of biomonitoring and validating non-invasive methods for metal analysis in reptiles, levels of lead (Pb) and mercury (Hg) in blood, faeces and carapace scales of freshwater turtles (Mauremys leprosa, n=86) from historical mining areas (i.e. Valle de Alcudia-Sierra Madrona district for Pb, Almadén district for Hg) in Central Spain were analysed. The highest levels of blood Pb were found in the animals collected from the area of Valle de Alcudia-Sierra Madrona, more specifically from Solana del Pino, with an average (±SD) of 5.59±3.66 μg/g dry weight (dw). Individuals from this location showed the clearest evidences of oxidative stress, as estimated from the highest values of malondialdehyde (biomarker of lipid peroxidation) and superoxide dismutase activity. However, no correlations were found between these biomarkers and blood Pb levels. Similarly, the highest levels of Hg were detected in the animals collected from the area of Almadén, specifically from Almadenejos, with an average (±SD) of 8.83±8.84 μg/g dw. These individuals showed no evidences of oxidative stress, but presented increased activity of glutathione peroxidase and reduced glutathione levels relative to the rest of populations, which would indicate that antioxidant system is preventing from Hg-derived oxidative damage to occur. Blood levels of these two elements were above those reported as susceptible to cause sub-lethal effects in reptiles for the vast majority of terrapins from the most contaminated sites (100% of terrapins from Solana del Pino with blood Pb levels > 15 μg/dl; 70.3% from Almadenejos with blood Hg levels > 2.76 μg/g dw). Faeces and carapace scales obtained non-invasively correlated significantly with blood levels for the case of Pb (R ≥ 0.705, P < 0.001), but not for Hg (R ≤ 0.362, P ≥ 0.127). Thus, these samples could be used as non-invasive methods for the analysis of Pb bioavailability in M.

leprosa, and by extension in reptiles, which will contribute to the development of

ecotoxicology in reptiles, a group very little studied in this regard.

MO044

An analysis of important life stages, exposure routes and test endpoints in amphibians and coverage by existing risk assessment regulatory requirements for plant protection products, part 1

A. Aldrich, Agroscope / Ecotoxicology; C. Berg, Uppsala universitet, Dept. of Environmental Toxicology / Dept of Environmental Toxicology; M. Ortiz Santaliestra, Institute for Game and Wildlife Research (IREC)

UCLM-CSIC-JCCM; S. Pieper, German Federal Environment Agency (UBA) / Plant Protection Products; S.M. Weir, Queens University of Charlotte / Department of Biology

Concerns have been raised that the current risk assessment of plant protection products (PPP) may not sufficiently cover the risk to amphibians and reptiles. To address these concerns, the European Food Safety Authority (EFSA) has published a Scientific Opinion addressing the state of the science regarding the risk to amphibians and reptiles exposed to pesticides (EFSA 2017). A central task of the EFSA working group was to understand how well important life stages, exposure

routes and endpoints are currently covered (or not) by the current risk assessment paradigm. We had four steps in this analysis: 1) Identify relevant life-stages, important exposure routes and possible endpoints with relevance to the specific protection goals (SPGs) defined for amphibians; 2) Evaluate the coverage by existing standard test guidelines with regard to these important

stages/routes/effects; 3) Explore whether tests with surrogate species could provide suitable information allowing for extrapolation to assess the hazard/risk for amphibians; 4) Identify proposed non-standardized test protocols suitable to address the gaps for specific protection goals defined for amphibians. Care was taken to address all relevant routes of exposure, life-stages, and short and long term effects. For amphibians, greater coverage or surrogacy exists for the aquatic larval stage and short term effects, with less coverage of the adult terrestrial stage, reproductive toxicity, and specifically for the potential effects via dermal exposure routes. To cover important life stages, exposure routes and effects, tests addressing dermal overspray and reproductive toxicity in amphibians may be needed. The analysis for reptiles will be presented in a separate poster. The concerns that the current risk assessment of pesticides may not sufficiently cover the risk to amphibians were supported after the analysis of the currently available data. The exercise provides a useful base for further research necessary to advance the ecotoxicological risk assessment of amphibians within the remit of the PPP authorization.

MO045

European common frog (Rana temporaria) larvae show subcellular responses under field-relevant Bacillus thuringiensis var. israelensis (Bti) exposure levels used in mosquito control

S. Allgeier, B. Frombold, University Koblenz-Landau; V. Mingo, Trier University / Biogeography; C. Bruehl, University of Koblenz-Landau / Institute for

Environmental Sciences

Bacillus thuringiensis var. israelensis (Bti) is presumed to be an environmental

friendly agent for use in either health-related mosquito control or the reduction of nuisance associated with mosquitoes coming from temporary flooded wetlands. Amphibians co-occurring with mosquito larvae in these wetlands may be exposed to Bti products several times during their breeding season. Up until now, information regarding effects on the non-targeted group of amphibians has to be considered rather inconsistent. On this account, we evaluated how repeated exposures to frequently used Bti formulations (VectoBac®_{12AS, VectoBac}®_{WG) in} field-relevant rates affect European common frog (Rana temporaria) larvae. In a laboratory approach, tadpoles experienced exposure conditions similar to realistic mosquito control in the Upper Rhine Valley (Germany). We assessed potential effects with regard to enzymatic biomarkers (glutathione-S-transferase, glutathione reductase, acetylcholine esterase), development, body condition and survival until the end of metamorphosis. Regardless of the formulation, delivery form or application rate, tadpole survival rates and time to metamorphosis were slightly reduced after repeated Bti exposures, while body condition was similar throughout the treatments. Furthermore, Bti induced significant increases of all enzymatic activities irrespectively of the applied field rate and formulation, indicating oxidative stress as well as unspecific neurotoxic effects. Repeatedly executed Bti applications, especially acting on early developmental stages, seem to increase the risk for adverse effects. The examination of several biochemical markers is needed to evaluate the ecotoxicological risk of Bti for amphibian populations, especially in the context of worldwide amphibian declines. Following the precautionary principle, the implementation of certain thresholds for application numbers and intervals should be considered in order to ensure environmentally friendly mosquito control programs, especially in areas originally designated for nature conservation.

MO046

Influence of salinity and temperature on tadpoles of Xenopus laevis

C. Monteiro, R. Alves, University of Aveiro / Department of Biology & CESAM, 3810-193 Aveiro; A.M. Soares, University of Aveiro / department of Biology & CESAM; I. Lopes, University of Aveiro / Department of Biology & CESAM, 3810-193 Aveiro

Worldwide coastal low-lying ecosystems are experiencing an increased salinity due to seawater intrusion. Considering the projections of the International Panel for Climate Changes regarding the increase of mean temperatures, until 2100, and consequent sea level rise, it is foreseen an increase in the number of costal ecosystems suffering from such salinization. Among coastal ecosystems that will be impacted with seawater intrusion are wetlands, which constitute hotspots of biodiversity and represent relevant ecosystems for amphibians. This class of vertebrates holds the highest proportion of endangered species and is considered very vulnerable to salinity changes. In this context, the present study aimed at evaluating the influence of temperature on the adverse effects that increase of salinity may cause to tadpoles of the amphibian species Xenopus laevis. To address this objective, X. laevis tadpoles (Gosner 25) were exposed to a range of 5 NaCl concentrations under three temperatures: 20, 23 and 26ºC. The following parameters were evaluated at the end of the test: feeding rate, body weight and growth rates. The differences reported for size between control and NaCl concentrations were mainly due to the tail length. For all concentrations, the feeding rate decreased with increasing NaCl concentration. As well, body weight decreased

with increasing salinity. Although significant interactions between temperature and salt concentration were registered, a pattern of influence in the temperature was not observed. Adverse effects were observed at the lowest tested salinity levels, which suggest that these species are highly vulnerable to small salinity increases and would be at high risk under seawater intrusion scenarios. Keywords: Salinity; toxicity; amphibia

MO047

EFFECTS OF THE EXPOSURE OF LARVAS OF Dendropsophus columbianus (ANURA: HYLIDAE) TO WATERS CONTAMINATED BY ANTHROPOGENIC ACTIVITIES IN A RIVER BASIN OF THE COLOMBIAN ANDES

V. Ramírez Castaño, Universidad de Caldas; B. Toro, Universidad de Caldas / Biological Sciences

The pollution generated by agriculture, livestock and mining have impacted the watersheds in the Colombian Andes. Amphibians have been used to evaluate this contamination due their biphasic lifecycle, which has made them ideal models in aquatic ecotoxicology. The objectives of this work were: 1) to determine if tadpoles of D. columbianus exposed to contaminants of agricultural, livestock and mining (with mercury: Hg, and with mercury and cyanide: Hg/CN) varied in the snout-vent length (SVL), tail length (TL), head width (HW), and body weight, and 2) to evaluate the effect of exposure on metamorphosis and behavior of the larvae. The AMPHITOX protocol was followed using ten larvae in each of the treatments and in the control, which were exposed from the moment of hatching to complete metamorphosis. Significant differences were found in the LRC between the larvae of the control and the Hg/CN mining treatment (Z= -28.92, p= 0.000) and between Hg/CN mining and agriculture treatments (Z= 25.325, p= 0.001) after 50 days of exposure. Differences in LC were found between the larvae of the control and the Hg/CN mining treatment (Z= -25.57, p= 0.001), and between Hg/CN mining and Hg mining treatments (Z= 21.525, p= 0.009) in the same time. The weight did not show differences. The majority of larvae of the control and the agriculture treatment showed similar development rates, reaching stage 46 between days 60 and 75 of exposure. While tadpoles exposed to Hg/CN mining did not complete the metamorphosis and reached stage 42 between days 50 and 55. Surface flotation was the activity that was most stable at the time of exposure, presenting percentages of individuals greater than 60% in the control (81%), and in the agriculture (70%) and Hg mining treatments (65%), between the last days of exposure evaluated: 22 to 28. It is shown that 1) the effects on growth of anuran larvae due to agricultural, livestock and mining contamination are similar. However, samples contaminated by mining produce smaller tadpoles than agriculture and livestock, and 2) tadpoles exposed to agricultural and livestock contamination, unlike other studies which report long times of metamorphosis, show a time of this process that approximates to the time that the species experimented in the control (134 days) and probably, to the time of this in situ.

MO048

Risks for amphibians and reptiles by dermal exposure to pesticides

F. Streissl, EFSA / Pesticides Unit; P.I. Adriaanse, Alterra Wageningen University and Research Centre; R. Sharp, EFSA - European Food Safety Authority / Pesticides Unit; K. Machera, Benaki Phytopathological Institute / Department of Pesticides Control & Phytopharmacy, Athens, Greece

Amphibian and reptilian species are found in agricultural landscapes. Some inhabit water bodies either permanently or during some time of their life cycle. Others use predominantly the terrestrial habitat. Some of those species migrate long distances through fields and some reside in fields and field margins where they can be exposed to pesticide residues in food items, water, soil and plant surfaces. A number of studies indicate that pesticide exposure can lead to severe impacts with up to 100% mortality from overspray at field rates for some pesticides. Dermal exposure was identified as an important exposure route in the EFSA opinion. The aim of the current study was to investigate different options to address the risk from dermal exposure by overspray and contact to soil and plant surfaces. Existing exposure models were reviewed with regard to their suitability for amphibian and reptilian risk assessment. This included comparison of parameters used in model calculations, comparison of sensitivity to dermal exposure and worst case exposure calculations to identify groups of amphibians and reptiles with greatest dermal exposure from overspray and to compare it with exposure from contact to soil and plant surfaces. In addition a novel approach was developed to estimate the uptake from soil and plant surfaces. Mammalian dermal toxicity related to local effects and dermal adsorption data may be used as surrogates for reptiles but not for

amphibians. The development of a test method investigating local effects on amphibian skin, as well as the estimation or measurement of body burden following exposure via the dermal route is needed. Comparison of body burden by overspray to body burden by exposure to treated soil or foliage showed that the maximal body burden by overspray was lower than the maximal body burden by passive or active dermal uptake from soil or by contact to foliage. An approach was suggested which can combine oral and all dermal exposure routes in one overall body burden which could form the basis for a realistic risk assessment.

MO049

Evaluating the Role of Fish as Surrogates for Amphibians in Ecological Risk

Assessment

S. Glaberman, University of South Alabama / Biology; J. Kiwiet, University of South Alabama; C. Aubee, US Environmental Protection Agency / Risk Assessment Division Office of Pollution Prevention and Toxics

Ecological risk of chemical exposure to aquatic-phase amphibians is historically evaluated using surrogate toxicity data from standard fish species. Recently published meta-analyses of fish and amphibian ecotoxicity data concluded that both groups are similarly sensitive to a range of chemicals. However, these analyses are limited because the amphibian data reported in the peer-reviewed literature are variable both with respect to experimental design and test species. In 2010, the U.S. Environmental Protection Agency began receiving ecotoxicity data for a standard amphibian test species (Xenopus laevis) as part of the Endocrine Disruptor Screening Program. Although these studies are primarily designed to inform a determination of potential thyroid interaction within the context of other endocrine screening studies, they also contain valuable data on survival and growth that can be compared to existing fish data for a given chemical. We used this dataset to compare no observed adverse effect concentration (NOAEC) values for survival, body weight, and length data between fish and amphibians for 45 different pesticide active ingredients. Overall, the results indicate that fish are a reasonably good predictor of amphibian toxicity as there were no statistically significant differences in NOAEC values between the two groups for the endpoints examined. However, toxicity endpoints were lower in amphibians as compared to fish approximately half the time across chemicals, challenging the notion that fish are consistently more sensitive than amphibians. Disclaimer: Disclaimer: The views expressed in

this presentation do not necessarily represent the views of the U.S. EPA or the United States.

MO050

Long-term survival of mancozeb exposed common vole populations from one to the following reproductive season

F. Von Blanckenhagen, J. Ludwigs, Rifcon GmbH

Rodent field effect studies relevant for pesticide risk assessment typically take place during crop development from spring to autumn and thus within a single reproductive season. However, animal survival covering multiple reproductive seasons has only rarely been considered. This includes also microtine rodents such as the common vole (Microtus arvalis), as they are considered as rather short-lived vertebrates. However, overwintering individuals are important for the survival of local populations, and potential pesticide exposure during maturation might impact survival during seasonal bottlenecks in winter from one to the next reproductive season. Furthermore, from a regulatory perspective and due to the

one-reproductive-season-duration of most field effect studies in wildlife, there is an increasing concern on potential long-term effects from exposure occurring delayed or that the reproduction in the following season might be affected by exposure during a previous application season. Against this background we monitored individually marked common vole populations from a long term effect study on spray applications of Dithane M-45 (Mancozeb 80% WP) during one reproductive season further on into the following reproductive season. The test item Dithane M-45 was applied four times in June according to Good Agricultural Practice at an application rate of 2 kg a.s./ha. Trapping and marking of voles in the same investigation plots was conducted until September, followed by further trapping until spring of the following year and the onset of the new reproductive cycle. Reproductive parameters recorded as indicators of potential long-term effects resulted in very similar patterns in treatment and control plots, and the data show no indication that common voles were negatively affected by multiple applications of the test item also in the following year. Furthermore and up to now not documented for wild living common voles, the data gives evidence that free-living common vole individuals can reproduce for more than one reproductive season.

MO051

An analysis of important life stages, exposure routes and test endpoints in reptiles with regard to coverage by existing risk assessment regulatory requirements for pesticides

A. Aldrich, Agroscope / Ecotoxicology; C. Berg, Uppsala universitet, Dept. of Environmental Toxicology / Dept of Environmental Toxicology; M. Ortiz Santaliestra, Institute for Game and Wildlife Research (IREC)

UCLM-CSIC-JCCM; S. Pieper, German Federal Environment Agency (UBA) / Plant Protection Products; S.M. Weir, Queens University of Charlotte / Biology Concerns have been raised that the current risk assessment of pesticides may not sufficiently cover the risk to amphibians and reptiles. To address these concerns, the