Towards simple tools to assess functional effects of contaminants on natural microbial and invertebrate sediment communities

Surface sediments can accumulate contaminants that affect microorganisms and invertebrates and disturb benthic ecological functions. However, effects of contaminants on ecological functions supported by sediment communities are understudied. Here, we tested the relevance of two simple tools to assess the ecotoxicological effects of metal contamination on natural sediment communities using particulate organic matter breakdown and decomposition as a functional descriptor. To this aim, we performed a 21-day laboratory microcosm experiment to assess the individual and combined effects of Cu and As (nominal concentration of 40 mg kg(-1) dw each) using the bait-lamina method (cellulose, bran flakes, and active coal in PVC strips) as well as artificial tablets (cellulose, bran flakes and active coal embedded in an agar matrix). Sediment toxicity was also evaluated using the standardized ostracod toxicity test. Both the bait-lamina and artificial tablet methods showed low effects of As on organic matter breakdown and decomposition but strong effects of Cu on this important ecological function. Both also showed that the presence of Cu and As in mixture in the sediment induced total inhibition of organic matter breakdown and decomposition. The ostracod toxicity test also showed high toxicity of Cu-spiked and Cu-plus-As-spiked sediments and low toxicity of As-spiked sediments. Besides confirming that artificial organic matter substrates are relevant and useful for assessing the functional effects of contaminants on sediment micro- and macro-organism communities, these results suggest that the proposed methods offer promising perspectives for developing tools for use in assessing functional ecotoxicology in the sediment compartment.

Fate of volatile fluorinated compounds in the subsurface

Christian Balsiger

Since their discovery around 1930, the synthetic compounds of the class of chlorofluorocarbons (CFCs) have been produced and used worldwide at very large scale. Their physico-chemical properties (non-flammability, chemical inertia, miscibility with water and oil, compatibility with many metals and plastics, solvent capacity) have made them ideal candidates for several applications, such as refrigerant fluids, aerosol propellants, foam blowing agents, or as solvents or intermediates of chemical synthesis. In the 70', it was demonstrated that following their release in the atmosphere, they are implicated in the process leading to the commonly called stratospheric ozone hole. The governments of several industrialised country have thus signed a protocol with the objective to stop the production and use of CFCs. CFCs have been increasingly replaced by compounds with similar structure, the hydrochlorofluorocarbons (HCFCs), but whose reactivity in the lower layers of the atmosphere is quite different from that of CFCs. Due to their chemical inertia, CFCs has been used as tracers for surface and groundwater datation. During such investigations, the biodegradability of these compounds and the existence of punctual pollution sources in soil and groundwater have been evidenced. It has been demonstrated that some of these compounds can be dechlorinated under anoxic conditions producing other compounds with sometimes higher toxicity and higher water solubility, and resulting in a risk linked to the consumption of natural groundwater. The goal of this thesis was to get indications about reductive transformations of CFCs and especially HCFCs in Swiss aquifers and landfills, on the potential of different microbial communities to reductively dechlorinate certain compounds, and under which redox conditions the transformations occur in aquifer systems. Data concerning the production, use, emission, regulation, as well as the biodegradability and toxicity of these compounds are presented in chapter 1 and in the appendix. The analytical methods developed and used during this work are presented in chapter 2. Particularly, a versatile analytical system allowing the measure of trace but also higher concentrations in different media, solid or aqueous, has been set up and used during this work. CFCs and HCFCs were analyzed at 3 sites in Switzerland. Samples were collected from a natural groundwater in which a chloroethene plume had been observed, from an old landfill closed in 1980, and from a more recent bioactive landfill constructed in horizontal layers. In addition, the behaviour of different CFCs and HCFCs was studied in microcosms inoculated with sludge originating from several sewage treatment plants in the canton of Vaud, and with a sediment originating from a contaminated aquifer in Holland. The studied CFCs and HCFCs were furthermore injected at the inlet of a 1.2 m height column filled with the aquifer sediment from Holland. In this quasi natural aquifer, the oxido-reductive conditions were modified during the experiment in order to investigate the conditions at which the biodegradation occurred. The field studies showed that CFCs and HCFCs are present in aquifers and landfills, and there were indications that their transformation also occurs there. Estimations of half-life times indicated that the reductive dechlorination occurred at slow rates. This could be due to the low temperatures of natural groundwater systems and other factors linked to the hydro-geological properties of the natural environment such as dispersion, sorption or partitionning. In the microcosms inoculated with sludge, CFC-11 was transformed to HCFC-21, the latter being further dechlorinated to HCFC-31. HCFC-123a and chlorotrifluoroethene (CTFE) were produced simultaneously from CFC-113. Inhibition of methanogesis with 2 bromoethane sulfonate decreased significantly the degradation rate of CFC-11, emphasizing the importance of the role of methanogens in the degradation process. In the microcosms inoculated with the contaminated sediment, the transformation of CFC-11 also occurred, but was not accompanied with methane production, as it was the case in the suldge microcosms. Moreover, the presence of acetate, lactate and butyrate in the microcosms containing CFC-11 suggested that methanogens were inhibited in the presence of CFC-11. CFC-113 was transformed in the same way as in the microcosms inoculated with the sludge; however, the produced CTFE was further transformed to trifluoroethene (TFE). Production of HCFC-151b from HCFC-141b was also observed. In the column filled with aquifer material, only the transformations of CFC-11 and CFC-113 were observed, which is probably due to the short residence time in the column (~10 days), during which the transformation of the other compounds was not sufficiently pronounced. The increasingly reducing conditions in the column were accompanied with an increase of the degradation rates of CFC-11 and CFC-113, with a maximum reached in presence of methanogenesis. The addition of sulfate in the artificial groundwater decreased the biodegradation rates of the two compounds, and the concentration of their transformation products was simultaneously increased. This suggests that these products become more persistent under sulfate-reducing conditions. Natural attenuation designates all the processes leading to the decrease of the concentration of organic compounds migrating from a pollution source throughout a natural environment. Whereas natural attenuation or enhanced natural attenuation seem adequate for several classes of compounds, the absence of defluorination reaction of CFCs and HCFCs reported in this work suggests the use of other remediation techniques for the decontamination of sites polluted with these compounds. Indeed, the transformation products of CFCs and HCFCs still contain a fluorine atom and are not less harmfull to the environment than the parent compounds. In addition, the use of CFCs as age-dating tracers in groundwater is not recommended under anoxic conditions, as it was demonstrated here that they are not stable in such environments. The HCFCs on the other hand seem to be more persistent, and it is primordial to decrease rapidly their production and release in the environment.

EPFL2004

Assessing toxic impacts on aquatic ecosystems in life cycle assessment (LCA)

Jérôme Payet

In the last decade, several Life Cycle Assessment (LCA) methods for assessing impact of products on living resources have been developed. Beyond the quantified assessments of impacts on living systems, it also checked the feasibility of the impact assessment on human health and ecosystems quality and helps to identify the limits of such methods. Among the different impact categories, that of toxic substances on ecosystems occupies an important place. The extent of these impacts has been stressed on many occasions and the necessity of preserving ecological areas and biodiversity has become a major issue on an international level. By focusing on aquatic ecosystems, this thesis aims at identifying constraints connected with assessment of the impact of chemical substances on ecosystems in LCA and setting up a method for assessing impacts of toxic substances on aquatic ecosystems which meets the requirements of a comparative approach like Life Cycle Assessment. The overall purpose of the thesis is to propose a comparative method for the Life Cycle Impact Assessment of toxics on aquatic ecosystems. With that aim, the dissertation is going throughout 6 major issues: The feasibility of the comparative impact assessment on ecosystems and the identification of associated constraints. The development of a statistical method for comparing impact on ecosystems; The review of the data availability for calculation of Effect Factors. The choice of the most relevant ecotoxicity measure (ECxs1, NOECs2 and LOECs3) for a comparative purpose. The development of best-estimate extrapolation factors for assessing chronic effects based on acute data. The analysis of the ecological realism of the comparative assessment method. These points are analysed throughout the 7 chapters of the thesis. Chapter 1 aims at introducing the thesis. A general presentation of Life Cycle Assessment is proposed, following by a detailed description of the Life Cycle Impact Assessment on ecosystems. This description covers the state of the art of researches and identifies the development needed. Therefore the scope of the thesis and the main points that must be addressed by this research are presented. Chapter 2 starts with a review of existing methods for Life Cycle Impact Assessment on ecosystems (LCIA), the chapter presents the parametric version of the AMI method (Assessment of the Mean Impact), which has been developed during the PhD for the assessment of impact on aquatic ecosystems. For this purpose, a framework and the main requirements for the development of this method are presented. For a comparative assessment, the Hazardous Concentration of a toxic affecting 50% of the species over their chronic EC50 (Effect Concentration affecting 50% of tested individuals), also called HC50EC50, is selected for the calculation of Effect Factors to be implemented in current LCIA methods. The Confidence Interval on the HC50EC50 is provided, enabling comparison between the impact values obtained as results of a Life Cycle Assessment study. The choice of EC50s is based on review of the main ecotoxicological databases, and analysis of the availability and reliability of test results. Moreover, bearing in mind that mostly acute data are available, while LCA deals mainly with chronic exposure, best-estimate extrapolation factors for the HC50EC50 and the associated uncertainty are provided for inorganics, nonpesticide organics, and pesticide organics. Concerning the method itself, in order to find the best methods for calculation of a toxicity indicator, several statistical estimators, parametric and non-parametric approaches are compared, identifying their properties and respective strengths for a comparative method. The analysis relates to both the reliability of the estimator and its Confidence Interval, especially in terms of statistical robustness and Effect Factor stability. Based on these findings, the AMI method is described in detail, and an example of application comparing two wheat crop scenarios differing by the pesticides used is presented. Chapter 3 presents in detail the four methods currently used for the development of Effect Factors for the Life Cycle Impact Assessment (LCIA) on Ecosystems: the parametric version of AMI (Assessment of the Mean Impact) based on HC50EC50s; the Eco-Indicator based on HC50NOECs; USES-LCA based on both HC5NOECs and the Most sensitive species, and the PNEC (Predicted No- Effect Concentration) based on the Most sensitive species. After presentation of the LCIA framework and its main divergences from Environmental Risk Assessment for chemical regulation, the four methods are detailed and applied for the calculation of Effect Factors for 83 substances, covering inorganics, non-pesticide organics, and pesticide organics. Each method is therefore analysed concerning three key points: applicability in the LCA framework, environmental relevance, and statistical reliability. Particular attention is paid to possible bias and the uncertainty, highlighting the following findings: (1) HC5NOECs are on average 50 times higher than the most sensitive species, and this difference in conservatism introduces a bias in the analyses for the method mixing HC5NOECs and most sensitive species. (2) Effect Factors based on the most sensitive species increase the relative weight of the most toxic chemicals by two orders of magnitude, depending on whether the study is based on US or European ecotoxicity databases. (3) the methods based on HC50EC50s and HC5NOECs are the only ones able to provide a Confidence Interval on the Effect Factor, but the Confidence Interval on the HC5NOECs can be more than 10 orders of magnitude greater than that of the HC50EC50s. (4) compared with the Confidence Interval on the HC50EC50s, the most sensitive species cannot be distinguished from HC50EC50s for chemicals characterised by fewer than 5 species, and the HC5NOECs cannot be distinguished from the HC50EC50s for chemicals characterised by fewer than 8 species. Chapter 4 compares two statistical estimators, aiming at calculate the average toxicity of substances on biological species. The two methods provide an estimation of the HC50EC50 and the associated Confidence Interval. On the one hand, parametric method using the geometric mean and a calculation of the confidence interval with Student is considered. On the other hand, a distribution-free method calculates the HC50EC50 based on the median response of species and the confidence interval based on bootstrap. In order to facilitate the use of the non-parametric method, a table linking the number of species tested and the size of the confidence interval is provided for samples from 5 to 500 species. The comparison is based on actual data concerning 191 substances covering inorganics, non-Pesticide organics, and Pesticide organics. The mean and width of the chronic EC50s samples for all the substances are presented. The Shapiro-Wilk test is performed for the 191 EC50s samples and the assumption of log-normality of the distribution failed in more than 20% of the cases. Two causes of this non Log-normality are identified; (1) the skewness, which is shown to be an important issue for the assessment of the average toxicity of chemicals while (2) the multi-modal distributions, which are not likely to influence considerably the final result. A detailed application of the two methods is done with the comparison of two herbicides, the Sulfosulfuron and the Prosulfuron, where the distribution-free method appears to be more powerful than the parametric for a substance-to-substance comparison. Nevertheless, the distribution-free method requires a minimum of 5 chronic EC50s, that cannot be satisfied in most cases. Chapter 5 aims at illustrating the previous chapter in using the nonparametric version of the AMI method for the comparative assessment of the impacts of metals on aquatic ecosystems. This chapter briefly describes the method, then it focuses on the comparative analysis of 9 metals sometimes tested with different salts and speciations. Two interesting results can be highlighted: (1) the toxicity of metals covers the whole range of toxicity of chemicals; (2) the confidence interval of the HC50EC50 for metals is on average twice as great for metals compared with other chemicals. This increase in the variability of ecotoxicological responses from species is likely to be due to the change in bioavailability of metals associated with a change of test conditions (e.g. pH, or Organic Matter). Chapter 6 reviews and analyses the reliability of existing aquatic toxicity databases which can be used for the calculation of Effect Factors for Life Cycle Impact Assessment (LCIA). For that purpose, the main LCIA methods are presented focusing on their data requirement. It concerns: EDIP4 (based on the PNEC); AMI5 (based on parametric HC50EC50); Eco-Indicator (based on the HC50NOEC); USES-LCA (based on the HC5NOEC). Moreover 6 ecotoxicity databases available in an electronic format are analysed: Aquire; Pesticide Ecotoxicity Database (PED); IUCLID; Acute Toxicity Database (ATD); Fathead Minnow database (FMD); and ECETOC Aquatic Toxicity Database (EAT). The analysis especially focuses on the identification of the substances and organisms, the definition of the tests conditions, and the control procedure of the database. A selection of tests is done, retaining a dataset of 128,864 tests results, acute, sub-chronic and chronic. A description of the data availability on the basis of the selected test is performed, considering the available EC50s (Effect Concentration affecting 50% of the individuals tested), LOECs and NOECs (Lowest or No Observed Effect Concentration). The number of covered substances is also analysed regarding the number of species or phyla considered. On that basis, an estimation of the maximum number of possible Effect Factors is performed. The results highlight the discrepancy between the large number of test results available (128,864), and the relatively restricted number of Effect Factors (betweeen 34 and 4959 depending on the method)) that can be calculated for a comparative purpose like LCIA. Chapter 7 provides a conclusion to the work, answering the points underlined in introduction. Then, the fey features of the AMI method are restated and the perspectives and output of the work are presented. In the last part of the thesis, the AMI HC50EC50 database is presented. It provides acute and chronic HC50EC50 data calculated with the parametric version of AMI (geometric mean of the EC50s and confidence interval based on Student) for 522 susbtances. ------------------------------ 1 ECx: concentration of susbtance that affects 50% of the individuals tested for a given effect. 2 NOEC: No Observable Effect Concentration 3 LOEC: Lowest Observable Effect Concentration 4 EDIP: Environmental Design for Industrial Product 5 AMI : Assessment of the Mean Impact

EPFL2004

Endocrine disruption in soil invertebrates

Sophie Campiche

In the past years, it has been observed that some compounds present in our environment can disturb the reproduction and development of animals like fishes, birds, or reptiles by interfering with their endocrine system. Indeed, these endocrine disrupting compounds (EDC) can mimic or antagonize the effects of hormones, alter the pattern of synthesis and metabolism of hormones or modify hormone receptor levels. These substances represent a risk for wildlife, and possibly for humans. Up to now, endocrine disruption was mainly evaluated for vertebrates and aquatic organisms and for oestrogeniclike substances. However, soil invertebrates, which play an important role in soil functioning, have rarely been considered. Moreover, as their endocrine system differs substantially from those of vertebrates (estrogens do not seem to regulate endocrine functions in invertebrates), other substances than can mimic invertebrate hormones should be taken into account. In this sense, insect growth regulators (IGR), which are third generation insecticides specially developed to interfere with insect endocrine system, are interesting compounds. These substances are supposed to have a high specificity for insect pest and a low toxicity for non-target organisms. In the first part of this study, the sublethal effects of six IGR (methoprene, fenoxycarb, precocene II, tebufenozide, hexaflumuron and teflubenzuron) were evaluated on the non-target soil arthropod Folsomia candida. The collembola F. candida represents an integral and beneficial part of the soil ecosystem. It is an euedaphic (subsurface) species which plays an important role in soil respiration and decomposition processes and is therefore vulnerable to the effects of soil contamination. This ecologically relevant organism is one of the most appropriate invertebrate test species for the assessment of environmental quality. It is recommended as test organism by the international standard ISO 11267. The 28-days reproduction tests conducted according to this protocol show that F. candida is affected by the chosen IGR. The most toxic compounds were the two chitin synthesis inhibitors, teflubenzuron and hexaflumuron, with an EC50 of 0.05 mg/kg (dw) for teflubenzuron and an EC50 of 0.6 mg/kg for hexaflumuron. These concentrations are probably environmentally relevant (toxicity/exposure ratios

EPFL2006

Endocrine disruption in soil invertebrates

Sophie Campiche

EPFL2006

Fate of volatile fluorinated compounds in the subsurface

Christian Balsiger

EPFL2004

Assessing toxic impacts on aquatic ecosystems in life cycle assessment (LCA)

Jérôme Payet

EPFL2004